U.S. patent application number 11/973344 was filed with the patent office on 2008-05-08 for genetic polymorphisms associated with coronary stenosis, methods of detection and uses thereof.
This patent application is currently assigned to APPLERA CORPORATION. Invention is credited to James J. Devlin, May Luke.
Application Number | 20080108081 11/973344 |
Document ID | / |
Family ID | 32686076 |
Filed Date | 2008-05-08 |
United States Patent
Application |
20080108081 |
Kind Code |
A1 |
Luke; May ; et al. |
May 8, 2008 |
Genetic polymorphisms associated with coronary stenosis, methods of
detection and uses thereof
Abstract
The present invention is based on the discovery of genetic
polymorphisms that are associated with coronary stenosis. In
particular, the present invention relates to nucleic acid molecules
containing the polymorphisms, variant proteins encoded by such
nucleic acid molecules, reagents for detecting the polymorphic
nucleic acid molecules and proteins, and methods of using the
nucleic acids and proteins as well as methods of using reagents for
their detection.
Inventors: |
Luke; May; (San Francisco,
CA) ; Devlin; James J.; (Lafayette, CA) |
Correspondence
Address: |
CELERA, AN APPLERA BUSINESS UNIT
1401 HARBOR BAY PARKWAY
ALAMEDA
CA
94502
US
|
Assignee: |
APPLERA CORPORATION
Norwalk
CT
06856-5435
|
Family ID: |
32686076 |
Appl. No.: |
11/973344 |
Filed: |
October 5, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10741601 |
Dec 22, 2003 |
7306913 |
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11973344 |
Oct 5, 2007 |
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60434741 |
Dec 20, 2002 |
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60453050 |
Mar 10, 2003 |
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60466437 |
Apr 30, 2003 |
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Current U.S.
Class: |
435/6.11 |
Current CPC
Class: |
C12Q 2600/156 20130101;
C12Q 1/6883 20130101 |
Class at
Publication: |
435/006 |
International
Class: |
C12Q 1/68 20060101
C12Q001/68 |
Claims
1. A method of identifying a human having an altered risk for
coronary stenosis, comprising detecting the presence of a single
nucleotide polymorphism (SNP) at position 101 of SEQ ID NO: 41 or
its complement thereof in said human's nucleic acids, wherein the
presence of G at position 101 of SEQ ID NO: 41 is indicative of an
increased risk for coronary stenosis, or the presence of A at
position 101 of SEQ ID NO: 41 is indicative of a decreased risk for
coronary stenosis.
2. The method of claim 1 wherein SEQ ID NO: 41 is a segment within
the genomic sequence of CD163 gene as represented by SEQ ID NO:
37.
3. The method of claim 1 wherein the SNP is located at position
24287 of SEQ ID NO: 37.
4. The method of claim 1 wherein said human's nucleic acids are
extracted from a biological sample therefrom.
5. The method of claim 1 wherein said human's nucleic acids are
amplified before being detected.
6. The method of claim 1 wherein the detecting is carried out by
using detection reagents comprising the nucleotide sequences of SEQ
ID NO: 51, SEQ ID NO: 52, and SEQ ID NO: 53.
7. The method of claim 1 in which the detecting is carried out by a
process selected from the group consisting of: allele-specific
probe hybridization, allele-specific primer extension,
allele-specific amplification, sequencing, 5' nuclease digestion,
molecular beacon assay, oligonucleotide ligation assay, size
analysis, and single-stranded conformation polymorphism.
8. A method of identifying a human having an increased risk for
coronary stenosis, comprising detecting the presence of a single
nucleotide polymorphism (SNP) at position 101 of SEQ ID NO: 41 or
its complement thereof in said human's nucleic acids, wherein the
presence of G at position 101 of SEQ ID NO: 41 is indicative of an
increased risk for coronary stenosis.
9. The method of claim 8 wherein SEQ ID NO: 41 is a segment within
the genomic sequence of CD163 gene as represented by SEQ ID NO:
37.
10. The method of claim 8 wherein the SNP is located at position
24287 of SEQ ID NO: 37.
11. The method of claim 8 wherein said human's nucleic acids are
extracted from a biological sample therefrom.
12. The method of claim 8 wherein said human's nucleic acids are
amplified before being detected.
13. The method of claim 8 wherein the detecting is carried out by
using detection reagents comprising the nucleotide sequences of SEQ
ID NO: 51, SEQ ID NO: 52, and SEQ ID NO: 53.
14. The method of claim 8 in which the detecting is carried out by
a process selected from the group consisting of: allele-specific
probe hybridization, allele-specific primer extension,
allele-specific amplification, sequencing, 5' nuclease digestion,
molecular beacon assay, oligonucleotide ligation assay, size
analysis, and single-stranded conformation polymorphism.
15. A method of identifying a human having a decreased risk for
coronary stenosis, comprising detecting the presence of a single
nucleotide polymorphism (SNP) at position 101 of SEQ ID NO: 41 or
its complement thereof in said human's nucleic acids, wherein the
presence of A at position 101 of SEQ ID NO: 41 is indicative of a
decreased risk for coronary stenosis.
16. The method of claim 15 wherein SEQ ID NO: 41 is a segment
within the genomic sequence of CD163 gene as represented by SEQ ID
NO: 37.
17. The method of claim 15 wherein the SNP is located at position
24287 of SEQ ID NO: 37.
18. The method of claim 15 wherein said human's nucleic acids are
extracted from a biological sample therefrom.
19. The method of claim 15 wherein said human's nucleic acids are
amplified before being detected.
20. The method of claim 15 wherein the detecting is carried out by
using detection reagents comprising the nucleotide sequences of SEQ
ID NO: 51, SEQ ID NO: 52, and SEQ ID NO: 53.
21. The method of claim 15 in which the detecting is carried out by
a process selected from the group consisting of: allele-specific
probe hybridization, allele-specific primer extension,
allele-specific amplification, sequencing, 5' nuclease digestion,
molecular beacon assay, oligonucleotide ligation assay, size
analysis, and single-stranded conformation polymorphism.
22. A method of determining a human's risk for developing coronary
stenosis, comprising detecting the presence of a single nucleotide
polymorphism (SNP) at position 101 of SEQ ID NO: 41 or its
complement thereof in said human's nucleic acids, wherein the
presence of G at position 101 of SEQ ID NO: 41 is indicative of an
increased risk for developing coronary stenosis in said human, or
the presence of A at position 101 of SEQ ID NO: 41 is indicative of
a decreased risk for developing coronary stenosis in said
human.
23. The method of claim 22 wherein said human's nucleic acids are
amplified before being detected.
24. The method of claim 22 wherein the detecting is carried out by
using detection reagents comprising the nucleotide sequences of SEQ
ID NO: 51, SEQ ID NO: 52, and SEQ ID NO: 53.
25. The method of claim 22 in which the detecting is carried out by
a process selected from the group consisting of: allele-specific
probe hybridization, allele-specific primer extension,
allele-specific amplification, sequencing, 5' nuclease digestion,
molecular beacon assay, oligonucleotide ligation assay, size
analysis, and single-stranded conformation polymorphism.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional application of U.S.
non-provisional application Ser. No. 10/741,601, filed Dec. 22,
2003, which claims priority to provisional application Ser. No.
60/434,741, filed Dec. 20, 2002, provisional application Ser. No.
60/453,050, filed Mar. 10, 2003, and provisional application Ser.
No. 60/466,437, filed Apr. 30, 2003, the contents of which are
hereby incorporated by reference in its entirety into this
application.
FIELD OF THE INVENTION
[0002] The present invention is in the field of stenosis diagnosis
and therapy. In particular, the present invention relates to
specific single nucleotide polymorphisms (SNPs) in the human
genome, and their association with stenosis and related
pathologies. Based on differences in allele frequencies in the
stenosis patient population relative to normal individuals, the
naturally-occurring SNPs disclosed herein can be used as targets
for the design of diagnostic reagents and the development of
therapeutic agents, as well as for disease association and linkage
analysis. In particular, the SNPs of the present invention are
useful for identifying an individual who is at an increased or
decreased risk of developing stenosis and for early detection of
the disease, for providing clinically important information for the
prevention and/or treatment of stenosis, and for screening and
selecting therapeutic agents. The SNPs disclosed herein are also
useful for human identification applications.
[0003] Methods, assays, kits, and reagents for detecting the
presence of these polymorphisms and their encoded products are
provided.
BACKGROUND OF THE INVENTION
[0004] Stenosis
[0005] Coronary stenosis is the narrowing of coronary arteries by
obstructive atherosclerotic plaques. The coronary arteries supply
oxygenated blood flow to the myocardium. Although mild and moderate
coronary stenosis do not impede resting coronary flow, stenosis
>30-45% starts to restrict maximal coronary flow. Severe
coronary stenosis (>70% reduction in luminal diameter) causes
stable angina (ischemic chest pain upon exertion). Significant
stenosis contributes, along with plaque rupture and thrombus
formation, coronary spasm, or inflammation/infection, to unstable
angina as well as myocardial infarction. Together with arrhythmia,
coronary stenosis is a major factor of sudden cardiac deaths, as
evidenced by its presence in two or more major coronary arteries in
90% of adult sudden cardiac death victims.
[0006] Coronary stenosis is a prevalent disease. Each year in the
United States, 440,000 new cases of stable angina and 150,000 new
cases of unstable angina occur. This year, an estimated 1.1 million
Americans will have a new or recurrent heart attack. These
incidences result in over six million individuals in the U.S.
living with stable or unstable angina pectoris, a debilitating
condition, and over seven million individuals in the U.S. living
with a history of myocardial infarction. Coronary stenosis is
frequently a deadly disease. It is a major underlying cause of
coronary heart disease (CHD), which is the single largest cause of
death in the U.S. Over half a million coronary deaths, including
250,000 sudden cardiac deaths, occur each year in U.S. Coronary
stenosis is also a costly disease. It is the major reason for 1.2
million cardiac catheterizations, 0.4 million angioplasties, and
0.6 million bypass surgeries, contributing to the estimated 110
billion dollar total costs of CHD in the U.S. for the year 2002.
Still, these statistics underestimate the true prevalence of the
disease since coronary stenosis often remains clinically
asymptomatic for decades, and only becomes symptomatic when the
disease has progressed to a severe, and sometime fatal, state.
[0007] There is therefore an unmet need in early diagnosis and
prognosis of asymptomatic coronary stenosis. This need is
particularly significant given that early diagnosis or prognosis
results can significantly influence the course of disease by
influencing treatment choices (for example, those with genetic
risks can be treated to modify risk factors such as hypertension,
diabetes, inactivity, dyslipidemia, etc.), thresholds (e.g., lipid
levels used to trigger the use of lipid-lowering drugs), and goals
(e.g., target blood pressure or lipid levels), and possibly enhance
compliance.
[0008] Diagnosis of coronary stenosis currently starts by assessing
if the risk profiles (e.g., hypertension, dyslipidemia, family
history, diabetes, etc.) and symptoms (e.g., angina) of patients
are consistent with coronary heart disease, followed most commonly
by resting and exercise EKGs. However, risk assessments and EKGs
are imperfect diagnostic tests for stenosis since they can be both
insensitive (giving false negatives) and non-specific (giving false
positives). Coronary arteriography is the definitive test for
assessing the severity of coronary stenosis, however, it is not
very sensitive in early detection of mild stenosis. It is also an
invasive procedure with a small risk of death due to the
catheterization procedure and the contrast dye. Because of this
risk, it is typically only used at a time when coronary stenosis is
considered likely from symptoms or other tests, which is hardly an
ideal time to start intervention.
[0009] Coronary stenosis risk is presumed to have a strong genetic
component. It is well known that several major risk factors of
coronary disease are heritable, e.g. serum lipid levels (Perusse L.
et. al., Arterioscler Thromb Vasc Biol (1997): 17(11) 3263-9) and
obesity (Rice T. et. al., Int J Obes Relat Metab Disord
(1997):21(11) 1024-31). Indeed, several known genetic defects are
individually sufficient to cause elevated serum LDL-cholesterol
(e.g., familial hypercholesterolemia) leading to premature coronary
disease (Goldstein and Brown, Science 292 (2001): 1310-12). In
addition, linkage studies in humans have replicated the findings of
the link of several chromosomal regions (quantitative trait loci)
to coronary heart disease and related diseases and risk factors
(Pajukanta P. et. al., Am J Hum Genet. 67 (2000):1481-93, Francke
S. et. al., Human Molecular Genetics (2001): (24) 2751-65).
Finally, a family history of premature coronary disease is a
significant factor in the risk assessment and diagnosis of coronary
disease (Braunwald E., Zipes D. and Libby P., Heart Disease,
6.sup.th ed. W.B. Saunders Company, 2001, 28).
[0010] Although many risk factors for coronary stenosis have been
identified, including age, diabetes, hypertension, high serum
cholesterol, smoking, etc., and genetic factors play significant
roles in several of these risk factors, significant genetic risk
factors are likely to exist which have not been identified to date.
In addition to the anecdotal coronary disease patients that exhibit
few traditional risk factors, a study of multiple existing risk
factors showed that only half of the "population-attributable risk"
was attributable to known risk factors (Change M. et. al., J Clin
Epidemiol (2001) 54 (6) 634-44). Therefore, the presently known
risk factors are inadequate for predicting coronary stenosis risk
in individuals. Given the magnitude of the disease, there is an
urgent need for genetic markers that are predictive of coronary
stenosis risk. Such genetic markers could increase the prognostic
ability of existing risk assessment methods and complement current
diagnostic methods such as exercise EKG, especially in early
detection of disease when intervention is most effective and should
ideally start.
[0011] SNPs
[0012] The genomes of all organisms undergo spontaneous mutation in
the course of their continuing evolution, generating variant forms
of progenitor genetic sequences (Gusella, Ann. Rev. Biochem. 55,
831-854 (1986)). A variant form may confer an evolutionary
advantage or disadvantage relative to a progenitor form or may be
neutral. In some instances, a variant form confers an evolutionary
advantage to the species and is eventually incorporated into the
DNA of many or most members of the species and effectively becomes
the progenitor form. Additionally, the effects of a variant form
may be both beneficial and detrimental, depending on the
circumstances. For example, a heterozygous sickle cell mutation
confers resistance to malaria, but a homozygous sickle cell
mutation is usually lethal. In many cases, both progenitor and
variant forms survive and co-exist in a species population. The
coexistence of multiple forms of a genetic sequence gives rise to
genetic polymorphisms, including SNPs.
[0013] Approximately 90% of all polymorphisms in the human genome
are SNPs. SNPs are single base positions in DNA at which different
alleles, or alternative nucleotides, exist in a population. The SNP
position (interchangeably referred to herein as SNP, SNP site, or
SNP locus) is usually preceded by and followed by highly conserved
sequences of the allele (e.g., sequences that vary in less than
1/100 or 1/1000 members of the populations). An individual may be
homozygous or heterozygous for an allele at each SNP position. A
SNP can, in some instances, be referred to as a "cSNP" to denote
that the nucleotide sequence containing the SNP is an amino acid
coding sequence.
[0014] A SNP may arise from a substitution of one nucleotide for
another at the polymorphic site. Substitutions can be transitions
or transversions. A transition is the replacement of one purine
nucleotide by another purine nucleotide, or one pyrimidine by
another pyrimidine. A transversion is the replacement of a purine
by a pyrimidine, or vice versa. A SNP may also be a single base
insertion or deletion variant referred to as an "indel" (Weber et
al., "Human diallelic insertion/deletion polymorphisms", Am J Hum
Genet. 2002 October; 71(4):854-62).
[0015] A synonymous codon change, or silent mutation/SNP (terms
such as "SNP", "polymorphism", "mutation", "mutant", "variation",
and "variant" are used herein interchangeably), is one that does
not result in a change of amino acid due to the degeneracy of the
genetic code. A substitution that changes a codon coding for one
amino acid to a codon coding for a different amino acid (i.e., a
non-synonymous codon change) is referred to as a missense mutation.
A nonsense mutation results in a type of non-synonymous codon
change in which a stop codon is formed, thereby leading to
premature termination of a polypeptide chain and a truncated
protein. A read-through mutation is another type of non-synonymous
codon change that causes the destruction of a stop codon, thereby
resulting in an extended polypeptide product. While SNPs can be
bi-, tri-, or tetra-allelic, the vast majority of the SNPs are
bi-allelic, and are thus often referred to as "bi-allelic markers",
or "di-allelic markers".
[0016] As used herein, references to SNPs and SNP genotypes include
individual SNPs and/or haplotypes, which are groups of SNPs that
are generally inherited together. Haplotypes can have stronger
correlations with diseases or other phenotypic effects compared
with individual SNPs, and therefore may provide increased
diagnostic accuracy in some cases (Stephens et al. Science 293,
489-493, 20 Jul. 2001).
[0017] Causative SNPs are those SNPs that produce alterations in
gene expression or in the expression, structure, and/or function of
a gene product, and therefore are most predictive of a possible
clinical phenotype. One such class includes SNPs falling within
regions of genes encoding a polypeptide product, i.e. cSNPs. These
SNPs may result in an alteration of the amino acid sequence of the
polypeptide product (i.e., non-synonymous codon changes) and give
rise to the expression of a defective or other variant protein.
Furthermore, in the case of nonsense mutations, a SNP may lead to
premature termination of a polypeptide product. Such variant
products can result in a pathological condition, e.g., genetic
disease. Examples of genes in which a SNP within a coding sequence
causes a genetic disease include sickle cell anemia and cystic
fibrosis.
[0018] Causative SNPs do not necessarily have to occur in coding
regions; causative SNPs can occur in, for example, any genetic
region that can ultimately affect the expression, structure, and/or
activity of the protein encoded by a nucleic acid. Such genetic
regions include, for example, those involved in transcription, such
as SNPs in transcription factor binding domains, SNPs in promoter
regions, in areas involved in transcript processing, such as SNPs
at intron-exon boundaries that may cause defective splicing, or
SNPs in mRNA processing signal sequences such as polyadenylation
signal regions. Some SNPs that are not causative SNPs nevertheless
are in close association with, and therefore segregate with, a
disease-causing sequence. In this situation, the presence of a SNP
correlates with the presence of, or predisposition to, or an
increased risk in developing the disease. These SNPs, although not
causative, are nonetheless also useful for diagnostics, disease
predisposition screening, and other uses.
[0019] An association study of a SNP and a specific disorder
involves determining the presence or frequency of the SNP allele in
biological samples from individuals with the disorder of interest,
such as stenosis, and comparing the information to that of controls
(i.e., individuals who do not have the disorder; controls may be
also referred to as "healthy" or "normal" individuals) who are
preferably of similar age and race. The appropriate selection of
patients and controls is important to the success of SNP
association studies. Therefore, a pool of individuals with
well-characterized phenotypes is extremely desirable.
[0020] A SNP may be screened in diseased tissue samples or any
biological sample obtained from a diseased individual, and compared
to control samples, and selected for its increased (or decreased)
occurrence in a specific pathological condition, such as
pathologies related to stenosis. Once a statistically significant
association is established between one or more SNP(s) and a
pathological condition (or other phenotype) of interest, then the
region around the SNP can optionally be thoroughly screened to
identify the causative genetic locus/sequence(s) (e.g., causative
SNP/mutation, gene, regulatory region, etc.) that influences the
pathological condition or phenotype. Association studies may be
conducted within the general population and are not limited to
studies performed on related individuals in affected families
(linkage studies).
[0021] Clinical trials have shown that patient response to
treatment with pharmaceuticals is often heterogeneous. There is a
continuing need to improve pharmaceutical agent design and therapy.
In that regard, SNPs can be used to identify patients most suited
to therapy with particular pharmaceutical agents (this is often
termed "pharmacogenomics"). Similarly, SNPs can be used to exclude
patients from certain treatment due to the patient's increased
likelihood of developing toxic side effects or their likelihood of
not responding to the treatment. Pharmacogenomics can also be used
in pharmaceutical research to assist the drug development and
selection process. (Linder et al. (1997), Clinical Chemistry, 43,
254; Marshall (1997), Nature Biotechnology, 15, 1249; International
Patent Application WO 97/40462, Spectra Biomedical; and Schafer et
al. (1998), Nature Biotechnology, 16, 3).
SUMMARY OF THE INVENTION
[0022] The present invention relates to the identification of novel
SNPs, unique combinations of such SNPs, and haplotypes of SNPs that
are associated with stenosis and related pathologies. The
polymorphisms disclosed herein are directly useful as targets for
the design of diagnostic reagents and the development of
therapeutic agents for use in the diagnosis and treatment of
stenosis and related pathologies.
[0023] Based on the identification of SNPs associated with
stenosis, the present invention also provides methods of detecting
these variants as well as the design and preparation of detection
reagents needed to accomplish this task. The invention specifically
provides novel SNPs in genetic sequences involved in stenosis,
variant proteins encoded by nucleic acid molecules containing such
SNPs, antibodies to the encoded variant proteins, computer-based
and data storage systems containing the novel SNP information,
methods of detecting these SNPs in a test sample, methods of
identifying individuals who have an altered (i.e., increased or
decreased) risk of developing stenosis based on the presence of a
SNP disclosed herein or its encoded product, methods of identifying
individuals who are more or less likely to respond to a treatment,
methods of screening for compounds useful in the treatment of a
disorder associated with a variant gene/protein, compounds
identified by these methods, methods of treating disorders mediated
by a variant gene/protein, and methods of using the novel SNPs of
the present invention for human identification.
[0024] In Tables 1-2, the present invention provides gene
information, transcript sequences (SEQ ID NOS:1-12), encoded amino
acid sequences (SEQ ID NOS:13-24), genomic sequences (SEQ ID
NOS:37-40), transcript-based context sequences (SEQ ID NOS:25-36)
and genomic-based context sequences (SEQ ID NOS:41-44) that contain
the SNPs of the present invention, and extensive SNP information
that includes observed alleles, allele frequencies,
populations/ethnic groups in which alleles have been observed,
information about the type of SNP and corresponding functional
effect, and, for cSNPs, information about the encoded polypeptide
product. The transcript sequences (SEQ ID NOS:1-12), amino acid
sequences (SEQ ID NOS:13-24), genomic sequences (SEQ ID NOS:37-40),
transcript-based SNP context sequences (SEQ ID NOS: 25-36), and
genomic-based SNP context sequences (SEQ ID NOS:41-44) are also
provided in the Sequence Listing.
[0025] In a specific embodiment of the present invention,
naturally-occurring SNPs in the human genome are provided. These
SNPs are associated with stenosis such that they can have a variety
of uses in the diagnosis and/or treatment of stenosis. One aspect
of the present invention relates to an isolated nucleic acid
molecule comprising a nucleotide sequence in which at least one
nucleotide is a SNP disclosed in Tables 3 and/or 4. In an
alternative embodiment, a nucleic acid of the invention is an
amplified polynucleotide, which is produced by amplification of a
SNP-containing nucleic acid template. In another embodiment, the
invention provides for a variant protein which is encoded by a
nucleic acid molecule containing a SNP disclosed herein.
[0026] In yet another embodiment of the invention, a reagent for
detecting a SNP in the context of its naturally-occurring flanking
nucleotide sequences (which can be, e.g., either DNA or mRNA) is
provided. In particular, such a reagent may be in the form of, for
example, a hybridization probe or an amplification primer that is
useful in the specific detection of a SNP of interest. In an
alternative embodiment, a protein detection reagent is used to
detect a variant protein which is encoded by a nucleic acid
molecule containing a SNP disclosed herein. A preferred embodiment
of a protein detection reagent is an antibody or an
antigen-reactive antibody fragment.
[0027] Also provided in the invention are kits comprising SNP
detection reagents, and methods for detecting the SNPs disclosed
herein by employing detection reagents. In a specific embodiment,
the present invention provides for a method of identifying an
individual having an increased or decreased risk of developing
stenosis by detecting the presence or absence of a SNP allele
disclosed herein. In another embodiment, a method for diagnosis of
stenosis by detecting the presence or absence of a SNP allele
disclosed herein is provided.
[0028] The nucleic acid molecules of the invention can be inserted
in an expression vector, such as to produce a variant protein in a
host cell. Thus, the present invention also provides for a vector
comprising a SNP-containing nucleic acid molecule,
genetically-engineered host cells containing the vector, and
methods for expressing a recombinant variant protein using such
host cells. In another specific embodiment, the host cells,
SNP-containing nucleic acid molecules, and/or variant proteins can
be used as targets in a method for screening and identifying
therapeutic agents or pharmaceutical compounds useful in the
treatment of stenosis.
[0029] An aspect of this invention is a method for treating
stenosis in a human subject wherein said human subject harbors a
gene, transcript, and/or encoded protein identified in Tables 1-2,
which method comprises administering to said human subject a
therapeutically or prophylactically effective amount of one or more
agents counteracting the effects of the disease, such as by
inhibiting (or stimulating) the activity of the gene, transcript,
and/or encoded protein identified in Tables 1-2.
[0030] Another aspect of this invention is a method for identifying
an agent useful in therapeutically or prophylactically treating
stenosis in a human subject wherein said human subject harbors a
gene, transcript, and/or encoded protein identified in Tables 1-2,
which method comprises contacting the gene, transcript, or encoded
protein with a candidate agent under conditions suitable to allow
formation of a binding complex between the gene, transcript, or
encoded protein and the candidate agent and detecting the formation
of the binding complex, wherein the presence of the complex
identifies said agent.
[0031] Another aspect of this invention is a method for treating
stenosis in a human subject, which method comprises:
[0032] (i) determining that said human subject harbors a gene,
transcript, and/or encoded protein identified in Tables 1-2,
and
[0033] (ii) administering to said subject a therapeutically or
prophylactically effective amount of one or more agents
counteracting the effects of the disease.
[0034] Many other uses and advantages of the present invention will
be apparent to those skilled in the art upon review of the detailed
description of the preferred embodiments herein. Solely for clarity
of discussion, the invention is described in the sections below by
way of non-limiting examples.
DESCRIPTION OF THE FILES CONTAINED ON THE CD-R NAMED CL1500DIV1
CDR
[0035] The CD-R named CL1500DIV1 CDR contains the following text
(ASCII) file:
[0036] 1) File SEQLIST_CL1500DIV1.txt provides the Sequence
Listing. The Sequence Listing provides the transcript sequences
(SEQ ID NOS:1-12) and protein sequences (SEQ ID NOS:13-24) as shown
in Table 1, and genomic sequences (SEQ ID NOS:37-40) as shown in
Table 2, for each stenosis-associated gene that contains one or
more SNPs of the present invention. Also provided in the Sequence
Listing are context sequences flanking each SNP, including both
transcript-based context sequences as shown in Table 1 (SEQ ID
NOS:25-36) and genomic-based context sequences as shown in Table 2
(SEQ ID NOS:41-44). The context sequences generally provide 100 bp
upstream (5') and 100 bp downstream (3') of each SNP, with the SNP
in the middle of the context sequence, for a total of 200 bp of
context sequence surrounding each SNP. File SEQLIST_CL1500DIV1.txt
is 307 KB in size.
[0037] The material contained on the CD-R labeled "CL 1500DIV1" is
hereby incorporated by reference pursuant to 37 CFR 1.77(b)(4).
DESCRIPTION OF TABLE 1 AND TABLE 2
[0038] Table 1 and Table 2 disclose the SNP and associated
gene/transcript/protein information of the present invention. For
each gene, Table 1 and Table 2 each provide a header containing
gene/transcript/protein information, followed by a transcript and
protein sequence (in Table 1) or genomic sequence (in Table 2), and
then SNP information regarding each SNP found in that
gene/transcript.
[0039] NOTE: SNPs may be included in both Table 1 and Table 2;
Table 1 presents the SNPs relative to their transcript sequences
and encoded protein sequences, whereas Table 2 presents the SNPs
relative to their genomic sequences (in some instances Table 2 may
also include, after the last gene sequence, genomic sequences of
one or more intergenic regions, as well as SNP context sequences
and other SNP information for any SNPs that lie within these
intergenic regions). SNPs can readily be cross-referenced between
Tables based on their hCV (or, in some instances, hDV)
identification numbers.
[0040] The gene/transcript/protein information includes: [0041] a
gene number (1 through n, where n=the total number of genes in the
Table) [0042] a Celera hCG and UID internal identification numbers
for the gene [0043] a Celera hCT and UID internal identification
numbers for the transcript (Table 1 only) [0044] a public Genbank
accession number (e.g., RefSeq NM number) for the transcript (Table
1 only) [0045] a Celera hCP and UID internal identification numbers
for the protein encoded by the hCT transcript (Table 1 only) [0046]
a public Genbank accession number (e.g., RefSeq NP number) for the
protein (Table 1 only) [0047] an art-known gene symbol [0048] an
art-known gene/protein name [0049] Celera genomic axis position
(indicating start nucleotide position-stop nucleotide position)
[0050] the chromosome number of the chromosome on which the gene is
located [0051] an OMIM (Online Mendelian Inheritance in Man; Johns
Hopkins University/NCBI) public reference number for obtaining
further information regarding the medical significance of each gene
[0052] alternative gene/protein name(s) and/or symbol(s) in the
OMIM entry
[0053] NOTE: Due to the presence of alternative splice forms,
multiple transcript/protein entries can be provided for a single
gene entry in Table 1; i.e., for a single Gene Number, multiple
entries may be provided in series that differ in their
transcript/protein information and sequences.
[0054] Following the gene/transcript/protein information is a
transcript sequence and protein sequence (in Table 1), or a genomic
sequence (in Table 2), for each gene, as follows: [0055] transcript
sequence (Table 1 only) (corresponding to SEQ ID NOS:1-12 of the
Sequence Listing), with SNPs identified by their IUB codes
(transcript sequences can include 5' UTR, protein coding, and 3'
UTR regions). (NOTE: If there are differences between the
nucleotide sequence of the hCT transcript and the corresponding
public transcript sequence identified by the Genbank accession
number, the hCT transcript sequence (and encoded protein) is
provided, unless the public sequence is a RefSeq transcript
sequence identified by an NM number, in which case the RefSeq NM
transcript sequence (and encoded protein) is provided. However,
whether the hCT transcript or RefSeq NM transcript is used as the
transcript sequence, the disclosed SNPs are represented by their
IUB codes within the transcript.) [0056] the encoded protein
sequence (Table 1 only) (corresponding to SEQ ID NOS:13-24 of the
Sequence Listing) [0057] the genomic sequence of the gene (Table 2
only), including 6 kb on each side of the gene boundaries (i.e., 6
kb on the 5' side of the gene plus 6 kb on the 3' side of the gene)
(corresponding to SEQ ID NOS:37-40 of the Sequence Listing).
[0058] After the last gene sequence, Table 2 may include additional
genomic sequences of intergenic regions (in such instances, these
sequences are identified as "Intergenic region:" followed by a
numerical identification number), as well as SNP context sequences
and other SNP information for any SNPs that lie within each
intergenic region (and such SNPs are identified as "INTERGENIC" for
SNP type).
[0059] NOTE: The transcript, protein, and transcript-based SNP
context sequences are provided in both Table 1 and in the Sequence
Listing. The genomic and genomic-based SNP context sequences are
provided in both Table 2 and in the Sequence Listing. SEQ ID NOS
are indicated in Table 1 for each transcript sequence (SEQ ID
NOS:1-12), protein sequence (SEQ ID NOS:13-24), and
transcript-based SNP context sequence (SEQ ID NOS:25-36), and SEQ
ID NOS are indicated in Table 2 for each genomic sequence (SEQ ID
NOS:37-40), and genomic-based SNP context sequence (SEQ ID
NOS:41-44).
[0060] The SNP information includes: [0061] context sequence (taken
from the transcript sequence in Table 1, and taken from the genomic
sequence in Table 2) with the SNP represented by its IUB code,
including 100 bp upstream (5') of the SNP position plus 100 bp
downstream (3') of the SNP position (the transcript-based SNP
context sequences in Table 1 are provided in the Sequence Listing
as SEQ ID NOS:25-36; the genomic-based SNP context sequences in
Table 2 are provided in the Sequence Listing as SEQ ID NOS:41-44).
[0062] Celera hCV internal identification number for the SNP (in
some instances, an "hDV" number is given instead of an "hCV"
number) [0063] SNP position [position of the SNP within the given
transcript sequence (Table 1) or within the given genomic sequence
(Table 2)] [0064] SNP source (may include any combination of one or
more of the following five codes, depending on which internal
sequencing projects and/or public databases the SNP has been
observed in: "Applera"=SNP observed during the re-sequencing of
genes and regulatory regions of 39 individuals, "Celera"=SNP
observed during shotgun sequencing and assembly of the Celera human
genome sequence, "Celera Diagnostics"=SNP observed during
re-sequencing of nucleic acid samples from individuals who have
stenosis or a related pathology, "dbSNP"=SNP observed in the dbSNP
public database, "HGBASE"=SNP observed in the HGBASE public
database, "HGMD"=SNP observed in the Human Gene Mutation Database
(HGMD) public database) (NOTE: multiple "Applera" source entries
for a single SNP indicate that the same SNP was covered by multiple
overlapping amplification products and the re-sequencing results
(e.g., observed allele counts) from each of these amplification
products is being provided) [0065] Population/allele/allele count
information in the format of
[population1(allele1,count|allele2,count) population2(allele
1,count|allele2,count) total (allele 1,total count|allele2,total
count)]. The information in this field includes populations/ethnic
groups in which particular SNP alleles have been observed
("cau"=Caucasian, "his"=Hispanic, "chn"=Chinese, and
"afr"=African-American, "jpn"=Japanese, "ind"=Indian,
"mex"=Mexican, "ain"="American Indian, "cra"=Celera donor,
"no_pop"=no population information available), identified SNP
alleles, and observed allele counts (within each population group
and total allele counts), where available ["-" in the allele field
represents a deletion allele of an insertion/deletion ("indel")
polymorphism (in which case the corresponding insertion allele,
which may be comprised of one or more nucleotides, is indicated in
the allele field on the opposite side of the "|"); "-" in the count
field indicates that allele count information is not
available].
[0066] NOTE: For SNPs of "Applera" SNP source, genes/regulatory
regions of 39 individuals (20 Caucasians and 19 African Americans)
were re-sequenced and, since each SNP position is represented by
two chromosomes in each individual (with the exception of SNPs on X
and Y chromosomes in males, for which each SNP position is
represented by a single chromosome), up to 78 chromosomes were
genotyped for each SNP position. Thus, the sum of the
African-American ("afr") allele counts is up to 38, the sum of the
Caucasian allele counts ("cau") is up to 40, and the total sum of
all allele counts is up to 78.
[0067] (NOTE: semicolons separate population/allele/count
information corresponding to each indicated SNP source; i.e., if
four SNP sources are indicated, such as "Celera", "dbSNP",
"HGBASE", and "HGMD", then population/allele/count information is
provided in four groups which are separated by semicolons and
listed in the same order as the listing of SNP sources, with each
population/allele/count information group corresponding to the
respective SNP source based on order; thus, in this example, the
first population/allele/count information group would correspond to
the first listed SNP source (Celera) and the third
population/allele/count information group separated by semicolons
would correspond to the third listed SNP source (HGBASE); if
population/allele/count information is not available for any
particular SNP source, then a pair of semicolons is still inserted
as a place-holder in order to maintain correspondence between the
list of SNP sources and the corresponding listing of
population/allele/count information) [0068] SNP type (e.g.,
location within gene/transcript and/or predicted functional effect)
["MIS-SENSE MUTATION"=SNP causes a change in the encoded amino acid
(i.e., a non-synonymous coding SNP); "SILENT MUTATION"=SNP does not
cause a change in the encoded amino acid (i.e., a synonymous coding
SNP); "STOP CODON MUTATION"=SNP is located in a stop codon;
"NONSENSE MUTATION"=SNP creates or destroys a stop codon; "UTR
5"=SNP is located in a 5' UTR of a transcript; "UTR 3"=SNP is
located in a 3' UTR of a transcript; "PUTATIVE UTR 5"=SNP is
located in a putative 5' UTR; "PUTATIVE UTR 3"=SNP is located in a
putative 3' UTR; "DONOR SPLICE SITE"=SNP is located in a donor
splice site (5' intron boundary); "ACCEPTOR SPLICE SITE"=SNP is
located in an acceptor splice site (3' intron boundary); "CODING
REGION"=SNP is located in a protein-coding region of the
transcript; "EXON"=SNP is located in an exon; "INTRON"=SNP is
located in an intron; "hmCS"=SNP is located in a human-mouse
conserved segment; "TFBS"=SNP is located in a transcription factor
binding site; "UNKNOWN"=SNP type is not defined; "INTERGENIC"=SNP
is intergenic, i.e., outside of any gene boundary] [0069] Protein
coding information (Table 1 only), where relevant, in the format of
[protein SEQ ID NO:#, amino acid position, (amino acid-1, codon1)
(amino acid-2, codon2)]. The information in this field includes SEQ
ID NO of the encoded protein sequence, position of the amino acid
residue within the protein identified by the SEQ ID NO that is
encoded by the codon containing the SNP, amino acids (represented
by one-letter amino acid codes) that are encoded by the alternative
SNP alleles (in the case of stop codons, "X" is used for the
one-letter amino acid code), and alternative codons containing the
alternative SNP nucleotides which encode the amino acid residues
(thus, for example, for missense mutation-type SNPs, at least two
different amino acids and at least two different codons are
generally indicated; for silent mutation-type SNPs, one amino acid
and at least two different codons are generally indicated, etc.).
In instances where the SNP is located outside of a protein-coding
region (e.g., in a UTR region), "None" is indicated following the
protein SEQ ID NO.
DESCRIPTION OF TABLE 3 AND TABLE 4
[0070] Tables 3 and 4 provide a list of a subset of SNPs from Table
1 (in the case of Table 3) or Table 2 (in the case of Table 4) for
which the SNP source falls into one of the following three
categories: 1) SNPs for which the SNP source is only "Applera" and
none other, 2) SNPs for which the SNP source is only "Celera" and
none other, and 3) SNPs for which the SNP source is both "Applera"
and "Celera" but none other.
[0071] These SNPs have not been observed in any of the public
databases (dbSNP, HGBASE, and HGMD), and were also not observed
during shotgun sequencing and assembly of the Celera human genome
sequence (i.e., "Celera" SNP source). Tables 3 and 4 provide the
hCV identification number (or hDV identification number for SNPs
having "Celera" SNP source) and the SEQ ID NO of the context
sequence for each of these SNPs.
DESCRIPTION OF TABLE 5
[0072] Table 5 provides sequences (SEQ ID NOS:45-56) of primers
that have been synthesized and used in the laboratory to carry out
allele-specific PCR reactions in order to assay the SNPs disclosed
in Tables 6-7 during the course of stenosis association
studies.
[0073] Table 5 provides the following: [0074] the column labeled
"hCV" provides an hCV identification number for each SNP site
[0075] the column labeled "Alleles" designates the two alternative
alleles at the SNP site identified by the hCV identification number
that are targeted by the allele-specific primers (the
allele-specific primers are shown as "Sequence A" and "Sequence B"
in each row) [0076] the column labeled "Sequence A (allele-specific
primer)" provides an allele-specific primer that is specific for
the first allele designated in the "Alleles" column [0077] the
column labeled "Sequence B (allele-specific primer)" provides an
allele-specific primer that is specific for the second allele
designated in the "Alleles" column [0078] the column labeled
"Sequence C (common primer)" provides a common primer that is used
in conjunction with each of the allele-specific primers (the
"Sequence A" primer and the "Sequence B" primer) and which
hybridizes at a site away from the SNP position.
[0079] All primer sequences are given in the 5' to 3'
direction.
[0080] Each of the alleles designated in the "Alleles" column
matches the 3' nucleotide of the allele-specific primer that is
specific for that allele. Thus, the first allele designated in the
"Alleles" column matches the 3' nucleotide of the "Sequence A"
primer, and the second allele designated in the "Alleles" column
matches the 3' nucleotide of the "Sequence B" primer.
DESCRIPTION OF TABLE 6 AND TABLE 7
[0081] Tables 6-7 provide results of statistical analyses for SNPs
disclosed in Tables 1-5 (SNPs can be cross-referenced between
Tables based on their hCV identification numbers). The statistical
results provide support for the association of these SNPs with
coronary stenosis.
[0082] NOTE: SNPs can be cross-referenced between Tables 1-7 based
on the hCV identification number of each SNP. However, six of the
SNPs that are included in Tables 1-7 possess two different hCV
identification numbers, as follows: [0083] hCV1129436 is equivalent
to hCV26581155 [0084] hCV15954277 is equivalent to hCV22272408
[0085] hCV16173091 is equivalent to hCV25473098 [0086] hCV16179628
is equivalent to hCV22272980 [0087] hCV16195242 is equivalent to
hCV22274712
[0088] hCV7482175 is equivalent to hCV26546221 TABLE-US-00001
Column heading Definition Marker Internal hCV identification number
for the tested SNP Gene HUGO gene symbol for the gene in which the
SNP Name resides Sample Sample set used in the analysis ("Sample
Set 1" or Set "Sample Set 2") p-value The result of the asymptotic
chi square test for allelic, dominant, or recessive (based on the
genotype reported in the "Mode" column) genotypic association, or
the results of Armitage trendtest for additive genotypic
association. For SNPs for which information is provided in an
"Adjust" column, it is the result of allelic, additive, dominant,
or recessive (based on the genotype reported in the "Mode" column)
p-value of the stratified analysis with Cochran Mantel Haenszel
test. OR Allelic, dominant, recessive, or additive (based on the
genotype reported in Mode column) odds ratio 95% CI 95% confidence
interval of the OR reported Case Frequency of Allele1, or genotype
containing Allele1, in Freq. the case group Cntrl Frequency of
Allele1, or genotype containing Allele1, in Freq. the control group
Allele1 Nucleotide (allele) of the tested SNP for which statistics
are being reported Mode Mode of inheritance for which p-values are
reported: Dom: dominant Rec: recessive Add: Additive Allelic Strata
Stratum in which the association study analysis was based All:
unstratified M: in male F: in female Age T1: age tertile 1 Age T2:
age tertile 2 Age T3: age tertile 3 Smoke+: people who are past or
current smoker Smoke-: people who never smoked MI-: people without
heart attack Adjust Adjustments that were done for the Cochran
Mantel Haenszel test (Indicates that the p-value was determined
using a Cochran Mantel Haenszel test that was adjusted for
confounders)
DETAILED DESCRIPTION OF THE INVENTION
[0089] The present invention provides SNPs associated with
stenosis, nucleic acid molecules containing SNPs, methods and
reagents for the detection of the SNPs disclosed herein, uses of
these SNPs for the development of detection reagents, and assays or
kits that utilize such reagents. The stenosis-associated SNPs
disclosed herein are useful for diagnosing, screening for, and
evaluating predisposition to stenosis and related pathologies in
humans. Furthermore, such SNPs and their encoded products are
useful targets for the development of therapeutic agents.
[0090] A large number of SNPs have been identified from
re-sequencing DNA from 39 individuals, and they are indicated as
"Applera" SNP source in Tables 1-2. Their allele frequencies
observed in each of the Caucasian and African-American ethnic
groups are provided. Additional SNPs included herein were
previously identified during shotgun sequencing and assembly of the
human genome, and they are indicated as "Celera" SNP source in
Tables 1-2. Furthermore, the information provided in Table 1-2,
particularly the allele frequency information obtained from 39
individuals and the identification of the precise position of each
SNP within each gene/transcript, allows haplotypes (i.e., groups of
SNPs that are co-inherited) to be readily inferred. The present
invention encompasses SNP haplotypes, as well as individual
SNPs.
[0091] Thus, the present invention provides individual SNPs
associated with stenosis, as well as combinations of SNPs and
haplotypes in genetic regions associated with stenosis,
polymorphic/variant transcript sequences (SEQ ID NOS:1-12) and
genomic sequences (SEQ ID NOS:37-40) containing SNPs, encoded amino
acid sequences (SEQ ID NOS:13-24), and both transcript-based SNP
context sequences (SEQ ID NOS: 25-36) and genomic-based SNP context
sequences (SEQ ID NOS:41-44) (transcript sequences, protein
sequences, and transcript-based SNP context sequences are provided
in Table 1 and the Sequence Listing; genomic sequences and
genomic-based SNP context sequences are provided in Table 2 and the
Sequence Listing), methods of detecting these polymorphisms in a
test sample, methods of determining the risk of an individual of
having or developing stenosis, methods of screening for compounds
useful for treating disorders associated with a variant
gene/protein such as stenosis, compounds identified by these
screening methods, methods of using the disclosed SNPs to select a
treatment strategy, methods of treating a disorder associated with
a variant gene/protein (i.e., therapeutic methods), and methods of
using the SNPs of the present invention for human
identification.
[0092] The present invention provides novel SNPs associated with
stenosis, as well as SNPs that were previously known in the art,
but were not previously known to be associated with stenosis.
Accordingly, the present invention provides novel compositions and
methods based on the novel SNPs disclosed herein, and also provides
novel methods of using the known, but previously unassociated, SNPs
in methods relating to stenosis (e.g., for diagnosing stenosis,
etc.). In Tables 1-2, known SNPs are identified based on the public
database in which they have been observed, which is indicated as
one or more of the following SNP types: "dbSNP"=SNP observed in
dbSNP, "HGBASE"=SNP observed in HGBASE, and "HGMD"=SNP observed in
the Human Gene Mutation Database (HGMD). Novel SNPs for which the
SNP source is only "Applera" and none other, i.e., those that have
not been observed in any public databases and which were also not
observed during shotgun sequencing and assembly of the Celera human
genome sequence (i.e., "Celera" SNP source), are indicated in
Tables 3-4.
[0093] Particular SNP alleles of the present invention can be
associated with either an increased risk of having or developing
stenosis, or a decreased risk of having or developing stenosis. SNP
alleles that are associated with a decreased risk of having or
developing stenosis may be referred to as "protective" alleles, and
SNP alleles that are associated with an increased risk of having or
developing stenosis may be referred to as "susceptibility" alleles
or "risk factors". Thus, whereas certain SNPs (or their encoded
products) can be assayed to determine whether an individual
possesses a SNP allele that is indicative of an increased risk of
having or developing stenosis (i.e., a susceptibility allele),
other SNPs (or their encoded products) can be assayed to determine
whether an individual possesses a SNP allele that is indicative of
a decreased risk of having or developing stenosis (i.e., a
protective allele). Similarly, particular SNP alleles of the
present invention can be associated with either an increased or
decreased likelihood of responding to a particular treatment or
therapeutic compound, or an increased or decreased likelihood of
experiencing toxic effects from a particular treatment or
therapeutic compound. The term "altered" may be used herein to
encompass either of these two possibilities (e.g., an increased or
a decreased risk/likelihood).
[0094] Those skilled in the art will readily recognize that nucleic
acid molecules may be double-stranded molecules and that reference
to a particular site on one strand refers, as well, to the
corresponding site on a complementary strand. In defining a SNP
position, SNP allele, or nucleotide sequence, reference to an
adenine, a thymine (uridine), a cytosine, or a guanine at a
particular site on one strand of a nucleic acid molecule also
defines the thymine (uridine), adenine, guanine, or cytosine
(respectively) at the corresponding site on a complementary strand
of the nucleic acid molecule. Thus, reference may be made to either
strand in order to refer to a particular SNP position, SNP allele,
or nucleotide sequence. Probes and primers, may be designed to
hybridize to either strand and SNP genotyping methods disclosed
herein may generally target either strand. Throughout the
specification, in identifying a SNP position, reference is
generally made to the protein-encoding strand, only for the purpose
of convenience.
[0095] References to variant peptides, polypeptides, or proteins of
the present invention include peptides, polypeptides, proteins, or
fragments thereof, that contain at least one amino acid residue
that differs from the corresponding amino acid sequence of the
art-known peptide/polypeptide/protein (the art-known protein may be
interchangeably referred to as the "wild-type", "reference", or
"normal" protein). Such variant peptides/polypeptides/proteins can
result from a codon change caused by a nonsynonymous nucleotide
substitution at a protein-coding SNP position (i.e., a missense
mutation) disclosed by the present invention. Variant
peptides/polypeptides/proteins of the present invention can also
result from a nonsense mutation, i.e. a SNP that creates a
premature stop codon, a SNP that generates a read-through mutation
by abolishing a stop codon, or due to any SNP disclosed by the
present invention that otherwise alters the structure,
function/activity, or expression of a protein, such as a SNP in a
regulatory region (e.g. a promoter or enhancer) or a SNP that leads
to alternative or defective splicing, such as a SNP in an intron or
a SNP at an exon/intron boundary. As used herein, the terms
"polypeptide", "peptide", and "protein" are used
interchangeably.
Isolated Nucleic Acid Molecules and SNP Detection Reagents &
Kits
[0096] Tables 1 and 2 provide a variety of information about each
SNP of the present invention that is associated with stenosis,
including the transcript sequences (SEQ ID NOS:1-12), genomic
sequences (SEQ ID NOS:37-40), and protein sequences (SEQ ID
NOS:13-24) of the encoded gene products (with the SNPs indicated by
IUB codes in the nucleic acid sequences). In addition, Tables 1 and
2 include SNP context sequences, which generally include 100
nucleotide upstream (5') plus 100 nucleotides downstream (3') of
each SNP position (SEQ ID NOS:25-36 correspond to transcript-based
SNP context sequences disclosed in Table 1, and SEQ ID NOS:41-44
correspond to genomic-based context sequences disclosed in Table
2), the alternative nucleotides (alleles) at each SNP position, and
additional information about the variant where relevant, such as
SNP type (coding, missense, splice site, UTR, etc.), human
populations in which the SNP was observed, observed allele
frequencies, information about the encoded protein, etc.
[0097] Isolated Nucleic Acid Molecules
[0098] The present invention provides isolated nucleic acid
molecules that contain one or more SNPs disclosed Table 1 and/or
Table 2. Preferred isolated nucleic acid molecules contain one or
more SNPs identified in Table 3 and/or Table 4. Isolated nucleic
acid molecules containing one or more SNPs disclosed in at least
one of Tables 1-4 may be interchangeably referred to throughout the
present text as "SNP-containing nucleic acid molecules". Isolated
nucleic acid molecules may optionally encode a full-length variant
protein or fragment thereof. The isolated nucleic acid molecules of
the present invention also include probes and primers (which are
described in greater detail below in the section entitled "SNP
Detection Reagents"), which may be used for assaying the disclosed
SNPs, and isolated full-length genes, transcripts, cDNA molecules,
and fragments thereof, which may be used for such purposes as
expressing an encoded protein.
[0099] As used herein, an "isolated nucleic acid molecule"
generally is one that contains a SNP of the present invention or
one that hybridizes to such molecule such as a nucleic acid with a
complementary sequence, and is separated from most other nucleic
acids present in the natural source of the nucleic acid molecule.
Moreover, an "isolated" nucleic acid molecule, such as a cDNA
molecule containing a SNP of the present invention, can be
substantially free of other cellular material, or culture medium
when produced by recombinant techniques, or chemical precursors or
other chemicals when chemically synthesized. A nucleic acid
molecule can be fused to other coding or regulatory sequences and
still be considered "isolated". Nucleic acid molecules present in
non-human transgenic animals, which do not naturally occur in the
animal, are also considered "isolated". For example, recombinant
DNA molecules contained in a vector are considered "isolated".
Further examples of "isolated" DNA molecules include recombinant
DNA molecules maintained in heterologous host cells, and purified
(partially or substantially) DNA molecules in solution. Isolated
RNA molecules include in vivo or in vitro RNA transcripts of the
isolated SNP-containing DNA molecules of the present invention.
Isolated nucleic acid molecules according to the present invention
further include such molecules produced synthetically.
[0100] Generally, an isolated SNP-containing nucleic acid molecule
comprises one or more SNP positions disclosed by the present
invention with flanking nucleotide sequences on either side of the
SNP positions. A flanking sequence can include nucleotide residues
that are naturally associated with the SNP site and/or heterologous
nucleotide sequences. Preferably the flanking sequence is up to
about 500, 300, 100, 60, 50, 30, 25, 20, 15, 10, 8, or 4
nucleotides (or any other length in-between) on either side of a
SNP position, or as long as the full-length gene or entire
protein-coding sequence (or any portion thereof such as an exon),
especially if the SNP-containing nucleic acid molecule is to be
used to produce a protein or protein fragment.
[0101] For full-length genes and entire protein-coding sequences, a
SNP flanking sequence can be, for example, up to about 5 KB, 4 KB,
3 KB, 2 KB, 1 KB on either side of the SNP. Furthermore, in such
instances, the isolated nucleic acid molecule comprises exonic
sequences (including protein-coding and/or non-coding exonic
sequences), but may also include intronic sequences. Thus, any
protein coding sequence may be either contiguous or separated by
introns. The important point is that the nucleic acid is isolated
from remote and unimportant flanking sequences and is of
appropriate length such that it can be subjected to the specific
manipulations or uses described herein such as recombinant protein
expression, preparation of probes and primers for assaying the SNP
position, and other uses specific to the SNP-containing nucleic
acid sequences.
[0102] An isolated SNP-containing nucleic acid molecule can
comprise, for example, a full-length gene or transcript, such as a
gene isolated from genomic DNA (e.g., by cloning or PCR
amplification), a cDNA molecule, or an mRNA transcript molecule.
Polymorphic transcript sequences are provided in Table 1 and in the
Sequence Listing (SEQ ID NOS: 1-12), and polymorphic genomic
sequences are provided in Table 2 and in the Sequence Listing (SEQ
ID NOS:37-40). Furthermore, fragments of such full-length genes and
transcripts that contain one or more SNPs disclosed herein are also
encompassed by the present invention, and such fragments may be
used, for example, to express any part of a protein, such as a
particular functional domain or an antigenic epitope.
[0103] Thus, the present invention also encompasses fragments of
the nucleic acid sequences provided in Tables 1-2 (transcript
sequences are provided in Table 1 as SEQ ID NOS:1-12, genomic
sequences are provided in Table 2 as SEQ ID NOS:37-40,
transcript-based SNP context sequences are provided in Table 1 as
SEQ ID NO:25-36, and genomic-based SNP context sequences are
provided in Table 2 as SEQ ID NO:41-44) and their complements. A
fragment typically comprises a contiguous nucleotide sequence at
least about 8 or more nucleotides, more preferably at least about
12 or more nucleotides, and even more preferably at least about 16
or more nucleotides. Further, a fragment could comprise at least
about 18, 20, 22, 25, 30, 40, 50, 60, 100, 250 or 500 (or any other
number in-between) nucleotides in length. The length of the
fragment will be based on its intended use. For example, the
fragment can encode epitope-bearing regions of a variant peptide or
regions of a variant peptide that differ from the normal/wild-type
protein, or can be useful as a polynucleotide probe or primer. Such
fragments can be isolated using the nucleotide sequences provided
in Table 1 and/or Table 2 for the synthesis of a polynucleotide
probe. A labeled probe can then be used, for example, to screen a
cDNA library, genomic DNA library, or mRNA to isolate nucleic acid
corresponding to the coding region. Further, primers can be used in
amplification reactions, such as for purposes of assaying one or
more SNPs sites or for cloning specific regions of a gene.
[0104] An isolated nucleic acid molecule of the present invention
further encompasses a SNP-containing polynucleotide that is the
product of any one of a variety of nucleic acid amplification
methods, which are used to increase the copy numbers of a
polynucleotide of interest in a nucleic acid sample. Such
amplification methods are well known in the art, and they include
but are not limited to, polymerase chain reaction (PCR) (U.S. Pat.
Nos. 4,683,195; and 4,683,202; PCR Technology: Principles and
Applications for DNA Amplification, ed. H.A. Erlich, Freeman Press,
NY, N.Y., 1992), ligase chain reaction (LCR) (Wu and Wallace,
Genomics 4:560, 1989; Landegren et al., Science 241:1077, 1988),
strand displacement amplification (SDA) (U.S. Pat. Nos. 5,270,184;
and 5,422,252), transcription-mediated amplification (TMA) (U.S.
Pat. No. 5,399,491), linked linear amplification (LLA) (U.S. Pat.
No. 6,027,923), and the like, and isothermal amplification methods
such as nucleic acid sequence based amplification (NASBA), and
self-sustained sequence replication (Guatelli et al., Proc. Natl.
Acad. Sci. USA 87: 1874, 1990). Based on such methodologies, a
person skilled in the art can readily design primers in any
suitable regions 5' and 3' to a SNP disclosed herein. Such primers
may be used to amplify DNA of any length so long that it contains
the SNP of interest in its sequence.
[0105] As used herein, an "amplified polynucleotide" of the
invention is a SNP-containing nucleic acid molecule whose amount
has been increased at least two fold by any nucleic acid
amplification method performed in vitro as compared to its starting
amount in a test sample. In other preferred embodiments, an
amplified polynucleotide is the result of at least ten fold, fifty
fold, one hundred fold, one thousand fold, or even ten thousand
fold increase as compared to its starting amount in a test sample.
In a typical PCR amplification, a polynucleotide of interest is
often amplified at least fifty thousand fold in amount over the
unamplified genomic DNA, but the precise amount of amplification
needed for an assay depends on the sensitivity of the subsequent
detection method used.
[0106] Generally, an amplified polynucleotide is at least about 16
nucleotides in length. More typically, an amplified polynucleotide
is at least about 20 nucleotides in length. In a preferred
embodiment of the invention, an amplified polynucleotide is at
least about 30 nucleotides in length. In a more preferred
embodiment of the invention, an amplified polynucleotide is at
least about 32, 40, 45, 50, or 60 nucleotides in length. In yet
another preferred embodiment of the invention, an amplified
polynucleotide is at least about 100, 200, or 300 nucleotides in
length. While the total length of an amplified polynucleotide of
the invention can be as long as an exon, an intron or the entire
gene where the SNP of interest resides, an amplified product is
typically no greater than about 1,000 nucleotides in length
(although certain amplification methods may generate amplified
products greater than 1000 nucleotides in length). More preferably,
an amplified polynucleotide is not greater than about 600
nucleotides in length. It is understood that irrespective of the
length of an amplified polynucleotide, a SNP of interest may be
located anywhere along its sequence.
[0107] In a specific embodiment of the invention, the amplified
product is at least about 201 nucleotides in length, comprises one
of the transcript-based context sequences or the genomic-based
context sequences shown in Tables 1-2. Such a product may have
additional sequences on its 5' end or 3' end or both. In another
embodiment, the amplified product is about 101 nucleotides in
length, and it contains a SNP disclosed herein. Preferably, the SNP
is located at the middle of the amplified product (e.g., at
position 101 in an amplified product that is 201 nucleotides in
length, or at position 51 in an amplified product that is 101
nucleotides in length), or within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
12, 15, or 20 nucleotides from the middle of the amplified product
(however, as indicated above, the SNP of interest may be located
anywhere along the length of the amplified product).
[0108] The present invention provides isolated nucleic acid
molecules that comprise, consist of, or consist essentially of one
or more polynucleotide sequences that contain one or more SNPs
disclosed herein, complements thereof, and SNP-containing fragments
thereof.
[0109] Accordingly, the present invention provides nucleic acid
molecules that consist of any of the nucleotide sequences shown in
Table 1 and/or Table 2 (transcript sequences are provided in Table
1 as SEQ ID NOS:1-12, genomic sequences are provided in Table 2 as
SEQ ID NOS:37-40, transcript-based SNP context sequences are
provided in Table 1 as SEQ ID NO:25-36, and genomic-based SNP
context sequences are provided in Table 2 as SEQ ID NO:41-44), or
any nucleic acid molecule that encodes any of the variant proteins
provided in Table 1 (SEQ ID NOS:13-24). A nucleic acid molecule
consists of a nucleotide sequence when the nucleotide sequence is
the complete nucleotide sequence of the nucleic acid molecule.
[0110] The present invention further provides nucleic acid
molecules that consist essentially of any of the nucleotide
sequences shown in Table 1 and/or Table 2 (transcript sequences are
provided in Table 1 as SEQ ID NOS:1-12, genomic sequences are
provided in Table 2 as SEQ ID NOS:37-40, transcript-based SNP
context sequences are provided in Table 1 as SEQ ID NO:25-36, and
genomic-based SNP context sequences are provided in Table 2 as SEQ
ID NO:41-44), or any nucleic acid molecule that encodes any of the
variant proteins provided in Table 1 (SEQ ID NOS:13-24). A nucleic
acid molecule consists essentially of a nucleotide sequence when
such a nucleotide sequence is present with only a few additional
nucleotide residues in the final nucleic acid molecule.
[0111] The present invention further provides nucleic acid
molecules that comprise any of the nucleotide sequences shown in
Table 1 and/or Table 2 or a SNP-containing fragment thereof
(transcript sequences are provided in Table 1 as SEQ ID NOS:1-12,
genomic sequences are provided in Table 2 as SEQ ID NOS:37-40,
transcript-based SNP context sequences are provided in Table 1 as
SEQ ID NO:25-36, and genomic-based SNP context sequences are
provided in Table 2 as SEQ ID NO:41-44), or any nucleic acid
molecule that encodes any of the variant proteins provided in Table
1 (SEQ ID NOS:13-24). A nucleic acid molecule comprises a
nucleotide sequence when the nucleotide sequence is at least part
of the final nucleotide sequence of the nucleic acid molecule. In
such a fashion, the nucleic acid molecule can be only the
nucleotide sequence or have additional nucleotide residues, such as
residues that are naturally associated with it or heterologous
nucleotide sequences. Such a nucleic acid molecule can have one to
a few additional nucleotides or can comprise many more additional
nucleotides. A brief description of how various types of these
nucleic acid molecules can be readily made and isolated is provided
below, and such techniques are well known to those of ordinary
skill in the art (Sambrook and Russell, 2000, Molecular Cloning: A
Laboratory Manual, Cold Spring Harbor Press, NY).
[0112] The isolated nucleic acid molecules can encode mature
proteins plus additional amino or carboxyl-terminal amino acids or
both, or amino acids interior to the mature peptide (when the
mature form has more than one peptide chain, for instance). Such
sequences may play a role in processing of a protein from precursor
to a mature form, facilitate protein trafficking, prolong or
shorten protein half-life, or facilitate manipulation of a protein
for assay or production. As generally is the case in situ, the
additional amino acids may be processed away from the mature
protein by cellular enzymes.
[0113] Thus, the isolated nucleic acid molecules include, but are
not limited to, nucleic acid molecules having a sequence encoding a
peptide alone, a sequence encoding a mature peptide and additional
coding sequences such as a leader or secretory sequence (e.g., a
pre-pro or pro-protein sequence), a sequence encoding a mature
peptide with or without additional coding sequences, plus
additional non-coding sequences, for example introns and non-coding
5' and 3' sequences such as transcribed but untranslated sequences
that play a role in, for example, transcription, mRNA processing
(including splicing and polyadenylation signals), ribosome binding,
and/or stability of mRNA. In addition, the nucleic acid molecules
may be fused to heterologous marker sequences encoding, for
example, a peptide that facilitates purification.
[0114] Isolated nucleic acid molecules can be in the form of RNA,
such as mRNA, or in the form DNA, including cDNA and genomic DNA,
which may be obtained, for example, by molecular cloning or
produced by chemical synthetic techniques or by a combination
thereof (Sambrook and Russell, 2000, Molecular Cloning: A
Laboratory Manual, Cold Spring Harbor Press, NY). Furthermore,
isolated nucleic acid molecules, particularly SNP detection
reagents such as probes and primers, can also be partially or
completely in the form of one or more types of nucleic acid
analogs, such as peptide nucleic acid (PNA) (U.S. Pat. Nos.
5,539,082; 5,527,675; 5,623,049; 5,714,331). The nucleic acid,
especially DNA, can be double-stranded or single-stranded.
Single-stranded nucleic acid can be the coding strand (sense
strand) or the complementary non-coding strand (anti-sense strand).
DNA, RNA, or PNA segments can be assembled, for example, from
fragments of the human genome (in the case of DNA or RNA) or single
nucleotides, short oligonucleotide linkers, or from a series of
oligonucleotides, to provide a synthetic nucleic acid molecule.
Nucleic acid molecules can be readily synthesized using the
sequences provided herein as a reference; oligonucleotide and PNA
oligomer synthesis techniques are well known in the art (see, e.g.,
Corey, "Peptide nucleic acids: expanding the scope of nucleic acid
recognition", Trends Biotechnol. 1997 June; 15(6):224-9, and Hyrup
et al., "Peptide nucleic acids (PNA): synthesis, properties and
potential applications", Bioorg Med. Chem. 1996 January;
4(1):5-23). Furthermore, large-scale automated oligonucleotide/PNA
synthesis (including synthesis on an array or bead surface or other
solid support) can readily be accomplished using commercially
available nucleic acid synthesizers, such as the Applied Biosystems
(Foster City, Calif.) 3900 High-Throughput DNA Synthesizer or
Expedite 8909 Nucleic Acid Synthesis System, and the sequence
information provided herein.
[0115] The present invention encompasses nucleic acid analogs that
contain modified, synthetic, or non-naturally occurring nucleotides
or structural elements or other alternative/modified nucleic acid
chemistries known in the art. Such nucleic acid analogs are useful,
for example, as detection reagents (e.g., primers/probes) for
detecting one or more SNPs identified in Table 1 and/or Table 2.
Furthermore, kits/systems (such as beads, arrays, etc.) that
include these analogs are also encompassed by the present
invention. For example, PNA oligomers that are based on the
polymorphic sequences of the present invention are specifically
contemplated. PNA oligomers are analogs of DNA in which the
phosphate backbone is replaced with a peptide-like backbone
(Lagriffoul et al., Bioorganic & Medicinal Chemistry Letters,
4: 1081-1082 (1994), Petersen et al., Bioorganic & Medicinal
Chemistry Letters, 6: 793-796 (1996), Kumar et al., Organic Letters
3(9): 1269-1272 (2001), WO96/04000). PNA hybridizes to
complementary RNA or DNA with higher affinity and specificity than
conventional oligonucleotides and oligonucleotide analogs. The
properties of PNA enable novel molecular biology and biochemistry
applications unachievable with traditional oligonucleotides and
peptides.
[0116] Additional examples of nucleic acid modifications that
improve the binding properties and/or stability of a nucleic acid
include the use of base analogs such as inosine, intercalators
(U.S. Pat. No. 4,835,263) and the minor groove binders (U.S. Pat.
No. 5,801,115). Thus, references herein to nucleic acid molecules,
SNP-containing nucleic acid molecules, SNP detection reagents
(e.g., probes and primers), oligonucleotides/polynucleotides
include PNA oligomers and other nucleic acid analogs. Other
examples of nucleic acid analogs and alternative/modified nucleic
acid chemistries known in the art are described in Current
Protocols in Nucleic Acid Chemistry, John Wiley & Sons, N.Y.
(2002).
[0117] The present invention further provides nucleic acid
molecules that encode fragments of the variant polypeptides
disclosed herein as well as nucleic acid molecules that encode
obvious variants of such variant polypeptides. Such nucleic acid
molecules may be naturally occurring, such as paralogs (different
locus) and orthologs (different organism), or may be constructed by
recombinant DNA methods or by chemical synthesis. Non-naturally
occurring variants may be made by mutagenesis techniques, including
those applied to nucleic acid molecules, cells, or organisms.
Accordingly, the variants can contain nucleotide substitutions,
deletions, inversions and insertions (in addition to the SNPs
disclosed in Tables 1-2). Variation can occur in either or both the
coding and non-coding regions. The variations can produce
conservative and/or non-conservative amino acid substitutions.
[0118] Further variants of the nucleic acid molecules disclosed in
Tables 1-2, such as naturally occurring allelic variants (as well
as orthologs and paralogs) and synthetic variants produced by
mutagenesis techniques, can be identified and/or produced using
methods well known in the art. Such further variants can comprise a
nucleotide sequence that shares at least 70-80%, 80-85%, 85-90%,
91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity
with a nucleic acid sequence disclosed in Table 1 and/or Table 2
(or a fragment thereof) and that includes a novel SNP allele
disclosed in Table 1 and/or Table 2. Further, variants can comprise
a nucleotide sequence that encodes a polypeptide that shares at
least 70-80%, 80-85%, 85-90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, or 99% sequence identity with a polypeptide sequence disclosed
in Table 1 (or a fragment thereof) and that includes a novel SNP
allele disclosed in Table 1 and/or Table 2. Thus, the present
invention specifically contemplates isolated nucleic acid molecule
that have a certain degree of sequence variation compared with the
sequences shown in Tables 1-2, but that contain a novel SNP allele
disclosed herein. In other words, as long as an isolated nucleic
acid molecule contains a novel SNP allele disclosed herein, other
portions of the nucleic acid molecule that flank the novel SNP
allele can vary to some degree from the specific transcript,
genomic, and context sequences shown in Tables 1-2, and can encode
a polypeptide that varies to some degree from the specific
polypeptide sequences shown in Table 1.
[0119] To determine the percent identity of two amino acid
sequences or two nucleotide sequences of two molecules that share
sequence homology, the sequences are aligned for optimal comparison
purposes (e.g., gaps can be introduced in one or both of a first
and a second amino acid or nucleic acid sequence for optimal
alignment and non-homologous sequences can be disregarded for
comparison purposes). In a preferred embodiment, at least 30%, 40%,
50%, 60%, 70%, 80%, or 90% or more of the length of a reference
sequence is aligned for comparison purposes. The amino acid
residues or nucleotides at corresponding amino acid positions or
nucleotide positions are then compared. When a position in the
first sequence is occupied by the same amino acid residue or
nucleotide as the corresponding position in the second sequence,
then the molecules are identical at that position (as used herein,
amino acid or nucleic acid "identity" is equivalent to amino acid
or nucleic acid "homology"). The percent identity between the two
sequences is a function of the number of identical positions shared
by the sequences, taking into account the number of gaps, and the
length of each gap, which need to be introduced for optimal
alignment of the two sequences.
[0120] The comparison of sequences and determination of percent
identity between two sequences can be accomplished using a
mathematical algorithm. (Computational Molecular Biology, Lesk, A.
M., ed., Oxford University Press, New York, 1988; Biocomputing:
Informatics and Genome Projects, Smith, D. W., ed., Academic Press,
New York, 1993; Computer Analysis of Sequence Data, Part 1,
Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey,
1994; Sequence Analysis in Molecular Biology, von Heinje, G.,
Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M.
and Devereux, J., eds., M Stockton Press, New York, 1991). In a
preferred embodiment, the percent identity between two amino acid
sequences is determined using the Needleman and Wunsch algorithm
(J. Mol. Biol. (48):444-453 (1970)) which has been incorporated
into the GAP program in the GCG software package, using either a
Blossom 62 matrix or a PAM250 matrix, and a gap weight of 16, 14,
12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.
[0121] In yet another preferred embodiment, the percent identity
between two nucleotide sequences is determined using the GAP
program in the GCG software package (Devereux, J., et al., Nucleic
Acids Res. 12(1):387 (1984)), using a NWSgapdna.CMP matrix and a
gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3,
4, 5, or 6. In another embodiment, the percent identity between two
amino acid or nucleotide sequences is determined using the
algorithm of E. Myers and W. Miller (CABIOS, 4:11-17 (1989)) which
has been incorporated into the ALIGN program (version 2.0), using a
PAM 120 weight residue table, a gap length penalty of 12, and a gap
penalty of 4.
[0122] The nucleotide and amino acid sequences of the present
invention can further be used as a "query sequence" to perform a
search against sequence databases to, for example, identify other
family members or related sequences. Such searches can be performed
using the NBLAST and XBLAST programs (version 2.0) of Altschul, et
al. (J. Mol. Biol. 215:403-10 (1990)). BLAST nucleotide searches
can be performed with the NBLAST program, score=100, wordlength=12
to obtain nucleotide sequences homologous to the nucleic acid
molecules of the invention. BLAST protein searches can be performed
with the XBLAST program, score=50, wordlength=3 to obtain amino
acid sequences homologous to the proteins of the invention. To
obtain gapped alignments for comparison purposes, Gapped BLAST can
be utilized as described in Altschul et al. (Nucleic Acids Res.
25(17):3389-3402 (1997)). When utilizing BLAST and gapped BLAST
programs, the default parameters of the respective programs (e.g.,
XBLAST and NBLAST) can be used. In addition to BLAST, examples of
other search and sequence comparison programs used in the art
include, but are not limited to, FASTA (Pearson, Methods Mol. Biol.
25, 365-389 (1994)) and KERR (Dufresne et al., Nat Biotechnol 2002
December; 20(12):1269-71). For further information regarding
bioinformatics techniques, see Current Protocols in Bioinformatics,
John Wiley & Sons, Inc., N.Y.
[0123] The present invention further provides non-coding fragments
of the nucleic acid molecules disclosed in Table 1 and/or Table 2.
Preferred non-coding fragments include, but are not limited to,
promoter sequences, enhancer sequences, intronic sequences, 5'
untranslated regions (UTRs), 3' untranslated regions, gene
modulating sequences and gene termination sequences. Such fragments
are useful, for example, in controlling heterologous gene
expression and in developing screens to identify gene-modulating
agents.
[0124] SNP Detection Reagents
[0125] In a specific aspect of the present invention, the SNPs
disclosed in Table 1 and/or Table 2, and their associated
transcript sequences (provided in Table 1 as SEQ ID NOS:1-12),
genomic sequences (provided in Table 2 as SEQ ID NOS:37-40), and
context sequences (transcript-based context sequences are provided
in Table 1 as SEQ ID NOS:25-36; genomic-based context sequences are
provided in Table 2 as SEQ ID NOS:41-44), can be used for the
design of SNP detection reagents. As used herein, a "SNP detection
reagent" is a reagent that specifically detects a specific target
SNP position disclosed herein, and that is preferably specific for
a particular nucleotide (allele) of the target SNP position (i.e.,
the detection reagent preferably can differentiate between
different alternative nucleotides at a target SNP position, thereby
allowing the identity of the nucleotide present at the target SNP
position to be determined). Typically, such detection reagent
hybridizes to a target SNP-containing nucleic acid molecule by
complementary base-pairing in a sequence specific manner, and
discriminates the target variant sequence from other nucleic acid
sequences such as an art-known form in a test sample. An example of
a detection reagent is a probe that hybridizes to a target nucleic
acid containing one or more of the SNPs provided in Table 1 and/or
Table 2. In a preferred embodiment, such a probe can differentiate
between nucleic acids having a particular nucleotide (allele) at a
target SNP position from other nucleic acids that have a different
nucleotide at the same target SNP position. In addition, a
detection reagent may hybridize to a specific region 5' and/or 3'
to a SNP position, particularly a region corresponding to the
context sequences provided in Table 1 and/or Table 2
(transcript-based context sequences are provided in Table 1 as SEQ
ID NOS:25-36; genomic-based context sequences are provided in Table
2 as SEQ ID NOS:41-44). Another example of a detection reagent is a
primer which acts as an initiation point of nucleotide extension
along a complementary strand of a target polynucleotide. The SNP
sequence information provided herein is also useful for designing
primers, e.g. allele-specific primers, to amplify (e.g., using PCR)
any SNP of the present invention.
[0126] In one preferred embodiment of the invention, a SNP
detection reagent is an isolated or synthetic DNA or RNA
polynucleotide probe or primer or PNA oligomer, or a combination of
DNA, RNA and/or PNA, that hybridizes to a segment of a target
nucleic acid molecule containing a SNP identified in Table 1 and/or
Table 2. A detection reagent in the form of a polynucleotide may
optionally contain modified base analogs, intercalators or minor
groove binders. Multiple detection reagents such as probes may be,
for example, affixed to a solid support (e.g., arrays or beads) or
supplied in solution (e.g., probe/primer sets for enzymatic
reactions such as PCR, RT-PCR, TaqMan assays, or primer-extension
reactions) to form a SNP detection kit.
[0127] A probe or primer typically is a substantially purified
oligonucleotide or PNA oligomer. Such oligonucleotide typically
comprises a region of complementary nucleotide sequence that
hybridizes under stringent conditions to at least about 8, 10, 12,
16, 18, 20, 22, 25, 30, 40, 50, 60, 100 (or any other number
in-between) or more consecutive nucleotides in a target nucleic
acid molecule. Depending on the particular assay, the consecutive
nucleotides can either include the target SNP position, or be a
specific region in close enough proximity 5' and/or 3' to the SNP
position to carry out the desired assay.
[0128] Other preferred primer and probe sequences can readily be
determined using the transcript sequences (SEQ ID NOS:1-12),
genomic sequences (SEQ ID NOS:37-40), and SNP context sequences
(transcript-based context sequences are provided in Table 1 as SEQ
ID NOS:25-36; genomic-based context sequences are provided in Table
2 as SEQ ID NOS:41-44) disclosed in the Sequence Listing and in
Tables 1-2. It will be apparent to one of skill in the art that
such primers and probes are directly useful as reagents for
genotyping the SNPs of the present invention, and can be
incorporated into any kit/system format.
[0129] In order to produce a probe or primer specific for a target
SNP-containing sequence, the gene/transcript and/or context
sequence surrounding the SNP of interest is typically examined
using a computer algorithm which starts at the 5' or at the 3' end
of the nucleotide sequence. Typical algorithms will then identify
oligomers of defined length that are unique to the gene/SNP context
sequence, have a GC content within a range suitable for
hybridization, lack predicted secondary structure that may
interfere with hybridization, and/or possess other desired
characteristics or that lack other undesired characteristics.
[0130] A primer or probe of the present invention is typically at
least about 8 nucleotides in length.
[0131] In one embodiment of the invention, a primer or a probe is
at least about 10 nucleotides in length. In a preferred embodiment,
a primer or a probe is at least about 12 nucleotides in length. In
a more preferred embodiment, a primer or probe is at least about
16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 nucleotides in length.
While the maximal length of a probe can be as long as the target
sequence to be detected, depending on the type of assay in which it
is employed, it is typically less than about 50, 60, 65, or 70
nucleotides in length. In the case of a primer, it is typically
less than about 30 nucleotides in length. In a specific preferred
embodiment of the invention, a primer or a probe is within the
length of about 18 and about 28 nucleotides. However, in other
embodiments, such as nucleic acid arrays and other embodiments in
which probes are affixed to a substrate, the probes can be longer,
such as on the order of 30-70, 75, 80, 90, 100, or more nucleotides
in length (see the section below entitled "SNP Detection Kits and
Systems").
[0132] For analyzing SNPs, it may be appropriate to use
oligonucleotides specific for alternative SNP alleles. Such
oligonucleotides which detect single nucleotide variations in
target sequences may be referred to by such terms as
"allele-specific oligonucleotides", "allele-specific probes", or
"allele-specific primers". The design and use of allele-specific
probes for analyzing polymorphisms is described in, e.g., Mutation
Detection A Practical Approach, ed. Cotton et al. Oxford University
Press, 1998; Saiki et al., Nature 324, 163-166 (1986); Dattagupta,
EP235,726; and Saiki, WO 89/11548.
[0133] While the design of each allele-specific primer or probe
depends on variables such as the precise composition of the
nucleotide sequences flanking a SNP position in a target nucleic
acid molecule, and the length of the primer or probe, another
factor in the use of primers and probes is the stringency of the
condition under which the hybridization between the probe or primer
and the target sequence is performed. Higher stringency conditions
utilize buffers with lower ionic strength and/or a higher reaction
temperature, and tend to require a more perfect match between
probe/primer and a target sequence in order to form a stable
duplex. If the stringency is too high, however, hybridization may
not occur at all. In contrast, lower stringency conditions utilize
buffers with higher ionic strength and/or a lower reaction
temperature, and permit the formation of stable duplexes with more
mismatched bases between a probe/primer and a target sequence. By
way of example and not limitation, exemplary conditions for high
stringency hybridization conditions using an allele-specific probe
are as follows: Prehybridization with a solution containing
5.times. standard saline phosphate EDTA (SSPE), 0.5% NaDodSO.sub.4
(SDS) at 55.degree. C., and incubating probe with target nucleic
acid molecules in the same solution at the same temperature,
followed by washing with a solution containing 2.times.SSPE, and
0.1% SDS at 55.degree. C. or room temperature.
[0134] Moderate stringency hybridization conditions may be used for
allele-specific primer extension reactions with a solution
containing, e.g., about 50 mM KCl at about 46.degree. C.
Alternatively, the reaction may be carried out at an elevated
temperature such as 60.degree. C. In another embodiment, a
moderately stringent hybridization condition suitable for
oligonucleotide ligation assay (OLA) reactions wherein two probes
are ligated if they are completely complementary to the target
sequence may utilize a solution of about 100 mM KCl at a
temperature of 46.degree. C.
[0135] In a hybridization-based assay, allele-specific probes can
be designed that hybridize to a segment of target DNA from one
individual but do not hybridize to the corresponding segment from
another individual due to the presence of different polymorphic
forms (e.g., alternative SNP alleles/nucleotides) in the respective
DNA segments from the two individuals. Hybridization conditions
should be sufficiently stringent that there is a significant
detectable difference in hybridization intensity between alleles,
and preferably an essentially binary response, whereby a probe
hybridizes to only one of the alleles or significantly more
strongly to one allele. While a probe may be designed to hybridize
to a target sequence that contains a SNP site such that the SNP
site aligns anywhere along the sequence of the probe, the probe is
preferably designed to hybridize to a segment of the target
sequence such that the SNP site aligns with a central position of
the probe (e.g., a position within the probe that is at least three
nucleotides from either end of the probe). This design of probe
generally achieves good discrimination in hybridization between
different allelic forms.
[0136] In another embodiment, a probe or primer may be designed to
hybridize to a segment of target DNA such that the SNP aligns with
either the 5' most end or the 3' most end of the probe or primer.
In a specific preferred embodiment which is particularly suitable
for use in a oligonucleotide ligation assay (U.S. Pat. No.
4,988,617), the 3' most nucleotide of the probe aligns with the SNP
position in the target sequence.
[0137] Oligonucleotide probes and primers may be prepared by
methods well known in the art. Chemical synthetic methods include,
but are limited to, the phosphotriester method described by Narang
et al., 1979, Methods in Enzymology 68:90; the phosphodiester
method described by Brown et al., 1979, Methods in Enzymology
68:109, the diethylphosphoamidate method described by Beaucage et
al., 1981, Tetrahedron Letters 22:1859; and the solid support
method described in U.S. Pat. No. 4,458,066.
[0138] Allele-specific probes are often used in pairs (or, less
commonly, in sets of 3 or 4, such as if a SNP position is known to
have 3 or 4 alleles, respectively, or to assay both strands of a
nucleic acid molecule for a target SNP allele), and such pairs may
be identical except for a one nucleotide mismatch that represents
the allelic variants at the SNP position. Commonly, one member of a
pair perfectly matches a reference form of a target sequence that
has a more common SNP allele (i.e., the allele that is more
frequent in the target population) and the other member of the pair
perfectly matches a form of the target sequence that has a less
common SNP allele (i.e., the allele that is rarer in the target
population). In the case of an array, multiple pairs of probes can
be immobilized on the same support for simultaneous analysis of
multiple different polymorphisms.
[0139] In one type of PCR-based assay, an allele-specific primer
hybridizes to a region on a target nucleic acid molecule that
overlaps a SNP position and only primes amplification of an allelic
form to which the primer exhibits perfect complementarity (Gibbs,
1989, Nucleic Acid Res. 17 2427-2448). Typically, the primer's
3'-most nucleotide is aligned with and complementary to the SNP
position of the target nucleic acid molecule. This primer is used
in conjunction with a second primer that hybridizes at a distal
site. Amplification proceeds from the two primers, producing a
detectable product that indicates which allelic form is present in
the test sample. A control is usually performed with a second pair
of primers, one of which shows a single base mismatch at the
polymorphic site and the other of which exhibits perfect
complementarity to a distal site. The single-base mismatch prevents
amplification or substantially reduces amplification efficiency, so
that either no detectable product is formed or it is formed in
lower amounts or at a slower pace. The method generally works most
effectively when the mismatch is at the 3'-most position of the
oligonucleotide (i.e., the 3'-most position of the oligonucleotide
aligns with the target SNP position) because this position is most
destabilizing to elongation from the primer (see, e.g., WO
93/22456). This PCR-based assay can be utilized as part of the
TaqMan assay, described below.
[0140] In a specific embodiment of the invention, a primer of the
invention contains a sequence substantially complementary to a
segment of a target SNP-containing nucleic acid molecule except
that the primer has a mismatched nucleotide in one of the three
nucleotide positions at the 3'-most end of the primer, such that
the mismatched nucleotide does not base pair with a particular
allele at the SNP site. In a preferred embodiment, the mismatched
nucleotide in the primer is the second from the last nucleotide at
the 3'-most position of the primer. In a more preferred embodiment,
the mismatched nucleotide in the primer is the last nucleotide at
the 3'-most position of the primer.
[0141] In another embodiment of the invention, a SNP detection
reagent of the invention is labeled with a fluorogenic reporter dye
that emits a detectable signal. While the preferred reporter dye is
a fluorescent dye, any reporter dye that can be attached to a
detection reagent such as an oligonucleotide probe or primer is
suitable for use in the invention. Such dyes include, but are not
limited to, Acridine, AMCA, BODIPY, Cascade Blue, Cy2, Cy3, Cy5,
Cy7, Dabcyl, Edans, Eosin, Erythrosin, Fluorescein, 6-Fam, Tet,
Joe, Hex, Oregon Green, Rhodamine, Rhodol Green, Tamra, Rox, and
Texas Red.
[0142] In yet another embodiment of the invention, the detection
reagent may be further labeled with a quencher dye such as Tamra,
especially when the reagent is used as a self-quenching probe such
as a TaqMan (U.S. Pat. Nos. 5,210,015 and 5,538,848) or Molecular
Beacon probe (U.S. Pat. Nos. 5,118,801 and 5,312,728), or other
stemless or linear beacon probe (Livak et al., 1995, PCR Method
Appl. 4:357-362; Tyagi et al., 1996, Nature Biotechnology 14:
303-308; Nazarenko et al., 1997, Nucl. Acids Res. 25:2516-2521;
U.S. Pat. Nos. 5,866,336 and 6,117,635).
[0143] The detection reagents of the invention may also contain
other labels, including but not limited to, biotin for streptavidin
binding, hapten for antibody binding, and oligonucleotide for
binding to another complementary oligonucleotide such as pairs of
zipcodes.
[0144] The present invention also contemplates reagents that do not
contain (or that are complementary to) a SNP nucleotide identified
herein but that are used to assay one or more SNPs disclosed
herein. For example, primers that flank, but do not hybridize
directly to a target SNP position provided herein are useful in
primer extension reactions in which the primers hybridize to a
region adjacent to the target SNP position (i.e., within one or
more nucleotides from the target SNP site). During the primer
extension reaction, a primer is typically not able to extend past a
target SNP site if a particular nucleotide (allele) is present at
that target SNP site, and the primer extension product can readily
be detected in order to determine which SNP allele is present at
the target SNP site. For example, particular ddNTPs are typically
used in the primer extension reaction to terminate primer extension
once a ddNTP is incorporated into the extension product (a primer
extension product which includes a ddNTP at the 3'-most end of the
primer extension product, and in which the ddNTP corresponds to a
SNP disclosed herein, is a composition that is encompassed by the
present invention). Thus, reagents that bind to a nucleic acid
molecule in a region adjacent to a SNP site, even though the bound
sequences do not necessarily include the SNP site itself, are also
encompassed by the present invention.
[0145] SNP Detection Kits and Systems
[0146] A person skilled in the art will recognize that, based on
the SNP and associated sequence information disclosed herein,
detection reagents can be developed and used to assay any SNP of
the present invention individually or in combination, and such
detection reagents can be readily incorporated into one of the
established kit or system formats which are well known in the art.
The terms "kits" and "systems", as used herein in the context of
SNP detection reagents, are intended to refer to such things as
combinations of multiple SNP detection reagents, or one or more SNP
detection reagents in combination with one or more other types of
elements or components (e.g., other types of biochemical reagents,
containers, packages such as packaging intended for commercial
sale, substrates to which SNP detection reagents are attached,
electronic hardware components, etc.). Accordingly, the present
invention further provides SNP detection kits and systems,
including but not limited to, packaged probe and primer sets (e.g.,
TaqMan probe/primer sets), arrays/microarrays of nucleic acid
molecules, and beads that contain one or more probes, primers, or
other detection reagents for detecting one or more SNPs of the
present invention. The kits/systems can optionally include various
electronic hardware components; for example, arrays ("DNA chips")
and microfluidic systems ("lab-on-a-chip" systems) provided by
various manufacturers typically comprise hardware components. Other
kits/systems (e.g., probe/primer sets) may not include electronic
hardware components, but may be comprised of, for example, one or
more SNP detection reagents (along with, optionally, other
biochemical reagents) packaged in one or more containers.
[0147] In some embodiments, a SNP detection kit typically contains
one or more detection reagents and other components (e.g., a
buffer, enzymes such as DNA polymerases or ligases, chain extension
nucleotides such as deoxynucleotide triphosphates, and in the case
of Sanger-type DNA sequencing reactions, chain terminating
nucleotides, positive control sequences, negative control
sequences, and the like) necessary t6 carry out an assay or
reaction, such as amplification and/or detection of a
SNP-containing nucleic acid molecule. A kit may further contain
means for determining the amount of a target nucleic acid, and
means for comparing the amount with a standard, and can comprise
instructions for using the kit to detect the SNP-containing nucleic
acid molecule of interest. In one embodiment of the present
invention, kits are provided which contain the necessary reagents
to carry out one or more assays to detect one or more SNPs
disclosed herein. In a preferred embodiment of the present
invention, SNP detection kits/systems are in the form of nucleic
acid arrays, or compartmentalized kits, including
microfluidic/lab-on-a-chip systems.
[0148] SNP detection kits/systems may contain, for example, one or
more probes, or pairs of probes, that hybridize to a nucleic acid
molecule at or near each target SNP position. Multiple pairs of
allele-specific probes may be included in the kit/system to
simultaneously assay large numbers of SNPs, at least one of which
is a SNP of the present invention. In some kits/systems, the
allele-specific probes are immobilized to a substrate such as an
array or bead. For example, the same substrate can comprise
allele-specific probes for detecting at least 1; 10; 100; 1000;
10,000; 100,000 (or any other number in-between) or substantially
all of the SNPs shown in Table 1 and/or Table 2.
[0149] The terms "arrays", "microarrays", and "DNA chips" are used
herein interchangeably to refer to an array of distinct
polynucleotides affixed to a substrate, such as glass, plastic,
paper, nylon or other type of membrane, filter, chip, or any other
suitable solid support. The polynucleotides can be synthesized
directly on the substrate, or synthesized separate from the
substrate and then affixed to the substrate. In one embodiment, the
microarray is prepared and used according to the methods described
in U.S. Pat. No. 5,837,832, Chee et al., PCT application WO95/11995
(Chee et al.), Lockhart, D. J. et al. (1996; Nat. Biotech. 14:
1675-1680) and Schena, M. et al. (1996; Proc. Natl. Acad. Sci. 93:
10614-10619), all of which are incorporated herein in their
entirety by reference. In other embodiments, such arrays are
produced by the methods described by Brown et al., U.S. Pat. No.
5,807,522.
[0150] Nucleic acid arrays are reviewed in the following
references: Zammatteo et al., "New chips for molecular biology and
diagnostics", Biotechnol Annu Rev. 2002; 8:85-101; Sosnowski et
al., "Active microelectronic array system for DNA hybridization,
genotyping and pharmacogenomic applications", Psychiatr Genet. 2002
December; 12(4):181-92; Heller, "DNA microarray technology:
devices, systems, and applications", Annu Rev Biomed Eng. 2002;
4:129-53. Epub 2002 Mar. 22; Kolchinsky et al., "Analysis of SNPs
and other genomic variations using gel-based chips", Hum Mutat.
2002 April; 19(4):343-60; and McGall et al., "High-density genechip
oligonucleotide probe arrays", Adv Biochem Eng Biotechnol. 2002;
77:21-42.
[0151] Any number of probes, such as allele-specific probes, may be
implemented in an array, and each probe or pair of probes can
hybridize to a different SNP position. In the case of
polynucleotide probes, they can be synthesized at designated areas
(or synthesized separately and then affixed to designated areas) on
a substrate using a light-directed chemical process. Each DNA chip
can contain, for example, thousands to millions of individual
synthetic polynucleotide probes arranged in a grid-like pattern and
miniaturized (e.g., to the size of a dime). Preferably, probes are
attached to a solid support in an ordered, addressable array.
[0152] A microarray can be composed of a large number of unique,
single-stranded polynucleotides, usually either synthetic antisense
polynucleotides or fragments of cDNAs, fixed to a solid support.
Typical polynucleotides are preferably about 6-60 nucleotides in
length, more preferably about 15-nucleotides in length, and most
preferably about 18-25 nucleotides in length. For certain types of
microarrays or other detection kits/systems, it may be preferable
to use oligonucleotides that are only about 7-20 nucleotides in
length. In other types of arrays, such as arrays used in
conjunction with chemiluminescent detection technology, preferred
probe lengths can be, for example, about 15-80 nucleotides in
length, preferably about 50-70 nucleotides in length, more
preferably about 55-65 nucleotides in length, and most preferably
about 60 nucleotides in length. The microarray or detection kit can
contain polynucleotides that cover the known 5' or 3' sequence of a
gene/transcript or target SNP site, sequential polynucleotides that
cover the full-length sequence of a gene/transcript; or unique
polynucleotides selected from particular areas along the length of
a target gene/transcript sequence, particularly areas corresponding
to one or more SNPs disclosed in Table 1 and/or Table 2.
Polynucleotides used in the microarray or detection kit can be
specific to a SNP or SNPs of interest (e.g., specific to a
particular SNP allele at a target SNP site, or specific to
particular SNP alleles at multiple different SNP sites), or
specific to a polymorphic gene/transcript or genes/transcripts of
interest.
[0153] Hybridization assays based on polynucleotide arrays rely on
the differences in hybridization stability of the probes to
perfectly matched and mismatched target sequence variants. For SNP
genotyping, it is generally preferable that stringency conditions
used in hybridization assays are high enough such that nucleic acid
molecules that differ from one another at as little as a single SNP
position can be differentiated (e.g., typical SNP hybridization
assays are designed so that hybridization will occur only if one
particular nucleotide is present at a SNP position, but will not
occur if an alternative nucleotide is present at that SNP
position). Such high stringency conditions may be preferable when
using, for example, nucleic acid arrays of allele-specific probes
for SNP detection. Such high stringency conditions are described in
the preceding section, and are well known to those skilled in the
art and can be found in, for example, Current Protocols in
Molecular Biology, John Wiley & Sons, N.Y. (1989),
6.3.1-6.3.6.
[0154] In other embodiments, the arrays are used in conjunction
with chemiluminescent detection technology. The following patents
and patent applications, which are all hereby incorporated by
reference, provide additional information pertaining to
chemiluminescent detection: U.S. patent application Ser. Nos.
10/620,332 and 10/620,333 describe chemiluminescent approaches for
microarray detection; U.S. Pat. Nos. 6,124,478, 6,107,024,
5,994,073, 5,981,768, 5,871,938, 5,843,681, 5,800,999, and
5,773,628 describe methods and compositions of dioxetane for
performing chemiluminescent detection; and U.S. published
application US2002/0110828 discloses methods and compositions for
microarray controls.
[0155] In one embodiment of the invention, a nucleic acid array can
comprise an array of probes of about 15-25 nucleotides in length.
In further embodiments, a nucleic acid array can comprise any
number of probes, in which at least one probe is capable of
detecting one or more SNPs disclosed in Table 1 and/or Table 2,
and/or at least one probe comprises a fragment of one of the
sequences selected from the group consisting of those disclosed in
Table 1, Table 2, the Sequence Listing, and sequences complementary
thereto, said fragment comprising at least about 8 consecutive
nucleotides, preferably 10, 12, 15, 16, 18, 20, more preferably 22,
25, 30, 40, 47, 50, 55, 60, 65, 70, 80, 90, 100, or more
consecutive nucleotides (or any other number in-between) and
containing (or being complementary to) a novel SNP allele disclosed
in Table 1 and/or Table 2. In some embodiments, the nucleotide
complementary to the SNP site is within 5, 4, 3, 2, or 1 nucleotide
from the center of the probe, more preferably at the center of said
probe.
[0156] A polynucleotide probe can be synthesized on the surface of
the substrate by using a chemical coupling procedure and an ink jet
application apparatus, as described in PCT application WO95/251116
(Baldeschweiler et al.) which is incorporated herein in its
entirety by reference. In another aspect, a "gridded" array
analogous to a dot (or slot) blot may be used to arrange and link
cDNA fragments or oligonucleotides to the surface of a substrate
using a vacuum system, thermal, UV, mechanical or chemical bonding
procedures. An array, such as those described above, may be
produced by hand or by using available devices (slot blot or dot
blot apparatus), materials (any suitable solid support), and
machines (including robotic instruments), and may contain 8, 24,
96, 384, 1536, 6144 or more polynucleotides, or any other number
which lends itself to the efficient use of commercially available
instrumentation.
[0157] Using such arrays or other kits/systems, the present
invention provides methods of identifying the SNPs disclosed herein
in a test sample. Such methods typically involve incubating a test
sample of nucleic acids with an array comprising one or more probes
corresponding to at least one SNP position of the present
invention, and assaying for binding of a nucleic acid from the test
sample with one or more of the probes. Conditions for incubating a
SNP detection reagent (or a kit/system that employs one or more
such SNP detection reagents) with a test sample vary. Incubation
conditions depend on such factors as the format employed in the
assay, the detection methods employed, and the type and nature of
the detection reagents used in the assay. One skilled in the art
will recognize that any one of the commonly available
hybridization, amplification and array assay formats can readily be
adapted to detect the SNPs disclosed herein.
[0158] A SNP detection kit/system of the present invention may
include components that are used to prepare nucleic acids from a
test sample for the subsequent amplification and/or detection of a
SNP-containing nucleic acid molecule. Such sample preparation
components can be used to produce nucleic acid extracts (including
DNA and/or RNA), proteins or membrane extracts from any bodily
fluids (such as blood, serum, plasma, urine, saliva, phlegm,
gastric juices, semen, tears, sweat, etc.), skin, hair, cells
(especially nucleated cells), biopsies, buccal swabs or tissue
specimens. The test samples used in the above-described methods
will vary based on such factors as the assay format, nature of the
detection method, and the specific tissues, cells or extracts used
as the test sample to be assayed. Methods of preparing nucleic
acids, proteins, and cell extracts are well known in the art and
can be readily adapted to obtain a sample that is compatible with
the system utilized. Automated sample preparation systems for
extracting nucleic acids from a test sample are commercially
available, and examples are Qiagen's BioRobot 9600, Applied
Biosystems' PRISM 6700, and Roche Molecular Systems'COBAS AmpliPrep
System.
[0159] Another form of kit contemplated by the present invention is
a compartmentalized kit. A compartmentalized kit includes any kit
in which reagents are contained in separate containers. Such
containers include, for example, small glass containers, plastic
containers, strips of plastic, glass or paper, or arraying material
such as silica. Such containers allow one to efficiently transfer
reagents from one compartment to another compartment such that the
test samples and reagents are not cross-contaminated, or from one
container to another vessel not included in the kit, and the agents
or solutions of each container can be added in a quantitative
fashion from one compartment to another or to another vessel. Such
containers may include, for example, one or more containers which
will accept the test sample, one or more containers which contain
at least one probe or other SNP detection reagent for detecting one
or more SNPs of the present invention, one or more containers which
contain wash reagents (such as phosphate buffered saline,
Tris-buffers, etc.), and one or more containers which contain the
reagents used to reveal the presence of the bound probe or other
SNP detection reagents. The kit can optionally further comprise
compartments and/or reagents for, for example, nucleic acid
amplification or other enzymatic reactions such as primer extension
reactions, hybridization, ligation, electrophoresis (preferably
capillary electrophoresis), mass spectrometry, and/or laser-induced
fluorescent detection. The kit may also include instructions for
using the kit. Exemplary compartmentalized kits include
microfluidic devices known in the art (see, e.g., Weigl et al.,
"Lab-on-a-chip for drug development", Adv Drug Deliv Rev. 2003 Feb.
24; 55(3):349-77). In such microfluidic devices, the containers may
be referred to as, for example, microfluidic "compartments",
"chambers", or "channels".
[0160] Microfluidic devices, which may also be referred to as
"lab-on-a-chip" systems, biomedical micro-electro-mechanical
systems (bioMEMs), or multicomponent integrated systems, are
exemplary kits/systems of the present invention for analyzing SNPs.
Such systems miniaturize and compartmentalize processes such as
probe/target hybridization, nucleic acid amplification, and
capillary electrophoresis reactions in a single functional device.
Such microfluidic devices typically utilize detection reagents in
at least one aspect of the system, and such detection reagents may
be used to detect one or more SNPs of the present invention. One
example of a microfluidic system is disclosed in U.S. Pat. No.
5,589,136, which describes the integration of PCR amplification and
capillary electrophoresis in chips. Exemplary microfluidic systems
comprise a pattern of microchannels designed onto a glass, silicon,
quartz, or plastic wafer included on a microchip. The movements of
the samples may be controlled by electric, electroosmotic or
hydrostatic forces applied across different areas of the microchip
to create functional microscopic valves and pumps with no moving
parts. Varying the voltage can be used as a means to control the
liquid flow at intersections between the micro-machined channels
and to change the liquid flow rate for pumping across different
sections of the microchip. See, for example, U.S. Pat. Nos.
6,153,073, Dubrow et al., and 6,156,181, Parce et al.
[0161] For genotyping SNPs, an exemplary microfluidic system may
integrate, for example, nucleic acid amplification, primer
extension, capillary electrophoresis, and a detection method such
as laser induced fluorescence detection. In a first step of an
exemplary process for using such an exemplary system, nucleic acid
samples are amplified, preferably by PCR. Then, the amplification
products are subjected to automated primer extension reactions
using ddNTPs (specific fluorescence for each ddNTP) and the
appropriate oligonucleotide primers to carry out primer extension
reactions which hybridize just upstream of the targeted SNP. Once
the extension at the 3' end is completed, the primers are separated
from the unincorporated fluorescent ddNTPs by capillary
electrophoresis. The separation medium used in capillary
electrophoresis can be, for example, polyacrylamide,
polyethyleneglycol or dextran. The incorporated ddNTPs in the
single nucleotide primer extension products are identified by
laser-induced fluorescence detection. Such an exemplary microchip
can be used to process, for example, at least 96 to 384 samples, or
more, in parallel.
[0162] Uses of Nucleic Acid Molecules
[0163] The nucleic acid molecules of the present invention have a
variety of uses, especially in the diagnosis and treatment of
stenosis. For example, the nucleic acid molecules are useful as
hybridization probes, such as for genotyping SNPs in messenger RNA,
transcript, cDNA, genomic DNA, amplified DNA or other nucleic acid
molecules, and for isolating full-length cDNA and genomic clones
encoding the variant peptides disclosed in Table 1 as well as their
orthologs.
[0164] A probe can hybridize to any nucleotide sequence along the
entire length of a nucleic acid molecule provided in Table 1 and/or
Table 2. Preferably, a probe of the present invention hybridizes to
a region of a target sequence that encompasses a SNP position
indicated in Table 1 and/or Table 2. More preferably, a probe
hybridizes to a SNP-containing target sequence in a
sequence-specific manner such that it distinguishes the target
sequence from other nucleotide sequences which vary from the target
sequence only by which nucleotide is present at the SNP site. Such
a probe is particularly useful for detecting the presence of a
SNP-containing nucleic acid in a test sample, or for determining
which nucleotide (allele) is present at a particular SNP site
(i.e., genotyping the SNP site).
[0165] A nucleic acid hybridization probe may be used for
determining the presence, level, form, and/or distribution of
nucleic acid expression. The nucleic acid whose level is determined
can be DNA or RNA. Accordingly, probes specific for the SNPs
described herein can be used to assess the presence, expression
and/or gene copy number in a given cell, tissue, or organism. These
uses are relevant for diagnosis of disorders involving an increase
or decrease in gene expression relative to normal levels. In vitro
techniques for detection of mRNA include, for example, Northern
blot hybridizations and in situ hybridizations. In vitro techniques
for detecting DNA include Southern blot hybridizations and in situ
hybridizations (Sambrook and Russell, 2000, Molecular Cloning: A
Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor,
N.Y.).
[0166] Probes can be used as part of a diagnostic test kit for
identifying cells or tissues in which a variant protein is
expressed, such as by measuring the level of a variant
protein-encoding nucleic acid (e.g., mRNA) in a sample of cells
from a subject or determining if a polynucleotide contains a SNP of
interest.
[0167] Thus, the nucleic acid molecules of the invention can be
used as hybridization probes to detect the SNPs disclosed herein,
thereby determining whether an individual with the polymorphisms is
at risk for stenosis or has developed early stage stenosis.
Detection of a SNP associated with a disease phenotype provides a
diagnostic tool for an active disease and/or genetic predisposition
to the disease.
[0168] The nucleic acid molecules of the invention are also useful
as primers to amplify any given region of a nucleic acid molecule,
particularly a region containing a SNP identified in Table 1 and/or
Table 2.
[0169] The nucleic acid molecules of the invention are also useful
for constructing recombinant vectors (described in greater detail
below). Such vectors include expression vectors that express a
portion of, or all of, any of the variant peptide sequences
provided in Table 1. Vectors also include insertion vectors, used
to integrate into another nucleic acid molecule sequence, such as
into the cellular genome, to alter in situ expression of a gene
and/or gene product. For example, an endogenous coding sequence can
be replaced via homologous recombination with all or part of the
coding region containing one or more specifically introduced
SNPs.
[0170] The nucleic acid molecules of the invention are also useful
for expressing antigenic portions of the variant proteins,
particularly antigenic portions that contain a variant amino acid
sequence (e.g., an amino acid substitution) caused by a SNP
disclosed in Table 1 and/or Table 2.
[0171] The nucleic acid molecules of the invention are also useful
for constructing vectors containing a gene regulatory region of the
nucleic acid molecules of the present invention.
[0172] The nucleic acid molecules of the invention are also useful
for designing ribozymes corresponding to all, or a part, of an mRNA
molecule expressed from a SNP-containing nucleic acid molecule
described herein.
[0173] The nucleic acid molecules of the invention are also useful
for constructing host cells expressing a part, or all, of the
nucleic acid molecules and variant peptides.
[0174] The nucleic acid molecules of the invention are also useful
for constructing transgenic animals expressing all, or a part, of
the nucleic acid molecules and variant peptides. The production of
recombinant cells and transgenic animals having nucleic acid
molecules which contain the SNPs disclosed in Table 1 and/or Table
2 allow, for example, effective clinical design of treatment
compounds and dosage regimens.
[0175] The nucleic acid molecules of the invention are also useful
in assays for drug screening to identify compounds that, for
example, modulate nucleic acid expression.
[0176] The nucleic acid molecules of the invention are also useful
in gene therapy in patients whose cells have aberrant gene
expression. Thus, recombinant cells, which include a patient's
cells that have been engineered ex vivo and returned to the
patient, can be introduced into an individual where the recombinant
cells produce the desired protein to treat the individual.
[0177] SNP Genotyping Methods
[0178] The process of determining which specific nucleotide (i.e.,
allele) is present at each of one or more SNP positions, such as a
SNP position in a nucleic acid molecule disclosed in Table 1 and/or
Table 2, is referred to as SNP genotyping. The present invention
provides methods of SNP genotyping, such as for use in screening
for stenosis or related pathologies, or determining predisposition
thereto, or determining responsiveness to a form of treatment, or
in genome mapping or SNP association analysis, etc.
[0179] Nucleic acid samples can be genotyped to determine which
allele(s) is/are present at any given genetic region (e.g., SNP
position) of interest by methods well known in the art. The
neighboring sequence can be used to design SNP detection reagents
such as oligonucleotide probes, which may optionally be implemented
in a kit format. Exemplary SNP genotyping methods are described in
Chen et al., "Single nucleotide polymorphism genotyping:
biochemistry, protocol, cost and throughput", Pharmacogenomics J.
2003; 3(2):77-96; Kwok et al., "Detection of single nucleotide
polymorphisms", Curr Issues Mol. Biol. 2003 April; 5(2):43-60; Shi,
"Technologies for individual genotyping: detection of genetic
polymorphisms in drug targets and disease genes", Am J.
Pharmacogenomics. 2002; 2(3):197-205; and Kwok, "Methods for
genotyping single nucleotide polymorphisms", Annu Rev Genomics Hum
Genet. 2001; 2:235-58. Exemplary techniques for high-throughput SNP
genotyping are described in Mamellos, "High-throughput SNP analysis
for genetic association studies", Curr Opin Drug Discov Devel. 2003
May; 6(3):317-21. Common SNP genotyping methods include, but are
not limited to, TaqMan assays, molecular beacon assays, nucleic
acid arrays, allele-specific primer extension, allele-specific PCR,
arrayed primer extension, homogeneous primer extension assays,
primer extension with detection by mass spectrometry,
pyrosequencing, multiplex primer extension sorted on genetic
arrays, ligation with rolling circle amplification, homogeneous
ligation, OLA (U.S. Pat. No. 4,988,167), multiplex ligation
reaction sorted on genetic arrays, restriction-fragment length
polymorphism, single base extension-tag assays, and the Invader
assay. Such methods may be used in combination with detection
mechanisms such as, for example, luminescence or chemiluminescence
detection, fluorescence detection, time-resolved fluorescence
detection, fluorescence resonance energy transfer, fluorescence
polarization, mass spectrometry, and electrical detection.
[0180] Various methods for detecting polymorphisms include, but are
not limited to, methods in which protection from cleavage agents is
used to detect mismatched bases in RNA/RNA or RNA/DNA duplexes
(Myers et al., Science 230:1242 (1985); Cotton et al., PNAS 85:4397
(1988); and Saleeba et al., Meth. Enzymol. 217:286-295 (1992)),
comparison of the electrophoretic mobility of variant and wild type
nucleic acid molecules (Orita et al., PNAS 86:2766 (1989); Cotton
et al., Mutat. Res. 285:125-144 (1993); and Hayashi et al., Genet.
Anal. Tech. Appl. 9:73-79 (1992)), and assaying the movement of
polymorphic or wild-type fragments in polyacrylamide gels
containing a gradient of denaturant using denaturing gradient gel
electrophoresis (DGGE) (Myers et al., Nature 313:495 (1985)).
Sequence variations at specific locations can also be assessed by
nuclease protection assays such as RNase and S1 protection or
chemical cleavage methods.
[0181] In a preferred embodiment, SNP genotyping is performed using
the TaqMan assay, which is also known as the 5' nuclease assay
(U.S. Pat. Nos. 5,210,015 and 5,538,848). The TaqMan assay detects
the accumulation of a specific amplified product during PCR. The
TaqMan assay utilizes an oligonucleotide probe labeled with a
fluorescent reporter dye and a quencher dye. The reporter dye is
excited by irradiation at an appropriate wavelength, it transfers
energy to the quencher dye in the same probe via a process called
fluorescence resonance energy transfer (FRET). When attached to the
probe, the excited reporter dye does not emit a signal. The
proximity of the quencher dye to the reporter dye in the intact
probe maintains a reduced fluorescence for the reporter. The
reporter dye and quencher dye may be at the 5' most and the 3' most
ends, respectively, or vice versa. Alternatively, the reporter dye
may be at the 5' or 3' most end while the quencher dye is attached
to an internal nucleotide, or vice versa. In yet another
embodiment, both the reporter and the quencher may be attached to
internal nucleotides at a distance from each other such that
fluorescence of the reporter is reduced.
[0182] During PCR, the 5' nuclease activity of DNA polymerase
cleaves the probe, thereby separating the reporter dye and the
quencher dye and resulting in increased fluorescence of the
reporter. Accumulation of PCR product is detected directly by
monitoring the increase in fluorescence of the reporter dye. The
DNA polymerase cleaves the probe between the reporter dye and the
quencher dye only if the probe hybridizes to the target
SNP-containing template which is amplified during PCR, and the
probe is designed to hybridize to the target SNP site only if a
particular SNP allele is present.
[0183] Preferred TaqMan primer and probe sequences can readily be
determined using the SNP and associated nucleic acid sequence
information provided herein. A number of computer programs, such as
Primer Express (Applied Biosystems, Foster City, Calif.), can be
used to rapidly obtain optimal primer/probe sets. It will be
apparent to one of skill in the art that such primers and probes
for detecting the SNPs of the present invention are useful in
diagnostic assays for stenosis and related pathologies, and can be
readily incorporated into a kit format. The present invention also
includes modifications of the Taqman assay well known in the art
such as the use of Molecular Beacon probes (U.S. Pat. Nos.
5,118,801 and 5,312,728) and other variant formats (U.S. Pat. Nos.
5,866,336 and 6,117,635).
[0184] Another preferred method for genotyping the SNPs of the
present invention is the use of two oligonucleotide probes in an
OLA (see, e.g., U.S. Pat. No. 4,988,617). In this method, one probe
hybridizes to a segment of a target nucleic acid with its 3' most
end aligned with the SNP site. A second probe hybridizes to an
adjacent segment of the target nucleic acid molecule directly 3' to
the first probe. The two juxtaposed probes hybridize to the target
nucleic acid molecule, and are ligated in the presence of a linking
agent such as a ligase if there is perfect complementarity between
the 3' most nucleotide of the first probe with the SNP site. If
there is a mismatch, ligation would not occur. After the reaction,
the ligated probes are separated from the target nucleic acid
molecule, and detected as indicators of the presence of a SNP.
[0185] The following patents, patent applications, and published
international patent applications, which are all hereby
incorporated by reference, provide additional information
pertaining to techniques for carrying out various types of OLA:
U.S. Pat. Nos. 6,027,889, 6,268,148, 5,494,810, 5,830,711, and
6054564 describe OLA strategies for performing SNP detection; WO
97/31256 and WO 00/56927 describe OLA strategies for performing SNP
detection using universal arrays, wherein a zipcode sequence can be
introduced into one of the hybridization probes, and the resulting
product, or amplified product, hybridized to a universal zip code
array; U.S. application US01/17329 (and 09/584,905) describes OLA
(or LDR) followed by PCR, wherein zipcodes are incorporated into
OLA probes, and amplified PCR products are determined by
electrophoretic or universal zipcode array readout; U.S.
applications 60/427,818, 60/445,636, and 60/445,494 describe SNPlex
methods and software for multiplexed SNP detection using OLA
followed by PCR, wherein zipcodes are incorporated into OLA probes,
and amplified PCR products are hybridized with a zipchute reagent,
and the identity of the SNP determined from electrophoretic readout
of the zipchute. In some embodiments, OLA is carried out prior to
PCR (or another method of nucleic acid amplification). In other
embodiments, PCR (or another method of nucleic acid amplification)
is carried out prior to OLA.
[0186] Another method for SNP genotyping is based on mass
spectrometry. Mass spectrometry takes advantage of the unique mass
of each of the four nucleotides of DNA. SNPs can be unambiguously
genotyped by mass spectrometry by measuring the differences in the
mass of nucleic acids having alternative SNP alleles. MALDI-TOF
(Matrix Assisted Laser Desorption Ionization--Time of Flight) mass
spectrometry technology is preferred for extremely precise
determinations of molecular mass, such as SNPs. Numerous approaches
to SNP analysis have been developed based on mass spectrometry.
Preferred mass spectrometry-based methods of SNP genotyping include
primer extension assays, which can also be utilized in combination
with other approaches, such as traditional gel-based formats and
microarrays.
[0187] Typically, the primer extension assay involves designing and
annealing a primer to a template PCR amplicon upstream (5') from a
target SNP position. A mix of dideoxynucleotide triphosphates
(ddNTPs) and/or deoxynucleotide triphosphates (dNTPs) are added to
a reaction mixture containing template (e.g., a SNP-containing
nucleic acid molecule which has typically been amplified, such as
by PCR), primer, and DNA polymerase. Extension of the primer
terminates at the first position in the template where a nucleotide
complementary to one of the ddNTPs in the mix occurs. The primer
can be either immediately adjacent (i.e., the nucleotide at the 3'
end of the primer hybridizes to the nucleotide next to the target
SNP site) or two or more nucleotides removed from the SNP position.
If the primer is several nucleotides removed from the target SNP
position, the only limitation is that the template sequence between
the 3' end of the primer and the SNP position cannot contain a
nucleotide of the same type as the one to be detected, or this will
cause premature termination of the extension primer. Alternatively,
if all four ddNTPs alone, with no dNTPs, are added to the reaction
mixture, the primer will always be extended by only one nucleotide,
corresponding to the target SNP position. In this instance, primers
are designed to bind one nucleotide upstream from the SNP position
(i.e., the nucleotide at the 3' end of the primer hybridizes to the
nucleotide that is immediately adjacent to the target SNP site on
the 5' side of the target SNP site). Extension by only one
nucleotide is preferable, as it minimizes the overall mass of the
extended primer, thereby increasing the resolution of mass
differences between alternative SNP nucleotides. Furthermore,
mass-tagged ddNTPs can be employed in the primer extension
reactions in place of unmodified ddNTPs. This increases the mass
difference between primers extended with these ddNTPs, thereby
providing increased sensitivity and accuracy, and is particularly
useful for typing heterozygous base positions. Mass-tagging also
alleviates the need for intensive sample-preparation procedures and
decreases the necessary resolving power of the mass
spectrometer.
[0188] The extended primers can then be purified and analyzed by
MALDI-TOF mass spectrometry to determine the identity of the
nucleotide present at the target SNP position. In one method of
analysis, the products from the primer extension reaction are
combined with light absorbing crystals that form a matrix. The
matrix is then hit with an energy source such as a laser to ionize
and desorb the nucleic acid molecules into the gas-phase. The
ionized molecules are then ejected into a flight tube and
accelerated down the tube towards a detector. The time between the
ionization event, such as a laser pulse, and collision of the
molecule with the detector is the time of flight of that molecule.
The time of flight is precisely correlated with the mass-to-charge
ratio (m/z) of the ionized molecule. Ions with smaller m/z travel
down the tube faster than ions with larger m/z and therefore the
lighter ions reach the detector before the heavier ions. The
time-of-flight is then converted into a corresponding, and highly
precise, m/z. In this manner, SNPs can be identified based on the
slight differences in mass, and the corresponding time of flight
differences, inherent in nucleic acid molecules having different
nucleotides at a single base position. For further information
regarding the use of primer extension assays in conjunction with
MALDI-TOF mass spectrometry for SNP genotyping, see, e.g., Wise et
al., "A standard protocol for single nucleotide primer extension in
the human genome using matrix-assisted laser desorption/ionization
time-of-flight mass spectrometry", Rapid Commun Mass Spectrom.
2003; 17(11):1195-202.
[0189] The following references provide further information
describing mass spectrometry-based methods for SNP genotyping:
Bocker, "SNP and mutation discovery using base-specific cleavage
and MALDI-TOF mass spectrometry", Bioinformatics. 2003 July; 19
Suppl 1:I44-I53; Storm et al., "MALDI-TOF mass spectrometry-based
SNP genotyping", Methods Mol. Biol. 2003; 212:241-62; Jurinke et
al., "The use of MassARRAY technology for high throughput
genotyping", Adv Biochem Eng Biotechnol. 2002; 77:57-74; and
Jurinke et al., "Automated genotyping using the DNA MassArray
technology", Methods Mol. Biol. 2002; 187:179-92.
[0190] SNPs can also be scored by direct DNA sequencing. A variety
of automated sequencing procedures can be utilized ((1995)
Biotechniques 19:448), including sequencing by mass spectrometry
(see, e.g., PCT International Publication No. WO94/16101; Cohen et
al., Adv. Chromatogr. 36:127-162 (1996); and Griffin et al., Appl.
Biochem. Biotechnol. 38:147-159 (1993)). The nucleic acid sequences
of the present invention enable one of ordinary skill in the art to
readily design sequencing primers for such automated sequencing
procedures. Commercial instrumentation, such as the Applied
Biosystems 377, 3100, 3700, 3730, and 3730x1 DNA Analyzers (Foster
City, Calif.), is commonly used in the art for automated
sequencing.
[0191] Other methods that can be used to genotype the SNPs of the
present invention include single-strand conformational polymorphism
(SSCP), and denaturing gradient gel electrophoresis (DGGE) (Myers
et al., Nature 313:495 (1985)). SSCP identifies base differences by
alteration in electrophoretic migration of single stranded PCR
products, as described in Orita et al., Proc. Nat. Acad.
Single-stranded PCR products can be generated by heating or
otherwise denaturing double stranded PCR products. Single-stranded
nucleic acids may refold or form secondary structures that are
partially dependent on the base sequence. The different
electrophoretic mobilities of single-stranded amplification
products are related to base-sequence differences at SNP positions.
DGGE differentiates SNP alleles based on the different
sequence-dependent stabilities and melting properties inherent in
polymorphic DNA and the corresponding differences in
electrophoretic migration patterns in a denaturing gradient gel
(Erlich, ed., PCR Technology, Principles and Applications for DNA
Amplification, W.H. Freeman and Co, New York, 1992, Chapter 7).
[0192] Sequence-specific ribozymes (U.S. Pat. No. 5,498,531) can
also be used to score SNPs based on the development or loss of a
ribozyme cleavage site. Perfectly matched sequences can be
distinguished from mismatched sequences by nuclease cleavage
digestion assays or by differences in melting temperature. If the
SNP affects a restriction enzyme cleavage site, the SNP can be
identified by alterations in restriction enzyme digestion patterns,
and the corresponding changes in nucleic acid fragment lengths
determined by gel electrophoresis
[0193] SNP genotyping can include the steps of, for example,
collecting a biological sample from a human subject (e.g., sample
of tissues, cells, fluids, secretions, etc.), isolating nucleic
acids (e.g., genomic DNA, mRNA or both) from the cells of the
sample, contacting the nucleic acids with one or more primers which
specifically hybridize to a region of the isolated nucleic acid
containing a target SNP under conditions such that hybridization
and amplification of the target nucleic acid region occurs, and
determining the nucleotide present at the SNP position of interest,
or, in some assays, detecting the presence or absence of an
amplification product (assays can be designed so that hybridization
and/or amplification will only occur if a particular SNP allele is
present or absent). In some assays, the size of the amplification
product is detected and compared to the length of a control sample;
for example, deletions and insertions can be detected by a change
in size of the amplified product compared to a normal genotype.
[0194] SNP genotyping is useful for numerous practical
applications, as described below. Examples of such applications
include, but are not limited to, SNP-disease association analysis,
disease predisposition screening, disease diagnosis, disease
prognosis, disease progression monitoring, determining therapeutic
strategies based on an individual's genotype ("pharmacogenomics"),
developing therapeutic agents based on SNP genotypes associated
with a disease or likelihood of responding to a drug, stratifying a
patient population for clinical trial for a treatment regimen,
predicting the likelihood that an individual will experience toxic
side effects from a therapeutic agent, and human identification
applications such as forensics.
[0195] Analysis of Genetic Association Between SNPs and Phenotypic
Traits
[0196] SNP genotyping for disease diagnosis, disease predisposition
screening, disease prognosis, determining drug responsiveness
(pharmacogenomics), drug toxicity screening, and other uses
described herein, typically relies on initially establishing a
genetic association between one or more specific SNPs and the
particular phenotypic traits of interest.
[0197] Different study designs may be used for genetic association
studies (Modern Epidemiology, Lippincott Williams & Wilkins
(1998), 609-622). Observational studies are most frequently carried
out in which the response of the patients is not interfered with.
The first type of observational study identifies a sample of
persons in whom the suspected cause of the disease is present and
another sample of persons in whom the suspected cause is absent,
and then the frequency of development of disease in the two samples
is compared. These sampled populations are called cohorts, and the
study is a prospective study. The other type of observational study
is case-control or a retrospective study. In typical case-control
studies, samples are collected from individuals with the phenotype
of interest (cases) such as certain manifestations of a disease,
and from individuals without the phenotype (controls) in a
population (target population) that conclusions are to be drawn
from. Then the possible causes of the disease are investigated
retrospectively. As the time and costs of collecting samples in
case-control studies are considerably less than those for
prospective studies, case-control studies are the more commonly
used study design in genetic association studies, at least during
the exploration and discovery stage.
[0198] In both types of observational studies, there may be
potential confounding factors that should be taken into
consideration. Confounding factors are those that are associated
with both the real cause(s) of the disease and the disease itself,
and they include demographic information such as age, gender,
ethnicity as well as environmental factors. When confounding
factors are not matched in cases and controls in a study, and are
not controlled properly, spurious association results can arise. If
potential confounding factors are identified, they should be
controlled for by analysis methods explained below.
[0199] In a genetic association study, the cause of interest to be
tested is a certain allele or a SNP or a combination of alleles or
a haplotype from several SNPs. Thus, tissue specimens (e.g., whole
blood) from the sampled individuals may be collected and genomic
DNA genotyped for the SNP(s) of interest. In addition to the
phenotypic trait of interest, other information such as demographic
(e.g., age, gender, ethnicity, etc.), clinical, and environmental
information that may influence the outcome of the trait can be
collected to further characterize and define the sample set. In
many cases, these factors are known to be associated with diseases
and/or SNP allele frequencies. There are likely gene-environment
and/or gene-gene interactions as well. Analysis methods to address
gene-environment and gene-gene interactions (for example, the
effects of the presence of both susceptibility alleles at two
different genes can be greater than the effects of the individual
alleles at two genes combined) are discussed below.
[0200] After all the relevant phenotypic and genotypic information
has been obtained, statistical analyses are carried out to
determine if there is any significant correlation between the
presence of an allele or a genotype with the phenotypic
characteristics of an individual. Preferably, data inspection and
cleaning are first performed before carrying out statistical tests
for genetic association. Epidemiological and clinical data of the
samples can be summarized by descriptive statistics with tables and
graphs. Data validation is preferably performed to check for data
completion, inconsistent entries, and outliers. Chi-squared tests
and t-tests (Wilcoxon rank-sum tests if distributions are not
normal) may then be used to check for significant differences
between cases and controls for discrete and continuous variables,
respectively. To ensure genotyping quality, Hardy-Weinberg
disequilibrium tests can be performed on cases and controls
separately. Significant deviation from Hardy-Weinberg equilibrium
(HWE) in both cases and controls for individual markers can be
indicative of genotyping errors. If HWE is violated in a majority
of markers, it is indicative of population substructure that should
be further investigated. Moreover, Hardy-Weinberg disequilibrium in
cases only can indicate genetic association of the markers with the
disease (Genetic Data Analysis, Weir B., Sinauer (1990)).
[0201] To test whether an allele of a single SNP is associated with
the case or control status of a phenotypic trait, one skilled in
the art can compare allele frequencies in cases and controls.
Standard chi-squared tests and Fisher exact tests can be carried
out on a 2.times.2 table (2 SNP alleles.times.2 outcomes in the
categorical trait of interest). To test whether genotypes of a SNP
are associated, chi-squared tests can be carried out on a 3.times.2
table (3 genotypes.times.2 outcomes). Score tests are also carried
out for genotypic association to contrast the three genotypic
frequencies (major homozygotes, heterozygotes and minor
homozygotes) in cases and controls, and to look for trends using 3
different modes of inheritance, namely dominant (with contrast
coefficients 2, -1, -1), additive (with contrast coefficients 1, 0,
-1) and recessive (with contrast coefficients 1, 1, -2). Odds
ratios for minor versus major alleles, and odds ratios for
heterozygote and homozygote variants versus the wild type genotypes
are calculated with the desired confidence limits, usually 95%.
[0202] In order to control for confounders and to test for
interaction and effect modifiers, stratified analyses may be
performed using stratified factors that are likely to be
confounding, including demographic information such as age,
ethnicity, and gender, or an interacting element or effect
modifier, such as a known major gene (e.g., APOE for Alzheimer's
disease or HLA genes for autoimmune diseases), or environmental
factors such as smoking in lung cancer. Stratified association
tests may be carried out using Cochran-Mantel-Haenszel tests that
take into account the ordinal nature of genotypes with 0, 1, and 2
variant alleles. Exact tests by StatXact may also be performed when
computationally possible. Another way to adjust for confounding
effects and test for interactions is to perform stepwise multiple
logistic regression analysis using statistical packages such as SAS
or R. Logistic regression is a model-building technique in which
the best fitting and most parsimonious model is built to describe
the relation between the dichotomous outcome (for instance, getting
a certain disease or not) and a set of independent variables (for
instance, genotypes of different associated genes, and the
associated demographic and environmental factors). The most common
model is one in which the logit transformation of the odds ratios
is expressed as a linear combination of the variables (main
effects) and their cross-product terms (interactions) (Applied
Logistic Regression, Hosmer and Lemeshow, Wiley (2000)). To test
whether a certain variable or interaction is significantly
associated with the outcome, coefficients in the model are first
estimated and then tested for statistical significance of their
departure from zero.
[0203] In addition to performing association tests one marker at a
time, haplotype association analysis may also be performed to study
a number of markers that are closely linked together. Haplotype
association tests can have better power than genotypic or allelic
association tests when the tested markers are not the
disease-causing mutations themselves but are in linkage
disequilibrium with such mutations. The test will even be more
powerful if the disease is indeed caused by a combination of
alleles on a haplotype (e.g., APOE is a haplotype formed by 2 SNPs
that are very close to each other). In order to perform haplotype
association effectively, marker-marker linkage disequilibrium
measures, both D' and R.sup.2, are typically calculated for the
markers within a gene to elucidate the haplotype structure. Recent
studies (Daly et al, Nature Genetics, 29, 232-235, 2001) in linkage
disequilibrium indicate that SNPs within a gene are organized in
block pattern, and a high degree of linkage disequilibrium exists
within blocks and very little linkage disequilibrium exists between
blocks. Haplotype association with the disease status can be
performed using such blocks once they have been elucidated.
[0204] Haplotype association tests can be carried out in a similar
fashion as the allelic and genotypic association tests. Each
haplotype in a gene is analogous to an allele in a multi-allelic
marker. One skilled in the art can either compare the haplotype
frequencies in cases and controls or test genetic association with
different pairs of haplotypes. It has been proposed (Schaid et al,
Am. J. Hum. Genet., 70, 425-434, 2002) that score tests can be done
on haplotypes using the program "haplo.score". In that method,
haplotypes are first inferred by EM algorithm and score tests are
carried out with a generalized linear model (GLM) framework that
allows the adjustment of other factors.
[0205] An important decision in the performance of genetic
association tests is the determination of the significance level at
which significant association can be declared when the p-value of
the tests reaches that level. In an exploratory analysis where
positive hits will be followed up in subsequent confirmatory
testing, an unadjusted p-value <0.1 (a significance level on the
lenient side) may be used for generating hypotheses for significant
association of a SNP with certain phenotypic characteristics of a
disease. It is preferred that a p-value <0.05 (a significance
level traditionally used in the art) is achieved in order for a SNP
to be considered to have an association with a disease. It is more
preferred that a p-value <0.01 (a significance level on the
stringent side) is achieved for an association to be declared. When
hits are followed up in confirmatory analyses in more samples of
the same source or in different samples from different sources,
adjustment for multiple testing will be performed as to avoid
excess number of hits while maintaining the experiment-wise error
rates at 0.05. While there are different methods to adjust for
multiple testing to control for different kinds of error rates, a
commonly used but rather conservative method is Bonferroni
correction to control the experiment-wise or family-wise error rate
(Multiple comparisons and multiple tests, Westfall et al, SAS
Institute (1999)). Permutation tests to control for the false
discovery rates, FDR, can be more powerful (Benjamini and Hochberg,
Journal of the Royal Statistical Society, Series B 57, 1289-1300,
1995, Resampling-based Multiple Testing, Westfall and Young, Wiley
(1993)). Such methods to control for multiplicity would be
preferred when the tests are dependent and controlling for false
discovery rates is sufficient as opposed to controlling for the
experiment-wise error rates.
[0206] In replication studies using samples from different
populations after statistically significant markers have been
identified in the exploratory stage, meta-analyses can then be
performed by combining evidence of different studies (Modern
Epidemiology, Lippincott Williams & Wilkins, 1998, 643-673). If
available, association results known in the art for the same SNPs
can be included in the meta-analyses.
[0207] Since both genotyping and disease status classification can
involve errors, sensitivity analyses may be performed to see how
odds ratios and p-values would change upon various estimates on
genotyping and disease classification error rates.
[0208] It has been well known that subpopulation-based sampling
bias between cases and controls can lead to spurious results in
case-control association studies (Ewens and Spielman, Am. J. Hum.
Genet. 62, 450-458, 1995) when prevalence of the disease is
associated with different subpopulation groups. Such bias can also
lead to a loss of statistical power in genetic association studies.
To detect population stratification, Pritchard and Rosenberg
(Pritchard et al. Am. J. Hum. Gen. 1999, 65:220-228) suggested
typing markers that are unlinked to the disease and using results
of association tests on those markers to determine whether there is
any population stratification. When stratification is detected, the
genomic control (GC) method as proposed by Devlin and Roeder
(Devlin et al. Biometrics 1999, 55:997-1004) can be used to adjust
for the inflation of test statistics due to population
stratification. GC method is robust to changes in population
structure levels as well as being applicable to DNA pooling designs
(Devlin et al. Genet. Epidem. 20001, 21:273-284).
[0209] While Pritchard's method recommended using 15-20 unlinked
microsatellite markers, it suggested using more than 30 biallelic
markers to get enough power to detect population stratification.
For the GC method, it has been shown (Bacanu et al. Am. J. Hum.
Genet. 2000, 66:1933-1944) that about 60-70 biallelic markers are
sufficient to estimate the inflation factor for the test statistics
due to population stratification. Hence, 70 intergenic SNPs can be
chosen in unlinked regions as indicated in a genome scan (Kehoe et
al. Hum. Mol. Genet. 1999, 8:237-245).
[0210] Once individual risk factors, genetic or non-genetic, have
been found for the predisposition to disease, the next step is to
set up a classification/prediction scheme to predict the category
(for instance, disease or no-disease) that an individual will be in
depending on his genotypes of associated SNPs and other non-genetic
risk factors. Logistic regression for discrete trait and linear
regression for continuous trait are standard techniques for such
tasks (Applied Regression Analysis, Draper and Smith, Wiley
(1998)). Moreover, other techniques can also be used for setting up
classification. Such techniques include, but are not limited to,
MART, CART, neural network, and discriminant analyses that are
suitable for use in comparing the performance of different methods
(The Elements of Statistical Learning, Hastie, Tibshirani &
Friedman, Springer (2002)).
[0211] Disease Diagnosis and Predisposition Screening
[0212] Information on association/correlation between genotypes and
disease-related phenotypes can be exploited in several ways. For
example, in the case of a highly statistically significant
association between one or more SNPs with predisposition to a
disease for which treatment is available, detection of such a
genotype pattern in an individual may justify immediate
administration of treatment, or at least the institution of regular
monitoring of the individual. Detection of the susceptibility
alleles associated with serious disease in a couple contemplating
having children may also be valuable to the couple in their
reproductive decisions. In the case of a weaker but still
statistically significant association between a SNP and a human
disease, immediate therapeutic intervention or monitoring may not
be justified after detecting the susceptibility allele or SNP.
Nevertheless, the subject can be motivated to begin simple
life-style changes (e.g., diet, exercise) that can be accomplished
at little or no cost to the individual but would confer potential
benefits in reducing the risk of developing conditions for which
that individual may have an increased risk by virtue of having the
susceptibility allele(s).
[0213] The SNPs of the invention may contribute to stenosis in an
individual in different ways. Some polymorphisms occur within a
protein coding sequence and contribute to disease phenotype by
affecting protein structure. Other polymorphisms occur in noncoding
regions but may exert phenotypic effects indirectly via influence
on, for example, replication, transcription, and/or translation. A
single SNP may affect more than one phenotypic trait. Likewise, a
single phenotypic trait may be affected by multiple SNPs in
different genes.
[0214] As used herein, the terms "diagnose", "diagnosis", and
"diagnostics" include, but are not limited to any of the following:
detection of stenosis that an individual may presently have,
predisposition screening (i.e., determining the increased risk of
an individual in developing stenosis in the future, or determining
whether an individual has a decreased risk of developing stenosis
in the future), determining a particular type or subclass of
stenosis in an individual known to have stenosis, confirming or
reinforcing a previously made diagnosis of stenosis,
pharmacogenomic evaluation of an individual to determine which
therapeutic strategy that individual is most likely to positively
respond to or to predict whether a patient is likely to respond to
a particular treatment, predicting whether a patient is likely to
experience toxic effects from a particular treatment or therapeutic
compound, and evaluating the future prognosis of an individual
having stenosis. Such diagnostic uses are based on the SNPs
individually or in a unique combination or SNP haplotypes of the
present invention.
[0215] Haplotypes are particularly useful in that, for example,
fewer SNPs can be genotyped to determine if a particular genomic
region harbors a locus that influences a particular phenotype, such
as in linkage disequilibrium-based SNP association analysis.
[0216] Linkage disequilibrium (LD) refers to the co-inheritance of
alleles (e.g., alternative nucleotides) at two or more different
SNP sites at frequencies greater than would be expected from the
separate frequencies of occurrence of each allele in a given
population. The expected frequency of co-occurrence of two alleles
that are inherited independently is the frequency of the first
allele multiplied by the frequency of the second allele. Alleles
that co-occur at expected frequencies are said to be in "linkage
equilibrium". In contrast, LD refers to any non-random genetic
association between allele(s) at two or more different SNP sites,
which is generally due to the physical proximity of the two loci
along a chromosome. LD can occur when two or more SNPs sites are in
close physical proximity to each other on a given chromosome and
therefore alleles at these SNP sites will tend to remain
unseparated for multiple generations with the consequence that a
particular nucleotide (allele) at one SNP site will show a
non-random association with a particular nucleotide (allele) at a
different SNP site located nearby. Hence, genotyping one of the SNP
sites will give almost the same information as genotyping the other
SNP site that is in LD.
[0217] For diagnostic purposes, if a particular SNP site is found
to be useful for diagnosing stenosis, then the skilled artisan
would recognize that other SNP sites which are in LD with this SNP
site would also be useful for diagnosing the condition. Various
degrees of LD can be encountered between two or more SNPs with the
result being that some SNPs are more closely associated (i.e., in
stronger LD) than others. Furthermore, the physical distance over
which LD extends along a chromosome differs between different
regions of the genome, and therefore the degree of physical
separation between two or more SNP sites necessary for LD to occur
can differ between different regions of the genome.
[0218] For diagnostic applications, polymorphisms (e.g., SNPs
and/or haplotypes) that are not the actual disease-causing
(causative) polymorphisms, but are in LD with such causative
polymorphisms, are also useful. In such instances, the genotype of
the polymorphism(s) that is/are in LD with the causative
polymorphism is predictive of the genotype of the causative
polymorphism and, consequently, predictive of the phenotype (e.g.,
stenosis) that is influenced by the causative SNP(s). Thus,
polymorphic markers that are in LD with causative polymorphisms are
useful as diagnostic markers, and are particularly useful when the
actual causative polymorphism(s) is/are unknown.
[0219] Linkage disequilibrium in the human genome is reviewed in:
Wall et al., "Haplotype blocks and linkage disequilibrium in the
human genome", Nat Rev Genet. 2003 August; 4(8):587-97; Garner et
al., "On selecting markers for association studies: patterns of
linkage disequilibrium between two and three diallelic loci", Genet
Epidemiol. 2003 January; 24(1):57-67; Ardlie et al., "Patterns of
linkage disequilibrium in the human genome", Nat Rev Genet. 2002
April; 3(4):299-309 (erratum in Nat Rev Genet. 2002 July;
3(7):566); and Remm et al., "High-density genotyping and linkage
disequilibrium in the human genome using chromosome 22 as a model";
Curr Opin Chem. Biol. 2002 February; 6(1):24-30.
[0220] The contribution or association of particular SNPs and/or
SNP haplotypes with disease phenotypes, such as stenosis, enables
the SNPs of the present invention to be used to develop superior
diagnostic tests capable of identifying individuals who express a
detectable trait, such as stenosis, as the result of a specific
genotype, or individuals whose genotype places them at an increased
or decreased risk of developing a detectable trait at a subsequent
time as compared to individuals who do not have that genotype. As
described herein, diagnostics may be based on a single SNP or a
group of SNPs. Combined detection of a plurality of SNPs (for
example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
18, 19, 20, 24, 25, 30, 32, 48, 50, 64, 96, 100, or any other
number in-between, or more, of the SNPs provided in Table 1 and/or
Table 2) typically increases the probability of an accurate
diagnosis. For example, the presence of a single SNP known to
correlate with stenosis might indicate a probability of 20% that an
individual has or is at risk of developing stenosis, whereas
detection of five SNPs, each of which correlates with stenosis,
might indicate a probability of 80% that an individual has or is at
risk of developing stenosis. To further increase the accuracy of
diagnosis or predisposition screening, analysis of the SNPs of the
present invention can be combined with that of other polymorphisms
or other risk factors of stenosis, such as disease symptoms,
pathological characteristics, family history, diet, environmental
factors or lifestyle factors.
[0221] It will, of course, be understood by practitioners skilled
in the treatment or diagnosis of stenosis that the present
invention generally does not intend to provide an absolute
identification of individuals who are at risk (or less at risk) of
developing stenosis, and/or pathologies related to stenosis, but
rather to indicate a certain increased (or decreased) degree or
likelihood of developing the disease based on statistically
significant association results. However, this information is
extremely valuable as it can be used to, for example, initiate
preventive treatments or to allow an individual carrying one or
more significant SNPs or SNP haplotypes to foresee warning signs
such as minor clinical symptoms, or to have regularly scheduled
physical exams to monitor for appearance of a condition in order to
identify and begin treatment of the condition at an early stage.
Particularly with diseases that are extremely debilitating or fatal
if not treated on time, the knowledge of a potential
predisposition, even if this predisposition is not absolute, would
likely contribute in a very significant manner to treatment
efficacy.
[0222] The diagnostic techniques of the present invention may
employ a variety of methodologies to determine whether a test
subject has a SNP or a SNP pattern associated with an increased or
decreased risk of developing a detectable trait or whether the
individual suffers from a detectable trait as a result of a
particular polymorphism/mutation, including, for example, methods
which enable the analysis of individual chromosomes for
haplotyping, family studies, single sperm DNA analysis, or somatic
hybrids. The trait analyzed using the diagnostics of the invention
may be any detectable trait that is commonly observed in
pathologies and disorders related to stenosis.
[0223] Another aspect of the present invention relates to a method
of determining whether an individual is at risk (or less at risk)
of developing one or more traits or whether an individual expresses
one or more traits as a consequence of possessing a particular
trait-causing or trait-influencing allele. These methods generally
involve obtaining a nucleic acid sample from an individual and
assaying the nucleic acid sample to determine which nucleotide(s)
is/are present at one or more SNP positions, wherein the assayed
nucleotide(s) is/are indicative of an increased or decreased risk
of developing the trait or indicative that the individual expresses
the trait as a result of possessing a particular trait-causing or
trait-influencing allele.
[0224] In another embodiment, the SNP detection reagents of the
present invention are used to determine whether an individual has
one or more SNP allele(s) affecting the level (e.g., the
concentration of mRNA or protein in a sample, etc.) or pattern
(e.g., the kinetics of expression, rate of decomposition, stability
profile, Km, Vmax, etc.) of gene expression (collectively, the
"gene response" of a cell or bodily fluid). Such a determination
can be accomplished by screening for mRNA or protein expression
(e.g., by using nucleic acid arrays, RT-PCR, TaqMan assays, or mass
spectrometry), identifying genes having altered expression in an
individual, genotyping SNPs disclosed in Table 1 and/or Table 2
that could affect the expression of the genes having altered
expression (e.g., SNPs that are in and/or around the gene(s) having
altered expression, SNPs in regulatory/control regions, SNPs in
and/or around other genes that are involved in pathways that could
affect the expression of the gene(s) having altered expression, or
all SNPs could be genotyped), and correlating SNP genotypes with
altered gene expression. In this manner, specific SNP alleles at
particular SNP sites can be identified that affect gene
expression.
[0225] Pharmacogenomics and Therapeutics/Drug Development
[0226] The present invention provides methods for assessing the
pharmacogenomics of a subject harboring particular SNP alleles or
haplotypes to a particular therapeutic agent or pharmaceutical
compound, or to a class of such compounds. Pharmacogenomics deals
with the roles which clinically significant hereditary variations
(e.g., SNPs) play in the response to drugs due to altered drug
disposition and/or abnormal action in affected persons. See, e.g.,
Roses, Nature 405, 857-865 (2000); Gould Rothberg, Nature
Biotechnology 19, 209-211 (2001); Eichelbaum, Clin. Exp. Pharmacol.
Physiol. 23(10-11):983-985 (1996); and Linder, Clin. Chem.
43(2):254-266 (1997). The clinical outcomes of these variations can
result in severe toxicity of therapeutic drugs in certain
individuals or therapeutic failure of drugs in certain individuals
as a result of individual variation in metabolism. Thus, the SNP
genotype of an individual can determine the way a therapeutic
compound acts on the body or the way the body metabolizes the
compound. For example, SNPs in drug metabolizing enzymes can affect
the activity of these enzymes, which in turn can affect both the
intensity and duration of drug action, as well as drug metabolism
and clearance.
[0227] The discovery of SNPs in drug metabolizing enzymes, drug
transporters, proteins for pharmaceutical agents, and other drug
targets has explained why some patients do not obtain the expected
drug effects, show an exaggerated drug effect, or experience
serious toxicity from standard drug dosages. SNPs can be expressed
in the phenotype of the extensive metabolizer and in the phenotype
of the poor metabolizer. Accordingly, SNPs may lead to allelic
variants of a protein in which one or more of the protein functions
in one population are different from those in another population.
SNPs and the encoded variant peptides thus provide targets to
ascertain a genetic predisposition that can affect treatment
modality. For example, in a ligand-based treatment, SNPs may give
rise to amino terminal extracellular domains and/or other
ligand-binding regions of a receptor that are more or less active
in ligand binding, thereby affecting subsequent protein activation.
Accordingly, ligand dosage would necessarily be modified to
maximize the therapeutic effect within a given population
containing particular SNP alleles or haplotypes.
[0228] As an alternative to genotyping, specific variant proteins
containing variant amino acid sequences encoded by alternative SNP
alleles could be identified. Thus, pharmacogenomic characterization
of an individual permits the selection of effective compounds and
effective dosages of such compounds for prophylactic or therapeutic
uses based on the individual's SNP genotype, thereby enhancing and
optimizing the effectiveness of the therapy. Furthermore, the
production of recombinant cells and transgenic animals containing
particular SNPs/haplotypes allow effective clinical design and
testing of treatment compounds and dosage regimens. For example,
transgenic animals can be produced that differ only in specific SNP
alleles in a gene that is orthologous to a human disease
susceptibility gene.
[0229] Pharmacogenomic uses of the SNPs of the present invention
provide several significant advantages for patient care,
particularly in treating stenosis. Pharmacogenomic characterization
of an individual, based on an individual's SNP genotype, can
identify those individuals unlikely to respond to treatment with a
particular medication and thereby allows physicians to avoid
prescribing the ineffective medication to those individuals. On the
other hand, SNP genotyping of an individual may enable physicians
to select the appropriate medication and dosage regimen that will
be most effective based on an individual's SNP genotype. This
information increases a physician's confidence in prescribing
medications and motivates patients to comply with their drug
regimens. Furthermore, pharmacogenomics may identify patients
predisposed to toxicity and adverse reactions to particular drugs
or drug dosages. Adverse drug reactions lead to more than 100,000
avoidable deaths per year in the United States alone and therefore
represent a significant cause of hospitalization and death, as well
as a significant economic burden on the healthcare system (Pfost
et. al., Trends in Biotechnology, August 2000.). Thus,
pharmacogenomics based on the SNPs disclosed herein has the
potential to both save lives and reduce healthcare costs
substantially.
[0230] Pharmacogenomics in general is discussed further in Rose et
al., "Pharmacogenetic analysis of clinically relevant genetic
polymorphisms", Methods Mol. Med. 2003; 85:225-37. Pharmacogenomics
as it relates to Alzheimer's disease and other neurodegenerative
disorders is discussed in Cacabelos, "Pharmacogenomics for the
treatment of dementia", Ann Med. 2002; 34(5):357-79, Maimone et
al., "Pharmacogenomics of neurodegenerative diseases", Eur J.
Pharmacol. 2001 Feb. 9; 413(1): 11-29, and Poirier, "Apolipoprotein
E: a pharmacogenetic target for the treatment of Alzheimer's
disease", Mol. Diagn. 1999 December; 4(4):335-41. Pharmacogenomics
as it relates to cardiovascular disorders is discussed in Siest et
al., "Pharmacogenomics of drugs affecting the cardiovascular
system", Clin Chem Lab Med. 2003 April; 41(4):590-9, Mukherjee et
al., "Pharmacogenomics in cardiovascular diseases", Prog Cardiovasc
Dis. 2002 May-June; 44(6):479-98, and Mooser et al.,
"Cardiovascular pharmacogenetics in the SNP era", J Thromb Haemost.
2003 July; 1(7): 1398-402. Pharmacogenomics as it relates to cancer
is discussed in McLeod et al., "Cancer pharmacogenomics: SNPs,
chips, and the individual patient", Cancer Invest. 2003;
21(4):630-40 and Watters et al., "Cancer pharmacogenomics: current
and future applications", Biochim Biophys Acta. 2003 Mar. 17;
1603(2):99-111.
[0231] The SNPs of the present invention also can be used to
identify novel therapeutic targets for stenosis. For example, genes
containing the disease-associated variants ("variant genes") or
their products, as well as genes or their products that are
directly or indirectly regulated by or interacting with these
variant genes or their products, can be targeted for the
development of therapeutics that, for example, treat the disease or
prevent or delay disease onset. The therapeutics may be composed
of, for example, small molecules, proteins, protein fragments or
peptides, antibodies, nucleic acids, or their derivatives or
mimetics which modulate the functions or levels of the target genes
or gene products.
[0232] The SNP-containing nucleic acid molecules disclosed herein,
and their complementary nucleic acid molecules, may be used as
antisense constructs to control gene expression in cells, tissues,
and organisms. Antisense technology is well established in the art
and extensively reviewed in Antisense Drug Technology. Principles,
Strategies, and Applications, Crooke (ed.), Marcel Dekker, Inc.:
New York (2001). An antisense nucleic acid molecule is generally
designed to be complementary to a region of mRNA expressed by a
gene so that the antisense molecule hybridizes to the mRNA and
thereby blocks translation of mRNA into protein. Various classes of
antisense oligonucleotides are used in the art, two of which are
cleavers and blockers. Cleavers, by binding to target RNAs,
activate intracellular nucleases (e.g., RNaseH or RNase L) that
cleave the target RNA. Blockers, which also bind to target RNAs,
inhibit protein translation through steric hindrance of ribosomes.
Exemplary blockers include peptide nucleic acids, morpholinos,
locked nucleic acids, and methylphosphonates (see, e.g., Thompson,
Drug Discovery Today, 7 (17): 912-917 (2002)). Antisense
oligonucleotides are directly useful as therapeutic agents, and are
also useful for determining and validating gene function (e.g., in
gene knock-out or knock-down experiments).
[0233] Antisense technology is further reviewed in: Lavery et al.,
"Antisense and RNAi: powerful tools in drug target discovery and
validation", Curr Opin Drug Discov Devel. 2003 July; 6(4):561-9;
Stephens et al., "Antisense oligonucleotide therapy in cancer",
Curr Opin Mol. Ther. 2003 April; 5(2): 118-22; Kurreck, "Antisense
technologies. Improvement through novel chemical modifications",
Eur J. Biochem. 2003 April; 270(8):1628-44; Dias et al., "Antisense
oligonucleotides: basic concepts and mechanisms", Mol Cancer Ther.
2002 March; 1(5):347-55; Chen, "Clinical development of antisense
oligonucleotides as anti-cancer therapeutics", Methods Mol. Med.
2003; 75:621-36; Wang et al., "Antisense anticancer oligonucleotide
therapeutics", Curr Cancer Drug Targets. 2001 November; 1 (3):
177-96; and Bennett, "Efficiency of antisense oligonucleotide drug
discovery", Antisense Nucleic Acid Drug Dev. 2002 June;
12(3):215-24.
[0234] The SNPs of the present invention are particularly useful
for designing antisense reagents that are specific for particular
nucleic acid variants. Based on the SNP information disclosed
herein, antisense oligonucleotides can be produced that
specifically target mRNA molecules that contain one or more
particular SNP nucleotides. In this manner, expression of mRNA
molecules that contain one or more undesired polymorphisms (e.g.,
SNP nucleotides that lead to a defective protein such as an amino
acid substitution in a catalytic domain) can be inhibited or
completely blocked. Thus, antisense oligonucleotides can be used to
specifically bind a particular polymorphic form (e.g., a SNP allele
that encodes a defective protein), thereby inhibiting translation
of this form, but which do not bind an alternative polymorphic form
(e.g., an alternative SNP nucleotide that encodes a protein having
normal function).
[0235] Antisense molecules can be used to inactivate mRNA in order
to inhibit gene expression and production of defective proteins.
Accordingly, these molecules can be used to treat a disorder, such
as stenosis, characterized by abnormal or undesired gene expression
or expression of certain defective proteins. This technique can
involve cleavage by means of ribozymes containing nucleotide
sequences complementary to one or more regions in the mRNA that
attenuate the ability of the mRNA to be translated. Possible mRNA
regions include, for example, protein-coding regions and
particularly protein-coding regions corresponding to catalytic
activities, substrate/ligand binding, or other functional
activities of a protein.
[0236] The SNPs of the present invention are also useful for
designing RNA interference reagents that specifically target
nucleic acid molecules having particular SNP variants. RNA
interference (RNAi), also referred to as gene silencing, is based
on using double-stranded RNA (dsRNA) molecules to turn genes off.
When introduced into a cell, dsRNAs are processed by the cell into
short fragments (generally about 21-22 bp in length) known as small
interfering RNAs (siRNAs) which the cell uses in a
sequence-specific manner to recognize and destroy complementary
RNAs (Thompson, Drug Discovery Today, 7 (17): 912-917 (2002)).
Thus, because RNAi molecules, including siRNAs, act in a
sequence-specific manner, the SNPs of the present invention can be
used to design RNAi reagents that recognize and destroy nucleic
acid molecules having specific SNP alleles/nucleotides (such as
deleterious alleles that lead to the production of defective
proteins), while not affecting nucleic acid molecules having
alternative SNP alleles (such as alleles that encode proteins
having normal function). As with antisense reagents, RNAi reagents
may be directly useful as therapeutic agents (e.g., for turning off
defective, disease-causing genes), and are also useful for
characterizing and validating gene function (e.g., in gene
knock-out or knock-down experiments).
[0237] The following references provide a further review of RNAi:
Agami, "RNAi and related mechanisms and their potential use for
therapy", Curr Opin Chem. Biol. 2002 December; 6(6):829-34; Lavery
et al., "Antisense and RNAi: powerful tools in drug target
discovery and validation", Curr Opin Drug Discov Devel. 2003 July;
6(4):561-9; Shi, "Mammalian RNAi for the masses", Trends Genet.
2003 January; 19(1):9-12), Shuey et al., "RNAi: gene-silencing in
therapeutic intervention", Drug Discovery Today 2002 October;
7(20):1040-1046; McManus et al., Nat Rev Genet. 2002 October;
3(10):737-47; Xia et al., Nat Biotechnol 2002 October;
20(10):1006-10; Plasterk et al., Curr Opin Genet Dev 2000 October;
10(5):562-7; Bosher et al., Nat Cell Biol 2000 February;
2(2):E31-6; and Hunter, Curr Biol 1999 Jun. 17; 9(12):R440-2).
[0238] A subject suffering from a pathological condition, such as
stenosis, ascribed to a SNP may be treated so as to correct the
genetic defect (see Kren et al., Proc. Natl. Acad. Sci. USA
96:10349-10354 (1999)). Such a subject can be identified by any
method that can detect the polymorphism in a biological sample
drawn from the subject. Such a genetic defect may be permanently
corrected by administering to such a subject a nucleic acid
fragment incorporating a repair sequence that supplies the
normal/wild-type nucleotide at the position of the SNP. This
site-specific repair sequence can encompass an RNA/DNA
oligonucleotide that operates to promote endogenous repair of a
subject's genomic DNA. The site-specific repair sequence is
administered in an appropriate vehicle, such as a complex with
polyethylenimine, encapsulated in anionic liposomes, a viral vector
such as an adenovirus, or other pharmaceutical composition that
promotes intracellular uptake of the administered nucleic acid. A
genetic defect leading to an inborn pathology may then be overcome,
as the chimeric oligonucleotides induce incorporation of the normal
sequence into the subject's genome. Upon incorporation, the normal
gene product is expressed, and the replacement is propagated,
thereby engendering a permanent repair and therapeutic enhancement
of the clinical condition of the subject.
[0239] In cases in which a cSNP results in a variant protein that
is ascribed to be the cause of, or a contributing factor to, a
pathological condition, a method of treating such a condition can
include administering to a subject experiencing the pathology the
wild-type/normal cognate of the variant protein. Once administered
in an effective dosing regimen, the wild-type cognate provides
complementation or remediation of the pathological condition.
[0240] The invention further provides a method for identifying a
compound or agent that can be used to treat stenosis. The SNPs
disclosed herein are useful as targets for the identification
and/or development of therapeutic agents. A method for identifying
a therapeutic agent or compound typically includes assaying the
ability of the agent or compound to modulate the activity and/or
expression of a SNP-containing nucleic acid or the encoded product
and thus identifying an agent or a compound that can be used to
treat a disorder characterized by undesired activity or expression
of the SNP-containing nucleic acid or the encoded product. The
assays can be performed in cell-based and cell-free systems.
Cell-based assays can include cells naturally expressing the
nucleic acid molecules of interest or recombinant cells genetically
engineered to express certain nucleic acid molecules.
[0241] Variant gene expression in a stenosis patient can include,
for example, either expression of a SNP-containing nucleic acid
sequence (for instance, a gene that contains a SNP can be
transcribed into an mRNA transcript molecule containing the SNP,
which can in turn be translated into a variant protein) or altered
expression of a normal/wild-type nucleic acid sequence due to one
or more SNPs (for instance, a regulatory/control region can contain
a SNP that affects the level or pattern of expression of a normal
transcript).
[0242] Assays for variant gene expression can involve direct assays
of nucleic acid levels (e.g., mRNA levels), expressed protein
levels, or of collateral compounds involved in a signal pathway.
Further, the expression of genes that are up- or down-regulated in
response to the signal pathway can also be assayed. In this
embodiment, the regulatory regions of these genes can be operably
linked to a reporter gene such as luciferase.
[0243] Modulators of variant gene expression can be identified in a
method wherein, for example, a cell is contacted with a candidate
compound/agent and the expression of mRNA determined. The level of
expression of mRNA in the presence of the candidate compound is
compared to the level of expression of mRNA in the absence of the
candidate compound. The candidate compound can then be identified
as a modulator of variant gene expression based on this comparison
and be used to treat a disorder such as stenosis that is
characterized by variant gene expression (e.g., either expression
of a SNP-containing nucleic acid or altered expression of a
normal/wild-type nucleic acid molecule due to one or more SNPs that
affect expression of the nucleic acid molecule) due to one or more
SNPs of the present invention. When expression of mRNA is
statistically significantly greater in the presence of the
candidate compound than in its absence, the candidate compound is
identified as a stimulator of nucleic acid expression. When nucleic
acid expression is statistically significantly less in the presence
of the candidate compound than in its absence, the candidate
compound is identified as an inhibitor of nucleic acid
expression.
[0244] The invention further provides methods of treatment, with
the SNP or associated nucleic acid domain (e.g., catalytic domain,
ligand/substrate-binding domain, regulatory/control region, etc.)
or gene, or the encoded mRNA transcript, as a target, using a
compound identified through drug screening as a gene modulator to
modulate variant nucleic acid expression. Modulation can include
either up-regulation (i.e., activation or agonization) or
down-regulation (i.e., suppression or antagonization) of nucleic
acid expression.
[0245] Expression of mRNA transcripts and encoded proteins, either
wild type or variant, may be altered in individuals with a
particular SNP allele in a regulatory/control element, such as a
promoter or transcription factor binding domain, that regulates
expression. In this situation, methods of treatment and compounds
can be identified, as discussed herein, that regulate or overcome
the variant regulatory/control element, thereby generating normal,
or healthy, expression levels of either the wild type or variant
protein.
[0246] The SNP-containing nucleic acid molecules of the present
invention are also useful for monitoring the effectiveness of
modulating compounds on the expression or activity of a variant
gene, or encoded product, in clinical trials or in a treatment
regimen. Thus, the gene expression pattern can serve as an
indicator for the continuing effectiveness of treatment with the
compound, particularly with compounds to which a patient can
develop resistance, as well as an indicator for toxicities. The
gene expression pattern can also serve as a marker indicative of a
physiological response of the affected cells to the compound.
Accordingly, such monitoring would allow either increased
administration of the compound or the administration of alternative
compounds to which the patient has not become resistant. Similarly,
if the level of nucleic acid expression falls below a desirable
level, administration of the compound could be commensurately
decreased.
[0247] In another aspect of the present invention, there is
provided a pharmaceutical pack comprising a therapeutic agent
(e.g., a small molecule drug, antibody, peptide, antisense or RNAi
nucleic acid molecule, etc.) and a set of instructions for
administration of the therapeutic agent to humans diagnostically
tested for one or more SNPs or SNP haplotypes provided by the
present invention.
[0248] The SNPs/haplotypes of the present invention are also useful
for improving many different aspects of the drug development
process. For example, individuals can be selected for clinical
trials based on their SNP genotype. Individuals with SNP genotypes
that indicate that they are most likely to respond to the drug can
be included in the trials and those individuals whose SNP genotypes
indicate that they are less likely to or would not respond to the
drug, or suffer adverse reactions, can be eliminated from the
clinical trials. This not only improves the safety of clinical
trials, but also will enhance the chances that the trial will
demonstrate statistically significant efficacy. Furthermore, the
SNPs of the present invention may explain why certain previously
developed drugs performed poorly in clinical trials and may help
identify a subset of the population that would benefit from a drug
that had previously performed poorly in clinical trials, thereby
"rescuing" previously developed drugs, and enabling the drug to be
made available to a particular stenosis patient population that can
benefit from it.
[0249] SNPs have many important uses in drug discovery, screening,
and development. A high probability exists that, for any
gene/protein selected as a potential drug target, variants of that
gene/protein will exist in a patient population. Thus, determining
the impact of gene/protein variants on the selection and delivery
of a therapeutic agent should be an integral aspect of the drug
discovery and development process. (Jazwinska, A Trends Guide to
Genetic Variation and Genomic Medicine, 2002 March; S30-S36).
[0250] Knowledge of variants (e.g., SNPs and any corresponding
amino acid polymorphisms) of a particular therapeutic target (e.g.,
a gene, mRNA transcript, or protein) enables parallel screening of
the variants in order to identify therapeutic candidates (e.g.,
small molecule compounds, antibodies, antisense or RNAi nucleic
acid compounds, etc.) that demonstrate efficacy across variants
(Rothberg, Nat Biotechnol 2001 March; 19(3):209-11). Such
therapeutic candidates would be expected to show equal efficacy
across a larger segment of the patient population, thereby leading
to a larger potential market for the therapeutic candidate.
[0251] Furthermore, identifying variants of a potential therapeutic
target enables the most common form of the target to be used for
selection of therapeutic candidates, thereby helping to ensure that
the experimental activity that is observed for the selected
candidates reflects the real activity expected in the largest
proportion of a patient population (Jazwinska, A Trends Guide to
Genetic Variation and Genomic Medicine, 2002 March; S30-S36).
[0252] Additionally, screening therapeutic candidates against all
known variants of a target can enable the early identification of
potential toxicities and adverse reactions relating to particular
variants. For example, variability in drug absorption,
distribution, metabolism and excretion (ADME) caused by, for
example, SNPs in therapeutic targets or drug metabolizing genes,
can be identified, and this information can be utilized during the
drug development process to minimize variability in drug
disposition and develop therapeutic agents that are safer across a
wider range of a patient population. The SNPs of the present
invention, including the variant proteins and encoding polymorphic
nucleic acid molecules provided in Tables 1-2, are useful in
conjunction with a variety of toxicology methods established in the
art, such as those set forth in Current Protocols in Toxicology,
John Wiley & Sons, Inc., N.Y.
[0253] Furthermore, therapeutic agents that target any art-known
proteins (or nucleic acid molecules, either RNA or DNA) may
cross-react with the variant proteins (or polymorphic nucleic acid
molecules) disclosed in Table 1, thereby significantly affecting
the pharmacokinetic properties of the drug. Consequently, the
protein variants and the SNP-containing nucleic acid molecules
disclosed in Tables 1-2 are useful in developing, screening, and
evaluating therapeutic agents that target corresponding art-known
protein forms (or nucleic acid molecules). Additionally, as
discussed above, knowledge of all polymorphic forms of a particular
drug target enables the design of therapeutic agents that are
effective against most or all such polymorphic forms of the drug
target.
[0254] Pharmaceutical Compositions and Administration Thereof
[0255] Any of the stenosis-associated proteins, and encoding
nucleic acid molecules, disclosed herein can be used as therapeutic
targets (or directly used themselves as therapeutic compounds) for
treating stenosis and related pathologies, and the present
disclosure enables therapeutic compounds (e.g., small molecules,
antibodies, therapeutic proteins, RNAi and antisense molecules,
etc.) to be developed that target (or are comprised of) any of
these therapeutic targets.
[0256] In general, a therapeutic compound will be administered in a
therapeutically effective amount by any of the accepted modes of
administration for agents that serve similar utilities. The actual
amount of the therapeutic compound of this invention, i.e., the
active ingredient, will depend upon numerous factors such as the
severity of the disease to be treated, the age and relative health
of the subject, the potency of the compound used, the route and
form of administration, and other factors.
[0257] Therapeutically effective amounts of therapeutic compounds
may range from, for example, approximately 0.01-50 mg per kilogram
body weight of the recipient per day; preferably about 0.1-20
mg/kg/day. Thus, as an example, for administration to a 70 kg
person, the dosage range would most preferably be about 7 mg to 1.4
g per day.
[0258] In general, therapeutic compounds will be administered as
pharmaceutical compositions by any one of the following routes:
oral, systemic (e.g., transdermal, intranasal, or by suppository),
or parenteral (e.g., intramuscular, intravenous, or subcutaneous)
administration. The preferred manner of administration is oral or
parenteral using a convenient daily dosage regimen, which can be
adjusted according to the degree of affliction. Oral compositions
can take the form of tablets, pills, capsules, semisolids, powders,
sustained release formulations, solutions, suspensions, elixirs,
aerosols, or any other appropriate compositions.
[0259] The choice of formulation depends on various factors such as
the mode of drug administration (e.g., for oral administration,
formulations in the form of tablets, pills, or capsules are
preferred) and the bioavailability of the drug substance. Recently,
pharmaceutical formulations have been developed especially for
drugs that show poor bioavailability based upon the principle that
bioavailability can be increased by increasing the surface area,
i.e., decreasing particle size. For example, U.S. Pat. No.
4,107,288 describes a pharmaceutical formulation having particles
in the size range from 10 to 1,000 nm in which the active material
is supported on a cross-linked matrix of macromolecules. U.S. Pat.
No. 5,145,684 describes the production of a pharmaceutical
formulation in which the drug substance is pulverized to
nanoparticles (average particle size of 400 nm) in the presence of
a surface modifier and then dispersed in a liquid medium to give a
pharmaceutical formulation that exhibits remarkably high
bioavailability.
[0260] Pharmaceutical compositions are comprised of, in general, a
therapeutic compound in combination with at least one
pharmaceutically acceptable excipient. Acceptable excipients are
non-toxic, aid administration, and do not adversely affect the
therapeutic benefit of the therapeutic compound. Such excipients
may be any solid, liquid, semi-solid or, in the case of an aerosol
composition, gaseous excipient that is generally available to one
skilled in the art.
[0261] Solid pharmaceutical excipients include starch, cellulose,
talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk,
silica gel, magnesium stearate, sodium stearate, glycerol
monostearate, sodium chloride, dried skim milk and the like. Liquid
and semisolid excipients may be selected from glycerol, propylene
glycol, water, ethanol and various oils, including those of
petroleum, animal, vegetable or synthetic origin, e.g., peanut oil,
soybean oil, mineral oil, sesame oil, etc. Preferred liquid
carriers, particularly for injectable solutions, include water,
saline, aqueous dextrose, and glycols.
[0262] Compressed gases may be used to disperse a compound of this
invention in aerosol form. Inert gases suitable for this purpose
are nitrogen, carbon dioxide, etc.
[0263] Other suitable pharmaceutical excipients and their
formulations are described in Remington's Pharmaceutical Sciences,
edited by E. W. Martin (Mack Publishing Company, 18th ed.,
1990).
[0264] The amount of the therapeutic compound in a formulation can
vary within the full range employed by those skilled in the art.
Typically, the formulation will contain, on a weight percent (wt %)
basis, from about 0.01-99.99 wt % of the therapeutic compound based
on the total formulation, with the balance being one or more
suitable pharmaceutical excipients. Preferably, the compound is
present at a level of about 1-80 wt %.
[0265] Therapeutic compounds can be administered alone or in
combination with other therapeutic compounds or in combination with
one or more other active ingredient(s). For example, an inhibitor
or stimulator of a stenosis-associated protein can be administered
in combination with another agent that inhibits or stimulates the
activity of the same or a different stenosis-associated protein to
thereby counteract the affects of stenosis.
[0266] For further information regarding pharmacology, see Current
Protocols in Pharmacology, John Wiley & Sons, Inc., N.Y.
[0267] Human Identification Applications
[0268] In addition to their diagnostic and therapeutic uses in
stenosis and related pathologies, the SNPs provided by the present
invention are also useful as human identification markers for such
applications as forensics, paternity testing, and biometrics (see,
e.g., Gill, "An assessment of the utility of single nucleotide
polymorphisms (SNPs) for forensic purposes", Int J Legal Med. 2001;
114(4-5):204-10). Genetic variations in the nucleic acid sequences
between individuals can be used as genetic markers to identify
individuals and to associate a biological sample with an
individual. Determination of which nucleotides occupy a set of SNP
positions in an individual identifies a set of SNP markers that
distinguishes the individual. The more SNP positions that are
analyzed, the lower the probability that the set of SNPs in one
individual is the same as that in an unrelated individual.
Preferably, if multiple sites are analyzed, the sites are unlinked
(i.e., inherited independently). Thus, preferred sets of SNPs can
be selected from among the SNPs disclosed herein, which may include
SNPs on different chromosomes, SNPs on different chromosome arms,
and/or SNPs that are dispersed over substantial distances along the
same chromosome arm.
[0269] Furthermore, among the SNPs disclosed herein, preferred SNPs
for use in certain forensic/human identification applications
include SNPs located at degenerate codon positions (i.e., the third
position in certain codons which can be one of two or more
alternative nucleotides and still encode the same amino acid),
since these SNPs do not affect the encoded protein. SNPs that do
not affect the encoded protein are expected to be under less
selective pressure and are therefore expected to be more
polymorphic in a population, which is typically an advantage for
forensic/human identification applications. However, for certain
forensics/human identification applications, such as predicting
phenotypic characteristics (e.g., inferring ancestry or inferring
one or more physical characteristics of an individual) from a DNA
sample, it may be desirable to utilize SNPs that affect the encoded
protein.
[0270] For many of the SNPs disclosed in Tables 1-2 (which are
identified as "Applera" SNP source), Tables 1-2 provide SNP allele
frequencies obtained by re-sequencing the DNA of chromosomes from
39 individuals (Tables 1-2 also provide allele frequency
information for "Celera" source SNPs and, where available, public
SNPs from dbEST, HGBASE, and/or HGMD). The allele frequencies
provided in Tables 1-2 enable these SNPs to be readily used for
human identification applications. Although any SNP disclosed in
Table 1 and/or Table 2 could be used for human identification, the
closer that the frequency of the minor allele at a particular SNP
site is to 50%, the greater the ability of that SNP to discriminate
between different individuals in a population since it becomes
increasingly likely that two randomly selected individuals would
have different alleles at that SNP site. Using the SNP allele
frequencies provided in Tables 1-2, one of ordinary skill in the
art could readily select a subset of SNPs for which the frequency
of the minor allele is, for example, at least 1%, 2%, 5%, 10%, 20%,
25%, 30%, 40%, 45%, or 50%, or any other frequency in-between.
Thus, since Tables 1-2 provide allele frequencies based on the
re-sequencing of the chromosomes from 39 individuals, a subset of
SNPs could readily be selected for human identification in which
the total allele count of the minor allele at a particular SNP site
is, for example, at least 1, 2, 4, 8, 10, 16, 20, 24, 30, 32, 36,
38, 39, 40, or any other number in-between.
[0271] Furthermore, Tables 1-2 also provide population group
(interchangeably referred to herein as ethnic or racial groups)
information coupled with the extensive allele frequency
information. For example, the group of 39 individuals whose DNA was
re-sequenced was made-up of 20 Caucasians and 19 African-Americans.
This population group information enables further refinement of SNP
selection for human identification. For example, preferred SNPs for
human identification can be selected from Tables 1-2 that have
similar allele frequencies in both the Caucasian and
African-American populations; thus, for example, SNPs can be
selected that have equally high discriminatory power in both
populations. Alternatively, SNPs can be selected for which there is
a statistically significant difference in allele frequencies
between the Caucasian and African-American populations (as an
extreme example, a particular allele may be observed only in either
the Caucasian or the African-American population group but not
observed in the other population group); such SNPs are useful, for
example, for predicting the race/ethnicity of an unknown
perpetrator from a biological sample such as a hair or blood stain
recovered at a crime scene. For a discussion of using SNPs to
predict ancestry from a DNA sample, including statistical methods,
see Frudakis et al., "A Classifier for the SNP-Based Inference of
Ancestry", Journal of Forensic Sciences 2003; 48(4):771-782.
[0272] SNPs have numerous advantages over other types of
polymorphic markers, such as short tandem repeats (STRs). For
example, SNPs can be easily scored and are amenable to automation,
making SNPs the markers of choice for large-scale forensic
databases. SNPs are found in much greater abundance throughout the
genome than repeat polymorphisms. Population frequencies of two
polymorphic forms can usually be determined with greater accuracy
than those of multiple polymorphic forms at multi-allelic loci.
SNPs are mutationaly more stable than repeat polymorphisms. SNPs
are not susceptible to artifacts such as stutter bands that can
hinder analysis. Stutter bands are frequently encountered when
analyzing repeat polymorphisms, and are particularly troublesome
when analyzing samples such as crime scene samples that may contain
mixtures of DNA from multiple sources. Another significant
advantage of SNP markers over STR markers is the much shorter
length of nucleic acid needed to score a SNP. For example, STR
markers are generally several hundred base pairs in length. A SNP,
on the other hand, comprises a single nucleotide, and generally a
short conserved region on either side of the SNP position for
primer and/or probe binding. This makes SNPs more amenable to
typing in highly degraded or aged biological samples that are
frequently encountered in forensic casework in which DNA may be
fragmented into short pieces.
[0273] SNPs also are not subject to microvariant and "off-ladder"
alleles frequently encountered when analyzing STR loci.
Microvariants are deletions or insertions within a repeat unit that
change the size of the amplified DNA product so that the amplified
product does not migrate at the same rate as reference alleles with
normal sized repeat units. When separated by size, such as by
electrophoresis on a polyacrylamide gel, microvariants do not align
with a reference allelic ladder of standard sized repeat units, but
rather migrate between the reference alleles. The reference allelic
ladder is used for precise sizing of alleles for allele
classification; therefore alleles that do not align with the
reference allelic ladder lead to substantial analysis problems.
Furthermore, when analyzing multi-allelic repeat polymorphisms,
occasionally an allele is found that consists of more or less
repeat units than has been previously seen in the population, or
more or less repeat alleles than are included in a reference
allelic ladder. These alleles will migrate outside the size range
of known alleles in a reference allelic ladder, and therefore are
referred to as "off-ladder" alleles. In extreme cases, the allele
may contain so few or so many repeats that it migrates well out of
the range of the reference allelic ladder. In this situation, the
allele may not even be observed, or, with multiplex analysis, it
may migrate within or close to the size range for another locus,
further confounding analysis.
[0274] SNP analysis avoids the problems of microvariants and
off-ladder alleles encountered in STR analysis. Importantly,
microvariants and off-ladder alleles may provide significant
problems, and may be completely missed, when using analysis methods
such as oligonucleotide hybridization arrays, which utilize
oligonucleotide probes specific for certain known alleles.
Furthermore, off-ladder alleles and microvariants encountered with
STR analysis, even when correctly typed, may lead to improper
statistical analysis, since their frequencies in the population are
generally unknown or poorly characterized, and therefore the
statistical significance of a matching genotype may be
questionable. All these advantages of SNP analysis are considerable
in light of the consequences of most DNA identification cases,
which may lead to life imprisonment for an individual, or
re-association of remains to the family of a deceased
individual.
[0275] DNA can be isolated from biological samples such as blood,
bone, hair, saliva, or semen, and compared with the DNA from a
reference source at particular SNP positions. Multiple SNP markers
can be assayed simultaneously in order to increase the power of
discrimination and the statistical significance of a matching
genotype. For example, oligonucleotide arrays can be used to
genotype a large number of SNPs simultaneously. The SNPs provided
by the present invention can be assayed in combination with other
polymorphic genetic markers, such as other SNPs known in the art or
STRs, in order to identify an individual or to associate an
individual with a particular biological sample.
[0276] Furthermore, the SNPs provided by the present invention can
be genotyped for inclusion in a database of DNA genotypes, for
example, a criminal DNA databank such as the FBI's Combined DNA
Index System (CODIS) database. A genotype obtained from a
biological sample of unknown source can then be queried against the
database to find a matching genotype, with the SNPs of the present
invention providing nucleotide positions at which to compare the
known and unknown DNA sequences for identity. Accordingly, the
present invention provides a database comprising novel SNPs or SNP
alleles of the present invention (e.g., the database can comprise
information indicating which alleles are possessed by individual
members of a population at one or more novel SNP sites of the
present invention), such as for use in forensics, biometrics, or
other human identification applications. Such a database typically
comprises a computer-based system in which the SNPs or SNP alleles
of the present invention are recorded on a computer readable medium
(see the section of the present specification entitled
"Computer-Related Embodiments").
[0277] The SNPs of the present invention can also be assayed for
use in paternity testing. The object of paternity testing is
usually to determine whether a male is the father of a child. In
most cases, the mother of the child is known and thus, the mother's
contribution to the child's genotype can be traced. Paternity
testing investigates whether the part of the child's genotype not
attributable to the mother is consistent with that of the putative
father. Paternity testing can be performed by analyzing sets of
polymorphisms in the putative father and the child, with the SNPs
of the present invention providing nucleotide positions at which to
compare the putative father's and child's DNA sequences for
identity. If the set of polymorphisms in the child attributable to
the father does not match the set of polymorphisms of the putative
father, it can be concluded, barring experimental error, that the
putative father is not the father of the child. If the set of
polymorphisms in the child attributable to the father match the set
of polymorphisms of the putative father, a statistical calculation
can be performed to determine the probability of coincidental
match, and a conclusion drawn as to the likelihood that the
putative father is the true biological father of the child.
[0278] In addition to paternity testing, SNPs are also useful for
other types of kinship testing, such as for verifying familial
relationships for immigration purposes, or for cases in which an
individual alleges to be related to a deceased individual in order
to claim an inheritance from the deceased individual, etc. For
further information regarding the utility of SNPs for paternity
testing and other types of kinship testing, including methods for
statistical analysis, see Krawczak, "Informativity assessment for
biallelic single nucleotide polymorphisms", Electrophoresis 1999
June; 20(8):1676-81.
[0279] The use of the SNPs of the present invention for human
identification further extends to various authentication systems,
commonly referred to as biometric systems, which typically convert
physical characteristics of humans (or other organisms) into
digital data. Biometric systems include various technological
devices that measure such unique anatomical or physiological
characteristics as finger, thumb, or palm prints; hand geometry;
vein patterning on the back of the hand; blood vessel patterning of
the retina and color and texture of the iris; facial
characteristics; voice patterns; signature and typing dynamics; and
DNA. Such physiological measurements can be used to verify identity
and, for example, restrict or allow access based on the
identification. Examples of applications for biometrics include
physical area security, computer and network security, aircraft
passenger check-in and boarding, financial transactions, medical
records access, government benefit distribution, voting, law
enforcement, passports, visas and immigration, prisons, various
military applications, and for restricting access to expensive or
dangerous items, such as automobiles or guns (see, for example,
O'Connor, Stanford Technology Law Review and U.S. Pat. No.
6,119,096).
[0280] Groups of SNPs, particularly the SNPs provided by the
present invention, can be typed to uniquely identify an individual
for biometric applications such as those described above. Such SNP
typing can readily be accomplished using, for example, DNA
chips/arrays. Preferably, a minimally invasive means for obtaining
a DNA sample is utilized. For example, PCR amplification enables
sufficient quantities of DNA for analysis to be obtained from
buccal swabs or fingerprints, which contain DNA-containing skin
cells and oils that are naturally transferred during contact.
[0281] Further information regarding techniques for using SNPs in
forensic/human identification applications can be found in, for
example, Current Protocols in Human Genetics, John Wiley &
Sons, N.Y. (2002), 14.1-14.7.
Variant Proteins, Antibodies, Vectors & Host Cells, & Uses
Thereof
[0282] Variant Proteins Encoded by SNP-Containing Nucleic Acid
Molecules
[0283] The present invention provides SNP-containing nucleic acid
molecules, many of which encode proteins having variant amino acid
sequences as compared to the art-known (i.e., wild-type) proteins.
Amino acid sequences encoded by the polymorphic nucleic acid
molecules of the present invention are provided as SEQ ID NOS:
13-24 in Table 1 and the Sequence Listing. These variants will
generally be referred to herein as variant
proteins/peptides/polypeptides, or polymorphic
proteins/peptides/polypeptides of the present invention. The terms
"protein", "peptide", and "polypeptide" are used herein
interchangeably.
[0284] A variant protein of the present invention may be encoded
by, for example, a nonsynonymous nucleotide substitution at any one
of the cSNP positions disclosed herein. In addition, variant
proteins may also include proteins whose expression, structure,
and/or function is altered by a SNP disclosed herein, such as a SNP
that creates or destroys a stop codon, a SNP that affects splicing,
and a SNP in control/regulatory elements, e.g. promoters,
enhancers, or transcription factor binding domains.
[0285] As used herein, a protein or peptide is said to be
"isolated" or "purified" when it is substantially free of cellular
material or chemical precursors or other chemicals. The variant
proteins of the present invention can be purified to homogeneity or
other lower degrees of purity. The level of purification will be
based on the intended use. The key feature is that the preparation
allows for the desired function of the variant protein, even if in
the presence of considerable amounts of other components.
[0286] As used herein, "substantially free of cellular material"
includes preparations of the variant protein having less than about
30% (by dry weight) other proteins (i.e., contaminating protein),
less than about 20% other proteins, less than about 10% other
proteins, or less than about 5% other proteins. When the variant
protein is recombinantly produced, it can also be substantially
free of culture medium, i.e., culture medium represents less than
about 20% of the volume of the protein preparation.
[0287] The language "substantially free of chemical precursors or
other chemicals" includes preparations of the variant protein in
which it is separated from chemical precursors or other chemicals
that are involved in its synthesis. In one embodiment, the language
"substantially free of chemical precursors or other chemicals"
includes preparations of the variant protein having less than about
30% (by dry weight) chemical precursors or other chemicals, less
than about 20% chemical precursors or other chemicals, less than
about 10% chemical precursors or other chemicals, or less than
about 5% chemical precursors or other chemicals.
[0288] An isolated variant protein may be purified from cells that
naturally express it, purified from cells that have been altered to
express it (recombinant host cells), or synthesized using known
protein synthesis methods. For example, a nucleic acid molecule
containing SNP(s) encoding the variant protein can be cloned into
an expression vector, the expression vector introduced into a host
cell, and the variant protein expressed in the host cell. The
variant protein can then be isolated from the cells by any
appropriate purification scheme using standard protein purification
techniques. Examples of these techniques are described in detail
below (Sambrook and Russell, 2000, Molecular Cloning: A Laboratory
Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor,
N.Y.).
[0289] The present invention provides isolated variant proteins
that comprise, consist of or consist essentially of amino acid
sequences that contain one or more variant amino acids encoded by
one or more codons which contain a SNP of the present
invention.
[0290] Accordingly, the present invention provides variant proteins
that consist of amino acid sequences that contain one or more amino
acid polymorphisms (or truncations or extensions due to creation or
destruction of a stop codon, respectively) encoded by the SNPs
provided in Table 1 and/or Table 2. A protein consists of an amino
acid sequence when the amino acid sequence is the entire amino acid
sequence of the protein.
[0291] The present invention further provides variant proteins that
consist essentially of amino acid sequences that contain one or
more amino acid polymorphisms (or truncations or extensions due to
creation or destruction of a stop codon, respectively) encoded by
the SNPs provided in Table 1 and/or Table 2. A protein consists
essentially of an amino acid sequence when such an amino acid
sequence is present with only a few additional amino acid residues
in the final protein.
[0292] The present invention further provides variant proteins that
comprise amino acid sequences that contain one or more amino acid
polymorphisms (or truncations or extensions due to creation or
destruction of a stop codon, respectively) encoded by the SNPs
provided in Table 1 and/or Table 2. A protein comprises an amino
acid sequence when the amino acid sequence is at least part of the
final amino acid sequence of the protein. In such a fashion, the
protein may contain only the variant amino acid sequence or have
additional amino acid residues, such as a contiguous encoded
sequence that is naturally associated with it or heterologous amino
acid residues. Such a protein can have a few additional amino acid
residues or can comprise many more additional amino acids. A brief
description of how various types of these proteins can be made and
isolated is provided below.
[0293] The variant proteins of the present invention can be
attached to heterologous sequences to form chimeric or fusion
proteins. Such chimeric and fusion proteins comprise a variant
protein operatively linked to a heterologous protein having an
amino acid sequence not substantially homologous to the variant
protein. "Operatively linked" indicates that the coding sequences
for the variant protein and the heterologous protein are ligated
in-frame. The heterologous protein can be fused to the N-terminus
or C-terminus of the variant protein. In another embodiment, the
fusion protein is encoded by a fusion polynucleotide that is
synthesized by conventional techniques including automated DNA
synthesizers. Alternatively, PCR amplification of gene fragments
can be carried out using anchor primers which give rise to
complementary overhangs between two consecutive gene fragments
which can subsequently be annealed and re-amplified to generate a
chimeric gene sequence (see Ausubel et al., Current Protocols in
Molecular Biology, 1992). Moreover, many expression vectors are
commercially available that already encode a fusion moiety (e.g., a
GST protein). A variant protein-encoding nucleic acid can be cloned
into such an expression vector such that the fusion moiety is
linked in-frame to the variant protein.
[0294] In many uses, the fusion protein does not affect the
activity of the variant protein. The fusion protein can include,
but is not limited to, enzymatic fusion proteins, for example,
beta-galactosidase fusions, yeast two-hybrid GAL fusions, poly-His
fusions, MYC-tagged, HI-tagged and Ig fusions. Such fusion
proteins, particularly poly-His fusions, can facilitate their
purification following recombinant expression. In certain host
cells (e.g., mammalian host cells), expression and/or secretion of
a protein can be increased by using a heterologous signal sequence.
Fusion proteins are further described in, for example, Terpe,
"Overview of tag protein fusions: from molecular and biochemical
fundamentals to commercial systems", Appl Microbiol Biotechnol.
2003 January; 60(5):523-33. Epub 2002 Nov. 7; Graddis et al.,
"Designing proteins that work using recombinant technologies", Curr
Pharm Biotechnol. 2002 December; 3(4):285-97; and Nilsson et al.,
"Affinity fusion strategies for detection, purification, and
immobilization of recombinant proteins", Protein Expr Purif. 1997
October; 11(1):1-16.
[0295] The present invention also relates to further obvious
variants of the variant polypeptides of the present invention, such
as naturally-occurring mature forms (e.g., alleleic variants),
non-naturally occurring recombinantly-derived variants, and
orthologs and paralogs of such proteins that share sequence
homology. Such variants can readily be generated using art-known
techniques in the fields of recombinant nucleic acid technology and
protein biochemistry. It is understood, however, that variants
exclude those known in the prior art before the present
invention.
[0296] Further variants of the variant polypeptides disclosed in
Table 1 can comprise an amino acid sequence that shares at least
70-80%, 80-85%, 85-90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or
99% sequence identity with an amino acid sequence disclosed in
Table 1 (or a fragment thereof) and that includes a novel amino
acid residue (allele) disclosed in Table 1 (which is encoded by a
novel SNP allele). Thus, the present invention specifically
contemplates polypeptides that have a certain degree of sequence
variation compared with the polypeptide sequences shown in Table 1,
but that contain a novel amino acid residue (allele) encoded by a
novel SNP allele disclosed herein.
[0297] In other words, as long as a polypeptide contains a novel
amino acid residue disclosed herein, other portions of the
polypeptide that flank the novel amino acid residue can vary to
some degree from the polypeptide sequences shown in Table 1.
[0298] Full-length pre-processed forms, as well as mature processed
forms, of proteins that comprise one of the amino acid sequences
disclosed herein can readily be identified as having complete
sequence identity to one of the variant proteins of the present
invention as well as being encoded by the same genetic locus as the
variant proteins provided herein.
[0299] Orthologs of a variant peptide can readily be identified as
having some degree of significant sequence homology/identity to at
least a portion of a variant peptide as well as being encoded by a
gene from another organism. Preferred orthologs will be isolated
from non-human mammals, preferably primates, for the development of
human therapeutic targets and agents. Such orthologs can be encoded
by a nucleic acid sequence that hybridizes to a variant
peptide-encoding nucleic acid molecule under moderate to stringent
conditions depending on the degree of relatedness of the two
organisms yielding the homologous proteins.
[0300] Variant proteins include, but are not limited to, proteins
containing deletions, additions and substitutions in the amino acid
sequence caused by the SNPs of the present invention. One class of
substitutions is conserved amino acid substitutions in which a
given amino acid in a polypeptide is substituted for another amino
acid of like characteristics. Typical conservative substitutions
are replacements, one for another, among the aliphatic amino acids
Ala, Val, Leu, and Ile; interchange of the hydroxyl residues Ser
and Thr; exchange of the acidic residues Asp and Glu; substitution
between the amide residues Asn and Gln; exchange of the basic
residues Lys and Arg; and replacements among the aromatic residues
Phe and Tyr. Guidance concerning which amino acid changes are
likely to be phenotypically silent are found in, for example, Bowie
et al., Science 247:1306-1310 (1990).
[0301] Variant proteins can be fully functional or can lack
function in one or more activities, e.g. ability to bind another
molecule, ability to catalyze a substrate, ability to mediate
signaling, etc. Fully functional variants typically contain only
conservative variations or variations in non-critical residues or
in non-critical regions. Functional variants can also contain
substitution of similar amino acids that result in no change or an
insignificant change in function. Alternatively, such substitutions
may positively or negatively affect function to some degree.
Non-functional variants typically contain one or more
non-conservative amino acid substitutions, deletions, insertions,
inversions, truncations or extensions, or a substitution,
insertion, inversion, or deletion of a critical residue or in a
critical region.
[0302] Amino acids that are essential for function of a protein can
be identified by methods known in the art, such as site-directed
mutagenesis or alanine-scanning mutagenesis (Cunningham et al.,
Science 244:1081-1085 (1989)), particularly using the amino acid
sequence and polymorphism information provided in Table 1. The
latter procedure introduces single alanine mutations at every
residue in the molecule. The resulting mutant molecules are then
tested for biological activity such as enzyme activity or in assays
such as an in vitro proliferative activity. Sites that are critical
for binding partner/substrate binding can also be determined by
structural analysis such as crystallization, nuclear magnetic
resonance or photoaffinity labeling (Smith et al., J. Mol. Biol.
224:899-904 (1992); de Vos et al. Science 255:306-312 (1992)).
[0303] Polypeptides can contain amino acids other than the 20 amino
acids commonly referred to as the 20 naturally occurring amino
acids. Further, many amino acids, including the terminal amino
acids, may be modified by natural processes, such as processing and
other post-translational modifications, or by chemical modification
techniques well known in the art. Accordingly, the variant proteins
of the present invention also encompass derivatives or analogs in
which a substituted amino acid residue is not one encoded by the
genetic code, in which a substituent group is included, in which
the mature polypeptide is fused with another compound, such as a
compound to increase the half-life of the polypeptide (e.g.,
polyethylene glycol), or in which additional amino acids are fused
to the mature polypeptide, such as a leader or secretory sequence
or a sequence for purification of the mature polypeptide or a
pro-protein sequence.
[0304] Known protein modifications include, but are not limited to,
acetylation, acylation, ADP-ribosylation, amidation, covalent
attachment of flavin, covalent attachment of a heme moiety,
covalent attachment of a nucleotide or nucleotide derivative,
covalent attachment of a lipid or lipid derivative, covalent
attachment of phosphotidylinositol, cross-linking, cyclization,
disulfide bond formation, demethylation, formation of covalent
crosslinks, formation of cystine, formation of pyroglutamate,
formylation, gamma carboxylation, glycosylation, GPI anchor
formation, hydroxylation, iodination, methylation, myristoylation,
oxidation, proteolytic processing, phosphorylation, prenylation,
racemization, selenoylation, sulfation, transfer-RNA mediated
addition of amino acids to proteins such as arginylation, and
ubiquitination.
[0305] Such protein modifications are well known to those of skill
in the art and have been described in great detail in the
scientific literature. Several particularly common modifications,
glycosylation, lipid attachment, sulfation, gamma-carboxylation of
glutamic acid residues, hydroxylation and ADP-ribosylation, for
instance, are described in most basic texts, such as
Proteins--Structure and Molecular Properties, 2nd Ed., T.E.
Creighton, W. H. Freeman and Company, New York (1993); Wold, F.,
Posttranslational Covalent Modification of Proteins, B. C. Johnson,
Ed., Academic Press, New York 1-12 (1983); Seifter et al., Meth.
Enzymol. 182: 626-646 (1990); and Rattan et al., Ann. N.Y Acad.
Sci. 663:48-62 (1992).
[0306] The present invention further provides fragments of the
variant proteins in which the fragments contain one or more amino
acid sequence variations (e.g., substitutions, or truncations or
extensions due to creation or destruction of a stop codon) encoded
by one or more SNPs disclosed herein. The fragments to which the
invention pertains, however, are not to be construed as
encompassing fragments that have been disclosed in the prior art
before the present invention.
[0307] As used herein, a fragment may comprise at least about 4, 8,
10, 12, 14, 16, 18, 20, 25, 30, 50, 100 (or any other number
in-between) or more contiguous amino acid residues from a variant
protein, wherein at least one amino acid residue is affected by a
SNP of the present invention, e.g., a variant amino acid residue
encoded by a nonsynonymous nucleotide substitution at a cSNP
position provided by the present invention. The variant amino acid
encoded by a cSNP may occupy any residue position along the
sequence of the fragment. Such fragments can be chosen based on the
ability to retain one or more of the biological activities of the
variant protein or the ability to perform a function, e.g., act as
an immunogen. Particularly important fragments are biologically
active fragments. Such fragments will typically comprise a domain
or motif of a variant protein of the present invention, e.g.,
active site, transmembrane domain, or ligand/substrate binding
domain. Other fragments include, but are not limited to, domain or
motif-containing fragments, soluble peptide fragments, and
fragments containing immunogenic structures. Predicted domains and
functional sites are readily identifiable by computer programs well
known to those of skill in the art (e.g., PROSITE analysis)
(Current Protocols in Protein Science, John Wiley & Sons, N.Y.
(2002)).
[0308] Uses of Variant Proteins
[0309] The variant proteins of the present invention can be used in
a variety of ways, including but not limited to, in assays to
determine the biological activity of a variant protein, such as in
a panel of multiple proteins for high-throughput screening; to
raise antibodies or to elicit another type of immune response; as a
reagent (including the labeled reagent) in assays designed to
quantitatively determine levels of the variant protein (or its
binding partner) in biological fluids; as a marker for cells or
tissues in which it is preferentially expressed (either
constitutively or at a particular stage of tissue differentiation
or development or in a disease state); as a target for screening
for a therapeutic agent; and as a direct therapeutic agent to be
administered into a human subject. Any of the variant proteins
disclosed herein may be developed into reagent grade or kit format
for commercialization as research products. Methods for performing
the uses listed above are well known to those skilled in the art
(see, e.g., Molecular Cloning: A Laboratory Manual, Cold Spring
Harbor Laboratory Press, Sambrook and Russell, 2000, and Methods in
Enzymology: Guide to Molecular Cloning Techniques, Academic Press,
Berger, S. L. and A. R. Kimmel eds., 1987).
[0310] In a specific embodiment of the invention, the methods of
the present invention include detection of one or more variant
proteins disclosed herein. Variant proteins are disclosed in Table
1 and in the Sequence Listing as SEQ ID NOS: 13-24. Detection of
such proteins can be accomplished using, for example, antibodies,
small molecule compounds, aptamers, ligands/substrates, other
proteins or protein fragments, or other protein-binding agents.
Preferably, protein detection agents are specific for a variant
protein of the present invention and can therefore discriminate
between a variant protein of the present invention and the
wild-type protein or another variant form. This can generally be
accomplished by, for example, selecting or designing detection
agents that bind to the region of a protein that differs between
the variant and wild-type protein, such as a region of a protein
that contains one or more amino acid substitutions that is/are
encoded by a non-synonymous cSNP of the present invention, or a
region of a protein that follows a nonsense mutation-type SNP that
creates a stop codon thereby leading to a shorter polypeptide, or a
region of a protein that follows a read-through mutation-type SNP
that destroys a stop codon thereby leading to a longer polypeptide
in which a portion of the polypeptide is present in one version of
the polypeptide but not the other.
[0311] In another specific aspect of the invention, the variant
proteins of the present invention are used as targets for
diagnosing stenosis or for determining predisposition to stenosis
in a human. Accordingly, the invention provides methods for
detecting the presence of, or levels of, one or more variant
proteins of the present invention in a cell, tissue, or organism.
Such methods typically involve contacting a test sample with an
agent (e.g., an antibody, small molecule compound, or peptide)
capable of interacting with the variant protein such that specific
binding of the agent to the variant protein can be detected. Such
an assay can be provided in a single detection format or a
multi-detection format such as an array, for example, an antibody
or aptamer array (arrays for protein detection may also be referred
to as "protein chips"). The variant protein of interest can be
isolated from a test sample and assayed for the presence of a
variant amino acid sequence encoded by one or more SNPs disclosed
by the present invention. The SNPs may cause changes to the protein
and the corresponding protein function/activity, such as through
non-synonymous substitutions in protein coding regions that can
lead to amino acid substitutions, deletions, insertions, and/or
rearrangements; formation or destruction of stop codons; or
alteration of control elements such as promoters. SNPs may also
cause inappropriate post-translational modifications.
[0312] One preferred agent for detecting a variant protein in a
sample is an antibody capable of selectively binding to a variant
form of the protein (antibodies are described in greater detail in
the next section). Such samples include, for example, tissues,
cells, and biological fluids isolated from a subject, as well as
tissues, cells and fluids present within a subject.
[0313] In vitro methods for detection of the variant proteins
associated with stenosis that are disclosed herein and fragments
thereof include, but are not limited to, enzyme linked
immunosorbent assays (ELISAs), radioimmunoassays (RIA), Western
blots, immunoprecipitations, immunofluorescence, and protein
arrays/chips (e.g., arrays of antibodies or aptamers). For further
information regarding immunoassays and related protein detection
methods, see Current Protocols in Immunology, John Wiley &
Sons, N.Y., and Hage, "Immunoassays", Anal Chem. 1999 Jun. 15;
71(12):294R-304R.
[0314] Additional analytic methods of detecting amino acid variants
include, but are not limited to, altered electrophoretic mobility,
altered tryptic peptide digest, altered protein activity in
cell-based or cell-free assay, alteration in ligand or
antibody-binding pattern, altered isoelectric point, and direct
amino acid sequencing.
[0315] Alternatively, variant proteins can be detected in vivo in a
subject by introducing into the subject a labeled antibody (or
other type of detection reagent) specific for a variant protein.
For example, the antibody can be labeled with a radioactive marker
whose presence and location in a subject can be detected by
standard imaging techniques.
[0316] Other uses of the variant peptides of the present invention
are based on the class or action of the protein. For example,
proteins isolated from humans and their mammalian orthologs serve
as targets for identifying agents (e.g., small molecule drugs or
antibodies) for use in therapeutic applications, particularly for
modulating a biological or pathological response in a cell or
tissue that expresses the protein. Pharmaceutical agents can be
developed that modulate protein activity.
[0317] As an alternative to modulating gene expression, therapeutic
compounds can be developed that modulate protein function. For
example, many SNPs disclosed herein affect the amino acid sequence
of the encoded protein (e.g., non-synonymous cSNPs and nonsense
mutation-type SNPs). Such alterations in the encoded amino acid
sequence may affect protein function, particularly if such amino
acid sequence variations occur in functional protein domains, such
as catalytic domains, ATP-binding domains, or ligand/substrate
binding domains. It is well established in the art that variant
proteins having amino acid sequence variations in functional
domains can cause or influence pathological conditions. In such
instances, compounds (e.g., small molecule drugs or antibodies) can
be developed that target the variant protein and modulate (e.g.,
up- or down-regulate) protein function/activity.
[0318] The therapeutic methods of the present invention further
include methods that target one or more variant proteins of the
present invention. Variant proteins can be targeted using, for
example, small molecule compounds, antibodies, aptamers,
ligands/substrates, other proteins, or other protein-binding
agents. Additionally, the skilled artisan will recognize that the
novel protein variants (and polymorphic nucleic acid molecules)
disclosed in Table 1 may themselves be directly used as therapeutic
agents by acting as competitive inhibitors of corresponding
art-known proteins (or nucleic acid molecules such as mRNA
molecules).
[0319] The variant proteins of the present invention are
particularly useful in drug screening assays, in cell-based or
cell-free systems. Cell-based systems can utilize cells that
naturally express the protein, a biopsy specimen, or cell cultures.
In one embodiment, cell-based assays involve recombinant host cells
expressing the variant protein. Cell-free assays can be used to
detect the ability of a compound to directly bind to a variant
protein or to the corresponding SNP-containing nucleic acid
fragment that encodes the variant protein.
[0320] A variant protein of the present invention, as well as
appropriate fragments thereof, can be used in high-throughput
screening assays to test candidate compounds for the ability to
bind and/or modulate the activity of the variant protein. These
candidate compounds can be further screened against a protein
having normal function (e.g., a wild-type/non-variant protein) to
further determine the effect of the compound on the protein
activity. Furthermore, these compounds can be tested in animal or
invertebrate systems to determine in vivo activity/effectiveness.
Compounds can be identified that activate (agonists) or inactivate
(antagonists) the variant protein, and different compounds can be
identified that cause various degrees of activation or inactivation
of the variant protein.
[0321] Further, the variant proteins can be used to screen a
compound for the ability to stimulate or inhibit interaction
between the variant protein and a target molecule that normally
interacts with the protein. The target can be a ligand, a substrate
or a binding partner that the protein normally interacts with (for
example, epinephrine or norepinephrine). Such assays typically
include the steps of combining the variant protein with a candidate
compound under conditions that allow the variant protein, or
fragment thereof, to interact with the target molecule, and to
detect the formation of a complex between the protein and the
target or to detect the biochemical consequence of the interaction
with the variant protein and the target, such as any of the
associated effects of signal transduction.
[0322] Candidate compounds include, for example, 1) peptides such
as soluble peptides, including Ig-tailed fusion peptides and
members of random peptide libraries (see, e.g., Lam et al., Nature
354:82-84 (1991); Houghten et al., Nature 354:84-86 (1991)) and
combinatorial chemistry-derived molecular libraries made of D-
and/or L-configuration amino acids; 2) phosphopeptides (e.g.,
members of random and partially degenerate, directed phosphopeptide
libraries, see, e.g., Songyang et al., Cell 72:767-778 (1993)); 3)
antibodies (e.g., polyclonal, monoclonal, humanized,
anti-idiotypic, chimeric, and single chain antibodies as well as
Fab, F(ab').sub.2, Fab expression library fragments, and
epitope-binding fragments of antibodies); and 4) small organic and
inorganic molecules (e.g., molecules obtained from combinatorial
and natural product libraries).
[0323] One candidate compound is a soluble fragment of the variant
protein that competes for ligand binding. Other candidate compounds
include mutant proteins or appropriate fragments containing
mutations that affect variant protein function and thus compete for
ligand. Accordingly, a fragment that competes for ligand, for
example with a higher affinity, or a fragment that binds ligand but
does not allow release, is encompassed by the invention.
[0324] The invention further includes other end point assays to
identify compounds that modulate (stimulate or inhibit) variant
protein activity. The assays typically involve an assay of events
in the signal transduction pathway that indicate protein activity.
Thus, the expression of genes that are up or down-regulated in
response to the variant protein dependent signal cascade can be
assayed. In one embodiment, the regulatory region of such genes can
be operably linked to a marker that is easily detectable, such as
luciferase. Alternatively, phosphorylation of the variant protein,
or a variant protein target, could also be measured. Any of the
biological or biochemical functions mediated by the variant protein
can be used as an endpoint assay. These include all of the
biochemical or biological events described herein, in the
references cited herein, incorporated by reference for these
endpoint assay targets, and other functions known to those of
ordinary skill in the art.
[0325] Binding and/or activating compounds can also be screened by
using chimeric variant proteins in which an amino terminal
extracellular domain or parts thereof, an entire transmembrane
domain or subregions, and/or the carboxyl terminal intracellular
domain or parts thereof, can be replaced by heterologous domains or
subregions. For example, a substrate-binding region can be used
that interacts with a different substrate than that which is
normally recognized by a variant protein. Accordingly, a different
set of signal transduction components is available as an end-point
assay for activation. This allows for assays to be performed in
other than the specific host cell from which the variant protein is
derived.
[0326] The variant proteins are also useful in competition binding
assays in methods designed to discover compounds that interact with
the variant protein. Thus, a compound can be exposed to a variant
protein under conditions that allow the compound to bind or to
otherwise interact with the variant protein. A binding partner,
such as ligand, that normally interacts with the variant protein is
also added to the mixture. If the test compound interacts with the
variant protein or its binding partner, it decreases the amount of
complex formed or activity from the variant protein. This type of
assay is particularly useful in screening for compounds that
interact with specific regions of the variant protein (Hodgson,
Bio/technology, 1992, Sep. 10(9), 973-80).
[0327] To perform cell-free drug screening assays, it is sometimes
desirable to immobilize either the variant protein or a fragment
thereof, or its target molecule, to facilitate separation of
complexes from uncomplexed forms of one or both of the proteins, as
well as to accommodate automation of the assay.
[0328] Any method for immobilizing proteins on matrices can be used
in drug screening assays. In one embodiment, a fusion protein
containing an added domain allows the protein to be bound to a
matrix. For example, glutathione-S-transferase/.sup.125I fusion
proteins can be adsorbed onto glutathione sepharose beads (Sigma
Chemical, St. Louis, Mo.) or glutathione derivatized microtitre
plates, which are then combined with the cell lysates (e.g.,
.sup.35S-labeled) and a candidate compound, such as a drug
candidate, and the mixture incubated under conditions conducive to
complex formation (e.g., at physiological conditions for salt and
pH). Following incubation, the beads can be washed to remove any
unbound label, and the matrix immobilized and radiolabel determined
directly, or in the supernatant after the complexes are
dissociated. Alternatively, the complexes can be dissociated from
the matrix, separated by SDS-PAGE, and the level of bound material
found in the bead fraction quantitated from the gel using standard
electrophoretic techniques.
[0329] Either the variant protein or its target molecule can be
immobilized utilizing conjugation of biotin and streptavidin.
Alternatively, antibodies reactive with the variant protein but
which do not interfere with binding of the variant protein to its
target molecule can be derivatized to the wells of the plate, and
the variant protein trapped in the wells by antibody conjugation.
Preparations of the target molecule and a candidate compound are
incubated in the variant protein-presenting wells and the amount of
complex trapped in the well can be quantitated. Methods for
detecting such complexes, in addition to those described above for
the GST-immobilized complexes, include immunodetection of complexes
using antibodies reactive with the protein target molecule, or
which are reactive with variant protein and compete with the target
molecule, and enzyme-linked assays that rely on detecting an
enzymatic activity associated with the target molecule.
[0330] Modulators of variant protein activity identified according
to these drug screening assays can be used to treat a subject with
a disorder mediated by the protein pathway, such as stenosis. These
methods of treatment typically include the steps of administering
the modulators of protein activity in a pharmaceutical composition
to a subject in need of such treatment.
[0331] The variant proteins, or fragments thereof, disclosed herein
can themselves be directly used to treat a disorder characterized
by an absence of, inappropriate, or unwanted expression or activity
of the variant protein. Accordingly, methods for treatment include
the use of a variant protein disclosed herein or fragments
thereof.
[0332] In yet another aspect of the invention, variant proteins can
be used as "bait proteins" in a two-hybrid assay or three-hybrid
assay (see, e.g., U.S. Pat. No. 5,283,317; Zervos et al. (1993)
Cell 72:223-232; Madura et al. (1993) J. Biol. Chem.
268:12046-12054; Bartel et al. (1993) Biotechniques 14:920-924;
Iwabuchi et al. (1993) Oncogene 8:1693-1696; and Brent WO94/10300)
to identify other proteins that bind to or interact with the
variant protein and are involved in variant protein activity. Such
variant protein-binding proteins are also likely to be involved in
the propagation of signals by the variant proteins or variant
protein targets as, for example, elements of a protein-mediated
signaling pathway. Alternatively, such variant protein-binding
proteins are inhibitors of the variant protein.
[0333] The two-hybrid system is based on the modular nature of most
transcription factors, which typically consist of separable
DNA-binding and activation domains. Briefly, the assay typically
utilizes two different DNA constructs. In one construct, the gene
that codes for a variant protein is fused to a gene encoding the
DNA binding domain of a known transcription factor (e.g., GAL-4).
In the other construct, a DNA sequence, from a library of DNA
sequences, that encodes an unidentified protein ("prey" or
"sample") is fused to a gene that codes for the activation domain
of the known transcription factor. If the "bait" and the "prey"
proteins are able to interact, in vivo, forming a variant
protein-dependent complex, the DNA-binding and activation domains
of the transcription factor are brought into close proximity. This
proximity allows transcription of a reporter gene (e.g., LacZ) that
is operably linked to a transcriptional regulatory site responsive
to the transcription factor. Expression of the reporter gene can be
detected, and cell colonies containing the functional transcription
factor can be isolated and used to obtain the cloned gene that
encodes the protein that interacts with the variant protein.
[0334] Antibodies Directed to Variant Proteins
[0335] The present invention also provides antibodies that
selectively bind to the variant proteins disclosed herein and
fragments thereof. Such antibodies may be used to quantitatively or
qualitatively detect the variant proteins of the present invention.
As used herein, an antibody selectively binds a target variant
protein when it binds the variant protein and does not
significantly bind to non-variant proteins, i.e., the antibody does
not significantly bind to normal, wild-type, or art-known proteins
that do not contain a variant amino acid sequence due to one or
more SNPs of the present invention (variant amino acid sequences
may be due to, for example, nonsynonymous cSNPs, nonsense SNPs that
create a stop codon, thereby causing a truncation of a polypeptide
or SNPs that cause read-through mutations resulting in an extension
of a polypeptide).
[0336] As used herein, an antibody is defined in terms consistent
with that recognized in the art: they are multi-subunit proteins
produced by an organism in response to an antigen challenge. The
antibodies of the present invention include both monoclonal
antibodies and polyclonal antibodies, as well as antigen-reactive
proteolytic fragments of such antibodies, such as Fab,
F(ab)'.sub.2, and Fv fragments. In addition, an antibody of the
present invention further includes any of a variety of engineered
antigen-binding molecules such as a chimeric antibody (U.S. Pat.
Nos. 4,816,567 and 4,816,397; Morrison et al., Proc. Natl. Acad.
Sci. USA, 81:6851, 1984; Neuberger et al., Nature 312:604, 1984), a
humanized antibody (U.S. Pat. Nos. 5,693,762; 5,585,089; and
5,565,332), a single-chain Fv (U.S. Pat. No. 4,946,778; Ward et
al., Nature 334:544, 1989), a bispecific antibody with two binding
specificities (Segal et al., J. Immunol. Methods 248:1, 2001;
Carter, J. Immunol. Methods 248:7, 2001), a diabody, a triabody,
and a tetrabody (Todorovska et al., J. Immunol. Methods, 248:47,
2001), as well as a Fab conjugate (dimer or trimer), and a
minibody.
[0337] Many methods are known in the art for generating and/or
identifying antibodies to a given target antigen (Harlow,
Antibodies, Cold Spring Harbor Press, (1989)). In general, an
isolated peptide (e.g., a variant protein of the present invention)
is used as an immunogen and is administered to a mammalian
organism, such as a rat, rabbit, hamster or mouse. Either a
full-length protein, an antigenic peptide fragment (e.g., a peptide
fragment containing a region that varies between a variant protein
and a corresponding wild-type protein), or a fusion protein can be
used. A protein used as an immunogen may be naturally-occurring,
synthetic or recombinantly produced, and may be administered in
combination with an adjuvant, including but not limited to,
Freund's (complete and incomplete), mineral gels such as aluminum
hydroxide, surface active substance such as lysolecithin, pluronic
polyols, polyanions, peptides, oil emulsions, keyhole limpet
hemocyanin, dinitrophenol, and the like.
[0338] Monoclonal antibodies can be produced by hybridoma
technology (Kohler and Milstein, Nature, 256:495, 1975), which
immortalizes cells secreting a specific monoclonal antibody. The
immortalized cell lines can be created in vitro by fusing two
different cell types, typically lymphocytes, and tumor cells. The
hybridoma cells may be cultivated in vitro or in vivo.
Additionally, fully human antibodies can be generated by transgenic
animals (He et al., J. Immunol., 169:595, 2002). Fd phage and Fd
phagemid technologies may be used to generate and select
recombinant antibodies in vitro (Hoogenboom and Chames, Immunol.
Today 21:371, 2000; Liu et al., J. Mol. Biol. 315:1063, 2002). The
complementarity-determining regions of an antibody can be
identified, and synthetic peptides corresponding to such regions
may be used to mediate antigen binding (U.S. Pat. No.
5,637,677).
[0339] Antibodies are preferably prepared against regions or
discrete fragments of a variant protein containing a variant amino
acid sequence as compared to the corresponding wild-type protein
(e.g., a region of a variant protein that includes an amino acid
encoded by a nonsynonymous cSNP, a region affected by truncation
caused by a nonsense SNP that creates a stop codon, or a region
resulting from the destruction of a stop codon due to read-through
mutation caused by a SNP). Furthermore, preferred regions will
include those involved in function/activity and/or protein/binding
partner interaction. Such fragments can be selected on a physical
property, such as fragments corresponding to regions that are
located on the surface of the protein, e.g., hydrophilic regions,
or can be selected based on sequence uniqueness, or based on the
position of the variant amino acid residue(s) encoded by the SNPs
provided by the present invention. An antigenic fragment will
typically comprise at least about 8-contiguous amino acid residues
in which at least one of the amino acid residues is an amino acid
affected by a SNP disclosed herein. The antigenic peptide can
comprise, however, at least 12, 14, 16, 20, 25, 50, 100 (or any
other number in-between) or more amino acid residues, provided that
at least one amino acid is affected by a SNP disclosed herein.
[0340] Detection of an antibody of the present invention can be
facilitated by coupling (i.e., physically linking) the antibody or
an antigen-reactive fragment thereof to a detectable substance.
Detectable substances include, but are not limited to, various
enzymes, prosthetic groups, fluorescent materials, luminescent
materials, bioluminescent materials, and radioactive materials.
Examples of suitable enzymes include horseradish peroxidase,
alkaline phosphatase, .beta.-galactosidase, or
acetylcholinesterase; examples of suitable prosthetic group
complexes include streptavidin/biotin and avidin/biotin; examples
of suitable fluorescent materials include umbelliferone,
fluorescein, fluorescein isothiocyanate, rhodamine,
dichlorotriazinylamine fluorescein, dansyl chloride or
phycoerythrin; an example of a luminescent material includes
luminol; examples of bioluminescent materials include luciferase,
luciferin, and aequorin, and examples of suitable radioactive
material include .sup.125I, .sup.131I, .sup.35S or .sup.3H.
[0341] Antibodies, particularly the use of antibodies as
therapeutic agents, are reviewed in: Morgan, "Antibody therapy for
Alzheimer's disease", Expert Rev Vaccines. 2003 February;
2(1):53-9; Ross et al., "Anticancer antibodies", Am J Clin Pathol.
2003 April; 119(4):472-85; Goldenberg, "Advancing role of
radiolabeled antibodies in the therapy of cancer", Cancer Immunol
Immunother. 2003 May; 52(5):281-96. Epub 2003 Mar. 11; Ross et al.,
"Antibody-based therapeutics in oncology", Expert Rev Anticancer
Ther. 2003 February; 3(1): 107-21; Cao et al., "Bispecific antibody
conjugates in therapeutics", Adv Drug Deliv Rev. 2003 Feb. 10;
55(2):171-97; von Mehren et al., "Monoclonal antibody therapy for
cancer", Annu Rev Med. 2003; 54:343-69. Epub 2001 Dec. 3; Hudson et
al., "Engineered antibodies", Nat. Med. 2003 January; 9(1):129-34;
Brekke et al., "Therapeutic antibodies for human diseases at the
dawn of the twenty-first century", Nat Rev Drug Discov. 2003
January; 2(1):52-62 (Erratum in: Nat Rev Drug Discov. 2003 March;
2(3):240); Houdebine, "Antibody manufacture in transgenic animals
and comparisons with other systems", Curr Opin Biotechnol. 2002
December; 13(6):625-9; Andreakos et al., "Monoclonal antibodies in
immune and inflammatory diseases", Curr Opin Biotechnol. 2002
December; 13(6):615-20; Kellermann et al., "Antibody discovery: the
use of transgenic mice to generate human monoclonal antibodies for
therapeutics", Curr Opin Biotechnol. 2002 December; 13(6):593-7;
Pini et al., "Phage display and colony filter screening for
high-throughput selection of antibody libraries", Comb Chem High
Throughput Screen. 2002 November; 5(7):503-10; Batra et al.,
"Pharmacokinetics and biodistribution of genetically engineered
antibodies", Curr Opin Biotechnol. 2002 December; 13(6):603-8; and
Tangri et al., "Rationally engineered proteins or antibodies with
absent or reduced immunogenicity", Curr Med. Chem. 2002 December;
9(24):2191-9.
[0342] Uses of Antibodies
[0343] Antibodies can be used to isolate the variant proteins of
the present invention from a natural cell source or from
recombinant host cells by standard techniques, such as affinity
chromatography or immunoprecipitation. In addition, antibodies are
useful for detecting the presence of a variant protein of the
present invention in cells or tissues to determine the pattern of
expression of the variant protein among various tissues in an
organism and over the course of normal development or disease
progression. Further, antibodies can be used to detect variant
protein in situ, in vitro, in a bodily fluid, or in a cell lysate
or supernatant in order to evaluate the amount and pattern of
expression. Also, antibodies can be used to assess abnormal tissue
distribution, abnormal expression during development, or expression
in an abnormal condition, such as stenosis. Additionally, antibody
detection of circulating fragments of the full-length variant
protein can be used to identify turnover.
[0344] Antibodies to the variant proteins of the present invention
are also useful in pharmacogenomic analysis. Thus, antibodies
against variant proteins encoded by alternative SNP alleles can be
used to identify individuals that require modified treatment
modalities.
[0345] Further, antibodies can be used to assess expression of the
variant protein in disease states such as in active stages of the
disease or in an individual with a predisposition to a disease
related to the protein's function, particularly stenosis.
Antibodies specific for a variant protein encoded by a
SNP-containing nucleic acid molecule of the present invention can
be used to assay for the presence of the variant protein, such as
to screen for predisposition to stenosis as indicated by the
presence of the variant protein.
[0346] Antibodies are also useful as diagnostic tools for
evaluating the variant proteins in conjunction with analysis by
electrophoretic mobility, isoelectric point, tryptic peptide
digest, and other physical assays well known in the art.
[0347] Antibodies are also useful for tissue typing. Thus, where a
specific variant protein has been correlated with expression in a
specific tissue, antibodies that are specific for this protein can
be used to identify a tissue type.
[0348] Antibodies can also be used to assess aberrant subcellular
localization of a variant protein in cells in various tissues. The
diagnostic uses can be applied, not only in genetic testing, but
also in monitoring a treatment modality. Accordingly, where
treatment is ultimately aimed at correcting the expression level or
the presence of variant protein or aberrant tissue distribution or
developmental expression of a variant protein, antibodies directed
against the variant protein or relevant fragments can be used to
monitor therapeutic efficacy.
[0349] The antibodies are also useful for inhibiting variant
protein function, for example, by blocking the binding of a variant
protein to a binding partner. These uses can also be applied in a
therapeutic context in which treatment involves inhibiting a
variant protein's function. An antibody can be used, for example,
to block or competitively inhibit binding, thus modulating
(agonizing or antagonizing) the activity of a variant protein.
Antibodies can be prepared against specific variant protein
fragments containing sites required for function or against an
intact variant protein that is associated with a cell or cell
membrane. For in vivo administration, an antibody may be linked
with an additional therapeutic payload such as a radionuclide, an
enzyme, an immunogenic epitope, or a cytotoxic agent. Suitable
cytotoxic agents include, but are not limited to, bacterial toxin
such as diphtheria, and plant toxin such as ricin. The in vivo
half-life of an antibody or a fragment thereof may be lengthened by
pegylation through conjugation to polyethylene glycol (Leong et
al., Cytokine 16:106, 2001).
[0350] The invention also encompasses kits for using antibodies,
such as kits for detecting the presence of a variant protein in a
test sample. An exemplary kit can comprise antibodies such as a
labeled or labelable antibody and a compound or agent for detecting
variant proteins in a biological sample; means for determining the
amount, or presence/absence of variant protein in the sample; means
for comparing the amount of variant protein in the sample with a
standard; and instructions for use.
[0351] Vectors and Host Cells
[0352] The present invention also provides vectors containing the
SNP-containing nucleic acid molecules described herein. The term
"vector" refers to a vehicle, preferably a nucleic acid molecule,
which can transport a SNP-containing nucleic acid molecule. When
the vector is a nucleic acid molecule, the SNP-containing nucleic
acid molecule can be covalently linked to the vector nucleic acid.
Such vectors include, but are not limited to, a plasmid, single or
double stranded phage, a single or double stranded RNA or DNA viral
vector, or artificial chromosome, such as a BAC, PAC, YAC, or
MAC.
[0353] A vector can be maintained in a host cell as an
extrachromosomal element where it replicates and produces
additional copies of the SNP-containing nucleic acid molecules.
Alternatively, the vector may integrate into the host cell genome
and produce additional copies of the SNP-containing nucleic acid
molecules when the host cell replicates.
[0354] The invention provides vectors for the maintenance (cloning
vectors) or vectors for expression (expression vectors) of the
SNP-containing nucleic acid molecules. The vectors can function in
prokaryotic or eukaryotic cells or in both (shuttle vectors).
[0355] Expression vectors typically contain cis-acting regulatory
regions that are operably linked in the vector to the
SNP-containing nucleic acid molecules such that transcription of
the SNP-containing nucleic acid molecules is allowed in a host
cell. The SNP-containing nucleic acid molecules can also be
introduced into the host cell with a separate nucleic acid molecule
capable of affecting transcription. Thus, the second nucleic acid
molecule may provide a trans-acting factor interacting with the
cis-regulatory control region to allow transcription of the
SNP-containing nucleic acid molecules from the vector.
Alternatively, a trans-acting factor may be supplied by the host
cell. Finally, a trans-acting factor can be produced from the
vector itself. It is understood, however, that in some embodiments,
transcription and/or translation of the nucleic acid molecules can
occur in a cell-free system.
[0356] The regulatory sequences to which the SNP-containing nucleic
acid molecules described herein can be operably linked include
promoters for directing mRNA transcription. These include, but are
not limited to, the left promoter from bacteriophage .lamda., the
lac, TRP, and TAC promoters from E. coli, the early and late
promoters from SV40, the CMV immediate early promoter, the
adenovirus early and late promoters, and retrovirus long-terminal
repeats.
[0357] In addition to control regions that promote transcription,
expression vectors may also include regions that modulate
transcription, such as repressor binding sites and enhancers.
Examples include the SV40 enhancer, the cytomegalovirus immediate
early enhancer, polyoma enhancer, adenovirus enhancers, and
retrovirus LTR enhancers.
[0358] In addition to containing sites for transcription initiation
and control, expression vectors can also contain sequences
necessary for transcription termination and, in the transcribed
region, a ribosome-binding site for translation. Other regulatory
control elements for expression include initiation and termination
codons as well as polyadenylation signals. A person of ordinary
skill in the art would be aware of the numerous regulatory
sequences that are useful in expression vectors (see, e.g.,
Sambrook and Russell, 2000, Molecular Cloning: A Laboratory Manual,
Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.).
[0359] A variety of expression vectors can be used to express a
SNP-containing nucleic acid molecule. Such vectors include
chromosomal, episomal, and virus-derived vectors, for example,
vectors derived from bacterial plasmids, from bacteriophage, from
yeast episomes, from yeast chromosomal elements, including yeast
artificial chromosomes, from viruses such as baculoviruses,
papovaviruses such as SV40, Vaccinia viruses, adenoviruses,
poxviruses, pseudorabies viruses, and retroviruses. Vectors can
also be derived from combinations of these sources such as those
derived from plasmid and bacteriophage genetic elements, e.g.,
cosmids and phagemids. Appropriate cloning and expression vectors
for prokaryotic and eukaryotic hosts are described in Sambrook and
Russell, 2000, Molecular Cloning: A Laboratory Manual, Cold Spring
Harbor Laboratory Press, Cold Spring Harbor, N.Y.
[0360] The regulatory sequence in a vector may provide constitutive
expression in one or more host cells (e.g., tissue specific
expression) or may provide for inducible expression in one or more
cell types such as by temperature, nutrient additive, or exogenous
factor, e.g., a hormone or other ligand. A variety of vectors that
provide constitutive or inducible expression of a nucleic acid
sequence in prokaryotic and eukaryotic host cells are well known to
those of ordinary skill in the art.
[0361] A SNP-containing nucleic acid molecule can be inserted into
the vector by methodology well-known in the art. Generally, the
SNP-containing nucleic acid molecule that will ultimately be
expressed is joined to an expression vector by cleaving the
SNP-containing nucleic acid molecule and the expression vector with
one or more restriction enzymes and then ligating the fragments
together. Procedures for restriction enzyme digestion and ligation
are well known to those of ordinary skill in the art.
[0362] The vector containing the appropriate nucleic acid molecule
can be introduced into an appropriate host cell for propagation or
expression using well-known techniques. Bacterial host cells
include, but are not limited to, E. coli, Streptomyces, and
Salmonella typhimurium. Eukaryotic host cells include, but are not
limited to, yeast, insect cells such as Drosophila, animal cells
such as COS and CHO cells, and plant cells.
[0363] As described herein, it may be desirable to express the
variant peptide as a fusion protein. Accordingly, the invention
provides fusion vectors that allow for the production of the
variant peptides. Fusion vectors can, for example, increase the
expression of a recombinant protein, increase the solubility of the
recombinant protein, and aid in the purification of the protein by
acting, for example, as a ligand for affinity purification. A
proteolytic cleavage site may be introduced at the junction of the
fusion moiety so that the desired variant peptide can ultimately be
separated from the fusion moiety. Proteolytic enzymes suitable for
such use include, but are not limited to, factor Xa, thrombin, and
enterokinase. Typical fusion expression vectors include pGEX (Smith
et al., Gene 67:31-40 (1988)), pMAL (New England Biolabs, Beverly,
Mass.) and pRIT5 (Pharmacia, Piscataway, N.J.) which fuse
glutathione S-transferase (GST), maltose E binding protein, or
protein A, respectively, to the target recombinant protein.
Examples of suitable inducible non-fusion E. coli expression
vectors include pTrc (Amann et al., Gene 69:301-315 (1988)) and pET
11d (Studier et al., Gene Expression Technology: Methods in
Enzymology 185:60-89 (1990)).
[0364] Recombinant protein expression can be maximized in a
bacterial host by providing a genetic background wherein the host
cell has an impaired capacity to proteolytically cleave the
recombinant protein (Gottesman, S., Gene Expression Technology.
Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990)
119-128). Alternatively, the sequence of the SNP-containing nucleic
acid molecule of interest can be altered to provide preferential
codon usage for a specific host cell, for example, E. coli (Wada et
al., Nucleic Acids Res. 20:2111-2118 (1992)).
[0365] The SNP-containing nucleic acid molecules can also be
expressed by expression vectors that are operative in yeast.
Examples of vectors for expression in yeast (e.g., S. cerevisiae)
include pYepSec1 (Baldari, et al., EMBO J. 6:229-234 (1987)), pMFa
(Kujan et al., Cell 30:933-943 (1982)), pJRY88 (Schultz et al.,
Gene 54:113-123 (1987)), and pYES2 (Invitrogen Corporation, San
Diego, Calif.).
[0366] The SNP-containing nucleic acid molecules can also be
expressed in insect cells using, for example, baculovirus
expression vectors. Baculovirus vectors available for expression of
proteins in cultured insect cells (e.g., Sf 9 cells) include the
pAc series (Smith et al., Mol. Cell. Biol. 3:2156-2165 (1983)) and
the pVL series (Lucklow et al., Virology 170:31-39 (1989)).
[0367] In certain embodiments of the invention, the SNP-containing
nucleic acid molecules described herein are expressed in mammalian
cells using mammalian expression vectors. Examples of mammalian
expression vectors include pCDM8 (Seed, B. Nature 329:840 (1987))
and pMT2PC (Kaufman et al., EMBO J. 6:187-195 (1987)).
[0368] The invention also encompasses vectors in which the
SNP-containing nucleic acid molecules described herein are cloned
into the vector in reverse orientation, but operably linked to a
regulatory sequence that permits transcription of antisense RNA.
Thus, an antisense transcript can be produced to the SNP-containing
nucleic acid sequences described herein, including both coding and
non-coding regions. Expression of this antisense RNA is subject to
each of the parameters described above in relation to expression of
the sense RNA (regulatory sequences, constitutive or inducible
expression, tissue-specific expression).
[0369] The invention also relates to recombinant host cells
containing the vectors described herein. Host cells therefore
include, for example, prokaryotic cells, lower eukaryotic cells
such as yeast, other eukaryotic cells such as insect cells, and
higher eukaryotic cells such as mammalian cells.
[0370] The recombinant host cells can be prepared by introducing
the vector constructs described herein into the cells by techniques
readily available to persons of ordinary skill in the art. These
include, but are not limited to, calcium phosphate transfection,
DEAE-dextran-mediated transfection, cationic lipid-mediated
transfection, electroporation, transduction, infection,
lipofection, and other techniques such as those described in
Sambrook and Russell, 2000, Molecular Cloning: A Laboratory Manual,
Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press,
Cold Spring Harbor, N.Y.).
[0371] Host cells can contain more than one vector. Thus, different
SNP-containing nucleotide sequences can be introduced in different
vectors into the same cell. Similarly, the SNP-containing nucleic
acid molecules can be introduced either alone or with other nucleic
acid molecules that are not related to the SNP-containing nucleic
acid molecules, such as those providing trans-acting factors for
expression vectors. When more than one vector is introduced into a
cell, the vectors can be introduced independently, co-introduced,
or joined to the nucleic acid molecule vector.
[0372] In the case of bacteriophage and viral vectors, these can be
introduced into cells as packaged or encapsulated virus by standard
procedures for infection and transduction. Viral vectors can be
replication-competent or replication-defective. In the case in
which viral replication is defective, replication can occur in host
cells that provide functions that complement the defects.
[0373] Vectors generally include selectable markers that enable the
selection of the subpopulation of cells that contain the
recombinant vector constructs. The marker can be inserted in the
same vector that contains the SNP-containing nucleic acid molecules
described herein or may be in a separate vector. Markers include,
for example, tetracycline or ampicillin-resistance genes for
prokaryotic host cells, and dihydrofolate reductase or neomycin
resistance genes for eukaryotic host cells. However, any marker
that provides selection for a phenotypic trait can be
effective.
[0374] While the mature variant proteins can be produced in
bacteria, yeast, mammalian cells, and other cells under the control
of the appropriate regulatory sequences, cell-free transcription
and translation systems can also be used to produce these variant
proteins using RNA derived from the DNA constructs described
herein.
[0375] Where secretion of the variant protein is desired, which is
difficult to achieve with multi-transmembrane domain containing
proteins such as G-protein-coupled receptors (GPCRs), appropriate
secretion signals can be incorporated into the vector. The signal
sequence can be endogenous to the peptides or heterologous to these
peptides.
[0376] Where the variant protein is not secreted into the medium,
the protein can be isolated from the host cell by standard
disruption procedures, including freeze/thaw, sonication,
mechanical disruption, use of lysing agents, and the like. The
variant protein can then be recovered and purified by well-known
purification methods including, for example, ammonium sulfate
precipitation, acid extraction, anion or cationic exchange
chromatography, phosphocellulose chromatography,
hydrophobic-interaction chromatography, affinity chromatography,
hydroxylapatite chromatography, lectin chromatography, or high
performance liquid chromatography.
[0377] It is also understood that, depending upon the host cell in
which recombinant production of the variant proteins described
herein occurs, they can have various glycosylation patterns, or may
be non-glycosylated, as when produced in bacteria. In addition, the
variant proteins may include an initial modified methionine in some
cases as a result of a host-mediated process.
[0378] For further information regarding vectors and host cells,
see Current Protocols in Molecular Biology, John Wiley & Sons,
N.Y.
[0379] Uses of Vectors and Host Cells, and Transgenic Animals
[0380] Recombinant host cells that express the variant proteins
described herein have a variety of uses. For example, the cells are
useful for producing a variant protein that can be further purified
into a preparation of desired amounts of the variant protein or
fragments thereof. Thus, host cells containing expression vectors
are useful for variant protein production.
[0381] Host cells are also useful for conducting cell-based assays
involving the variant protein or variant protein fragments, such as
those described above as well as other formats known in the art.
Thus, a recombinant host cell expressing a variant protein is
useful for assaying compounds that stimulate or inhibit variant
protein function. Such an ability of a compound to modulate variant
protein function may not be apparent from assays of the compound on
the native/wild-type protein, or from cell-free assays of the
compound. Recombinant host cells are also useful for assaying
functional alterations in the variant proteins as compared with a
known function.
[0382] Genetically-engineered host cells can be further used to
produce non-human transgenic animals. A transgenic animal is
preferably a non-human mammal, for example, a rodent, such as a rat
or mouse, in which one or more of the cells of the animal include a
transgene. A transgene is exogenous DNA containing a SNP of the
present invention which is integrated into the genome of a cell
from which a transgenic animal develops and which remains in the
genome of the mature animal in one or more of its cell types or
tissues. Such animals are useful for studying the function of a
variant protein in vivo, and identifying and evaluating modulators
of variant protein activity. Other examples of transgenic animals
include, but are not limited to, non-human primates, sheep, dogs,
cows, goats, chickens, and amphibians. Transgenic non-human mammals
such as cows and goats can be used to produce variant proteins
which can be secreted in the animal's milk and then recovered.
[0383] A transgenic animal can be produced by introducing a
SNP-containing nucleic acid molecule into the male pronuclei of a
fertilized oocyte, e.g., by microinjection or retroviral infection,
and allowing the oocyte to develop in a pseudopregnant female
foster animal. Any nucleic acid molecules that contain one or more
SNPs of the present invention can potentially be introduced as a
transgene into the genome of a non-human animal.
[0384] Any of the regulatory or other sequences useful in
expression vectors can form part of the transgenic sequence. This
includes intronic sequences and polyadenylation signals, if not
already included. A tissue-specific regulatory sequence(s) can be
operably linked to the transgene to direct expression of the
variant protein in particular cells or tissues.
[0385] Methods for generating transgenic animals via embryo
manipulation and microinjection, particularly animals such as mice,
have become conventional in the art and are described in, for
example, U.S. Pat. Nos. 4,736,866 and 4,870,009, both by Leder et
al., U.S. Pat. No. 4,873,191 by Wagner et al., and in Hogan, B.,
Manipulating the Mouse Embryo, (Cold Spring Harbor Laboratory
Press, Cold Spring Harbor, N.Y., 1986). Similar methods are used
for production of other transgenic animals. A transgenic founder
animal can be identified based upon the presence of the transgene
in its genome and/or expression of transgenic mRNA in tissues or
cells of the animals. A transgenic founder animal can then be used
to breed additional animals carrying the transgene. Moreover,
transgenic animals carrying a transgene can further be bred to
other transgenic animals carrying other transgenes. A transgenic
animal also includes a non-human animal in which the entire animal
or tissues in the animal have been produced using the homologously
recombinant host cells described herein.
[0386] In another embodiment, transgenic non-human animals can be
produced which contain selected systems that allow for regulated
expression of the transgene. One example of such a system is the
cre/loxP recombinase system of bacteriophage P1 (Lakso et al. PNAS
89:6232-6236 (1992)). Another example of a recombinase system is
the FLP recombinase system of S. cerevisiae (O'Gorman et al.
Science 251:1351-1355 (1991)). If a cre/loxP recombinase system is
used to regulate expression of the transgene, animals containing
transgenes encoding both the Cre recombinase and a selected protein
are generally needed. Such animals can be provided through the
construction of "double" transgenic animals, e.g., by mating two
transgenic animals, one containing a transgene encoding a selected
variant protein and the other containing a transgene encoding a
recombinase.
[0387] Clones of the non-human transgenic animals described herein
can also be produced according to the methods described in, for
example, Wilmut, I. et al. Nature 385:810-813 (1997) and PCT
International Publication Nos. WO 97/07668 and WO 97/07669. In
brief, a cell (e.g., a somatic cell) from the transgenic animal can
be isolated and induced to exit the growth cycle and enter Go
phase. The quiescent cell can then be fused, e.g., through the use
of electrical pulses, to an enucleated oocyte from an animal of the
same species from which the quiescent cell is isolated. The
reconstructed oocyte is then cultured such that it develops to
morula or blastocyst and then transferred to pseudopregnant female
foster animal. The offspring born of this female foster animal will
be a clone of the animal from which the cell (e.g., a somatic cell)
is isolated.
[0388] Transgenic animals containing recombinant cells that express
the variant proteins described herein are useful for conducting the
assays described herein in an in vivo context. Accordingly, the
various physiological factors that are present in vivo and that
could influence ligand or substrate binding, variant protein
activation, signal transduction, or other processes or
interactions, may not be evident from in vitro cell-free or
cell-based assays. Thus, non-human transgenic animals of the
present invention may be used to assay in vivo variant protein
function as well as the activities of a therapeutic agent or
compound that modulates variant protein function/activity or
expression. Such animals are also suitable for assessing the
effects of null mutations (i.e., mutations that substantially or
completely eliminate one or more variant protein functions).
[0389] For further information regarding transgenic animals, see
Houdebine, "Antibody manufacture in transgenic animals and
comparisons with other systems", Curr Opin Biotechnol. 2002
December; 13(6):625-9; Petters et al., "Transgenic animals as
models for human disease", Transgenic Res. 2000; 9(4-5):347-51;
discussion 345-6; Wolf et al., "Use of transgenic animals in
understanding molecular mechanisms of toxicity", J Pharm Pharmacol.
1998 June; 50(6):567-74; Echelard, "Recombinant protein production
in transgenic animals", Curr Opin Biotechnol. 1996 October;
7(5):536-40; Houdebine, "Transgenic animal bioreactors", Transgenic
Res. 2000; 9(4-5):305-20; Pirity et al., "Embryonic stem cells,
creating transgenic animals", Methods Cell Biol. 1998; 57:279-93;
and Robl et al., "Artificial chromosome vectors and expression of
complex proteins in transgenic animals", Theriogenology. 2003 Jan.
1; 59(1):107-13.
EXAMPLES
Statistical Analysis of SNP Association with Coronary Stenosis
[0390] A case-control genetic study to determine the association of
SNPs in the human genome with coronary stenosis was carried out
using genomic DNA extracted from 2 independently collected
case-control sample sets. A sample set from the University of
California, San Francisco (UCSF), referred to as "Sample Set 1",
consisted of DNA from 1,654 Caucasian patients with varying degrees
of coronary artery stenosis as evidenced by coronary angiography. A
sample set from the Cleveland Clinic (CCF), referred to as "Sample
Set 2", consisted of DNA from 1,501 Caucasian patients with very
little or severe coronary artery stenosis, also evidenced by
coronary angiography. All samples were obtained from people between
the ages of 18 to 75. All individuals who were included in each
study had signed a written informed consent form. The study
protocols were IRB approved.
[0391] DNA was extracted from blood samples using conventional DNA
extraction methods such as the QIA-amp kit from Qiagen. SNP markers
in the extracted DNA samples were analyzed by genotyping. While
some samples were individually genotyped, the same samples were
also used for pooling studies, in which DNA samples from about 50
individuals were pooled, and allele frequencies were determined in
pooled DNA. Genotypes and pool allele frequencies were obtained
using a PRISM 7900HT sequence detection PCR system (Applied
Biosystems, Foster City, Calif.) by allele-specific PCR, similar to
the method described by Germer et al (Germer S., Holland M. J.,
Higuchi R. 2000, Genome Res. 10: 258-266). Primers for the
allele-specific PCR reactions are provided in Table 5.
[0392] Genotype or allele frequency results from 287 SNPs in Sample
Set 1 and 177 SNPs in Sample Set 2 were analyzed for association
with coronary stenosis. Analysis of the Sample Set 1 data was
performed by placing individuals into groups based on quartiles of
the sum score and using the two extreme quartiles as a binary
endpoint. For Sample Set 2 samples, analysis was done in two ways.
One method placed individuals into groups based on quartiles of the
sum scores and used the two extreme quartiles as a binary endpoint.
The other method placed individuals into groups of sum score "0"
(the control group) or the sum score ">0" (the case group). The
"sum score" is a measure of the overall extent of coronary artery
stenosis and was determined by summing measures of percent stenosis
from various arterial locations. The percent stenosis was
determined from images obtained using coronary angiography. The
case and control groups were further stratified by sex (F, M), age
(tertile 1, 2, or 3) and smoking status ("never smoked" and "ever
smoked"). The allele or genotype frequencies for the tested SNPs
were obtained, and compared between cases and controls. No multiple
testing corrections were made.
[0393] Several tests of association were calculated for both
unstratified and stratified settings: 1) Fisher's exact test or
asymptotic chi-square test for allelic association, 2) asymptotic
chi-square test of genotypic association of two different modes of
inheritance: dominant and recessive, and 3) Armitage trendtest for
the additive mode of genotypic association.
[0394] Effect sizes were estimated through genotypic or allelic
odds ratios, including 95% confidence intervals. The reported
allele (Allele1) or genotype may be under-represented in cases
(with a lower frequency in cases than in controls, indicating that
the reported allele or genotype is associated with decreased risk
and the other allele or genotype is a risk factor for disease) or
over-represented in cases (indicating that the reported allele or
genotype is a risk factor for disease).
[0395] The replicated coronary stenosis markers are reported in
Table 6. A SNP is considered a replicated marker if the association
analyses in two studies showed that the risk allele is the same,
the p-values are each less than or equal to 0.05, and the
significant association is seen in either the same stratum or in a
stratum and its substratum. Table 6 also includes SNPs for which
Cochran Mantel Haenszel test showed that the adjusted p-value of
the meta analysis was less than 0.05, although the p-value for the
individual sample sets might be greater than 0.05. Table 7 provides
SNPs having a significant association (p-value of less than or
equal to 0.05) with coronary stenosis in either the Sample Set 1 or
the Sample Set 2 study.
[0396] An example of a replicated marker, where the homozygous
reported allele is associated with a decreased risk for coronary
stenosis is hCV1608777 (Table 6). Individuals in the second age
tertile (Age T2) with 2 copies of the reported allele of HCV1608777
(Mode Rec) showed significant association (p-values 0.0344 and
0.0076) with decreased risk (odds ratios of 0.51 and 0.19 times of
the reference) when compared to those carrying one or none of the
reported allele (heterozygotes and major homozygotes) in both the
Sample Set 1 and the Sample Set 2 studies.
[0397] An example of a replicated marker, where the reported allele
is associated with increased risk for coronary stenosis is
hCV16165996 (Table 6). Among the whole study population
(Strata=ALL), carriers of one or two copies of the reported allele
(Mode Dom) of HCV16165996 showed significant association (p-values
of 0.0306 and 0.0037) with coronary stenosis with an increased risk
(odds ratios of 1.31 and 1.69) compared with those carrying none of
the reported allele (major homozygotes) in both the Sample Set 1
and the Sample Set 2 studies.
[0398] All publications and patents cited in this specification are
herein incorporated by reference in their entirety. Various
modifications and variations of the described compositions, methods
and systems of the invention will be apparent to those skilled in
the art without departing from the scope and spirit of the
invention. Although the invention has been described in connection
with specific preferred embodiments and certain working examples,
it should be understood that the invention as claimed should not be
unduly limited to such specific embodiments. Indeed, various
modifications of the above-described modes for carrying out the
invention that are obvious to those skilled in the field of
molecular biology, genetics and related fields are intended to be
within the scope of the following claims. TABLE-US-00002 TABLE 1
Gene Number: 1 Celera Gene: hCG1811758 - 63000132574665 Celera
Transcript: hCT1954383 - 63000132574666 Public Transcript
Accession: Celera Protein: hCP1766736 - 197000069463822 Public
Protein Accession: Gene Symbol: CD163 Protein Name: CD163 antigen
Celera Genomic Axis: GA_x5YUV32W234 (5436647 . . . 5489410)
Chromosome: 12 OMIM NUMBER: 605545 OMIM Information: Transcript
Sequence (SEQ ID NO: 1): Protein Sequence (SEQ ID NO: 13): SNP
Information Context (SEQ ID NO: 25):
GGCTGTGCAGACAAAGGGAAAATCAACCCTGCATCTTTAGACAAGGCCAT
GTCCATTCCCATGTGGGTGGACAATGTTCAGTGTCCAAAAGGACCTGACA Y
GCTGTGGCAGTGCCCATCATCTCCATGGGAGAAGAGACTGGCCAGCCCCT
CGGAGGAGACCTGGATCACATGTGACAACAAGATAAGACTTCAGGAAGGA Celera SNP ID:
hCV25591528 Public SNP ID: SNP in Transcript Sequence SEQ ID NO: 1
SNP Position Transcript: 2788 SNP Source: Applera Population
(Allele, Count): Caucasian (T,8|C,32) african american (T,1|C,37)
total (T,9|C,69) SNP Type: Missense Mutation Protein Coding: SEQ ID
NO: 13, at position 901, (T,ACG)(M,ATG) Gene Number: 1 Celera Gene:
hCG1811758 - 63000132574665 Celera Transcript: hCT2286904 -
63000132574686 Public Transcript Accession: Celera Protein:
hCP1900559 - 197000069463823 Public Protein Accession: Gene Symbol:
CD163 Protein Name: CD163 antigen Celera Genomic Axis:
GA_x5YUV32W234 (5436647 . . . 5489410) Chromosome: 12 OMIM NUMBER:
605545 OMIM Information: Transcript Sequence (SEQ ID NO: 2):
Protein Sequence (SEQ ID NO: 14): SNP Information Context (SEQ ID
NO: 26): GGCTGTGCAGACAAAGGGAAAATCAACCCTGCATCTTTAGACAAGGCCAT
GTCCATTCCCATGTGGGTGGACAATGTTCAGTGTCCAAAAGGACCTGACA Y
GCTGTGGCAGTGCCCATCATCTCCATGGGAGAAGAGACTGGCCAGCCCCT
CGGAGGAGACCTGGATCACATGTGACAACAAGATAAGACTTCAGGAAGGA Celera SNP ID:
hCV25591528 Public SNP ID: SNP in Transcript Sequence SEQ ID NO: 2
SNP Position Transcript: 2788 SNP Source: Applera Population
(Allele, Count): caucasian (T,8|C,32) african american (T,1|C,37)
total (T,9|C,69) SNP Type: Missense Mutation Protein Coding: SEQ ID
NO: 14, at position 901, (T,ACG)(M,ATG) Gene Number: 1 Celera Gene:
hCG1811758 - 63000132574665 Celera Transcript: hCT2286906 -
63000132574728 Public Transcript Accession: Celera Protein:
hCP1900561 - 197000069463825 Public Protein Accession: Gene Symbol:
CD163 Protein Name: CD163 antigen Celera Genomic Axis:
GA_x5YUV32W234 (5436647 . . . 5489410) Chromosome: 12 OMIM NUMBER:
605545 OMIM Information: Transcript Sequence (SEQ ID NO: 3):
Protein Sequence (SEQ ID NO: 15): SNP Information Context (SEQ ID
NO: 27): GGCTGTGCAGACAAAGGGAAAATCAACCCTGCATCTTTAGACAAGGCCAT
GTCCATTCCCATGTGGGTGGACAATGTTCAGTGTCCAAAAGGACCTGACA Y
GCTGTGGCAGTGCCCATCATCTCCATGGGAGAAGAGACTGGCCAGCCCCT
CGGAGGAGACCTGGATCACATGTGACAACAAGATAAGACTTCAGGAAGGA Celera SNP ID:
hCV25591528 Public SNP ID: SNP in Transcript Sequence SEQ ID NO: 3
SNP Position Transcript: 2887 SNP Source: Applera Population
(Allele, Count): caucasian (T,8|C,32) african american (T,1|C,37)
total (T,9|C,69) SNP Type: Missense Mutation Protein Coding: SEQ ID
NO: 15, at position 934, (T,ACG)(M,ATG) Gene Number: 1 Celera Gene:
hCG1811758 - 63000132574665 Celera Transcript: hCT2286907 -
63000132574707 Public Transcript Accession: Celera Protein:
hCP1900560 - 197000069463824 Public Protein Accession: Gene Symbol:
CD163 Protein Name: CD163 antigen Celera Genomic Axis:
GA_x5YUV32W234 (5436647 . . . 5489410) Chromosome: 12 OMIM NUMBER:
605545 OMIM Information: Transcript Sequence (SEQ ID NO: 4):
Protein Sequence (SEQ ID NO: 16): SNP Information Context (SEQ ID
NO: 28): GGCTGTGCAGACAAAGGGAAAATCAACCCTGCATCTTTAGACAAGGCCAT
GTCCATTCCCATGTGGGTGGACAATGTTCAGTGTCCAAAAGGACCTGACA Y
GCTGTGGCAGTGCCCATCATCTCCATGGGAGAAGAGACTGGCCAGCCCCT
CGGAGGAGACCTGGATCACATGTGACAACAAGATAAGACTTCAGGAAGGA Celera SNP ID:
hCV25591528 Public SNP ID: SNP in Transcript Sequence SEQ ID NO: 4
SNP Position Transcript: 2788 SNP Source: Applera Population
(Allele, Count): caucasian (T,8|C,32) african american (T,1|C,37)
total (T,9|C,69) SNP Type: Missense Mutation Protein Coding: SEQ ID
NO: 16, at position 901, (T,ACG)(M,ATG) Gene Number: 2 Celera Gene:
hCG19417 - 79000075877786 Celera Transcript: hCT10488 -
79000075877787 Public Transcript Accession: NM_001278 Celera
Protein: hCP37140 - 197000069450514 Public Protein Accession:
NP_001269 Gene Symbol: CHUK Protein Name: conserved
helix-loop-helix ubiquitous kinase Celera Genomic Axis:
GA_x54KRFTF114 (32119433 . . . 32180761) Chromosome: 10 OMIM
NUMBER: 600664 OMIM Information: Transcript Sequence (SEQ ID NO:
5): Protein Sequence (SEQ ID NO: 17): SNP Information Context (SEQ
ID NO: 29): TAAGAAGAAGGATCCAAAGTGTATATTTGCATGTGAAGAGATGTCAGGAG
AAGTTCGGTTTAGTAGCCATTTACCTCAACCAAATAGCCTTTGTAGTTTA R
TAGTAGAACCCATGGAAAACTGGCTACAGTTGATGTTGAATTGGGACCCT
CAGCAGAGAGGAGGACCTGTTGACCTTACTTTGAAGCAGCCAAGATGTTT Celera SNP ID:
hCV1345898 Public SNP ID: rs2230804 SNP in Transcript Sequence SEQ
ID NO: 5 SNP Position Transcript: 877 SNP Source: Celera; HGBASE;
dbSNP Population (Allele, Count): no_pop (G,-|A,-) SNP Type:
Missense Mutation Protein Coding: SEQ ID NO: 17, at position 268,
(V,GTA)(I,ATA) Gene Number: 3 Celera Gene: hCG27811 -
66000116063099 Celera Transcript: hCT18953 - 66000116063100 Public
Transcript Accession: NM_001168 Celera Protein: hCP43533 -
197000064921779 Public Protein Accession: NP_001159 Gene Symbol:
BIRC5 Protein Name: baculoviral IAP repeat-containing 5 (survivin)
Celera Genomic Axis: GA_x5YUV32W262 (13301753 . . . 13333146)
Chromosome: 17 OMIM NUMBER: OMIM Information: Transcript Sequence
(SEQ ID NO: 6): Protein Sequence (SEQ ID NO: 18): SNP Information
Context (SEQ ID NO: 30):
ATTAACCCTTGGTGAATTTTTGAAACTGGACAGAGAAAGAGCCAAGAACA
AAATTGCAAAGGAAACCAACAATAAGAAGAAAGAATTTGAGGAAACTGCG R
AGAAAGTGCGCCGTGCCATCGAGCAGCTGGCTGCCATGGATTGAGGCCTC
TGGCCGGAGCTGCCTGGTCCCAGAGTGGCTGCACCACTTCCAGGGTTTAT Celera SNP ID:
hCV16266313 Public SNP ID: rs2071214 SNP in Transcript Sequence SEQ
ID NO: 6 SNP Position Transcript: 459 SNP Source: Applera
Population (Allele, Count): caucasian (A,35|G,3) african american
(A,30|G,0) total (A,65|G,3) SNP Type: Missense Mutation Protein
Coding: SEQ ID NO: 18, at position 129, (K,AAG)(E,GAG) SNP Source:
dbSNP; HapMap; HGBASE Population (Allele, Count: caucasian
(G,6|A,114) SNP Type: Missense Mutation Protein Coding: SEQ ID NO:
18, at position 129, (K,AAG)(E,GAG) Gene Number: 3 Celera Gene:
hCG27811 - 66000116063099 Celera Transcript: hCT1958629
-66000116063116 Public Transcript Accession: NM_001012271 Celera
Protein: hCP1766717 - 197000064921781 Public Protein Accession:
NP_001012271 Gene Symbol: BIRC5 Protein Name: baculoviral IAP
repeat-containing 5 (survivin) Celera Genomic Axis: GA_x5YUV32W262
(13301753 . . . 13333146) Chromosome: 17 OMIM NUMBER: OMIM
Information: Transcript Sequence (SEQ ID NO: 7): Protein Sequence
(SEQ ID NO: 19): SNP Information Context (SEQ ID NO: 31):
ATTAACCCTTGGTGAATTTTTGAAACTGGACAGAGAAAGAGCCAAGAACA
AAATTGCAAAGGAAACCAACAATAAGAAGAAAGAATTTGAGGAAACTGCG R
AGAAAGTGCGCCGTGCCATCGAGCAGCTGGCTGCCATGGATTGAGGCCTC
TGGCCGGAGCTGCCTGGTCCCAGAGTGGCTGCACCACTTCCAGGGTTTAT Celera SNP ID:
hCV16266313 Public SNP ID: rs2071214
SNP in Transcript Sequence SEQ ID NO: 7 SNP Position Transcript:
528 SNP Source: Applera Population (Allele, Count): caucasian
(A,35|G,3) african american (A,30|G,0) total (A,65|G,3) SNP Type:
Missense Mutation Protein Coding: SEQ ID NO: 19, at position 152,
(K,AAG)(E,GAG) SNP Source: dbSNP; HapMap; HGBASE Population
(Allele, Count): caucasian (G,6|A,114) SNP Type: Missense Mutation
Protein Coding: SEQ ID NO: 19, at position 152, (K,AAG)(E,GAG) Gene
Number: 3 Celera Gene: hCG27811 - 66000116063099 Celera Transcript:
hCT1962326 - 66000116063125 Public Transcript Accession:
NM_001012270 Celera Protein: hCP1778153 - 197000064921782 Public
Protein Accession: NP_001012270 Gene Symbol: BIRC5 Protein Name:
baculoviral IAP repeat-containing 5 (survivin) Celera Genomic Axis:
GA_x5YUV32W262 (13301753 . . . 13333146) Chromosome: 17 OMIM
NUMBER: OMIM Information: Transcript Sequence (SEQ ID NO: 8):
Protein Sequence (SEQ ID NO: 20): SNP Information Context (SEQ ID
NO: 32): AGTGTTTCTTCTGCTTCAAGGAGCTGGAAGGCTGGGAGCCAGATGACGAC
CCCATGCAAAGGAAACCAACAATAAGAAGAAAGAATTTGAGGAAACTGCG R
AGAAAGTGCGCCGTGCCATCGAGCAGCTGGCTGCCATGGATTGAGGCCTC
TGGCCGGAGCTGCCTGGTCCCAGAGTGGCTGCACCACTTCCAGGGTTTAT Celera SNP ID:
hCV16266313 Public SNP ID: rs2071214 SNP in Transcript Sequence SEQ
ID NO: 8 SNP Position Transcript: 341 SNP Source: Applera
Population (Allele, Count): caucasian (A,35|G,3) african american
(A,30|G,0) total (A,65|G,3) SNP Type: Silent Mutation Protein
Coding: SEQ ID NO: 20, at position 89, (R,CGA)(R,CGG) SNP Source:
dbSNP; HapMap; HGBASE Population (Allele, Count): caucasian
(G,6|A,114) SNP Type: Silent Mutation Protein Coding: SEQ ID NO:
20, at position 89, (R,CGA)(R,CGG) Gene Number: 3 Celera Gene:
hCG27811 - 66000116063099 Celera Transcript: hCT1967440 -
66000116063140 Public Transcript Accession: NM_001168 Celera
Protein: hCP1781899 - 197000064921785 Public Protein Accession:
NP_001159 Gene Symbol: BIRC5 Protein Name: baculoviral IAP
repeat-containing 5 (survivin) Celera Genomic Axis: GA_x5YUV32W262
(13301753 . . . 13333146) Chromosome: 17 OMIM NUMBER: OMIM
Information: Transcript Sequence (SEQ ID NO: 9): Protein Sequence
(SEQ ID NO: 21): SNP Information Context (SEQ ID NO: 33):
ATTAACCCTTGGTGAATTTTTGAAACTGGACAGAGAAAGAGCCAAGAACA
AAATTGCAAAGGAAACCAACAATAAGAAGAAAGAATTTGAGGAAACTGCG R
AGAAAGTGCGCCGTGCCATCGAGCAGCTGGCTGCCATGGATTGAGGCCTC
TGGCCGGAGCTGCCTGGTCCCAGAGTGGCTGCACCACTTCCAGGGTTTAT Celera SNP ID:
hCV16266313 Public SNP ID: rs2071214 SNP in Transcript Sequence SEQ
ID NO: 9 SNP Position Transcript: 459 SNP Source: Applera
Population (Allele, Count): caucasian (A,35|G,3) african american
(A,30|G,0) total (A,65|G,3) SNP Type: Missense Mutation Protein
Coding: SEQ ID NO: 21, at position 129, (K,AAG)(E,GAG) SNP Source:
dbSNP; HapMap; HGBASE Population (Allele, Count): caucasian
(G,6|A,114) SNP Type: Missense Mutation Protein Coding: SEQ ID NO:
21, at position 129, (K,AAG)(E,GAG) Gene Number: 3 Celera Gene:
hCG27811 - 66000116063099 Celera Transcript: hCT2336835 -
66000116063149 Public Transcript Accession: NM_001012271 Celera
Protein: hCP1789145 - 197000064921786 Public Protein Accession:
NP_001012271 Gene Symbol: BIRC5 Protein Name: baculoviral IAP
repeat-containing 5 (survivin) Celera Genomic Axis: GA_x5YUV32W262
(13301753 . . . 13333146) Chromosome: 17 OMIM NUMBER: OMIM
Information: Transcript Sequence (SEQ ID NO: 10): Protein Sequence
(SEQ ID NO: 22): SNP Information Context (SEQ ID NO: 34):
ATTAACCCTTGGTGAATTTTTGAAACTGGACAGAGAAAGAGCCAAGAACA
AAATTGCAAAGGAAACCAACAATAAGAAGAAAGAATTTGAGGAAACTGCG R
AGAAAGTGCGCCGTGCCATCGAGCAGCTGGCTGCCATGGATTGAGGCCTC
TGGCCGGAGCTGCCTGGTCCCAGAGTGGCTGCACCACTTCCAGGGTTTAT Celera SNP ID:
hCV16266313 Public SNP ID: rs2071214 SNP in Transcript Sequence SEQ
ID NO: 10 SNP Position Transcript: 528 SNP Source: Applera
Population (Allele, Count): caucasian (A,35|G,3) african american
(A,30|G,0) total (A,65|G,3) SNP Type: Missense Mutation Protein
Coding: SEQ ID NO: 22, at position 152, (K,AAG)(E,GAG) SNP Source:
dbSNP; HapMap; HGBASE Population (Allele, Count): caucasian
(G,6|A,114) SNP Type: Missense Mutation Protein Coding: SEQ ID NO:
22, at position 152, (K,AAG)(E,GAG) Gene Number: 3 Celera Gene:
hCG27811 - 66000116063099 Celera Transcript: hCT2336837 -
66000116063132 Public Transcript Accession: NM_001012270 Celera
Protein: hCP1789143 - 197000064921784 Public Protein Accession:
NP_001012270 Gene Symbol: BIRC5 Protein Name: baculoviral IAP
repeat-containing 5 (survivin) Celera Genomic Axis: GA_x5YUV32W262
(13301753 . . . 13333146) Chromosome: 17 OMIM NUMBER: OMIM
Information: Transcript Sequence (SEQ ID NO: 11): Protein Sequence
(SEQ ID NO: 23): SNP Information Context (SEQ ID NO: 35):
AGTGTTTCTTCTGCTTCAAGGAGCTGGAAGGCTGGGAGCCAGATGACGAC
CCCATGCAAAGGAAACCAACAATAAGAAGAAAGAATTTGAGGAAACTGCG R
AGAAAGTGCGCCGTGCCATCGAGCAGCTGGCTGCCATGGATTGAGGCCTC
TGGCCGGAGCTGCCTGGTCCCAGAGTGGCTGCACCACTTCCAGGGTTTAT Celera SNP ID:
hCV16266313 Public SNP ID: rs2071214 SNP in Transcript Sequence SEQ
ID NO: 11 SNP Position Transcript: 341 SNP Source: Applera
Population (Allele, Count): caucasian (A,35|G,3) african american
(A,30|G,0) total (A,65|G,3) SNP Type: Silent Mutation Protein
Coding: SEQ ID NO: 23, at position 89, (R,CGA)(R,CGG) SNP Source:
dbSNP; HapMap; HGBASE Population (Allele, Count): caucasian
(G,6|A,114) SNP Type: Silent Mutation Protein Coding: SEQ ID NO:
23, at position 89, (R,CGA)(R,CGG) Gene Number: 4 Celera Gene:
hCG38633 -226000018878110 Celera Transcript: hCT29876 -
226000018878111 Public Transcript Accession: NM_000134 Celera
Protein: hCP48455 - 197000069464429 Public Protein Accession:
NP_000125 Gene Symbol: FABP2 Protein Name: fatty acid binding
protein 2, intestinal Celera Genomic Axis: GA_x5YUV32VYAM (759337 .
. . 784251) Chromosome: 4 OMIM NUMBER: 134640 OMIM Information:
Transcript Sequence (SEQ ID NO: 12): Protein Sequence (SEQ ID NO:
24): SNP Information Context (SEQ ID NO: 36):
AATGGGTGTTAATATAGTGAAAAGGAAGCTTGCAGCTCATGACAATTTGA
AGCTGACAATTACACAAGAAGGAAATAAATTCACAGTCAAAGAATCAAGC R
CTTTTCGAAACATTGAAGTTGTTTTTGAACTTGGTGTCACCTTTAATTAC
AATCTAGCAGACGGAACTGAACTCAGGGGGACCTGGAGCCTTGAGGGAAA Celera SNP ID:
hCV761961 Public SNP ID: rs1799883 SNP in Transcript Sequence SEQ
ID NO: 12 SNP Position Transcript: 224 SNP Source: HGMD; dbSNP;
HapMap; HGBASE Population (Allele, Count): caucasian (A,44|G,74)
SNP Type: Missense Mutation Protein Coding: SEQ ID NO: 24, at
position 55, (A,GCT)(T,ACT)
[0399] TABLE-US-00003 TABLE 2 Gene Number: 1 Celera Gene:
hCG1811758 -63000132574665 Gene Symbol: CD163 Protein Name: CD163
antigen Celera Genomic Axis: GA_x5YUV32W234 (5436647 . . . 5489410)
Chromosome: 12 OMIM NUMBER: 605545 OMIM Information: Genomic
Sequence (SEQ ID NO: 37): SNP Information Context (SEQ ID NO: 41):
CATATAGGTCGATGGATACTCACTGTCACATGTGATCCAGGTCTCCTCCG
AGGGGCTGGCCAGTCTCTTCTCCCATGGAGATGATGGGCACTGCCACAGC R
TGTCAGGTCCTTTTGGACACTGAACATTGTCCACCCACATGGGAATGGAC
ATGGCCTTGTCTAAAGATGCAGGGTTGATTTTCCCTTTGTCTGCACAGCC Celera SNP ID:
hCV25591528 Public SNP ID: SNP in Genomic Sequence: SEQ ID NO: 37
SNP Position Genomic: 24287 SNP Source: Applera Population (Allele,
Count): caucasian (A,8|G,32) african american (A,1|G,37) total
(A,9|G,69) SNP Type: MISSENSE MUTATION; HUMAN-MOUSE SYNTENIC REGION
Gene Number: 2 Celera Gene: hCG19417 - 79000075877786 Gene Symbol:
CHUK Protein Name: conserved helix-loop-helix ubiquitous kinase
Celera Genomic Axis: GA_x54KRFTF114 (32119433 . . . 32180761)
Chromosome: 10 OMIM NUMBER: 600664 OMIM Information: Genomic
Sequence (SEQ ID NO: 38): SNP Information Context (SEQ ID NO: 42):
AAACATCTTGGCTGCTTCAAAGTAAGGTCAACAGGTCCTCCTCTCTGCTG
AGGGTCCCAATTCAACATCAACTGTAGCCAGTTTTCCATGGGTTCTACTA Y
TAAACTAGAAAACATACAAAATAGGGTGAAAATCAAATCATTATGTTCCA
ATTTCCCTTTATACTGTTAGAAAGGTAATTTTGCAGGTTGTCCATTTTCT Celera SNP ID:
hCV1345898 Public SNP ID: rs2230804 SNP in Genomic Sequence: SEQ ID
NO: 38 SNP Position Genomic: 39847 SNP Source: Celera; HGBASE;
dbSNP Population (Allele, Count): no_pop (C,-|T,-) SNP Type:
MISSENSE MUTATION; HUMAN-MOUSE SYNTENIC REGION; SILENT MUTATION
Gene Number: 3 Celera Gene: hCG27811 - 66000116063099 Gene Symbol:
BIRC5 Protein Name: baculoviral IAP repeat-containing 5 (survivin)
Celera Genomic Axis: GA_x5YUV32W262 (13301753 . . . 13333146)
Chromosome: 17 OMIM NUMBER: OMIM Information: Genomic Sequence (SEQ
ID NO: 39): SNP Information Context (SEQ ID NO: 43):
GGATGTGACTGGGAAGCTCTGGTTTCAGTGTCATGTGTCTATTCTTTATT
TCCAGGCAAAGGAAACCAACAATAAGAAGAAAGAATTTGAGGAAACTGCG R
AGAAAGTGCGCCGTGCCATCGAGCAGCTGGCTGCCATGGATTGAGGCCTC
TGGCCGGAGCTGCCTGGTCCCAGAGTGGCTGCACCACTTCCAGGGTTTAT Celera SNP ID:
hCV16266313 Public SNP ID: rs2071214 SNP in Genomic Sequence: SEQ
ID NO: 39 SNP Position Genomic: 19267 SNP Source: Applera
Population (Allele, Count): caucasian (A,35|G,3) african american
(A,30|G,0) total (A,65|G,3) SNP Type: MISSENSE MUTATION;
HUMAN-MOUSE SYNTENIC REGION; SILENT MUTATION SNP Source: dbSNP;
HapMap; HGBASE Population (Allele, Count): caucasian (G,6|A,114)
SNP Type: MISSENSE MUTATION; HUMAN-MOUSE SYNTENIC REGION; SILENT
MUTATION Gene Number: 4 Celera Gene: hCG38633 - 226000018878110
Gene Symbol: FABP2 Protein Name: fatty acid binding protein 2,
intestinal Celera Genomic Axis: GA_x5YUV32VYAM (759337 . . .
784251) Chromosome: 4 OMIM NUMBER: 134640 OMIM Information: Genomic
Sequence (SEQ ID NO: 40): SNP Information Context (SEQ ID NO: 44):
CTCATAAAAAAAAAAATTCTTACCCTGAGTTCAGTTCCGTCTGCTAGATT
GTAATTAAAGGTGACACCAAGTTCAAAAACAACTTCAATGTTTCGAAAAG Y
GCTTGATTCTTTGACTGTGAATTTATTTCCTTCTTGTGTAATTGTCAGCT
TCAAATTGTCATGAGCTGCAAGCTTCCTTTTCACTATATTAACACCTGTA Celera SNP ID:
hCV761961 Public SNP ID: rs1799883 SNP in Genomic Sequence: SEQ ID
NO: 40 SNP Position Genomic: 13498 SNP Source: HGMD; dbSNP; HapMap;
HGBASE Population (Allele, Count): caucasian (T,44|C,74) SNP Type:
MISSENSE MUTATION; HUMAN-MOUSE SYNTENIC REGION
[0400] TABLE-US-00004 TABLE 3 hCV25591528 SEQ ID NO: 25 hCV25591528
SEQ ID NO: 26 hCV25591528 SEQ ID NO: 27 hCV25591528 SEQ ID NO:
28
[0401] TABLE-US-00005 TABLE 4 hCV25591528 SEQ ID NO: 41
[0402] TABLE-US-00006 TABLE 5 Primers Sequence A Sequence B
Sequence C hCV Alleles (Allele-specific Primer) (Allele-specific
Primer) (Common Primer) hCV1345898 C/T CAGTTTTCCATGGGTTCTACTAC
CAGTTTTCCATGGGTTCTACTAT TTATGAAATGGTACAGACAAGTGAT (SEQ ID NO:45)
(SEQ ID NO:46) (SEQ ID NO:47) hCV16266313 A/G CACGGCGCACTTTCTT
CACGGCGCACTTTCTC TGTTTTTTCCTTTGTCATCTTATCTA (SEQ ID NO:48) (SEQ ID
NO:49) (SEQ ID NO:50) hCV25591528 A/G TCCAAAAGGACCTGACAT
TCCAAAAGGACCTGACAC GGCTGCAGAATGGAATTT (SEQ ID NO:51) (SEQ ID NO:52)
(SEQ ID NO:53) hCV761961 C/T CACAGTCAAAGAATCAAGCG
TCACAGTCAAAGAATCAAGCA AAATTCTTACCCTGAGTTCAGTTC (SEQ ID NO:54) (SEQ
ID NO:55) (SEQ ID NO:56)
[0403] TABLE-US-00007 TABLE 6 Gene Case Cntrl Marker Name Sample
Set p-value OR 95% CI Freq. Freq. Allele1 Mode Strata Adjust
hCV1608777 PLSCR1 Sample Set 2 0.0344 0.51 0.27-0.96 5.1 9.6 T Rec
Age T2 hCV1608777 PLSCR1 Sample Set 1 0.0076 0.19 0.05-0.72 2.8
13.2 T Rec Age T2 hCV16165996 LRP2 Sample Set 2 0.0306 1.31
1.03-1.68 49.0 42.3 T Dom ALL hCV16165996 LRP2 Sample Set 2 0.0281
1.31 1.02-1.68 28.3 23.2 T Add Smoke+ hCV16165996 LRP2 Sample Set 1
0.0037 1.69 1.18-2.4 47.9 35.2 T Dom ALL hCV16165996 LRP2 Sample
Set 1 0.0321 1.67 1.04-2.7 48.3 35.9 T Dom Smoke+ hCV16181123 ACADL
Sample Set 2 0.0182 1.74 1.09-2.77 19.3 12.1 G Rec MI- hCV16181123
ACADL Sample Set 1 0.0072 1.87 1.18-2.96 66.9 52.0 G Dom MI-
hCV2143203 CD44 Sample Set 2 0.0394 4.00 0.97-16.49 6.7 1.8 T Rec
Age T1 hCV2143203 CD44 Sample Set 2 0.0326 2.08 1.05-4.11 26.0 14.5
T Add MI-Age T1 hCV2143203 CD44 Sample Set 1 0.0274 6.00 0.7-50.8
5.5 0.0 T Rec Age T1 hCV2143203 CD44 Sample Set 1 0.0163 8.90
0.8-88.9 6.7 0.0 T Rec MI-Age T1 hCV2143205 CD44 Sample Set 2
0.0090 1.68 1.12-2.52 39.8 28.3 A Add MI-Smoke- hCV2143205 CD44
Sample Set 1 0.0123 3.88 1.3-12 18.2 5.4 A Rec MI-Smoke-
hCV22273419 GP6 Sample Set 2 0.0332 0.81 0.67-0.99 15.5 18.4 C Add
ALL hCV22273419 GP6 Sample Set 1 0.0229 0.51 0.29-0.92 27.7 42.7 C
Dom Smoke- hCV25591528 CD163 Sample Set 2 0.0285 0.73 0.55-0.97 9.7
12.8 A Add ALL hCV25591528 CD163 Sample Set 2 0.0356 0.20 0-1.5 0.0
1.9 A Rec Male hCV25591528 CD163 Sample Set 2 0.0105 0.51 0.3-0.88
7.3 13.4 A Add Smoke- hCV25591528 CD163 Sample Set 1 0.0096 0.10
0-1 0.0 2.2 A Rec ALL hCV25591528 CD163 Sample Set 1 0.0328 0.20
0-1.4 0.0 2.5 A Rec Male hCV25591528 CD163 Sample Set 1 0.0457 0.52
0.28-0.99 7.8 14.0 A Allelic Smoke- hCV25603879 SCNN1A Sample Set 2
0.0330 2.24 1.05-4.76 11.1 5.3 T Dom Smoke- hCV25603879 SCNN1A
Sample Set 2 0.0243 2.71 1.08-6.8 6.7 2.6 T Add MI-Age T1
hCV25603879 SCNN1A Sample Set 2 0.0429 1.65 1.01-2.68 9.5 6.0 T Dom
MI- hCV25603879 SCNN1A Sample Set 2 0.0100 2.26 1.18-4.32 4.8 2.2 T
Add MI-Male hCV25603879 SCNN1A Sample Set 2 0.0474 1.83 1-3.33 10.4
6.0 T Dom MI- hCV25603879 SCNN1A Sample Set 2 0.0054 3.22 1.36-7.62
15.3 5.3 T Dom MI-Smoke- hCV25603879 SCNN1A Sample Set 1 0.0147
3.67 1.2-11.5 6.4 1.8 T Add Smoke- hCV25603879 SCNN1A Sample Set 1
0.0201 5.99 1.07-33.6 6.7 1.2 T Add MI-Age T1 hCV25603879 SCNN1A
Sample Set 1 0.0035 3.62 1.43-9.2 5.4 1.6 T Add MI- hCV25603879
SCNN1A Sample Set 1 0.0165 3.93 1.2-13.3 6.3 1.7 T Add MI-Male
hCV25603879 SCNN1A Sample Set 1 0.0035 3.62 1.43-9.2 5.4 1.6 T Add
MI- hCV25603879 SCNN1A Sample Set 1 0.0020 5.53 1.6-18.9 9.3 1.8 T
Add MI-Smoke- hCV25627634 SMTN Sample Set 2 0.0396 1.82 1.02-3.24
64.4 49.8 G Dom MI-Smoke- hCV25627634 SMTN Sample Set 1 0.0068 2.62
1.28-5.4 15.7 6.6 G Rec MI- hCV3084793 APOE Sample Set 2 0.0487
1.62 1-2.64 30.5 21.3 C Dom MI-Female hCV3084793 APOE Sample Set 2
0.0299 1.82 1.06-3.14 38.5 25.6 C Dom MI-Age T2 hCV3084793 APOE
Sample Set 1 0.0069 2.25 1.2-4.2 23.2 11.8 C Add MI-Female
hCV3084793 APOE Sample Set 1 0.0137 8.40 0.9-78.2 7.9 0.0 C Rec
MI-Age T2 hCV5478 TCF1 Sample Set 2 0.0100 3.07 1.24-7.57 3.5 1.3 T
Allelic Age T2 hCV5478 TCF1 Sample Set 1 0.0152 12.21 1.18-126.50
4.6 0.6 T Allelic Age T2 hCV598677 EDN1 Sample Set 2 0.0065 1.82
1.18-2.79 45.8 31.7 T Dom Age T1 hCV598677 EDN1 Sample Set 2 0.0048
3.23 1.38-7.53 60.0 31.7 T Dom MI-Age T1 hCV598677 EDN1 Sample Set
1 0.0372 4.60 0.97-22 9.9 2.3 T Rec Age T1 hCV598677 EDN1 Sample
Set 1 0.0048 8.31 1.5-45.4 16.7 2.4 T Rec MI-Age T1 hCV7482175
HLA-A Sample Set 2 0.0163 0.66 0.47-0.93 25.7 34.4 A Dom Smoke+
hCV7482175 HLA-A Sample Set 1 0.0132 0.61 0.4-0.9 14.3 21.6 A Add
Smoke+ hCV7530616 WRN Sample Set 2 0.0027 14.10 1.46-136.45 1.9 0.1
A Rec MI- hCV7530616 WRN Sample Set 1 0.0492 5.90 0.5-57.9 3.6 0.0
A Rec MI-Female hCV7530616 WRN Sample Set 1 0.0230 2.43 1.1-5.2
11.7 5.2 A Add MI-Smoke+ hCV7584364 PTGS1 Sample Set 2 0.0006 9.32
1.97-44.18 2.7 0.3 T Rec MI- hCV7584364 PTGS1 Sample Set 2 0.0051
11.90 1.2-117 3.6 0.0 T Rec MI-Age T1 hCV7584364 PTGS1 Sample Set 1
0.0209 1.99 1.10-3.6 10.0 5.3 T Add MI- hCV7584364 PTGS1 Sample Set
1 0.0282 3.11 1.1-8.7 13.3 4.7 T Add MI-Age T1 hCV761961 FABP2
Sample Set 2 0.0360 1.48 1.03-2.14 50.0 40.3 T Dom Age T3 hCV761961
FABP2 Sample Set 1 0.0397 6.73 0.84-53.9 9.5 1.5 T Rec Age T3
hCV790057 CETP Sample Set 2 0.0121 0.78 0.64-0.95 26.3 31.4 G
Allelic ALL hCV790057 CETP Sample Set 2 0.0443 0.61 0.37-0.99 6.9
11.0 G Rec MI- hCV790057 CETP Sample Set 1 0.0033 0.27 0.11-0.67
30.0 61.2 G Dom MI-Age T1 hCV790057 CETP Sample Set 1 0.0186 0.49
0.27-0.89 44.0 61.6 G Dom Age T1 hCV790057 CETP Sample Set 1 0.0482
0.49 0.24-1.0 37.2 55.0 G Dom MI-Smoke- hCV8705506 KLK1 Sample Set
2 0.0122 0.58 0.38-0.89 45.6 59.2 C Dom Smoke- hCV8705506 KLK1
Sample Set 1 0.0388 0.57 0.34-0.98 9.2 15.0 C Rec ALL hCV8921288
GAPD Sample Set 2 0.0130 1.38 1.07-1.77 40.9 33.5 G Dom ALL
hCV8921288 GAPD Sample Set 1 0.0486 6.45 0.78-53.3 5.8 1.0 G Rec
Female hCV9458082 NOS2A Sample Set 2 0.0092 1.47 1.1-1.96 17.8 12.9
T Rec ALL hCV9458082 NOS2A Sample Set 1 0.0406 1.87 1.02-3.4 66.4
51.3 T Dom Age T2 hCV9458082 NOS2A Sample Set 1 0.0200 1.49
1.06-2.1 41.3 32.1 T Add Smoke+ hCV9458082 NOS2A Sample Set 1
0.0228 1.74 1.08-2.8 68.0 55.0 T Dom Male hCV9506149 FN1 Sample Set
2 0.0058 1.32 1.08-1.6 28.7 23.4 T Add ALL hCV9506149 FN1 Sample
Set 1 0.0152 1.60 1.08-2.4 27.3 19.0 T Add Male hCV1552900 ALOX12
Sample Set 2 0.1180 1.38 0.92-2.06 50.0 42.1 A Allelic Smoke-
gender, age_group_lt54_le64 hCV1552900 ALOX12 Sample Set 1 0.2066
1.40 0.83-2.33 50.0 42.5 A Allelic Smoke- gender,
age_group_lt54_le64 hCV1552900 ALOX12 Meta 0.0445 1.38 1.01-1.90
50.0 42.2 A Allelic Smoke- source, gender, age_group_lt54_le6
hCV16266313 BIRC5 Sample Set 2 0.3686 1.37 0.69-2.72 6.1 4.9 G
Allelic Age T3 gender, smoke hCV16266313 BIRC5 Sample Set 1 0.0280
3.21 1.08-9.60 9.5 3.1 G Allelic Age T3 gender, smoke hCV16266313
BIRC5 Meta 0.0359 1.83 1.04-3.22 7.7 4.5 G Allelic Age T3 source,
gender, smoke hCV1985480 AGT Sample Set 2 0.2513 0.84 0.63-1.13
11.8 14.2 A Allelic ALL gender, age_group_lt54_le64, smoke
hCV1985480 AGT Sample Set 1 0.0447 0.57 0.32-0.99 7.5 12.1 A
Allelic ALL gender, age_group_lt54_le64, smoke hCV1985480 AGT Meta
0.0478 0.77 0.59-1.00 10.6 13.7 A Allelic ALL source, gender,
age_group_lt54_le64, smoke hCV22274624 HDLBP Sample Set 2 0.0348
0.67 0.47-0.97 40.1 48.7 C Dom ALL gender, age_group_lt54_le64
hCV22274624 HDLBP Sample Set 1 0.4815 0.85 0.54-1.34 38.8 43.3 C
Dom ALL gender, age_group_lt54_le64 hCV22274624 HDLBP Meta 0.0369
0.74 0.55-0.98 39.6 47.4 C Dom ALL source, gender,
age_group_lt54_le64 hCV2548962 PON1 Sample Set 2 0.1393 0.85
0.69-1.05 27.2 30.4 C Allelic Smoke+ gender, age_group_lt54_le64,
smoke hCV2548962 PON1 Sample Set 1 0.0545 0.71 0.50-1.01 28.1 35.9
C Allelic Smoke+ gender, age_group_lt54_le64, smoke hCV2548962 PON1
Meta 0.0238 0.81 0.68-0.97 27.4 31.7 C Allelic Smoke+ source,
gender, age_group_lt54_le64, smoke hCV11660791 MTHFD1 Sample Set 2
0.01 1.5 1.1-2.1 86 81 C Allelic ALL hCV11660791 MTHFD1 Sample Set
1 0.0364 0.58 0.3-1.0 14 21 T Allelic Age T3
[0404] TABLE-US-00008 TABLE 7 Gene Case Cntrl Marker Name Sample
Set p-value OR 95% CI Freq. Freq. Allele1 Mode Strata hCV11975277
SELP Sample Set 1 0.039 n/a* G Rec Age T3 hCV1575287 IL8RA Sample
Set 1 0.027 n/a* G Dom Older hCV1575287 IL8RA Sample Set 1 0.046
n/a* G Dom Smoke- hCV1603656 HSPG2 Sample Set 1 0.037 1.13 T
Allelic Female hCV1639938 F13A1 Sample Set 1 0.04 n/a* A Dom Age T2
hCV3216553 APOB Sample Set 1 0.047 n/a* A Dom Age T2 hCV3216553
APOB Sample Set 1 0.034 1.55 A Allelic Age T1 hCV7582933 PLA2G7
Sample Set 1 0.019 5.95 T Rec Age T3 hCV1129436 APOL3 Sample Set 1
0.02463 C Allelic Smoke+ hCV1129436 APOL3 Sample Set 1 0.02409 C
Allelic male/Age T2 hCV1129436 APOL3 Sample Set 1 0.02811 C Allelic
male/no hypertension hCV1129436 APOL3 Sample Set 1 0.00455 C
Allelic MaleSmoke+ hCV11623862 TBXAS1 Sample Set 1 0.04724 T
Allelic All hCV11972321 F13A1 Sample Set 1 0.04190 G Allelic Smoke+
hCV11972321 F13A1 Sample Set 1 0.04755 G Allelic female/no
hypertension hCV11972321 F13A1 Sample Set 1 0.02064 G Allelic
male/Age T1 hCV11972321 F13A1 Sample Set 1 0.01487 G Allelic
MaleSMoke+ hCV1202883 MTHFR Sample Set 1 0.01357 A Allelic MI+
hCV1202883 MTHFR Sample Set 1 0.03654 A Allelic MaleMI+ hCV1345898
CHUK Sample Set 1 0.02992 C Allelic male/Age T3 hCV15954277 PRKCQ
Sample Set 1 0.03260 A Allelic female/no hypertension hCV15963704
ITGA3 Sample Set 1 0.03398 A Allelic Hypertension hCV15963704 ITGA3
Sample Set 1 0.02997 A Allelic male/hypertension hCV16170982 SREBF2
Sample Set 1 0.04406 C Allelic female/Age T2 hCV16170982 SREBF2
Sample Set 1 0.04529 C Allelic FemaleMI- hCV16170993 SELPLG Sample
Set 1 0.04733 G Allelic Smoke+ hCV16172262 FABP6 Sample Set 1
0.04577 G Allelic MI- hCV16172262 FABP6 Sample Set 1 0.02549 G
Allelic No hypertension hCV16172262 FABP6 Sample Set 1 0.04855 G
Allelic MaleMI- hCV16172262 FABP6 Sample Set 1 0.03227 G Allelic
male/no hypertension hCV16179628 ABCC2 Sample Set 1 0.03322 T
Allelic female/Age T2 hCV16195242 MTP Sample Set 1 0.00814 G
Allelic MI- hCV16195242 MTP Sample Set 1 0.01728 G Allelic Smoke+
hCV16195242 MTP Sample Set 1 0.00212 G Allelic MaleMI- hCV16195242
MTP Sample Set 1 0.02587 G Allelic MaleSmoke+ hCV1932478 P2RY12
Sample Set 1 0.04012 T Allelic MI- hCV2213764 MMP11 Sample Set 1
0.02493 C Allelic Smoke+ hCV2213764 MMP11 Sample Set 1 0.02596 C
Allelic FemaleMI+ hCV22271841 PDGFRA Sample Set 1 0.04281 C Allelic
MI+ hCV22271841 PDGFRA Sample Set 1 0.03142 C Allelic
female/hypertension hCV22272408 PRKCQ Sample Set 1 0.02252 A
Allelic female/no hypertension hCV22272408 PRKCQ Sample Set 1
0.04503 A Allelic MaleSmoke- hCV22272567 ABCC2 Sample Set 1 0.03614
A Allelic MI- hCV22272567 ABCC2 Sample Set 1 0.01204 A Allelic
MaleMI- hCV22272567 ABCC2 Sample Set 1 0.01309 A Allelic
male/hypertension hCV22274307 MTP Sample Set 1 0.03426 C Allelic
age T2 hCV22274307 MTP Sample Set 1 0.04299 C Allelic MI-
hCV22274307 MTP Sample Set 1 0.02673 C Allelic male/age T2
hCV25472673 NPC1 Sample Set 1 0.02505 C Allelic MI+ hCV25591743
ABCC2 Sample Set 1 0.02414 T Allelic MI- hCV25591743 ABCC2 Sample
Set 1 0.03752 T Allelic male/hypertension hCV25614474 PLG Sample
Set 1 0.01528 A Allelic female hCV25614474 PLG Sample Set 1 0.02338
A Allelic MI- hCV25614474 PLG Sample Set 1 0.00122 A Allelic
FemaleMI- hCV25614474 PLG Sample Set 1 0.02906 A Allelic
female/hypertension hCV25629888 TIMP2 Sample Set 1 0.01835 G
Allelic age T 1 hCV25638153 APOA5 Sample Set 1 0.03379 G Allelic
age T3 hCV25646316 LRP2 Sample Set 1 0.04595 G Allelic male/no
hypertension hCV25652767 LRP1 Sample Set 1 0.02443 A Allelic MI+
hCV25652767 LRP1 Sample Set 1 0.00243 A Allelic MI- hCV2705229
ITGA10 Sample Set 1 0.03914 T Allelic Hypertension hCV2705229
ITGA10 Sample Set 1 0.00468 T Allelic male/hypertension hCV2822674
CUBN Sample Set 1 0.02925 T Allelic female hCV2822674 CUBN Sample
Set 1 0.00076 T Allelic female/age T3 hCV2822674 CUBN Sample Set 1
0.03335 T Allelic FemaleMI+ hCV3135085 CUBN Sample Set 1 0.00245 T
Allelic No hypertension hCV3135085 CUBN Sample Set 1 0.01122 T
Allelic female/no hypertension hCV3135085 CUBN Sample Set 1 0.02797
T Allelic male/age T3 hCV342590 F5 Sample Set 1 0.04203 C Allelic
FemaleSmoke- hCV342590 F5 Sample Set 1 0.03999 C Allelic MaleSmoke+
hCV5687 CX3CR1 Sample Set 1 0.03556 T Allelic male hCV7490135 NPC1
Sample Set 1 0.01191 G Allelic age T2 hCV7490135 NPC1 Sample Set 1
0.04298 G Allelic MI+ hCV7490135 NPC1 Sample Set 1 0.02428 G
Allelic male/age T2 hCV7900503 CX3CR1 Sample Set 1 0.02118 C
Allelic female/age T1 hCV7900503 CX3CR1 Sample Set 1 0.03188 C
Allelic FemaleSmoke+ hCV7900503 CX3CR1 Sample Set 1 0.02491 C
Allelic MaleMI+ hCV1361979 ACAT2 Sample Set 2 0.0027 1.71388
1.2-2.4 0.7342 0.6171 A Dom Age T2 hCV1361979 ACAT2 Sample Set 2
0.0298 1.37325 1-1.8 0.7114 0.6422 A Dom Male hCV1361979 ACAT2
Sample Set 2 0.0152 1.2601 1-1.5 0.4429 0.3869 A Add Smoke+
hCV1361979 ACAT2 Sample Set 2 0.0198 1.1936 1-1.4 0.4431 0.4 A Add
ALL hCV1361979 ACAT2 Sample Set 2 0.00753 1.73944 1.2-2.6 0.7371
0.6171 A Dom Age T2 hCV1361979 ACAT2 Sample Set 2 0.0301 1.47134
1-2.1 0.7253 0.6422 A Dom Male hCV1361979 ACAT2 Sample Set 2 0.0181
1.49327 1.1-2.1 0.7153 0.6272 A Dom Smoke+ hCV1361979 ACAT2 Sample
Set 2 0.0358 1.32885 1-1.7 0.7 0.6371 A Dom ALL hCV1361979 ACAT2
Sample Set 2 0.0126 2.1026 1.2-3.8 0.7722 0.6171 A Dom MI-AgeT2
hCV1361979 ACAT2 Sample Set 2 0.00369 1.9409 1.2-3 0.7578 0.6171 A
Dom MI-AgeT2 hCV1361979 ACAT2 Sample Set 2 0.0373 1.2953 1-1.7
0.4497 0.3869 A Add MI-Smoke+ hCV22274712 MTP Sample Set 2 0.0403
1.41589 1-2 0.6321 0.5482 G Dom Age T2 hCV22274712 MTP Sample Set 2
0.0369 1.43566 1-2 0.6596 0.5745 G Dom Female hCV22274712 MTP
Sample Set 2 0.0498 1.4498 1-2.1 0.6733 0.5871 G Dom MI- hCV2531431
THBD Sample Set 2 0.0111 0.22613 0.1-0.8 0.0149 0.0625 T Rec Age T3
hCV2531431 THBD Sample Set 2 0.0299 0.42323 0.2-0.9 0.0202 0.0464 T
Rec Male hCV2531431 THBD Sample Set 2 0.00295 0.5156 0.3-0.8 0.1186
0.207 T Allelic Age T3 hCV25608818 SLC10A2 Sample Set 2 0.00737
6.0214 1.3-27 0.021 0.0035 A Add Female hCV25608818 SLC10A2 Sample
Set 2 0.0363 3.5574 1.1-11.6 0.022 0.0063 A Add Smoke- hCV25608818
SLC10A2 Sample Set 2 0.0212 5.3694 1.1-26.8 0.0188 0.0035 A Add
Female hCV25608818 SLC10A2 Sample Set 2 0.0263 4.208 1.2-15 0.0259
0.0063 A Allelic Smoke- hCV25608818 SLC10A2 Sample Set 2 0.0101
7.4602 1.2-45.2 0.0259 0.0035 A Add MI-Female hCV25608818 SLC10A2
Sample Set 2 0.0341 -- -- 0.0104 0 A Rec MI-Male hCV25608818
SLC10A2 Sample Set 2 0.0156 5.5614 1.4-22.6 0.0339 0.0063 A Add
MI-Smoke- hCV25608818 SLC10A2 Sample Set 2 0.0314 -- -- 0.0065 0 A
Rec MI- hCV25608818 SLC10A2 Sample Set 2 0.00132 8.5895 1.8-41.7
0.0297 0.0035 A Add MI-Female hCV25608818 SLC10A2 Sample Set 2
0.0239 4.3227 1.2-15.5 0.0265 0.0063 A Allelic MI-Smoke-
hCV25653599 WDR12 Sample Set 2 0.00488 1.77148 1.2-2.6 0.2868 0.185
C Dom Age T2 hCV25653599 WDR12 Sample Set 2 0.0125 1.4324 1.1-1.9
0.148 0.1081 C Add Male hCV25653599 WDR12 Sample Set 2 0.0011
2.06754 1.3-3.2 0.3194 0.185 C Dom Age T2 hCV25653599 WDR12 Sample
Set 2 0.00925 1.5412 1.1-2.1 0.1574 0.1081 C Add Male hCV25653599
WDR12 Sample Set 2 0.0449 1.33735 1-1.8 0.2716 0.218 C Dom ALL
hCV25653599 WDR12 Sample Set 2 0.00797 2.1608 1.2-3.8 0.3291 0.185
C Dom MI-AgeT2 hCV25653599 WDR12 Sample Set 2 0.00341 2.0357
1.3-3.3 0.3438 0.2047 C Dom MI-Male hCV25653599 WDR12 Sample Set 2
0.0204 1.5756 1.1-2.3 0.3052 0.218 C Dom MI- hCV25653599 WDR12
Sample Set 2 0.00334 1.6595 1.2-2.3 0.1675 0.1081 C Add MI-Male
hCV25653599 WDR12 Sample Set 2 0.0106 1.4203 1.1-1.9 0.1571 0.116 C
Add MI- hCV25653599 WDR12 Sample Set 2 0.0257 1.7171 1.1-2.8 0.2805
0.185 C Dom MI-AgeT2 hCV25653599 WDR12 Sample Set 2 0.0289 1.4654
1-2.1 0.1559 0.112 C Add MI-Smoke+ hCV25963638 SRPX Sample Set 2
0.0333 1.51 1-2.2 0.079 0.0538 A Allelic Smoke+ hCV25963638 SRPX
Sample Set 2 0.0246 1.9735 1.1-3.5 0.1392 0.0758 A Allelic MI-AgeT2
hCV25963638 SRPX Sample Set 2 0.00997 2.0235 1.2-3.4 0.1237 0.0652
A Allelic MI-Male hCV25963638 SRPX Sample Set 2 0.0465 1.8005 1-3.2
0.0928 0.0538 A Allelic MI-Smoke+ hCV25963638 SRPX Sample Set 2
0.048 1.9943 1-4 0.0769 0.0401 A Rec MI- hCV25963638 SRPX Sample
Set 2 0.00373 1.8731 1.2-2.8 0.1156 0.0652 A Allelic MI-Male
hCV25963638 SRPX Sample Set 2 0.00569 1.893 1.2-3 0.0971 0.0538 A
Allelic MI-Smoke+ hCV25963638 SRPX Sample Set 2 0.0174 1.9469
1.1-3.4 0.0752 0.0401 A Rec MI- hCV25963638 SRPX Sample Set 2
0.0204 1.7792 1.1-2.9 0.1273 0.0758 A Allelic MI-AgeT2 hCV2782570
PTGIS Sample Set 2 0.0202 0.7282 0.6-0.9 0.3184 0.3908 G Allelic
Smoke- hCV2782570 PTGIS Sample Set 2 0.0439 0.3075 0.1-1 0.0526
0.153 G Rec MI-Female hCV2782570 PTGIS Sample Set 2 0.0345 1.4814
1-2.1 0.697 0.6082 G Dom MI-Smoke+ hCV2782570 PTGIS Sample Set 2
0.0428 0.4656 0.2-1 0.0776 0.153 G Rec MI-Female hCV2908485 none
Sample Set 2 0.00678 0.6959 0.5-0.9 0.3621 0.4492 G Add Age T3
hCV2908485 none Sample Set 2 0.00146 0.63584 0.5-0.8 0.5955 0.6984
G Dom Male hCV2908485 none Sample Set 2 0.00761 0.7751 0.6-0.9
0.3935 0.4557 G Allelic Smoke+ hCV2908485 none Sample Set 2 0.015
0.76396 0.6-0.9 0.6211 0.6821 G Dom ALL hCV2908485 none Sample Set
2 0.00553 0.6407 0.5-0.9 0.3432 0.4492 G Add Age T3 hCV2908485 none
Sample Set 2 0.00236 0.59936 0.4-0.8 0.5812 0.6984 G Dom Male
hCV2908485 none Sample Set 2 0.0142 0.66644 0.5-0.9 0.6187 0.7089 G
Dom Smoke+ hCV2908485 none Sample Set 2 0.0181 0.73421 0.6-0.9
0.6117 0.6821 G Dom ALL hCV2908485 none Sample Set 2 0.0473 1.9907
1-4 0.2414 0.1378 G Rec MI-Female hCV2908485 none Sample Set 2
0.0102 0.5553 0.4-0.9 0.5625 0.6984 G Dom MI-Male hCV2908485 none
Sample Set 2 0.00378 2.5953 1.3-5 0.2712 0.1254 G Rec MI-Smoke-
hCV2908485 none Sample Set 2 0.00978 0.6506 0.5-0.9 0.3526 0.4557 G
Add MI-Smoke+ hCV2908485 none Sample Set 2 0.0432 0.6917 0.5-1
0.5974 0.6821 G Dom MI- hCV2908485 none Sample Set 2 0.00684 0.6135
0.4-0.9 0.599 0.7089 G Dom MI-Smoke+ hCV2908485 none Sample Set 2
0.007 0.6185 0.4-0.9 0.5888 0.6984 G Dom MI-Male hCV2908485 none
Sample Set 2 0.00798 0.69 0.5-0.9 0.5968 0.6821 G Dom MI-
hCV2908485 none Sample Set 2 0.0178 0.5526 0.3-0.9 0.5761 0.7109 G
Dom MI-AgeT3 hCV2908485 none Sample Set 2 0.0431 1.7825 1-3.1
0.2035 0.1254 G Rec MI-Smoke- hCV783138 F7 Sample Set 2 0.0245 0 --
0 0.0177 A Rec Female hCV11592758 BDNF Sample Set 2 0.003689
1.418521 0.85736073 0.8090617 C Allelic ALL hCV11592758 BDNF Sample
Set 2 0.015578 1.442573 0.85240696 0.8001411 C Allelic Male
hCV11592758 BDNF Sample Set 2 0.048839 1.410359 0.85028022
0.8010637 C Allelic Younger hCV2531086 CD22 Sample Set 2 0.016484
1.45531 0.80197937 0.7356524 G Allelic Younger hCV25474320 LCP1
Sample Set 2 0.003797 0.421814 0.04477473 0.1000101 T Allelic
Female hCV25474320 LCP1 Sample Set 2 0.040595 0.609457 0.05770926
0.0913129 T Allelic Older hCV25608809 TLR5 Sample Set 2 0.033066
1.352714 0.58624226 0.5115828 A Allelic Female hCV25608809 TLR5
Sample Set 2 0.001546 1.482234 0.59272859 0.4954267 A Allelic Older
hCV25608809 TLR5 Sample Set 2 0.037258 1.249806 0.56389988
0.5085033 A Allelic Smoke+ hCV2676030 EDG2 Sample Set 2 0.047657
1.202427 0.35389562 0.3129634 G Allelic ALL hCV2676030 EDG2 Sample
Set 2 0.047791 1.250291 0.359831 0.310138 G Allelic Smoke+
hCV7490119 NPC1 Sample Set 2 8.66E-05 0.684841 0.27083028 0.3516384
C Allelic ALL hCV7490119 NPC1 Sample Set 2 0.020383 0.703492
0.28775573 0.3647954 C Allelic Female hCV7490119 NPC1 Sample Set 2
0.001487 0.672101 0.25987035 0.3431481 C
Allelic Male hCV7490119 NPC1 Sample Set 2 0.000415 0.502755
0.23248743 0.3759751 C Allelic Smoke- hCV7490119 NPC1 Sample Set 2
0.005119 0.690612 0.28847403 0.3699039 C Allelic Older hCV7490119
NPC1 Sample Set 2 0.002153 0.640858 0.24675175 0.3382584 C Allelic
Younger hCV7490119 NPC1 Sample Set 2 0.039905 0.787741 0.28126535
0.3318993 C Allelic Smoke+ hCV8827241 SPARCL1 Sample Set 2 0.011283
0.716295 0.57830575 0.6568944 C Allelic Younger hCV8932279 SERPINB5
Sample Set 2 0.026358 1.465232 0.48857258 0.3946682 G Allelic
Smoke- hCV9482394 KIAA1608 Sample Set 2 0.04301 1.74935 0.12036092
0.0725434 A Allelic Smoke- hCV9482394 KIAA1608 Sample Set 2 0.02777
0.641196 0.06323883 0.0952556 A Allelic Smoke+ hCV9578831 TNFRSF6
Sample Set 2 0.034212 0.615467 0.07409659 0.115064 T Allelic
Younger hCV1309246 PPP1R12A Sample Set 2 0.03147 0.339874
0.01228995 0.0353173 C Allelic Male hCV1309246 PPP1R12A Sample Set
2 0.032389 0.187827 0.00861413 0.0442151 C Allelic Smoke-
hCV1403468 IL12A Sample Set 2 0.032773 0.663131 0.13769986
0.1940753 G Allelic Male hCV1487384 CD33 Sample Set 2 0.008942
1.597195 0.83948062 0.7660464 G Allelic Male hCV16173091 FABP1
Sample Set 2 0.04212 0.66867 0.27076982 0.3570351 C Allelic Female
hCV16173091 FABP1 Sample Set 2 0.040581 0.738459 0.29386913
0.3604351 C Allelic Smoke+ hCV25594815 LRP3 Sample Set 2 0.003239
1.690589 0.14376135 0.0903416 A Allelic ALL hCV25594815 LRP3 Sample
Set 2 0.002488 1.960164 0.16132547 0.0893639 A Allelic Male
hCV25594815 LRP3 Sample Set 2 0.019092 1.672316 0.14861088 0.094512
A Allelic Smoke+ hCV25944011 EGLN2 Sample Set 2 0.047039 1.264556
0.37561049 0.322361 A Allelic ALL hCV25944011 EGLN2 Sample Set 2
0.036898 1.366324 0.39094994 0.319636 A Allelic Male hCV313778
GOLGA5 Sample Set 2 0.022064 0.751072 0.2535017 0.3113598 A Allelic
ALL hCV313778 GOLGA5 Sample Set 2 0.002363 0.52557 0.19889103
0.3208284 A Allelic Female hCV313778 GOLGA5 Sample Set 2 0.019467
0.696256 0.24191677 0.314285 A Allelic Smoke+ Note: * Odds Ratio
(OR) not applicable because sum score is a continuous end point
[0405]
Sequence CWU 1
1
56 1 5313 DNA Homo sapien 1 gaattcttag ttgttttctt tagaagaaca
tttctaggga ataatacaag aagatttagg 60 aatcattgaa gttataaatc
tttggaatga gcaaactcag aatggtgcta cttgaagact 120 ctggatctgc
tgacttcaga agacattttg tcaacctgag tcccttcacc attactgtgg 180
tcttacttct cagtgcctgt tttgtcacca gttctcttgg aggaacagac aaggagctga
240 ggctagtgga tggtgaaaac aagtgtagcg ggagagtgga agtgaaagtc
caggaggagt 300 ggggaacggt gtgtaataat ggctggagca tggaagcggt
ctctgtgatt tgtaaccagc 360 tgggatgtcc aactgctatc aaagcccctg
gatgggctaa ttccagtgca ggttctggac 420 gcatttggat ggatcatgtt
tcttgtcgtg ggaatgagtc agctctttgg gattgcaaac 480 atgatggatg
gggaaagcat agtaactgta ctcaccaaca agatgctgga gtgacctgct 540
cagatggatc caatttggaa atgaggctga cgcgtggagg gaatatgtgt tctggaagaa
600 tagagatcaa attccaagga cggtggggaa cagtgtgtga tgataacttc
aacatagatc 660 atgcatctgt catttgtaga caacttgaat gtggaagtgc
tgtcagtttc tctggttcat 720 ctaattttgg agaaggctct ggaccaatct
ggtttgatga tcttatatgc aacggaaatg 780 agtcagctct ctggaactgc
aaacatcaag gatggggaaa gcataactgt gatcatgctg 840 aggatgctgg
agtgatttgc tcaaagggag cagatctgag cctgagactg gtagatggag 900
tcactgaatg ttcaggaaga ttagaagtga gattccaagg ggaatggggg acaatatgtg
960 atgacggctg ggacagttac gatgctgctg tggcatgcaa gcaactggga
tgtccaactg 1020 ccgtcacagc cattggtcga gttaacgcca gtaagggatt
tggacacatc tggcttgaca 1080 gcgtttcttg ccagggacat gaacctgctg
tctggcaatg taaacaccat gaatggggaa 1140 agcattattg caatcacaat
gaagatgctg gcgtgacatg ttctgatgga tcagatctgg 1200 agctaagact
tagaggtgga ggcagccgct gtgctgggac agttgaggtg gagattcaga 1260
gactgttagg gaaggtgtgt gacagaggct ggggactgaa agaagctgat gtggtttgca
1320 ggcagctggg atgtggatct gcactcaaaa catcttatca agtgtactcc
aaaatccagg 1380 caacaaacac atggctgttt ctaagtagct gtaacggaaa
tgaaacttct ctttgggact 1440 gcaagaactg gcaatggggt ggacttacct
gtgatcacta tgaagaagcc aaaattacct 1500 gctcagccca cagggaaccc
agactggttg gaggggacat tccctgttct ggacgtgttg 1560 aagtgaagca
tggtgacacg tggggctcca tctgtgattc ggacttctct ctggaagctg 1620
ccagcgttct atgcagggaa ttacagtgtg gcacagttgt ctctatcctg gggggagctc
1680 actttggaga gggaaatgga cagatctggg ctgaagaatt ccagtgtgag
ggacatgagt 1740 cccatctttc actctgccca gtagcacccc gcccagaagg
aacttgtagc cacagcaggg 1800 atgttggagt agtctgctca agatacacag
aaattcgctt ggtgaatggc aagaccccgt 1860 gtgagggcag agtggagctc
aaaacgcttg gtgcctgggg atccctctgt aactctcact 1920 gggacataga
agatgcccat gttctttgcc agcagcttaa atgtggagtt gccctttcta 1980
ccccaggagg agcacgtttt ggaaaaggaa atggtcagat ctggaggcat atgtttcact
2040 gcactgggac tgagcagcac atgggagatt gtcctgtaac tgctctaggt
gcttcattat 2100 gtccttcaga gcaagtggcc tctgtaatct gctcaggaaa
ccagtcccaa acactgtcct 2160 cgtgcaattc atcgtctttg ggcccaacaa
ggcctaccat tccagaagaa agtgctgtgg 2220 cctgcataga gagtggtcaa
cttcgcctgg taaatggagg aggtcgctgt gctgggagag 2280 tagagatcta
tcatgagggc tcctggggca ccatctgtga tgacagctgg gacctgagtg 2340
atgcccacgt ggtttgcaga cagctgggct gtggagaggc cattaatgcc actggttctg
2400 ctcattttgg ggaaggaaca gggcccatct ggctggatga gatgaaatgc
aatggaaaag 2460 aatcccgcat ttggcagtgc cattcacacg gctgggggca
gcaaaattgc aggcacaagg 2520 aggatgcggg agttatctgc tcagaattca
tgtctctgag actgaccagt gaagccagca 2580 gagaggcctg tgcagggcgt
ctggaagttt tttacaatgg agcttggggc actgttggca 2640 agagtagcat
gtctgaaacc actgtgggtg tggtgtgcag gcagctgggc tgtgcagaca 2700
aagggaaaat caaccctgca tctttagaca aggccatgtc cattcccatg tgggtggaca
2760 atgttcagtg tccaaaagga cctgacacgc tgtggcagtg cccatcatct
ccatgggaga 2820 agagactggc cagcccctcg gaggagacct ggatcacatg
tgacaacaag ataagacttc 2880 aggaaggacc cacttcctgt tctggacgtg
tggagatctg gcatggaggt tcctggggga 2940 cagtgtgtga tgactcttgg
gacttggacg atgctcaggt ggtgtgtcaa caacttggct 3000 gtggtccagc
tttgaaagca ttcaaagaag cagagtttgg tcaggggact ggacccgata 3060
tggctcaatg aagtgaagtg caaagggaat gagtcttcct tgtgggattg tcctgccaga
3120 cgctggggcc atagtgagtg tgggcacaag gaagacgctg cagtgaattg
cacagatatt 3180 tcagtgcaga aaaccccaca aaaagccaca acaggtcgct
catcccgtca gtcatccttt 3240 attgcagtcg ggatccttgg ggttgttctg
ttggccattt tcgtcgcatt attcttcttg 3300 actaaaaagc gaagacagag
acagcggctt gcagtttcct caagaggaga gaacttagtc 3360 caccaaattc
aataccggga gatgaattct tgcctgaatg cagatgatct ggacctaatg 3420
aattcctcag gtctgtgggt tcttggaggg tctattgccc aggggttcag atcagtggct
3480 gcagttgagg cacagacatt ctactttgat aaacagttaa aaaagtctaa
aaatgtaata 3540 ggaagcttag atgcatataa tggacaagaa tgactgaaaa
ttattcttgg agaatatcaa 3600 aattgcaatc atagggaggc ctttagctta
agaggcctgt gattattcct gatagaggta 3660 tggaaagaac catgcagagg
aatattatga cttggacctc attttattaa aacagaaatt 3720 aatcttacaa
aagattgtca taagtgacag tttaactttt ttctttaaat tttgttgtgt 3780
atatttaagg tatacaacat gattttatgg gatgtatata gatagtaaaa agcttactaa
3840 agcaaagcaa atgaacacac ccatcatctg acatagttac ccttttttgt
gttgttcttg 3900 tggcaagagc agctaaaacc tactcactta gcatgaatcc
tacatacagc acaatgttat 3960 tacctataat cctcatgttg tacattagac
ctctagactg gttcattcta cgtatctgct 4020 actttgtatc ctctgaccta
catacgtctt tcacagtttc ttccattccc atttcctgtc 4080 attttttttc
tctagcttga tatttattat atttttccct aaaagtctaa aaccttaaac 4140
tttcaatatc tttattgcat gagaagccat acaaatccac agaactagcc ttatttctca
4200 tcacatcatg ctgttttatc cttgaacttc tatttagcac cagtgcacta
attctgcatc 4260 tgggcaggat gactttactg ggttggaaga aatatcccaa
aacccattgt ctttactcca 4320 tgaagggtcc ctgaccttct gagaggggcc
tgcctcactt cttccatcca aagaattatg 4380 catctgctac tgtgtcaggg
aacatattta aggaacatgt actgttactg tgtcaggaaa 4440 catatttaag
aaataggaaa gactttctct gccccttaaa tcacacatgc ttttcttcct 4500
agttatgggt ggtgttttta gttgctcaaa gagcctcaca gttacgtgag aagaggtctg
4560 gtttatttcc cagtaattat tttcttcctt tcagaaaatt cccatgagtc
agctgatttc 4620 agtgctgctg aactaatttc tgtgtctaaa tttcttccta
tttctggaat ggaaaaggag 4680 gccattctga gccacactga aaaggaaaat
gggaatttat aacccagtga gttcagcctt 4740 taagatacct tgatgaagac
ctggactatt gaatggagca gaaattcacc tctctcactg 4800 actattacag
ttgcattttt atggagttct tcttctccta ggattcctaa gactgctgct 4860
gaatttataa aaattaagtt tgtgaatgtg actacttagt ggtgtatatg agactttcaa
4920 gggaattaaa taaataaata agaatgttat tgatttgagt ttgctttaat
tacttgtcct 4980 taattctatt aatttctaaa cgggcttcct aattttttgt
agagtttcct agatgtatta 5040 taatgtgttt tatttgacag tgtttcaatt
tgcatataca gtactgtata ttttttctta 5100 tttggtttga ataattttcc
tattaccaaa taaaaataaa tttattttta ctttagtttt 5160 tctaagacag
gaaaagttaa tgatattgaa gggtctgtaa ataatatatg gctaacttta 5220
taaggcatga ctcacaacga ttctttaact gctttttgtt actgtaattc tgttcactag
5280 aataaaatgc agagccacac ctggtgaggg cac 5313 2 4066 DNA Homo
sapien 2 gaattcttag ttgttttctt tagaagaaca tttctaggga ataatacaag
aagatttagg 60 aatcattgaa gttataaatc tttggaatga gcaaactcag
aatggtgcta cttgaagact 120 ctggatctgc tgacttcaga agacattttg
tcaacctgag tcccttcacc attactgtgg 180 tcttacttct cagtgcctgt
tttgtcacca gttctcttgg aggaacagac aaggagctga 240 ggctagtgga
tggtgaaaac aagtgtagcg ggagagtgga agtgaaagtc caggaggagt 300
ggggaacggt gtgtaataat ggctggagca tggaagcggt ctctgtgatt tgtaaccagc
360 tgggatgtcc aactgctatc aaagcccctg gatgggctaa ttccagtgca
ggttctggac 420 gcatttggat ggatcatgtt tcttgtcgtg ggaatgagtc
agctctttgg gattgcaaac 480 atgatggatg gggaaagcat agtaactgta
ctcaccaaca agatgctgga gtgacctgct 540 cagatggatc caatttggaa
atgaggctga cgcgtggagg gaatatgtgt tctggaagaa 600 tagagatcaa
attccaagga cggtggggaa cagtgtgtga tgataacttc aacatagatc 660
atgcatctgt catttgtaga caacttgaat gtggaagtgc tgtcagtttc tctggttcat
720 ctaattttgg agaaggctct ggaccaatct ggtttgatga tcttatatgc
aacggaaatg 780 agtcagctct ctggaactgc aaacatcaag gatggggaaa
gcataactgt gatcatgctg 840 aggatgctgg agtgatttgc tcaaagggag
cagatctgag cctgagactg gtagatggag 900 tcactgaatg ttcaggaaga
ttagaagtga gattccaagg ggaatggggg acaatatgtg 960 atgacggctg
ggacagttac gatgctgctg tggcatgcaa gcaactggga tgtccaactg 1020
ccgtcacagc cattggtcga gttaacgcca gtaagggatt tggacacatc tggcttgaca
1080 gcgtttcttg ccagggacat gaacctgctg tctggcaatg taaacaccat
gaatggggaa 1140 agcattattg caatcacaat gaagatgctg gcgtgacatg
ttctgatgga tcagatctgg 1200 agctaagact tagaggtgga ggcagccgct
gtgctgggac agttgaggtg gagattcaga 1260 gactgttagg gaaggtgtgt
gacagaggct ggggactgaa agaagctgat gtggtttgca 1320 ggcagctggg
atgtggatct gcactcaaaa catcttatca agtgtactcc aaaatccagg 1380
caacaaacac atggctgttt ctaagtagct gtaacggaaa tgaaacttct ctttgggact
1440 gcaagaactg gcaatggggt ggacttacct gtgatcacta tgaagaagcc
aaaattacct 1500 gctcagccca cagggaaccc agactggttg gaggggacat
tccctgttct ggacgtgttg 1560 aagtgaagca tggtgacacg tggggctcca
tctgtgattc ggacttctct ctggaagctg 1620 ccagcgttct atgcagggaa
ttacagtgtg gcacagttgt ctctatcctg gggggagctc 1680 actttggaga
gggaaatgga cagatctggg ctgaagaatt ccagtgtgag ggacatgagt 1740
cccatctttc actctgccca gtagcacccc gcccagaagg aacttgtagc cacagcaggg
1800 atgttggagt agtctgctca agatacacag aaattcgctt ggtgaatggc
aagaccccgt 1860 gtgagggcag agtggagctc aaaacgcttg gtgcctgggg
atccctctgt aactctcact 1920 gggacataga agatgcccat gttctttgcc
agcagcttaa atgtggagtt gccctttcta 1980 ccccaggagg agcacgtttt
ggaaaaggaa atggtcagat ctggaggcat atgtttcact 2040 gcactgggac
tgagcagcac atgggagatt gtcctgtaac tgctctaggt gcttcattat 2100
gtccttcaga gcaagtggcc tctgtaatct gctcaggaaa ccagtcccaa acactgtcct
2160 cgtgcaattc atcgtctttg ggcccaacaa ggcctaccat tccagaagaa
agtgctgtgg 2220 cctgcataga gagtggtcaa cttcgcctgg taaatggagg
aggtcgctgt gctgggagag 2280 tagagatcta tcatgagggc tcctggggca
ccatctgtga tgacagctgg gacctgagtg 2340 atgcccacgt ggtttgcaga
cagctgggct gtggagaggc cattaatgcc actggttctg 2400 ctcattttgg
ggaaggaaca gggcccatct ggctggatga gatgaaatgc aatggaaaag 2460
aatcccgcat ttggcagtgc cattcacacg gctgggggca gcaaaattgc aggcacaagg
2520 aggatgcggg agttatctgc tcagaattca tgtctctgag actgaccagt
gaagccagca 2580 gagaggcctg tgcagggcgt ctggaagttt tttacaatgg
agcttggggc actgttggca 2640 agagtagcat gtctgaaacc actgtgggtg
tggtgtgcag gcagctgggc tgtgcagaca 2700 aagggaaaat caaccctgca
tctttagaca aggccatgtc cattcccatg tgggtggaca 2760 atgttcagtg
tccaaaagga cctgacacgc tgtggcagtg cccatcatct ccatgggaga 2820
agagactggc cagcccctcg gaggagacct ggatcacatg tgacaacaag ataagacttc
2880 aggaaggacc cacttcctgt tctggacgtg tggagatctg gcatggaggt
tcctggggga 2940 cagtgtgtga tgactcttgg gacttggacg atgctcaggt
ggtgtgtcaa caacttggct 3000 gtggtccagc tttgaaagca ttcaaagaag
cagagtttgg tcaggggact ggacccgata 3060 tggctcaatg aagtgaagtg
caaagggaat gagtcttcct tgtgggattg tcctgccaga 3120 cgctggggcc
atagtgagtg tgggcacaag gaagacgctg cagtgaattg cacagatatt 3180
tcagtgcaga aaaccccaca aaaagccaca acaggtcgct catcccgtca gtcatccttt
3240 attgcagtcg ggatccttgg ggttgttctg ttggccattt tcgtcgcatt
attcttcttg 3300 actaaaaagc gaagacagag acagcggctt gcagtttcct
caagaggaga gaacttagtc 3360 caccaaattc aataccggga gatgaattct
tgcctgaatg cagatgatct ggacctaatg 3420 aattcctcag gaggccattc
tgagccacac tgaaaaggaa aatgggaatt tataacccag 3480 tgagttcagc
ctttaagata ccttgatgaa gacctggact attgaatgga gcagaaattc 3540
acctctctca ctgactatta cagttgcatt tttatggagt tcttcttctc ctaggattcc
3600 taagactgct gctgaattta taaaaattaa gtttgtgaat gtgactactt
agtggtgtat 3660 atgagacttt caagggaatt aaataaataa ataagaatgt
tattgatttg agtttgcttt 3720 aattacttgt ccttaattct attaatttct
aaacgggctt cctaattttt tgtagagttt 3780 cctagatgta ttataatgtg
ttttatttga cagtgtttca atttgcatat acagtactgt 3840 atattttttc
ttatttggtt tgaataattt tcctattacc aaataaaaat aaatttattt 3900
ttactttagt ttttctaaga caggaaaagt taatgatatt gaagggtctg taaataatat
3960 atggctaact ttataaggca tgactcacaa cgattcttta actgcttttt
gttactgtaa 4020 ttctgttcac tagaataaaa tgcagagcca cacctggtga gggcac
4066 3 4248 DNA Homo sapien 3 gaattcttag ttgttttctt tagaagaaca
tttctaggga ataatacaag aagatttagg 60 aatcattgaa gttataaatc
tttggaatga gcaaactcag aatggtgcta cttgaagact 120 ctggatctgc
tgacttcaga agacattttg tcaacctgag tcccttcacc attactgtgg 180
tcttacttct cagtgcctgt tttgtcacca gttctcttgg aggaacagac aaggagctga
240 ggctagtgga tggtgaaaac aagtgtagcg ggagagtgga agtgaaagtc
caggaggagt 300 ggggaacggt gtgtaataat ggctggagca tggaagcggt
ctctgtgatt tgtaaccagc 360 tgggatgtcc aactgctatc aaagcccctg
gatgggctaa ttccagtgca ggttctggac 420 gcatttggat ggatcatgtt
tcttgtcgtg ggaatgagtc agctctttgg gattgcaaac 480 atgatggatg
gggaaagcat agtaactgta ctcaccaaca agatgctgga gtgacctgct 540
cagatggatc caatttggaa atgaggctga cgcgtggagg gaatatgtgt tctggaagaa
600 tagagatcaa attccaagga cggtggggaa cagtgtgtga tgataacttc
aacatagatc 660 atgcatctgt catttgtaga caacttgaat gtggaagtgc
tgtcagtttc tctggttcat 720 ctaattttgg agaaggctct ggaccaatct
ggtttgatga tcttatatgc aacggaaatg 780 agtcagctct ctggaactgc
aaacatcaag gatggggaaa gcataactgt gatcatgctg 840 aggatgctgg
agtgatttgc tcaaagggag cagatctgag cctgagactg gtagatggag 900
tcactgaatg ttcaggaaga ttagaagtga gattccaagg ggaatggggg acaatatgtg
960 atgacggctg ggacagttac gatgctgctg tggcatgcaa gcaactggga
tgtccaactg 1020 ccgtcacagc cattggtcga gttaacgcca gtaagggatt
tggacacatc tggcttgaca 1080 gcgtttcttg ccagggacat gaacctgctg
tctggcaatg taaacaccat gaatggggaa 1140 agcattattg caatcacaat
gaagatgctg gcgtgacatg ttctgatgga tcagatctgg 1200 agctaagact
tagaggtgga ggcagccgct gtgctgggac agttgaggtg gagattcaga 1260
gactgttagg gaaggtgtgt gacagaggct ggggactgaa agaagctgat gtggtttgca
1320 ggcagctggg atgtggatct gcactcaaaa catcttatca agtgtactcc
aaaatccagg 1380 caacaaacac atggctgttt ctaagtagct gtaacggaaa
tgaaacttct ctttgggact 1440 gcaagaactg gcaatggggt ggacttacct
gtgatcacta tgaagaagcc aaaattacct 1500 gctcagccca cagggaaccc
agactggttg gaggggacat tccctgttct ggacgtgttg 1560 aagtgaagca
tggtgacacg tggggctcca tctgtgattc ggacttctct ctggaagctg 1620
ccagcgttct atgcagggaa ttacagtgtg gcacagttgt ctctatcctg gggggagctc
1680 actttggaga gggaaatgga cagatctggg ctgaagaatt ccagtgtgag
ggacatgagt 1740 cccatctttc actctgccca gtagcacccc gcccagaagg
aacttgtagc cacagcaggg 1800 atgttggagt agtctgctca agtaagaccc
agaaaacatc tttaattggt tctcatactg 1860 tgaaagggac agggttaggg
agtcatagct gtctttttct aaagccctgt ctccttccag 1920 gatacacaga
aattcgcttg gtgaatggca agaccccgtg tgagggcaga gtggagctca 1980
aaacgcttgg tgcctgggga tccctctgta actctcactg ggacatagaa gatgcccatg
2040 ttctttgcca gcagcttaaa tgtggagttg ccctttctac cccaggagga
gcacgttttg 2100 gaaaaggaaa tggtcagatc tggaggcata tgtttcactg
cactgggact gagcagcaca 2160 tgggagattg tcctgtaact gctctaggtg
cttcattatg tccttcagag caagtggcct 2220 ctgtaatctg ctcaggaaac
cagtcccaaa cactgtcctc gtgcaattca tcgtctttgg 2280 gcccaacaag
gcctaccatt ccagaagaaa gtgctgtggc ctgcatagag agtggtcaac 2340
ttcgcctggt aaatggagga ggtcgctgtg ctgggagagt agagatctat catgagggct
2400 cctggggcac catctgtgat gacagctggg acctgagtga tgcccacgtg
gtttgcagac 2460 agctgggctg tggagaggcc attaatgcca ctggttctgc
tcattttggg gaaggaacag 2520 ggcccatctg gctggatgag atgaaatgca
atggaaaaga atcccgcatt tggcagtgcc 2580 attcacacgg ctgggggcag
caaaattgca ggcacaagga ggatgcggga gttatctgct 2640 cagaattcat
gtctctgaga ctgaccagtg aagccagcag agaggcctgt gcagggcgtc 2700
tggaagtttt ttacaatgga gcttggggca ctgttggcaa gagtagcatg tctgaaacca
2760 ctgtgggtgt ggtgtgcagg cagctgggct gtgcagacaa agggaaaatc
aaccctgcat 2820 ctttagacaa ggccatgtcc attcccatgt gggtggacaa
tgttcagtgt ccaaaaggac 2880 ctgacacgct gtggcagtgc ccatcatctc
catgggagaa gagactggcc agcccctcgg 2940 aggagacctg gatcacatgt
gacaacaaga taagacttca ggaaggaccc acttcctgtt 3000 ctggacgtgt
ggagatctgg catggaggtt cctgggggac agtgtgtgat gactcttggg 3060
acttggacga tgctcaggtg gtgtgtcaac aacttggctg tggtccagct ttgaaagcat
3120 tcaaagaagc agagtttggt caggggactg gacccgatat ggctcaatga
agtgaagtgc 3180 aaagggaatg agtcttcctt gtgggattgt cctgccagac
gctggggcca tagtgagtgt 3240 gggcacaagg aagacgctgc agtgaattgc
acagatattt cagtgcagaa aaccccacaa 3300 aaagccacaa caggtcgctc
atcccgtcag tcatccttta ttgcagtcgg gatccttggg 3360 gttgttctgt
tggccatttt cgtcgcatta ttcttcttga ctaaaaagcg aagacagaga 3420
cagcggcttg cagtttcctc aagaggagag aacttagtcc accaaattca ataccgggag
3480 atgaattctt gcctgaatgc agatgatctg gacctaatga attcctcaga
aaattcccat 3540 gagtcagctg atttcagtgc tgctgaacta atttctgtgt
ctaaatttct tcctatttct 3600 ggaatggaaa aggaggccat tctgagccac
actgaaaagg aaaatgggaa tttataaccc 3660 agtgagttca gcctttaaga
taccttgatg aagacctgga ctattgaatg gagcagaaat 3720 tcacctctct
cactgactat tacagttgca tttttatgga gttcttcttc tcctaggatt 3780
cctaagactg ctgctgaatt tataaaaatt aagtttgtga atgtgactac ttagtggtgt
3840 atatgagact ttcaagggaa ttaaataaat aaataagaat gttattgatt
tgagtttgct 3900 ttaattactt gtccttaatt ctattaattt ctaaacgggc
ttcctaattt tttgtagagt 3960 ttcctagatg tattataatg tgttttattt
gacagtgttt caatttgcat atacagtact 4020 gtatattttt tcttatttgg
tttgaataat tttcctatta ccaaataaaa ataaatttat 4080 ttttacttta
gtttttctaa gacaggaaaa gttaatgata ttgaagggtc tgtaaataat 4140
atatggctaa ctttataagg catgactcac aacgattctt taactgcttt ttgttactgt
4200 aattctgttc actagaataa aatgcagagc cacacctggt gagggcac 4248 4
4149 DNA Homo sapien 4 gaattcttag ttgttttctt tagaagaaca tttctaggga
ataatacaag aagatttagg 60 aatcattgaa gttataaatc tttggaatga
gcaaactcag aatggtgcta cttgaagact 120 ctggatctgc tgacttcaga
agacattttg tcaacctgag tcccttcacc attactgtgg 180 tcttacttct
cagtgcctgt tttgtcacca gttctcttgg aggaacagac aaggagctga 240
ggctagtgga tggtgaaaac aagtgtagcg ggagagtgga agtgaaagtc caggaggagt
300 ggggaacggt gtgtaataat ggctggagca tggaagcggt ctctgtgatt
tgtaaccagc 360 tgggatgtcc aactgctatc aaagcccctg gatgggctaa
ttccagtgca ggttctggac 420 gcatttggat ggatcatgtt tcttgtcgtg
ggaatgagtc agctctttgg gattgcaaac 480 atgatggatg gggaaagcat
agtaactgta ctcaccaaca agatgctgga gtgacctgct 540 cagatggatc
caatttggaa atgaggctga cgcgtggagg gaatatgtgt tctggaagaa 600
tagagatcaa attccaagga cggtggggaa cagtgtgtga tgataacttc aacatagatc
660 atgcatctgt catttgtaga caacttgaat gtggaagtgc tgtcagtttc
tctggttcat 720 ctaattttgg agaaggctct ggaccaatct ggtttgatga
tcttatatgc aacggaaatg 780 agtcagctct ctggaactgc aaacatcaag
gatggggaaa gcataactgt gatcatgctg 840 aggatgctgg agtgatttgc
tcaaagggag cagatctgag cctgagactg gtagatggag 900 tcactgaatg
ttcaggaaga ttagaagtga gattccaagg ggaatggggg acaatatgtg 960
atgacggctg ggacagttac gatgctgctg tggcatgcaa gcaactggga tgtccaactg
1020 ccgtcacagc cattggtcga gttaacgcca gtaagggatt tggacacatc
tggcttgaca 1080 gcgtttcttg ccagggacat gaacctgctg tctggcaatg
taaacaccat gaatggggaa 1140 agcattattg caatcacaat gaagatgctg
gcgtgacatg ttctgatgga tcagatctgg 1200 agctaagact tagaggtgga
ggcagccgct gtgctgggac agttgaggtg gagattcaga 1260 gactgttagg
gaaggtgtgt
gacagaggct ggggactgaa agaagctgat gtggtttgca 1320 ggcagctggg
atgtggatct gcactcaaaa catcttatca agtgtactcc aaaatccagg 1380
caacaaacac atggctgttt ctaagtagct gtaacggaaa tgaaacttct ctttgggact
1440 gcaagaactg gcaatggggt ggacttacct gtgatcacta tgaagaagcc
aaaattacct 1500 gctcagccca cagggaaccc agactggttg gaggggacat
tccctgttct ggacgtgttg 1560 aagtgaagca tggtgacacg tggggctcca
tctgtgattc ggacttctct ctggaagctg 1620 ccagcgttct atgcagggaa
ttacagtgtg gcacagttgt ctctatcctg gggggagctc 1680 actttggaga
gggaaatgga cagatctggg ctgaagaatt ccagtgtgag ggacatgagt 1740
cccatctttc actctgccca gtagcacccc gcccagaagg aacttgtagc cacagcaggg
1800 atgttggagt agtctgctca agatacacag aaattcgctt ggtgaatggc
aagaccccgt 1860 gtgagggcag agtggagctc aaaacgcttg gtgcctgggg
atccctctgt aactctcact 1920 gggacataga agatgcccat gttctttgcc
agcagcttaa atgtggagtt gccctttcta 1980 ccccaggagg agcacgtttt
ggaaaaggaa atggtcagat ctggaggcat atgtttcact 2040 gcactgggac
tgagcagcac atgggagatt gtcctgtaac tgctctaggt gcttcattat 2100
gtccttcaga gcaagtggcc tctgtaatct gctcaggaaa ccagtcccaa acactgtcct
2160 cgtgcaattc atcgtctttg ggcccaacaa ggcctaccat tccagaagaa
agtgctgtgg 2220 cctgcataga gagtggtcaa cttcgcctgg taaatggagg
aggtcgctgt gctgggagag 2280 tagagatcta tcatgagggc tcctggggca
ccatctgtga tgacagctgg gacctgagtg 2340 atgcccacgt ggtttgcaga
cagctgggct gtggagaggc cattaatgcc actggttctg 2400 ctcattttgg
ggaaggaaca gggcccatct ggctggatga gatgaaatgc aatggaaaag 2460
aatcccgcat ttggcagtgc cattcacacg gctgggggca gcaaaattgc aggcacaagg
2520 aggatgcggg agttatctgc tcagaattca tgtctctgag actgaccagt
gaagccagca 2580 gagaggcctg tgcagggcgt ctggaagttt tttacaatgg
agcttggggc actgttggca 2640 agagtagcat gtctgaaacc actgtgggtg
tggtgtgcag gcagctgggc tgtgcagaca 2700 aagggaaaat caaccctgca
tctttagaca aggccatgtc cattcccatg tgggtggaca 2760 atgttcagtg
tccaaaagga cctgacacgc tgtggcagtg cccatcatct ccatgggaga 2820
agagactggc cagcccctcg gaggagacct ggatcacatg tgacaacaag ataagacttc
2880 aggaaggacc cacttcctgt tctggacgtg tggagatctg gcatggaggt
tcctggggga 2940 cagtgtgtga tgactcttgg gacttggacg atgctcaggt
ggtgtgtcaa caacttggct 3000 gtggtccagc tttgaaagca ttcaaagaag
cagagtttgg tcaggggact ggacccgata 3060 tggctcaatg aagtgaagtg
caaagggaat gagtcttcct tgtgggattg tcctgccaga 3120 cgctggggcc
atagtgagtg tgggcacaag gaagacgctg cagtgaattg cacagatatt 3180
tcagtgcaga aaaccccaca aaaagccaca acaggtcgct catcccgtca gtcatccttt
3240 attgcagtcg ggatccttgg ggttgttctg ttggccattt tcgtcgcatt
attcttcttg 3300 actaaaaagc gaagacagag acagcggctt gcagtttcct
caagaggaga gaacttagtc 3360 caccaaattc aataccggga gatgaattct
tgcctgaatg cagatgatct ggacctaatg 3420 aattcctcag aaaattccca
tgagtcagct gatttcagtg ctgctgaact aatttctgtg 3480 tctaaatttc
ttcctatttc tggaatggaa aaggaggcca ttctgagcca cactgaaaag 3540
gaaaatggga atttataacc cagtgagttc agcctttaag ataccttgat gaagacctgg
3600 actattgaat ggagcagaaa ttcacctctc tcactgacta ttacagttgc
atttttatgg 3660 agttcttctt ctcctaggat tcctaagact gctgctgaat
ttataaaaat taagtttgtg 3720 aatgtgacta cttagtggtg tatatgagac
tttcaaggga attaaataaa taaataagaa 3780 tgttattgat ttgagtttgc
tttaattact tgtccttaat tctattaatt tctaaacggg 3840 cttcctaatt
ttttgtagag tttcctagat gtattataat gtgttttatt tgacagtgtt 3900
tcaatttgca tatacagtac tgtatatttt ttcttatttg gtttgaataa ttttcctatt
3960 accaaataaa aataaattta tttttacttt agtttttcta agacaggaaa
agttaatgat 4020 attgaagggt ctgtaaataa tatatggcta actttataag
gcatgactca caacgattct 4080 ttaactgctt tttgttactg taattctgtt
cactagaata aaatgcagag ccacacctgg 4140 tgagggcac 4149 5 3618 DNA
Homo sapien 5 gacgcgcgag catcggcggc ccggaaccgg ccttggaaca
actgtggaac ctgaggccgc 60 ttgccctccc gccccatgga gcggcccccg
gggctgcggc cgggcgcggg cgggccctgg 120 gagatgcggg agcggctggg
caccggcggc ttcgggaacg tctgtctgta ccagcatcgg 180 gaacttgatc
tcaaaatagc aattaagtct tgtcgcctag agctaagtac caaaaacaga 240
gaacgatggt gccatgaaat ccagattatg aagaagttga accatgccaa tgttgtaaag
300 gcctgtgatg ttcctgaaga attgaatatt ttgattcatg atgtgcctct
tctagcaatg 360 gaatactgtt ctggaggaga tctccgaaag ctgctcaaca
aaccagaaaa ttgttgtgga 420 cttaaagaaa gccagatact ttctttacta
agtgatatag ggtctgggat tcgatatttg 480 catgaaaaca aaattataca
tcgagatcta aaacctgaaa acatagttct tcaggatgtt 540 ggtggaaaga
taatacataa aataattgat ctgggatatg ccaaagatgt tgatcaagga 600
agtctgtgta catcttttgt gggaacactg cagtatctgg ccccagagct ctttgagaat
660 aagccttaca cagccactgt tgattattgg agctttggga ccatggtatt
tgaatgtatt 720 gctggatata ggcctttttt gcatcatctg cagccattta
cctggcatga gaagattaag 780 aagaaggatc caaagtgtat atttgcatgt
gaagagatgt caggagaagt tcggtttagt 840 agccatttac ctcaaccaaa
tagcctttgt agtttagtag tagaacccat ggaaaactgg 900 ctacagttga
tgttgaattg ggaccctcag cagagaggag gacctgttga ccttactttg 960
aagcagccaa gatgttttgt attaatggat cacattttga atttgaagat agtacacatc
1020 ctaaatatga cttctgcaaa gataatttct tttctgttac cacctgatga
aagtcttcat 1080 tcactacagt ctcgtattga gcgtgaaact ggaataaata
ctggttctca agaacttctt 1140 tcagagacag gaatttctct ggatcctcgg
aaaccagcct ctcaatgtgt tctagatgga 1200 gttagaggct gtgatagcta
tatggtttat ttgtttgata aaagtaaaac tgtatatgaa 1260 gggccatttg
cttccagaag tttatctgat tgtgtaaatt atattgtaca ggacagcaaa 1320
atacagcttc caattataca gctgcgtaaa gtgtgggctg aagcagtgca ctatgtgtct
1380 ggactaaaag aagactatag caggctcttt cagggacaaa gggcagcaat
gttaagtctt 1440 cttagatata atgctaactt aacaaaaatg aagaacactt
tgatctcagc atcacaacaa 1500 ctgaaagcta aattggagtt ttttcacaaa
agcattcagc ttgacttgga gagatacagc 1560 gagcagatga cgtatgggat
atcttcagaa aaaatgctaa aagcatggaa agaaatggaa 1620 gaaaaggcca
tccactatgc tgaggttggt gtcattggat acctggagga tcagattatg 1680
tctttgcatg ctgaaatcat ggagctacag aagagcccct atggaagacg tcagggagac
1740 ttgatggaat ctctggaaca gcgtgccatt gatctatata agcagttaaa
acacagacct 1800 tcagatcact cctacagtga cagcacagag atggtgaaaa
tcattgtgca cactgtgcag 1860 agtcaggacc gtgtgctcaa ggagctgttt
ggtcatttga gcaagttgtt gggctgtaag 1920 cagaagatta ttgatctact
ccctaaggtg gaagtggccc tcagtaatat caaagaagct 1980 gacaatactg
tcatgttcat gcagggaaaa aggcagaaag aaatatggca tctccttaaa 2040
attgcctgta cacagagttc tgcccggtcc cttgtaggat ccagtctaga aggtgcagta
2100 acccctcaga catcagcatg gctgcccccg acttcagcag aacatgatca
ttctctgtca 2160 tgtgtggtaa ctcctcaaga tggggagact tcagcacaaa
tgatagaaga aaatttgaac 2220 tgccttggcc atttaagcac tattattcat
gaggcaaatg aggaacaggg caatagtatg 2280 atgaatcttg attggagttg
gttaacagaa tgagttgtca cttgttcact gtccccaaac 2340 ctatggaagt
tgttgctata catgttggaa atgtgttttt cccccatgaa accattcttc 2400
agacatcagt caatggaaga aatggctatg aacagaaact acatttctac tatgatcaga
2460 agaacatgat tttacaagta taacagtttt gagtaattca agcctctaaa
cagacaggaa 2520 tttagaaaaa gtcaatgtac ttgtttgaat atttgtttta
ataccacagc tatttagaag 2580 catcatcacg acacatttgc cttcagtctt
ggtaaaacat tacttattta actgattaaa 2640 aataccttct atgtattagt
gtcaactttt aacttttggg cgtaagacca aatgtagttt 2700 tgtatacaga
gaagaaaacc tcaagtaata ggcattttaa gtaaaagtct acctgtgttt 2760
ttttctaaaa aggctgctca caagttctat ttcttgaaga ataaattcta cctccttgtg
2820 ttgcactgaa caggttctct tcctggcatc ataaggagtt ggtgtaatca
ttttaaattc 2880 cactgaaaat ttaacagtat ccccttctca tcgaagggat
tgtgtatctg tgcttctaat 2940 attagttggc tttcataaat catgttgttg
tgtgtatatg tatttaagat gtacatttaa 3000 taatatcaaa gagaagatgc
ctgttaattt ataatgtatt tgaaaattac atgttttttc 3060 atttgtaaaa
atgagtcatt tgtttaaaca atctttcatg tcttgtcata caaatttata 3120
aaggtctgca ctcctttatc tgtaattgta attccaaaat ccaaaaagct ctgaaaacaa
3180 ggtttccata agcttggtga caaaattcat ttgcttgcaa tctaatctga
actgaccttg 3240 aatcttttta tcccatttag tgtgaatatt cctttatttt
gctgcttgat gatgagaggg 3300 agggctgctg ccacagactg tggtgagggc
tggttaatgt agtatggtat atgcacaaaa 3360 ctacttttct aaaatctaaa
atttcataat tctgaaacaa cttgccccaa gggtttcaga 3420 gaaaggactg
tggacctcta tcatctgcta agtaatttag aagatattat ttgtcttaaa 3480
aaatgtgaaa tgcttttata ttctaatagt ttttcacttt gtgtattaaa tggtttttaa
3540 attactttct tgatctctat tcattataaa aatcagatta taataaaaca
gttgaatatg 3600 gcttaggaaa atatgaag 3618 6 2585 DNA Homo sapien 6
gacatgcccc gcggcgcgcc attaaccgcc agatttgaat cgcgggaccc gttggcagag
60 gtggcggcgg cggcatgggt gccccgacgt tgccccctgc ctggcagccc
tttctcaagg 120 accaccgcat ctctacattc aagaactggc ccttcttgga
gggctgcgcc tgcaccccgg 180 agcggatggc cgaggctggc ttcatccact
gccccactga gaacgagcca gacttggccc 240 agtgtttctt ctgcttcaag
gagctggaag gctgggagcc agatgacgac cccatagagg 300 aacataaaaa
gcattcgtcc ggttgcgctt tcctttctgt caagaagcag tttgaagaat 360
taacccttgg tgaatttttg aaactggaca gagaaagagc caagaacaaa attgcaaagg
420 aaaccaacaa taagaagaaa gaatttgagg aaactgcgaa gaaagtgcgc
cgtgccatcg 480 agcagctggc tgccatggat tgaggcctct ggccggagct
gcctggtccc agagtggctg 540 caccacttcc agggtttatt ccctggtgcc
accagccttc ctgtgggccc cttagcaatg 600 tcttaggaaa ggagatcaac
attttcaaat tagatgtttc aactgtgctc ttgttttgtc 660 ttgaaagtgg
caccagaggt gcttctgcct gtgcagcggg tgctgctggt aacagtggct 720
gcttctctct ctctctctct tttttggggg ctcatttttg ctgttttgat tcccgggctt
780 accaggtgag aagtgaggga ggaagaaggc agtgtccctt ttgctagagc
tgacagcttt 840 gttcgcgtgg gcagagcctt ccacagtgaa tgtgtctgga
cctcatgttg ttgaggctgt 900 cacagtcctg agtgtggact tggcaggtgc
ctgttgaatc tgagctgcag gttccttatc 960 tgtcacacct gtgcctcctc
agaggacagt ttttttgttg ttgtgttttt ttgttttttt 1020 ttttttggta
gatgcatgac ttgtgtgtga tgagagaatg gagacagagt ccctggctcc 1080
tctactgttt aacaacatgg ctttcttatt ttgtttgaat tgttaattca cagaatagca
1140 caaactacaa ttaaaactaa gcacaaagcc attctaagtc attggggaaa
cggggtgaac 1200 ttcaggtgga tgaggagaca gaatagagtg ataggaagcg
tctggcagat actccttttg 1260 ccactgctgt gtgattagac aggcccagtg
agccgcgggg cacatgctgg ccgctcctcc 1320 ctcagaaaaa ggcagtggcc
taaatccttt ttaaatgact tggctcgatg ctgtggggga 1380 ctggctgggc
tgctgcaggc cgtgtgtctg tcagcccaac cttcacatct gtcacgttct 1440
ccacacgggg gagagacgca gtccgcccag gtccccgctt tctttggagg cagcagctcc
1500 cgcagggctg aagtctggcg taagatgatg gatttgattc gccctcctcc
ctgtcataga 1560 gctgcagggt ggattgttac agcttcgctg gaaacctctg
gaggtcatct cggctgttcc 1620 tgagaaataa aaagcctgtc atttcaaaca
ctgctgtgga ccctactggg tttttaaaat 1680 attgtcagtt tttcatcgtc
gtccctagcc tgccaacagc catctgccca gacagccgca 1740 gtgaggatga
gcgtcctggc agagacgcag ttgtctctgg gcgcttgcca gagccacgaa 1800
ccccagacct gtttgtatca tccgggctcc ttccgggcag aaacaactga aaatgcactt
1860 cagacccact tatttctgcc acatctgagt cggcctgaga tagacttttc
cctctaaact 1920 gggagaatat cacagtggtt tttgttagca gaaaatgcac
tccagcctct gtactcatct 1980 aagctgctta tttttgatat ttgtgtcagt
ctgtaaatgg atacttcact ttaataactg 2040 ttgcttagta attggctttg
tagagaagct ggaaaaaaat ggttttgtct tcaactcctt 2100 tgcatgccag
gcggtgatgt ggatctcggc ttctgtgagc ctgtgctgtg ggcagggctg 2160
agctggagcc gcccctctca gcccgcctgc cacggccttt ccttaaaggc catccttaaa
2220 accagaccct catggctacc agcacctgaa agcttcctcg acatctgtta
ataaagccgt 2280 aggcccttgt ctaagtgcaa ccgcctagac tttctttcag
atacatgtcc acatgtccat 2340 ttttcaggtt ctctaagttg gagtggagtc
tgggaagggt tgtgaatgag gcttctgggc 2400 tatgggtgag gttccaatgg
caggttagag cccctcgggc caactgccat cctggaaagt 2460 agagacagca
gtgcccgctg cccagaagag accagcaagc caaactggag cccccattgc 2520
aggctgtcgc catgtggaaa gagtaactca caattgccaa taaagtctca tgtggtttta
2580 tctac 2585 7 2654 DNA Homo sapien 7 gacatgcccc gcggcgcgcc
attaaccgcc agatttgaat cgcgggaccc gttggcagag 60 gtggcggcgg
cggcatgggt gccccgacgt tgccccctgc ctggcagccc tttctcaagg 120
accaccgcat ctctacattc aagaactggc ccttcttgga gggctgcgcc tgcaccccgg
180 agcggatggc cgaggctggc ttcatccact gccccactga gaacgagcca
gacttggccc 240 agtgtttctt ctgcttcaag gagctggaag gctgggagcc
agatgacgac cccattgggc 300 cgggcacggt ggcttacgcc tgtaatacca
gcactttggg aggccgaggc gggcggatca 360 cgagagagga acataaaaag
cattcgtccg gttgcgcttt cctttctgtc aagaagcagt 420 ttgaagaatt
aacccttggt gaatttttga aactggacag agaaagagcc aagaacaaaa 480
ttgcaaagga aaccaacaat aagaagaaag aatttgagga aactgcgaag aaagtgcgcc
540 gtgccatcga gcagctggct gccatggatt gaggcctctg gccggagctg
cctggtccca 600 gagtggctgc accacttcca gggtttattc cctggtgcca
ccagccttcc tgtgggcccc 660 ttagcaatgt cttaggaaag gagatcaaca
ttttcaaatt agatgtttca actgtgctct 720 tgttttgtct tgaaagtggc
accagaggtg cttctgcctg tgcagcgggt gctgctggta 780 acagtggctg
cttctctctc tctctctctt ttttgggggc tcatttttgc tgttttgatt 840
cccgggctta ccaggtgaga agtgagggag gaagaaggca gtgtcccttt tgctagagct
900 gacagctttg ttcgcgtggg cagagccttc cacagtgaat gtgtctggac
ctcatgttgt 960 tgaggctgtc acagtcctga gtgtggactt ggcaggtgcc
tgttgaatct gagctgcagg 1020 ttccttatct gtcacacctg tgcctcctca
gaggacagtt tttttgttgt tgtgtttttt 1080 tgtttttttt tttttggtag
atgcatgact tgtgtgtgat gagagaatgg agacagagtc 1140 cctggctcct
ctactgttta acaacatggc tttcttattt tgtttgaatt gttaattcac 1200
agaatagcac aaactacaat taaaactaag cacaaagcca ttctaagtca ttggggaaac
1260 ggggtgaact tcaggtggat gaggagacag aatagagtga taggaagcgt
ctggcagata 1320 ctccttttgc cactgctgtg tgattagaca ggcccagtga
gccgcggggc acatgctggc 1380 cgctcctccc tcagaaaaag gcagtggcct
aaatcctttt taaatgactt ggctcgatgc 1440 tgtgggggac tggctgggct
gctgcaggcc gtgtgtctgt cagcccaacc ttcacatctg 1500 tcacgttctc
cacacggggg agagacgcag tccgcccagg tccccgcttt ctttggaggc 1560
agcagctccc gcagggctga agtctggcgt aagatgatgg atttgattcg ccctcctccc
1620 tgtcatagag ctgcagggtg gattgttaca gcttcgctgg aaacctctgg
aggtcatctc 1680 ggctgttcct gagaaataaa aagcctgtca tttcaaacac
tgctgtggac cctactgggt 1740 ttttaaaata ttgtcagttt ttcatcgtcg
tccctagcct gccaacagcc atctgcccag 1800 acagccgcag tgaggatgag
cgtcctggca gagacgcagt tgtctctggg cgcttgccag 1860 agccacgaac
cccagacctg tttgtatcat ccgggctcct tccgggcaga aacaactgaa 1920
aatgcacttc agacccactt atttctgcca catctgagtc ggcctgagat agacttttcc
1980 ctctaaactg ggagaatatc acagtggttt ttgttagcag aaaatgcact
ccagcctctg 2040 tactcatcta agctgcttat ttttgatatt tgtgtcagtc
tgtaaatgga tacttcactt 2100 taataactgt tgcttagtaa ttggctttgt
agagaagctg gaaaaaaatg gttttgtctt 2160 caactccttt gcatgccagg
cggtgatgtg gatctcggct tctgtgagcc tgtgctgtgg 2220 gcagggctga
gctggagccg cccctctcag cccgcctgcc acggcctttc cttaaaggcc 2280
atccttaaaa ccagaccctc atggctacca gcacctgaaa gcttcctcga catctgttaa
2340 taaagccgta ggcccttgtc taagtgcaac cgcctagact ttctttcaga
tacatgtcca 2400 catgtccatt tttcaggttc tctaagttgg agtggagtct
gggaagggtt gtgaatgagg 2460 cttctgggct atgggtgagg ttccaatggc
aggttagagc ccctcgggcc aactgccatc 2520 ctggaaagta gagacagcag
tgcccgctgc ccagaagaga ccagcaagcc aaactggagc 2580 ccccattgca
ggctgtcgcc atgtggaaag agtaactcac aattgccaat aaagtctcat 2640
gtggttttat ctac 2654 8 2467 DNA Homo sapien 8 gacatgcccc gcggcgcgcc
attaaccgcc agatttgaat cgcgggaccc gttggcagag 60 gtggcggcgg
cggcatgggt gccccgacgt tgccccctgc ctggcagccc tttctcaagg 120
accaccgcat ctctacattc aagaactggc ccttcttgga gggctgcgcc tgcaccccgg
180 agcggatggc cgaggctggc ttcatccact gccccactga gaacgagcca
gacttggccc 240 agtgtttctt ctgcttcaag gagctggaag gctgggagcc
agatgacgac cccatgcaaa 300 ggaaaccaac aataagaaga aagaatttga
ggaaactgcg aagaaagtgc gccgtgccat 360 cgagcagctg gctgccatgg
attgaggcct ctggccggag ctgcctggtc ccagagtggc 420 tgcaccactt
ccagggttta ttccctggtg ccaccagcct tcctgtgggc cccttagcaa 480
tgtcttagga aaggagatca acattttcaa attagatgtt tcaactgtgc tcttgttttg
540 tcttgaaagt ggcaccagag gtgcttctgc ctgtgcagcg ggtgctgctg
gtaacagtgg 600 ctgcttctct ctctctctct cttttttggg ggctcatttt
tgctgttttg attcccgggc 660 ttaccaggtg agaagtgagg gaggaagaag
gcagtgtccc ttttgctaga gctgacagct 720 ttgttcgcgt gggcagagcc
ttccacagtg aatgtgtctg gacctcatgt tgttgaggct 780 gtcacagtcc
tgagtgtgga cttggcaggt gcctgttgaa tctgagctgc aggttcctta 840
tctgtcacac ctgtgcctcc tcagaggaca gtttttttgt tgttgtgttt ttttgttttt
900 ttttttttgg tagatgcatg acttgtgtgt gatgagagaa tggagacaga
gtccctggct 960 cctctactgt ttaacaacat ggctttctta ttttgtttga
attgttaatt cacagaatag 1020 cacaaactac aattaaaact aagcacaaag
ccattctaag tcattgggga aacggggtga 1080 acttcaggtg gatgaggaga
cagaatagag tgataggaag cgtctggcag atactccttt 1140 tgccactgct
gtgtgattag acaggcccag tgagccgcgg ggcacatgct ggccgctcct 1200
ccctcagaaa aaggcagtgg cctaaatcct ttttaaatga cttggctcga tgctgtgggg
1260 gactggctgg gctgctgcag gccgtgtgtc tgtcagccca accttcacat
ctgtcacgtt 1320 ctccacacgg gggagagacg cagtccgccc aggtccccgc
tttctttgga ggcagcagct 1380 cccgcagggc tgaagtctgg cgtaagatga
tggatttgat tcgccctcct ccctgtcata 1440 gagctgcagg gtggattgtt
acagcttcgc tggaaacctc tggaggtcat ctcggctgtt 1500 cctgagaaat
aaaaagcctg tcatttcaaa cactgctgtg gaccctactg ggtttttaaa 1560
atattgtcag tttttcatcg tcgtccctag cctgccaaca gccatctgcc cagacagccg
1620 cagtgaggat gagcgtcctg gcagagacgc agttgtctct gggcgcttgc
cagagccacg 1680 aaccccagac ctgtttgtat catccgggct ccttccgggc
agaaacaact gaaaatgcac 1740 ttcagaccca cttatttctg ccacatctga
gtcggcctga gatagacttt tccctctaaa 1800 ctgggagaat atcacagtgg
tttttgttag cagaaaatgc actccagcct ctgtactcat 1860 ctaagctgct
tatttttgat atttgtgtca gtctgtaaat ggatacttca ctttaataac 1920
tgttgcttag taattggctt tgtagagaag ctggaaaaaa atggttttgt cttcaactcc
1980 tttgcatgcc aggcggtgat gtggatctcg gcttctgtga gcctgtgctg
tgggcagggc 2040 tgagctggag ccgcccctct cagcccgcct gccacggcct
ttccttaaag gccatcctta 2100 aaaccagacc ctcatggcta ccagcacctg
aaagcttcct cgacatctgt taataaagcc 2160 gtaggccctt gtctaagtgc
aaccgcctag actttctttc agatacatgt ccacatgtcc 2220 atttttcagg
ttctctaagt tggagtggag tctgggaagg gttgtgaatg aggcttctgg 2280
gctatgggtg aggttccaat ggcaggttag agcccctcgg gccaactgcc atcctggaaa
2340 gtagagacag cagtgcccgc tgcccagaag agaccagcaa gccaaactgg
agcccccatt 2400 gcaggctgtc gccatgtgga aagagtaact cacaattgcc
aataaagtct catgtggttt 2460 tatctac 2467 9 2570 DNA Homo sapien 9
gacatgcccc gcggcgcgcc attaaccgcc agatttgaat cgcgggaccc gttggcagag
60 gtggcggcgg cggcatgggt gccccgacgt tgccccctgc ctggcagccc
tttctcaagg 120 accaccgcat ctctacattc aagaactggc ccttcttgga
gggctgcgcc tgcaccccgg 180 agcggatggc cgaggctggc ttcatccact
gccccactga gaacgagcca gacttggccc 240 agtgtttctt ctgcttcaag
gagctggaag gctgggagcc agatgacgac cccatagagg 300 aacataaaaa
gcattcgtcc ggttgcgctt tcctttctgt caagaagcag tttgaagaat 360
taacccttgg tgaatttttg aaactggaca gagaaagagc caagaacaaa attgcaaagg
420 aaaccaacaa taagaagaaa gaatttgagg aaactgcgaa gaaagtgcgc
cgtgccatcg 480 agcagctggc
tgccatggat tgaggcctct ggccggagct gcctggtccc agagtggctg 540
caccacttcc agggtttatt ccctggtgcc accagccttc ctgtgggccc cttagcaatg
600 tcttaggaaa ggagatcaac attttcaaat tagatgtttc aactgtgctc
ttgttttgtc 660 ttgaaagtgg caccagaggt gcttctgcct gtgcagcggg
tgctgctggt aacagtggct 720 gcttctctct ctctctctct tttttggggg
ctcatttttg ctgttttgat tcccgggctt 780 accaggtgag aagtgaggga
ggaagaaggc agtgtccctt ttgctagagc tgacagcttt 840 gttcgcgtgg
gcagagcctt ccacagtgaa tgtgtctgga cctcatgttg ttgaggctgt 900
cacagtcctg agtgtggact tggcaggtgc ctgttgaatc tgagctgcag gttccttatc
960 tgtcacacct gtgcctcctc agaggacatg tttttttgtt tttttttttt
tggtagatgc 1020 atgacttgtg tgtgatgaga gaatggagac agagtccctg
gctcctctac tgtttaacaa 1080 catggctttc ttattttgtt tgaattgtta
attcacagaa tagcacaaac tacaattaaa 1140 actaagcaca aagccattct
aagtcattgg ggaaacgggg tgaacttcag gtggatgagg 1200 agacagaata
gagtgatagg aagcgtctgg cagatactcc ttttgccact gctgtgtgat 1260
tagacaggcc cagtgagccg cggggcacat gctggccgct cctccctcag aaaaaggcag
1320 tggcctaaat cctttttaaa tgacttggct cgatgctgtg ggggactggc
tgggctgctg 1380 caggccgtgt gtctgtcagc ccaaccttca catctgtcac
gttctccaca cgggggagag 1440 acgcagtccg cccaggtccc cgctttcttt
ggaggcagca gctcccgcag ggctgaagtc 1500 tggcgtaaga tgatggattt
gattcgccct cctccctgtc atagagctgc agggtggatt 1560 gttacagctt
cgctggaaac ctctggaggt catctcggct gttcctgaga aataaaaagc 1620
ctgtcatttc aaacactgct gtggacccta ctgggttttt aaaatattgt cagtttttca
1680 tcgtcgtccc tagcctgcca acagccatct gcccagacag ccgcagtgag
gatgagcgtc 1740 ctggcagaga cgcagttgtc tctgggcgct tgccagagcc
acgaacccca gacctgtttg 1800 tatcatccgg gctccttccg ggcagaaaca
actgaaaatg cacttcagac ccacttattt 1860 ctgccacatc tgagtcggcc
tgagatagac ttttccctct aaactgggag aatatcacag 1920 tggtttttgt
tagcagaaaa tgcactccag cctctgtact catctaagct gcttattttt 1980
gatatttgtg tcagtctgta aatggatact tcactttaat aactgttgct tagtaattgg
2040 ctttgtagag aagctggaaa aaaatggttt tgtcttcaac tcctttgcat
gccaggcggt 2100 gatgtggatc tcggcttctg tgagcctgtg ctgtgggcag
ggctgagctg gagccgcccc 2160 tctcagcccg cctgccacgg cctttcctta
aaggccatcc ttaaaaccag accctcatgg 2220 ctaccagcac ctgaaagctt
cctcgacatc tgttaataaa gccgtaggcc cttgtctaag 2280 tgcaaccgcc
tagactttct ttcagataca tgtccacatg tccatttttc aggttctcta 2340
agttggagtg gagtctggga agggttgtga atgaggcttc tgggctatgg gtgaggttcc
2400 aatggcaggt tagagcccct cgggccaact gccatcctgg aaagtagaga
cagcagtgcc 2460 cgctgcccag aagagaccag caagccaaac tggagccccc
attgcaggct gtcgccatgt 2520 ggaaagagta actcacaatt gccaataaag
tctcatgtgg ttttatctac 2570 10 2639 DNA Homo sapien 10 gacatgcccc
gcggcgcgcc attaaccgcc agatttgaat cgcgggaccc gttggcagag 60
gtggcggcgg cggcatgggt gccccgacgt tgccccctgc ctggcagccc tttctcaagg
120 accaccgcat ctctacattc aagaactggc ccttcttgga gggctgcgcc
tgcaccccgg 180 agcggatggc cgaggctggc ttcatccact gccccactga
gaacgagcca gacttggccc 240 agtgtttctt ctgcttcaag gagctggaag
gctgggagcc agatgacgac cccattgggc 300 cgggcacggt ggcttacgcc
tgtaatacca gcactttggg aggccgaggc gggcggatca 360 cgagagagga
acataaaaag cattcgtccg gttgcgcttt cctttctgtc aagaagcagt 420
ttgaagaatt aacccttggt gaatttttga aactggacag agaaagagcc aagaacaaaa
480 ttgcaaagga aaccaacaat aagaagaaag aatttgagga aactgcgaag
aaagtgcgcc 540 gtgccatcga gcagctggct gccatggatt gaggcctctg
gccggagctg cctggtccca 600 gagtggctgc accacttcca gggtttattc
cctggtgcca ccagccttcc tgtgggcccc 660 ttagcaatgt cttaggaaag
gagatcaaca ttttcaaatt agatgtttca actgtgctct 720 tgttttgtct
tgaaagtggc accagaggtg cttctgcctg tgcagcgggt gctgctggta 780
acagtggctg cttctctctc tctctctctt ttttgggggc tcatttttgc tgttttgatt
840 cccgggctta ccaggtgaga agtgagggag gaagaaggca gtgtcccttt
tgctagagct 900 gacagctttg ttcgcgtggg cagagccttc cacagtgaat
gtgtctggac ctcatgttgt 960 tgaggctgtc acagtcctga gtgtggactt
ggcaggtgcc tgttgaatct gagctgcagg 1020 ttccttatct gtcacacctg
tgcctcctca gaggacatgt ttttttgttt tttttttttt 1080 ggtagatgca
tgacttgtgt gtgatgagag aatggagaca gagtccctgg ctcctctact 1140
gtttaacaac atggctttct tattttgttt gaattgttaa ttcacagaat agcacaaact
1200 acaattaaaa ctaagcacaa agccattcta agtcattggg gaaacggggt
gaacttcagg 1260 tggatgagga gacagaatag agtgatagga agcgtctggc
agatactcct tttgccactg 1320 ctgtgtgatt agacaggccc agtgagccgc
ggggcacatg ctggccgctc ctccctcaga 1380 aaaaggcagt ggcctaaatc
ctttttaaat gacttggctc gatgctgtgg gggactggct 1440 gggctgctgc
aggccgtgtg tctgtcagcc caaccttcac atctgtcacg ttctccacac 1500
gggggagaga cgcagtccgc ccaggtcccc gctttctttg gaggcagcag ctcccgcagg
1560 gctgaagtct ggcgtaagat gatggatttg attcgccctc ctccctgtca
tagagctgca 1620 gggtggattg ttacagcttc gctggaaacc tctggaggtc
atctcggctg ttcctgagaa 1680 ataaaaagcc tgtcatttca aacactgctg
tggaccctac tgggttttta aaatattgtc 1740 agtttttcat cgtcgtccct
agcctgccaa cagccatctg cccagacagc cgcagtgagg 1800 atgagcgtcc
tggcagagac gcagttgtct ctgggcgctt gccagagcca cgaaccccag 1860
acctgtttgt atcatccggg ctccttccgg gcagaaacaa ctgaaaatgc acttcagacc
1920 cacttatttc tgccacatct gagtcggcct gagatagact tttccctcta
aactgggaga 1980 atatcacagt ggtttttgtt agcagaaaat gcactccagc
ctctgtactc atctaagctg 2040 cttatttttg atatttgtgt cagtctgtaa
atggatactt cactttaata actgttgctt 2100 agtaattggc tttgtagaga
agctggaaaa aaatggtttt gtcttcaact cctttgcatg 2160 ccaggcggtg
atgtggatct cggcttctgt gagcctgtgc tgtgggcagg gctgagctgg 2220
agccgcccct ctcagcccgc ctgccacggc ctttccttaa aggccatcct taaaaccaga
2280 ccctcatggc taccagcacc tgaaagcttc ctcgacatct gttaataaag
ccgtaggccc 2340 ttgtctaagt gcaaccgcct agactttctt tcagatacat
gtccacatgt ccatttttca 2400 ggttctctaa gttggagtgg agtctgggaa
gggttgtgaa tgaggcttct gggctatggg 2460 tgaggttcca atggcaggtt
agagcccctc gggccaactg ccatcctgga aagtagagac 2520 agcagtgccc
gctgcccaga agagaccagc aagccaaact ggagccccca ttgcaggctg 2580
tcgccatgtg gaaagagtaa ctcacaattg ccaataaagt ctcatgtggt tttatctac
2639 11 2452 DNA Homo sapien 11 gacatgcccc gcggcgcgcc attaaccgcc
agatttgaat cgcgggaccc gttggcagag 60 gtggcggcgg cggcatgggt
gccccgacgt tgccccctgc ctggcagccc tttctcaagg 120 accaccgcat
ctctacattc aagaactggc ccttcttgga gggctgcgcc tgcaccccgg 180
agcggatggc cgaggctggc ttcatccact gccccactga gaacgagcca gacttggccc
240 agtgtttctt ctgcttcaag gagctggaag gctgggagcc agatgacgac
cccatgcaaa 300 ggaaaccaac aataagaaga aagaatttga ggaaactgcg
aagaaagtgc gccgtgccat 360 cgagcagctg gctgccatgg attgaggcct
ctggccggag ctgcctggtc ccagagtggc 420 tgcaccactt ccagggttta
ttccctggtg ccaccagcct tcctgtgggc cccttagcaa 480 tgtcttagga
aaggagatca acattttcaa attagatgtt tcaactgtgc tcttgttttg 540
tcttgaaagt ggcaccagag gtgcttctgc ctgtgcagcg ggtgctgctg gtaacagtgg
600 ctgcttctct ctctctctct cttttttggg ggctcatttt tgctgttttg
attcccgggc 660 ttaccaggtg agaagtgagg gaggaagaag gcagtgtccc
ttttgctaga gctgacagct 720 ttgttcgcgt gggcagagcc ttccacagtg
aatgtgtctg gacctcatgt tgttgaggct 780 gtcacagtcc tgagtgtgga
cttggcaggt gcctgttgaa tctgagctgc aggttcctta 840 tctgtcacac
ctgtgcctcc tcagaggaca tgtttttttg tttttttttt tttggtagat 900
gcatgacttg tgtgtgatga gagaatggag acagagtccc tggctcctct actgtttaac
960 aacatggctt tcttattttg tttgaattgt taattcacag aatagcacaa
actacaatta 1020 aaactaagca caaagccatt ctaagtcatt ggggaaacgg
ggtgaacttc aggtggatga 1080 ggagacagaa tagagtgata ggaagcgtct
ggcagatact ccttttgcca ctgctgtgtg 1140 attagacagg cccagtgagc
cgcggggcac atgctggccg ctcctccctc agaaaaaggc 1200 agtggcctaa
atccttttta aatgacttgg ctcgatgctg tgggggactg gctgggctgc 1260
tgcaggccgt gtgtctgtca gcccaacctt cacatctgtc acgttctcca cacgggggag
1320 agacgcagtc cgcccaggtc cccgctttct ttggaggcag cagctcccgc
agggctgaag 1380 tctggcgtaa gatgatggat ttgattcgcc ctcctccctg
tcatagagct gcagggtgga 1440 ttgttacagc ttcgctggaa acctctggag
gtcatctcgg ctgttcctga gaaataaaaa 1500 gcctgtcatt tcaaacactg
ctgtggaccc tactgggttt ttaaaatatt gtcagttttt 1560 catcgtcgtc
cctagcctgc caacagccat ctgcccagac agccgcagtg aggatgagcg 1620
tcctggcaga gacgcagttg tctctgggcg cttgccagag ccacgaaccc cagacctgtt
1680 tgtatcatcc gggctccttc cgggcagaaa caactgaaaa tgcacttcag
acccacttat 1740 ttctgccaca tctgagtcgg cctgagatag acttttccct
ctaaactggg agaatatcac 1800 agtggttttt gttagcagaa aatgcactcc
agcctctgta ctcatctaag ctgcttattt 1860 ttgatatttg tgtcagtctg
taaatggata cttcacttta ataactgttg cttagtaatt 1920 ggctttgtag
agaagctgga aaaaaatggt tttgtcttca actcctttgc atgccaggcg 1980
gtgatgtgga tctcggcttc tgtgagcctg tgctgtgggc agggctgagc tggagccgcc
2040 cctctcagcc cgcctgccac ggcctttcct taaaggccat ccttaaaacc
agaccctcat 2100 ggctaccagc acctgaaagc ttcctcgaca tctgttaata
aagccgtagg cccttgtcta 2160 agtgcaaccg cctagacttt ctttcagata
catgtccaca tgtccatttt tcaggttctc 2220 taagttggag tggagtctgg
gaagggttgt gaatgaggct tctgggctat gggtgaggtt 2280 ccaatggcag
gttagagccc ctcgggccaa ctgccatcct ggaaagtaga gacagcagtg 2340
cccgctgccc agaagagacc agcaagccaa actggagccc ccattgcagg ctgtcgccat
2400 gtggaaagag taactcacaa ttgccaataa agtctcatgt ggttttatct ac 2452
12 2252 DNA Homo sapien 12 aattctcgcc caaggacaga cctgaatctc
tagctgccta gaggctgact caactgaaat 60 catggcgttt gacagcactt
ggaaggtaga ccggagtgaa aactatgaca agttcatgga 120 aaaaatgggt
gttaatatag tgaaaaggaa gcttgcagct catgacaatt tgaagctgac 180
aattacacaa gaaggaaata aattcacagt caaagaatca agcgcttttc gaaacattga
240 agttgttttt gaacttggtg tcacctttaa ttacaatcta gcagacggaa
ctgaactcag 300 ggggacctgg agccttgagg gaaataaact tattggaaaa
ttcaaacgga cagacaatgg 360 aaacgaactg aatactgtcc gagaaattat
aggtgatgaa ctagtccaga cttatgtgta 420 tgaaggagta gaagccaaaa
ggatctttaa aaaggattga gcattattct tggcgcacag 480 tccaaaatac
aaattggaca gaagatctat attgtaccag aactatttat ttcaccccat 540
caagtataag gttactgatt gattggtcct tttataaaca ttggtatatt tccattcatg
600 ccaaagcaaa agaagtaaaa gctaattagg atttaatttg ttttatattc
tctaagatat 660 atatttacta aaagaatttg tgacatttta aaaaacaaaa
ataaatattg cgtccatgtt 720 gctttatatg tagccttgcc ttttaaaaga
aaaagtatgt gaatatgaat tgacagactg 780 ttttcgtaga gagagggtct
tactctttca ctcaggctgg aatgtagtgg agagatcata 840 gctcactgta
acctcaaact cctggactca tgcaatcttc ctgcctcagg cttctgagta 900
gctaggacta tgggtacatt ccacagtgcc cagctaattt ttgttttgtt ttctttttat
960 tttttttaga gatggggtct tgctatattg cccaggctgg tcttgaaccc
ctggcctcaa 1020 gcaatcctcc tgcctcagcc tctcaagttg ttttttttct
ttacatttga taaactaaaa 1080 gcataggctg catatgagtc tttaacatct
tgaactggtt gtgaataatt ttctggcact 1140 ggttgtaagt aatatctatt
attataaaaa taatatatgc tcaaccagaa aacttagaaa 1200 taagaaacac
aaatgtaaaa taagtatttc cataactcat aatccagaga taattgccat 1260
tctgattttg atagatatcc tctcagctct cttccctggg ggcagatatt tcccaataca
1320 taccactttg aataggatga taggaaataa atgatgtact acattaaatt
aaattattgt 1380 attacatttt tgtacacatc agtcattccc acgcttggct
gaaaatcagg atcatctgag 1440 aaacttaaac aatttctgca ttcttaatct
ccactgttat tctattatat cagaatcgct 1500 aatagaacca agaattctag
aaaatttcct ggtgattctg atgcagcctg tccactaact 1560 tgtctttgag
aacaatggag atatcagtta tcaatgttat ttttaaccac ccccttcttt 1620
tttgttgttg tgggtttttt cacataaaca catacattgc caattttcca gggctcaaaa
1680 agtttccttt tattcatatt ttcacatgac gtaaaatttt atgtgcttca
cataattgta 1740 ttttagcagg gtacatatta ggggatggag agggggtaca
atttttaagc atttgcagct 1800 gcaactctat agacttttga caaattcttt
ttcacactga tgtagataac agcttaacat 1860 ttacatagtt cttactactt
gccacacact gttctaagtg gtatatacat atgacatata 1920 tattaaaatg
taggaagaaa aatgtttgag tacctgatga aaatgaatag agagtatgta 1980
atctttgaaa gctgaatact gcgtgttctc acttataagt gggagctaaa taatgtatac
2040 acatggacac accagagtgt agaataatag acactggaga cttggaagag
ttggagggtt 2100 ggcagggggc gatgataatt tacttaatgg gtacaatgta
cattattcag ggggtgatgg 2160 ttacagccca gactttacca ctatgcaata
tatcaatgta acaaaactat acttgtactc 2220 cttaaattta tataaaaata
ccttaaatct at 2252 13 994 PRT Homo sapien 13 Met Ser Lys Leu Arg
Met Val Leu Leu Glu Asp Ser Gly Ser Ala Asp 1 5 10 15 Phe Arg Arg
His Phe Val Asn Leu Ser Pro Phe Thr Ile Thr Val Val 20 25 30 Leu
Leu Leu Ser Ala Cys Phe Val Thr Ser Ser Leu Gly Gly Thr Asp 35 40
45 Lys Glu Leu Arg Leu Val Asp Gly Glu Asn Lys Cys Ser Gly Arg Val
50 55 60 Glu Val Lys Val Gln Glu Glu Trp Gly Thr Val Cys Asn Asn
Gly Trp 65 70 75 80 Ser Met Glu Ala Val Ser Val Ile Cys Asn Gln Leu
Gly Cys Pro Thr 85 90 95 Ala Ile Lys Ala Pro Gly Trp Ala Asn Ser
Ser Ala Gly Ser Gly Arg 100 105 110 Ile Trp Met Asp His Val Ser Cys
Arg Gly Asn Glu Ser Ala Leu Trp 115 120 125 Asp Cys Lys His Asp Gly
Trp Gly Lys His Ser Asn Cys Thr His Gln 130 135 140 Gln Asp Ala Gly
Val Thr Cys Ser Asp Gly Ser Asn Leu Glu Met Arg 145 150 155 160 Leu
Thr Arg Gly Gly Asn Met Cys Ser Gly Arg Ile Glu Ile Lys Phe 165 170
175 Gln Gly Arg Trp Gly Thr Val Cys Asp Asp Asn Phe Asn Ile Asp His
180 185 190 Ala Ser Val Ile Cys Arg Gln Leu Glu Cys Gly Ser Ala Val
Ser Phe 195 200 205 Ser Gly Ser Ser Asn Phe Gly Glu Gly Ser Gly Pro
Ile Trp Phe Asp 210 215 220 Asp Leu Ile Cys Asn Gly Asn Glu Ser Ala
Leu Trp Asn Cys Lys His 225 230 235 240 Gln Gly Trp Gly Lys His Asn
Cys Asp His Ala Glu Asp Ala Gly Val 245 250 255 Ile Cys Ser Lys Gly
Ala Asp Leu Ser Leu Arg Leu Val Asp Gly Val 260 265 270 Thr Glu Cys
Ser Gly Arg Leu Glu Val Arg Phe Gln Gly Glu Trp Gly 275 280 285 Thr
Ile Cys Asp Asp Gly Trp Asp Ser Tyr Asp Ala Ala Val Ala Cys 290 295
300 Lys Gln Leu Gly Cys Pro Thr Ala Val Thr Ala Ile Gly Arg Val Asn
305 310 315 320 Ala Ser Lys Gly Phe Gly His Ile Trp Leu Asp Ser Val
Ser Cys Gln 325 330 335 Gly His Glu Pro Ala Val Trp Gln Cys Lys His
His Glu Trp Gly Lys 340 345 350 His Tyr Cys Asn His Asn Glu Asp Ala
Gly Val Thr Cys Ser Asp Gly 355 360 365 Ser Asp Leu Glu Leu Arg Leu
Arg Gly Gly Gly Ser Arg Cys Ala Gly 370 375 380 Thr Val Glu Val Glu
Ile Gln Arg Leu Leu Gly Lys Val Cys Asp Arg 385 390 395 400 Gly Trp
Gly Leu Lys Glu Ala Asp Val Val Cys Arg Gln Leu Gly Cys 405 410 415
Gly Ser Ala Leu Lys Thr Ser Tyr Gln Val Tyr Ser Lys Ile Gln Ala 420
425 430 Thr Asn Thr Trp Leu Phe Leu Ser Ser Cys Asn Gly Asn Glu Thr
Ser 435 440 445 Leu Trp Asp Cys Lys Asn Trp Gln Trp Gly Gly Leu Thr
Cys Asp His 450 455 460 Tyr Glu Glu Ala Lys Ile Thr Cys Ser Ala His
Arg Glu Pro Arg Leu 465 470 475 480 Val Gly Gly Asp Ile Pro Cys Ser
Gly Arg Val Glu Val Lys His Gly 485 490 495 Asp Thr Trp Gly Ser Ile
Cys Asp Ser Asp Phe Ser Leu Glu Ala Ala 500 505 510 Ser Val Leu Cys
Arg Glu Leu Gln Cys Gly Thr Val Val Ser Ile Leu 515 520 525 Gly Gly
Ala His Phe Gly Glu Gly Asn Gly Gln Ile Trp Ala Glu Glu 530 535 540
Phe Gln Cys Glu Gly His Glu Ser His Leu Ser Leu Cys Pro Val Ala 545
550 555 560 Pro Arg Pro Glu Gly Thr Cys Ser His Ser Arg Asp Val Gly
Val Val 565 570 575 Cys Ser Arg Tyr Thr Glu Ile Arg Leu Val Asn Gly
Lys Thr Pro Cys 580 585 590 Glu Gly Arg Val Glu Leu Lys Thr Leu Gly
Ala Trp Gly Ser Leu Cys 595 600 605 Asn Ser His Trp Asp Ile Glu Asp
Ala His Val Leu Cys Gln Gln Leu 610 615 620 Lys Cys Gly Val Ala Leu
Ser Thr Pro Gly Gly Ala Arg Phe Gly Lys 625 630 635 640 Gly Asn Gly
Gln Ile Trp Arg His Met Phe His Cys Thr Gly Thr Glu 645 650 655 Gln
His Met Gly Asp Cys Pro Val Thr Ala Leu Gly Ala Ser Leu Cys 660 665
670 Pro Ser Glu Gln Val Ala Ser Val Ile Cys Ser Gly Asn Gln Ser Gln
675 680 685 Thr Leu Ser Ser Cys Asn Ser Ser Ser Leu Gly Pro Thr Arg
Pro Thr 690 695 700 Ile Pro Glu Glu Ser Ala Val Ala Cys Ile Glu Ser
Gly Gln Leu Arg 705 710 715 720 Leu Val Asn Gly Gly Gly Arg Cys Ala
Gly Arg Val Glu Ile Tyr His 725 730 735 Glu Gly Ser Trp Gly Thr Ile
Cys Asp Asp Ser Trp Asp Leu Ser Asp 740 745 750 Ala His Val Val Cys
Arg Gln Leu Gly Cys Gly Glu Ala Ile Asn Ala 755 760 765 Thr Gly Ser
Ala His Phe Gly Glu Gly Thr Gly Pro Ile Trp Leu Asp 770 775 780 Glu
Met Lys Cys Asn Gly Lys Glu Ser Arg Ile Trp Gln Cys His Ser 785 790
795 800 His Gly Trp Gly Gln Gln Asn Cys Arg His Lys Glu Asp Ala Gly
Val 805 810 815 Ile Cys Ser Glu Phe Met Ser Leu Arg Leu Thr Ser Glu
Ala Ser Arg 820 825 830 Glu Ala Cys Ala Gly Arg Leu Glu Val Phe Tyr
Asn Gly Ala Trp Gly 835 840 845 Thr Val Gly Lys Ser Ser Met Ser Glu
Thr Thr Val Gly Val Val Cys 850 855 860 Arg Gln Leu Gly Cys Ala Asp
Lys Gly Lys Ile Asn Pro Ala Ser Leu 865 870 875 880 Asp Lys Ala Met
Ser Ile Pro Met Trp Val Asp Asn Val Gln Cys Pro
885 890 895 Lys Gly Pro Asp Thr Leu Trp Gln Cys Pro Ser Ser Pro Trp
Glu Lys 900 905 910 Arg Leu Ala Ser Pro Ser Glu Glu Thr Trp Ile Thr
Cys Asp Asn Lys 915 920 925 Ile Arg Leu Gln Glu Gly Pro Thr Ser Cys
Ser Gly Arg Val Glu Ile 930 935 940 Trp His Gly Gly Ser Trp Gly Thr
Val Cys Asp Asp Ser Trp Asp Leu 945 950 955 960 Asp Asp Ala Gln Val
Val Cys Gln Gln Leu Gly Cys Gly Pro Ala Leu 965 970 975 Lys Ala Phe
Lys Glu Ala Glu Phe Gly Gln Gly Thr Gly Pro Asp Met 980 985 990 Ala
Gln 14 994 PRT Homo sapien 14 Met Ser Lys Leu Arg Met Val Leu Leu
Glu Asp Ser Gly Ser Ala Asp 1 5 10 15 Phe Arg Arg His Phe Val Asn
Leu Ser Pro Phe Thr Ile Thr Val Val 20 25 30 Leu Leu Leu Ser Ala
Cys Phe Val Thr Ser Ser Leu Gly Gly Thr Asp 35 40 45 Lys Glu Leu
Arg Leu Val Asp Gly Glu Asn Lys Cys Ser Gly Arg Val 50 55 60 Glu
Val Lys Val Gln Glu Glu Trp Gly Thr Val Cys Asn Asn Gly Trp 65 70
75 80 Ser Met Glu Ala Val Ser Val Ile Cys Asn Gln Leu Gly Cys Pro
Thr 85 90 95 Ala Ile Lys Ala Pro Gly Trp Ala Asn Ser Ser Ala Gly
Ser Gly Arg 100 105 110 Ile Trp Met Asp His Val Ser Cys Arg Gly Asn
Glu Ser Ala Leu Trp 115 120 125 Asp Cys Lys His Asp Gly Trp Gly Lys
His Ser Asn Cys Thr His Gln 130 135 140 Gln Asp Ala Gly Val Thr Cys
Ser Asp Gly Ser Asn Leu Glu Met Arg 145 150 155 160 Leu Thr Arg Gly
Gly Asn Met Cys Ser Gly Arg Ile Glu Ile Lys Phe 165 170 175 Gln Gly
Arg Trp Gly Thr Val Cys Asp Asp Asn Phe Asn Ile Asp His 180 185 190
Ala Ser Val Ile Cys Arg Gln Leu Glu Cys Gly Ser Ala Val Ser Phe 195
200 205 Ser Gly Ser Ser Asn Phe Gly Glu Gly Ser Gly Pro Ile Trp Phe
Asp 210 215 220 Asp Leu Ile Cys Asn Gly Asn Glu Ser Ala Leu Trp Asn
Cys Lys His 225 230 235 240 Gln Gly Trp Gly Lys His Asn Cys Asp His
Ala Glu Asp Ala Gly Val 245 250 255 Ile Cys Ser Lys Gly Ala Asp Leu
Ser Leu Arg Leu Val Asp Gly Val 260 265 270 Thr Glu Cys Ser Gly Arg
Leu Glu Val Arg Phe Gln Gly Glu Trp Gly 275 280 285 Thr Ile Cys Asp
Asp Gly Trp Asp Ser Tyr Asp Ala Ala Val Ala Cys 290 295 300 Lys Gln
Leu Gly Cys Pro Thr Ala Val Thr Ala Ile Gly Arg Val Asn 305 310 315
320 Ala Ser Lys Gly Phe Gly His Ile Trp Leu Asp Ser Val Ser Cys Gln
325 330 335 Gly His Glu Pro Ala Val Trp Gln Cys Lys His His Glu Trp
Gly Lys 340 345 350 His Tyr Cys Asn His Asn Glu Asp Ala Gly Val Thr
Cys Ser Asp Gly 355 360 365 Ser Asp Leu Glu Leu Arg Leu Arg Gly Gly
Gly Ser Arg Cys Ala Gly 370 375 380 Thr Val Glu Val Glu Ile Gln Arg
Leu Leu Gly Lys Val Cys Asp Arg 385 390 395 400 Gly Trp Gly Leu Lys
Glu Ala Asp Val Val Cys Arg Gln Leu Gly Cys 405 410 415 Gly Ser Ala
Leu Lys Thr Ser Tyr Gln Val Tyr Ser Lys Ile Gln Ala 420 425 430 Thr
Asn Thr Trp Leu Phe Leu Ser Ser Cys Asn Gly Asn Glu Thr Ser 435 440
445 Leu Trp Asp Cys Lys Asn Trp Gln Trp Gly Gly Leu Thr Cys Asp His
450 455 460 Tyr Glu Glu Ala Lys Ile Thr Cys Ser Ala His Arg Glu Pro
Arg Leu 465 470 475 480 Val Gly Gly Asp Ile Pro Cys Ser Gly Arg Val
Glu Val Lys His Gly 485 490 495 Asp Thr Trp Gly Ser Ile Cys Asp Ser
Asp Phe Ser Leu Glu Ala Ala 500 505 510 Ser Val Leu Cys Arg Glu Leu
Gln Cys Gly Thr Val Val Ser Ile Leu 515 520 525 Gly Gly Ala His Phe
Gly Glu Gly Asn Gly Gln Ile Trp Ala Glu Glu 530 535 540 Phe Gln Cys
Glu Gly His Glu Ser His Leu Ser Leu Cys Pro Val Ala 545 550 555 560
Pro Arg Pro Glu Gly Thr Cys Ser His Ser Arg Asp Val Gly Val Val 565
570 575 Cys Ser Arg Tyr Thr Glu Ile Arg Leu Val Asn Gly Lys Thr Pro
Cys 580 585 590 Glu Gly Arg Val Glu Leu Lys Thr Leu Gly Ala Trp Gly
Ser Leu Cys 595 600 605 Asn Ser His Trp Asp Ile Glu Asp Ala His Val
Leu Cys Gln Gln Leu 610 615 620 Lys Cys Gly Val Ala Leu Ser Thr Pro
Gly Gly Ala Arg Phe Gly Lys 625 630 635 640 Gly Asn Gly Gln Ile Trp
Arg His Met Phe His Cys Thr Gly Thr Glu 645 650 655 Gln His Met Gly
Asp Cys Pro Val Thr Ala Leu Gly Ala Ser Leu Cys 660 665 670 Pro Ser
Glu Gln Val Ala Ser Val Ile Cys Ser Gly Asn Gln Ser Gln 675 680 685
Thr Leu Ser Ser Cys Asn Ser Ser Ser Leu Gly Pro Thr Arg Pro Thr 690
695 700 Ile Pro Glu Glu Ser Ala Val Ala Cys Ile Glu Ser Gly Gln Leu
Arg 705 710 715 720 Leu Val Asn Gly Gly Gly Arg Cys Ala Gly Arg Val
Glu Ile Tyr His 725 730 735 Glu Gly Ser Trp Gly Thr Ile Cys Asp Asp
Ser Trp Asp Leu Ser Asp 740 745 750 Ala His Val Val Cys Arg Gln Leu
Gly Cys Gly Glu Ala Ile Asn Ala 755 760 765 Thr Gly Ser Ala His Phe
Gly Glu Gly Thr Gly Pro Ile Trp Leu Asp 770 775 780 Glu Met Lys Cys
Asn Gly Lys Glu Ser Arg Ile Trp Gln Cys His Ser 785 790 795 800 His
Gly Trp Gly Gln Gln Asn Cys Arg His Lys Glu Asp Ala Gly Val 805 810
815 Ile Cys Ser Glu Phe Met Ser Leu Arg Leu Thr Ser Glu Ala Ser Arg
820 825 830 Glu Ala Cys Ala Gly Arg Leu Glu Val Phe Tyr Asn Gly Ala
Trp Gly 835 840 845 Thr Val Gly Lys Ser Ser Met Ser Glu Thr Thr Val
Gly Val Val Cys 850 855 860 Arg Gln Leu Gly Cys Ala Asp Lys Gly Lys
Ile Asn Pro Ala Ser Leu 865 870 875 880 Asp Lys Ala Met Ser Ile Pro
Met Trp Val Asp Asn Val Gln Cys Pro 885 890 895 Lys Gly Pro Asp Thr
Leu Trp Gln Cys Pro Ser Ser Pro Trp Glu Lys 900 905 910 Arg Leu Ala
Ser Pro Ser Glu Glu Thr Trp Ile Thr Cys Asp Asn Lys 915 920 925 Ile
Arg Leu Gln Glu Gly Pro Thr Ser Cys Ser Gly Arg Val Glu Ile 930 935
940 Trp His Gly Gly Ser Trp Gly Thr Val Cys Asp Asp Ser Trp Asp Leu
945 950 955 960 Asp Asp Ala Gln Val Val Cys Gln Gln Leu Gly Cys Gly
Pro Ala Leu 965 970 975 Lys Ala Phe Lys Glu Ala Glu Phe Gly Gln Gly
Thr Gly Pro Asp Met 980 985 990 Ala Gln 15 1027 PRT Homo sapien 15
Met Ser Lys Leu Arg Met Val Leu Leu Glu Asp Ser Gly Ser Ala Asp 1 5
10 15 Phe Arg Arg His Phe Val Asn Leu Ser Pro Phe Thr Ile Thr Val
Val 20 25 30 Leu Leu Leu Ser Ala Cys Phe Val Thr Ser Ser Leu Gly
Gly Thr Asp 35 40 45 Lys Glu Leu Arg Leu Val Asp Gly Glu Asn Lys
Cys Ser Gly Arg Val 50 55 60 Glu Val Lys Val Gln Glu Glu Trp Gly
Thr Val Cys Asn Asn Gly Trp 65 70 75 80 Ser Met Glu Ala Val Ser Val
Ile Cys Asn Gln Leu Gly Cys Pro Thr 85 90 95 Ala Ile Lys Ala Pro
Gly Trp Ala Asn Ser Ser Ala Gly Ser Gly Arg 100 105 110 Ile Trp Met
Asp His Val Ser Cys Arg Gly Asn Glu Ser Ala Leu Trp 115 120 125 Asp
Cys Lys His Asp Gly Trp Gly Lys His Ser Asn Cys Thr His Gln 130 135
140 Gln Asp Ala Gly Val Thr Cys Ser Asp Gly Ser Asn Leu Glu Met Arg
145 150 155 160 Leu Thr Arg Gly Gly Asn Met Cys Ser Gly Arg Ile Glu
Ile Lys Phe 165 170 175 Gln Gly Arg Trp Gly Thr Val Cys Asp Asp Asn
Phe Asn Ile Asp His 180 185 190 Ala Ser Val Ile Cys Arg Gln Leu Glu
Cys Gly Ser Ala Val Ser Phe 195 200 205 Ser Gly Ser Ser Asn Phe Gly
Glu Gly Ser Gly Pro Ile Trp Phe Asp 210 215 220 Asp Leu Ile Cys Asn
Gly Asn Glu Ser Ala Leu Trp Asn Cys Lys His 225 230 235 240 Gln Gly
Trp Gly Lys His Asn Cys Asp His Ala Glu Asp Ala Gly Val 245 250 255
Ile Cys Ser Lys Gly Ala Asp Leu Ser Leu Arg Leu Val Asp Gly Val 260
265 270 Thr Glu Cys Ser Gly Arg Leu Glu Val Arg Phe Gln Gly Glu Trp
Gly 275 280 285 Thr Ile Cys Asp Asp Gly Trp Asp Ser Tyr Asp Ala Ala
Val Ala Cys 290 295 300 Lys Gln Leu Gly Cys Pro Thr Ala Val Thr Ala
Ile Gly Arg Val Asn 305 310 315 320 Ala Ser Lys Gly Phe Gly His Ile
Trp Leu Asp Ser Val Ser Cys Gln 325 330 335 Gly His Glu Pro Ala Val
Trp Gln Cys Lys His His Glu Trp Gly Lys 340 345 350 His Tyr Cys Asn
His Asn Glu Asp Ala Gly Val Thr Cys Ser Asp Gly 355 360 365 Ser Asp
Leu Glu Leu Arg Leu Arg Gly Gly Gly Ser Arg Cys Ala Gly 370 375 380
Thr Val Glu Val Glu Ile Gln Arg Leu Leu Gly Lys Val Cys Asp Arg 385
390 395 400 Gly Trp Gly Leu Lys Glu Ala Asp Val Val Cys Arg Gln Leu
Gly Cys 405 410 415 Gly Ser Ala Leu Lys Thr Ser Tyr Gln Val Tyr Ser
Lys Ile Gln Ala 420 425 430 Thr Asn Thr Trp Leu Phe Leu Ser Ser Cys
Asn Gly Asn Glu Thr Ser 435 440 445 Leu Trp Asp Cys Lys Asn Trp Gln
Trp Gly Gly Leu Thr Cys Asp His 450 455 460 Tyr Glu Glu Ala Lys Ile
Thr Cys Ser Ala His Arg Glu Pro Arg Leu 465 470 475 480 Val Gly Gly
Asp Ile Pro Cys Ser Gly Arg Val Glu Val Lys His Gly 485 490 495 Asp
Thr Trp Gly Ser Ile Cys Asp Ser Asp Phe Ser Leu Glu Ala Ala 500 505
510 Ser Val Leu Cys Arg Glu Leu Gln Cys Gly Thr Val Val Ser Ile Leu
515 520 525 Gly Gly Ala His Phe Gly Glu Gly Asn Gly Gln Ile Trp Ala
Glu Glu 530 535 540 Phe Gln Cys Glu Gly His Glu Ser His Leu Ser Leu
Cys Pro Val Ala 545 550 555 560 Pro Arg Pro Glu Gly Thr Cys Ser His
Ser Arg Asp Val Gly Val Val 565 570 575 Cys Ser Ser Lys Thr Gln Lys
Thr Ser Leu Ile Gly Ser His Thr Val 580 585 590 Lys Gly Thr Gly Leu
Gly Ser His Ser Cys Leu Phe Leu Lys Pro Cys 595 600 605 Leu Leu Pro
Gly Tyr Thr Glu Ile Arg Leu Val Asn Gly Lys Thr Pro 610 615 620 Cys
Glu Gly Arg Val Glu Leu Lys Thr Leu Gly Ala Trp Gly Ser Leu 625 630
635 640 Cys Asn Ser His Trp Asp Ile Glu Asp Ala His Val Leu Cys Gln
Gln 645 650 655 Leu Lys Cys Gly Val Ala Leu Ser Thr Pro Gly Gly Ala
Arg Phe Gly 660 665 670 Lys Gly Asn Gly Gln Ile Trp Arg His Met Phe
His Cys Thr Gly Thr 675 680 685 Glu Gln His Met Gly Asp Cys Pro Val
Thr Ala Leu Gly Ala Ser Leu 690 695 700 Cys Pro Ser Glu Gln Val Ala
Ser Val Ile Cys Ser Gly Asn Gln Ser 705 710 715 720 Gln Thr Leu Ser
Ser Cys Asn Ser Ser Ser Leu Gly Pro Thr Arg Pro 725 730 735 Thr Ile
Pro Glu Glu Ser Ala Val Ala Cys Ile Glu Ser Gly Gln Leu 740 745 750
Arg Leu Val Asn Gly Gly Gly Arg Cys Ala Gly Arg Val Glu Ile Tyr 755
760 765 His Glu Gly Ser Trp Gly Thr Ile Cys Asp Asp Ser Trp Asp Leu
Ser 770 775 780 Asp Ala His Val Val Cys Arg Gln Leu Gly Cys Gly Glu
Ala Ile Asn 785 790 795 800 Ala Thr Gly Ser Ala His Phe Gly Glu Gly
Thr Gly Pro Ile Trp Leu 805 810 815 Asp Glu Met Lys Cys Asn Gly Lys
Glu Ser Arg Ile Trp Gln Cys His 820 825 830 Ser His Gly Trp Gly Gln
Gln Asn Cys Arg His Lys Glu Asp Ala Gly 835 840 845 Val Ile Cys Ser
Glu Phe Met Ser Leu Arg Leu Thr Ser Glu Ala Ser 850 855 860 Arg Glu
Ala Cys Ala Gly Arg Leu Glu Val Phe Tyr Asn Gly Ala Trp 865 870 875
880 Gly Thr Val Gly Lys Ser Ser Met Ser Glu Thr Thr Val Gly Val Val
885 890 895 Cys Arg Gln Leu Gly Cys Ala Asp Lys Gly Lys Ile Asn Pro
Ala Ser 900 905 910 Leu Asp Lys Ala Met Ser Ile Pro Met Trp Val Asp
Asn Val Gln Cys 915 920 925 Pro Lys Gly Pro Asp Thr Leu Trp Gln Cys
Pro Ser Ser Pro Trp Glu 930 935 940 Lys Arg Leu Ala Ser Pro Ser Glu
Glu Thr Trp Ile Thr Cys Asp Asn 945 950 955 960 Lys Ile Arg Leu Gln
Glu Gly Pro Thr Ser Cys Ser Gly Arg Val Glu 965 970 975 Ile Trp His
Gly Gly Ser Trp Gly Thr Val Cys Asp Asp Ser Trp Asp 980 985 990 Leu
Asp Asp Ala Gln Val Val Cys Gln Gln Leu Gly Cys Gly Pro Ala 995
1000 1005 Leu Lys Ala Phe Lys Glu Ala Glu Phe Gly Gln Gly Thr Gly
Pro Asp 1010 1015 1020 Met Ala Gln 1025 16 994 PRT Homo sapien 16
Met Ser Lys Leu Arg Met Val Leu Leu Glu Asp Ser Gly Ser Ala Asp 1 5
10 15 Phe Arg Arg His Phe Val Asn Leu Ser Pro Phe Thr Ile Thr Val
Val 20 25 30 Leu Leu Leu Ser Ala Cys Phe Val Thr Ser Ser Leu Gly
Gly Thr Asp 35 40 45 Lys Glu Leu Arg Leu Val Asp Gly Glu Asn Lys
Cys Ser Gly Arg Val 50 55 60 Glu Val Lys Val Gln Glu Glu Trp Gly
Thr Val Cys Asn Asn Gly Trp 65 70 75 80 Ser Met Glu Ala Val Ser Val
Ile Cys Asn Gln Leu Gly Cys Pro Thr 85 90 95 Ala Ile Lys Ala Pro
Gly Trp Ala Asn Ser Ser Ala Gly Ser Gly Arg 100 105 110 Ile Trp Met
Asp His Val Ser Cys Arg Gly Asn Glu Ser Ala Leu Trp 115 120 125 Asp
Cys Lys His Asp Gly Trp Gly Lys His Ser Asn Cys Thr His Gln 130 135
140 Gln Asp Ala Gly Val Thr Cys Ser Asp Gly Ser Asn Leu Glu Met Arg
145 150 155 160 Leu Thr Arg Gly Gly Asn Met Cys Ser Gly Arg Ile Glu
Ile Lys Phe 165 170 175 Gln Gly Arg Trp Gly Thr Val Cys Asp Asp Asn
Phe Asn Ile Asp His 180 185 190 Ala Ser Val Ile Cys Arg Gln Leu Glu
Cys Gly Ser Ala Val Ser Phe 195 200 205 Ser Gly Ser Ser Asn Phe Gly
Glu Gly Ser Gly Pro Ile Trp Phe Asp 210 215 220 Asp Leu Ile Cys Asn
Gly Asn Glu Ser Ala Leu Trp Asn Cys Lys His 225 230 235 240 Gln Gly
Trp Gly Lys His Asn Cys Asp His Ala Glu Asp Ala Gly Val 245 250 255
Ile Cys Ser Lys Gly Ala Asp Leu Ser Leu Arg Leu Val Asp Gly Val 260
265 270 Thr Glu Cys Ser Gly Arg Leu Glu Val Arg Phe Gln Gly Glu Trp
Gly 275 280 285 Thr Ile Cys Asp Asp Gly Trp Asp Ser Tyr Asp Ala Ala
Val Ala Cys 290 295 300 Lys Gln Leu Gly Cys Pro Thr Ala Val Thr Ala
Ile Gly Arg Val Asn 305 310 315 320 Ala Ser
Lys Gly Phe Gly His Ile Trp Leu Asp Ser Val Ser Cys Gln 325 330 335
Gly His Glu Pro Ala Val Trp Gln Cys Lys His His Glu Trp Gly Lys 340
345 350 His Tyr Cys Asn His Asn Glu Asp Ala Gly Val Thr Cys Ser Asp
Gly 355 360 365 Ser Asp Leu Glu Leu Arg Leu Arg Gly Gly Gly Ser Arg
Cys Ala Gly 370 375 380 Thr Val Glu Val Glu Ile Gln Arg Leu Leu Gly
Lys Val Cys Asp Arg 385 390 395 400 Gly Trp Gly Leu Lys Glu Ala Asp
Val Val Cys Arg Gln Leu Gly Cys 405 410 415 Gly Ser Ala Leu Lys Thr
Ser Tyr Gln Val Tyr Ser Lys Ile Gln Ala 420 425 430 Thr Asn Thr Trp
Leu Phe Leu Ser Ser Cys Asn Gly Asn Glu Thr Ser 435 440 445 Leu Trp
Asp Cys Lys Asn Trp Gln Trp Gly Gly Leu Thr Cys Asp His 450 455 460
Tyr Glu Glu Ala Lys Ile Thr Cys Ser Ala His Arg Glu Pro Arg Leu 465
470 475 480 Val Gly Gly Asp Ile Pro Cys Ser Gly Arg Val Glu Val Lys
His Gly 485 490 495 Asp Thr Trp Gly Ser Ile Cys Asp Ser Asp Phe Ser
Leu Glu Ala Ala 500 505 510 Ser Val Leu Cys Arg Glu Leu Gln Cys Gly
Thr Val Val Ser Ile Leu 515 520 525 Gly Gly Ala His Phe Gly Glu Gly
Asn Gly Gln Ile Trp Ala Glu Glu 530 535 540 Phe Gln Cys Glu Gly His
Glu Ser His Leu Ser Leu Cys Pro Val Ala 545 550 555 560 Pro Arg Pro
Glu Gly Thr Cys Ser His Ser Arg Asp Val Gly Val Val 565 570 575 Cys
Ser Arg Tyr Thr Glu Ile Arg Leu Val Asn Gly Lys Thr Pro Cys 580 585
590 Glu Gly Arg Val Glu Leu Lys Thr Leu Gly Ala Trp Gly Ser Leu Cys
595 600 605 Asn Ser His Trp Asp Ile Glu Asp Ala His Val Leu Cys Gln
Gln Leu 610 615 620 Lys Cys Gly Val Ala Leu Ser Thr Pro Gly Gly Ala
Arg Phe Gly Lys 625 630 635 640 Gly Asn Gly Gln Ile Trp Arg His Met
Phe His Cys Thr Gly Thr Glu 645 650 655 Gln His Met Gly Asp Cys Pro
Val Thr Ala Leu Gly Ala Ser Leu Cys 660 665 670 Pro Ser Glu Gln Val
Ala Ser Val Ile Cys Ser Gly Asn Gln Ser Gln 675 680 685 Thr Leu Ser
Ser Cys Asn Ser Ser Ser Leu Gly Pro Thr Arg Pro Thr 690 695 700 Ile
Pro Glu Glu Ser Ala Val Ala Cys Ile Glu Ser Gly Gln Leu Arg 705 710
715 720 Leu Val Asn Gly Gly Gly Arg Cys Ala Gly Arg Val Glu Ile Tyr
His 725 730 735 Glu Gly Ser Trp Gly Thr Ile Cys Asp Asp Ser Trp Asp
Leu Ser Asp 740 745 750 Ala His Val Val Cys Arg Gln Leu Gly Cys Gly
Glu Ala Ile Asn Ala 755 760 765 Thr Gly Ser Ala His Phe Gly Glu Gly
Thr Gly Pro Ile Trp Leu Asp 770 775 780 Glu Met Lys Cys Asn Gly Lys
Glu Ser Arg Ile Trp Gln Cys His Ser 785 790 795 800 His Gly Trp Gly
Gln Gln Asn Cys Arg His Lys Glu Asp Ala Gly Val 805 810 815 Ile Cys
Ser Glu Phe Met Ser Leu Arg Leu Thr Ser Glu Ala Ser Arg 820 825 830
Glu Ala Cys Ala Gly Arg Leu Glu Val Phe Tyr Asn Gly Ala Trp Gly 835
840 845 Thr Val Gly Lys Ser Ser Met Ser Glu Thr Thr Val Gly Val Val
Cys 850 855 860 Arg Gln Leu Gly Cys Ala Asp Lys Gly Lys Ile Asn Pro
Ala Ser Leu 865 870 875 880 Asp Lys Ala Met Ser Ile Pro Met Trp Val
Asp Asn Val Gln Cys Pro 885 890 895 Lys Gly Pro Asp Thr Leu Trp Gln
Cys Pro Ser Ser Pro Trp Glu Lys 900 905 910 Arg Leu Ala Ser Pro Ser
Glu Glu Thr Trp Ile Thr Cys Asp Asn Lys 915 920 925 Ile Arg Leu Gln
Glu Gly Pro Thr Ser Cys Ser Gly Arg Val Glu Ile 930 935 940 Trp His
Gly Gly Ser Trp Gly Thr Val Cys Asp Asp Ser Trp Asp Leu 945 950 955
960 Asp Asp Ala Gln Val Val Cys Gln Gln Leu Gly Cys Gly Pro Ala Leu
965 970 975 Lys Ala Phe Lys Glu Ala Glu Phe Gly Gln Gly Thr Gly Pro
Asp Met 980 985 990 Ala Gln 17 745 PRT Homo sapien 17 Met Glu Arg
Pro Pro Gly Leu Arg Pro Gly Ala Gly Gly Pro Trp Glu 1 5 10 15 Met
Arg Glu Arg Leu Gly Thr Gly Gly Phe Gly Asn Val Cys Leu Tyr 20 25
30 Gln His Arg Glu Leu Asp Leu Lys Ile Ala Ile Lys Ser Cys Arg Leu
35 40 45 Glu Leu Ser Thr Lys Asn Arg Glu Arg Trp Cys His Glu Ile
Gln Ile 50 55 60 Met Lys Lys Leu Asn His Ala Asn Val Val Lys Ala
Cys Asp Val Pro 65 70 75 80 Glu Glu Leu Asn Ile Leu Ile His Asp Val
Pro Leu Leu Ala Met Glu 85 90 95 Tyr Cys Ser Gly Gly Asp Leu Arg
Lys Leu Leu Asn Lys Pro Glu Asn 100 105 110 Cys Cys Gly Leu Lys Glu
Ser Gln Ile Leu Ser Leu Leu Ser Asp Ile 115 120 125 Gly Ser Gly Ile
Arg Tyr Leu His Glu Asn Lys Ile Ile His Arg Asp 130 135 140 Leu Lys
Pro Glu Asn Ile Val Leu Gln Asp Val Gly Gly Lys Ile Ile 145 150 155
160 His Lys Ile Ile Asp Leu Gly Tyr Ala Lys Asp Val Asp Gln Gly Ser
165 170 175 Leu Cys Thr Ser Phe Val Gly Thr Leu Gln Tyr Leu Ala Pro
Glu Leu 180 185 190 Phe Glu Asn Lys Pro Tyr Thr Ala Thr Val Asp Tyr
Trp Ser Phe Gly 195 200 205 Thr Met Val Phe Glu Cys Ile Ala Gly Tyr
Arg Pro Phe Leu His His 210 215 220 Leu Gln Pro Phe Thr Trp His Glu
Lys Ile Lys Lys Lys Asp Pro Lys 225 230 235 240 Cys Ile Phe Ala Cys
Glu Glu Met Ser Gly Glu Val Arg Phe Ser Ser 245 250 255 His Leu Pro
Gln Pro Asn Ser Leu Cys Ser Leu Val Val Glu Pro Met 260 265 270 Glu
Asn Trp Leu Gln Leu Met Leu Asn Trp Asp Pro Gln Gln Arg Gly 275 280
285 Gly Pro Val Asp Leu Thr Leu Lys Gln Pro Arg Cys Phe Val Leu Met
290 295 300 Asp His Ile Leu Asn Leu Lys Ile Val His Ile Leu Asn Met
Thr Ser 305 310 315 320 Ala Lys Ile Ile Ser Phe Leu Leu Pro Pro Asp
Glu Ser Leu His Ser 325 330 335 Leu Gln Ser Arg Ile Glu Arg Glu Thr
Gly Ile Asn Thr Gly Ser Gln 340 345 350 Glu Leu Leu Ser Glu Thr Gly
Ile Ser Leu Asp Pro Arg Lys Pro Ala 355 360 365 Ser Gln Cys Val Leu
Asp Gly Val Arg Gly Cys Asp Ser Tyr Met Val 370 375 380 Tyr Leu Phe
Asp Lys Ser Lys Thr Val Tyr Glu Gly Pro Phe Ala Ser 385 390 395 400
Arg Ser Leu Ser Asp Cys Val Asn Tyr Ile Val Gln Asp Ser Lys Ile 405
410 415 Gln Leu Pro Ile Ile Gln Leu Arg Lys Val Trp Ala Glu Ala Val
His 420 425 430 Tyr Val Ser Gly Leu Lys Glu Asp Tyr Ser Arg Leu Phe
Gln Gly Gln 435 440 445 Arg Ala Ala Met Leu Ser Leu Leu Arg Tyr Asn
Ala Asn Leu Thr Lys 450 455 460 Met Lys Asn Thr Leu Ile Ser Ala Ser
Gln Gln Leu Lys Ala Lys Leu 465 470 475 480 Glu Phe Phe His Lys Ser
Ile Gln Leu Asp Leu Glu Arg Tyr Ser Glu 485 490 495 Gln Met Thr Tyr
Gly Ile Ser Ser Glu Lys Met Leu Lys Ala Trp Lys 500 505 510 Glu Met
Glu Glu Lys Ala Ile His Tyr Ala Glu Val Gly Val Ile Gly 515 520 525
Tyr Leu Glu Asp Gln Ile Met Ser Leu His Ala Glu Ile Met Glu Leu 530
535 540 Gln Lys Ser Pro Tyr Gly Arg Arg Gln Gly Asp Leu Met Glu Ser
Leu 545 550 555 560 Glu Gln Arg Ala Ile Asp Leu Tyr Lys Gln Leu Lys
His Arg Pro Ser 565 570 575 Asp His Ser Tyr Ser Asp Ser Thr Glu Met
Val Lys Ile Ile Val His 580 585 590 Thr Val Gln Ser Gln Asp Arg Val
Leu Lys Glu Leu Phe Gly His Leu 595 600 605 Ser Lys Leu Leu Gly Cys
Lys Gln Lys Ile Ile Asp Leu Leu Pro Lys 610 615 620 Val Glu Val Ala
Leu Ser Asn Ile Lys Glu Ala Asp Asn Thr Val Met 625 630 635 640 Phe
Met Gln Gly Lys Arg Gln Lys Glu Ile Trp His Leu Leu Lys Ile 645 650
655 Ala Cys Thr Gln Ser Ser Ala Arg Ser Leu Val Gly Ser Ser Leu Glu
660 665 670 Gly Ala Val Thr Pro Gln Thr Ser Ala Trp Leu Pro Pro Thr
Ser Ala 675 680 685 Glu His Asp His Ser Leu Ser Cys Val Val Thr Pro
Gln Asp Gly Glu 690 695 700 Thr Ser Ala Gln Met Ile Glu Glu Asn Leu
Asn Cys Leu Gly His Leu 705 710 715 720 Ser Thr Ile Ile His Glu Ala
Asn Glu Glu Gln Gly Asn Ser Met Met 725 730 735 Asn Leu Asp Trp Ser
Trp Leu Thr Glu 740 745 18 142 PRT Homo sapien 18 Met Gly Ala Pro
Thr Leu Pro Pro Ala Trp Gln Pro Phe Leu Lys Asp 1 5 10 15 His Arg
Ile Ser Thr Phe Lys Asn Trp Pro Phe Leu Glu Gly Cys Ala 20 25 30
Cys Thr Pro Glu Arg Met Ala Glu Ala Gly Phe Ile His Cys Pro Thr 35
40 45 Glu Asn Glu Pro Asp Leu Ala Gln Cys Phe Phe Cys Phe Lys Glu
Leu 50 55 60 Glu Gly Trp Glu Pro Asp Asp Asp Pro Ile Glu Glu His
Lys Lys His 65 70 75 80 Ser Ser Gly Cys Ala Phe Leu Ser Val Lys Lys
Gln Phe Glu Glu Leu 85 90 95 Thr Leu Gly Glu Phe Leu Lys Leu Asp
Arg Glu Arg Ala Lys Asn Lys 100 105 110 Ile Ala Lys Glu Thr Asn Asn
Lys Lys Lys Glu Phe Glu Glu Thr Ala 115 120 125 Lys Lys Val Arg Arg
Ala Ile Glu Gln Leu Ala Ala Met Asp 130 135 140 19 165 PRT Homo
sapien 19 Met Gly Ala Pro Thr Leu Pro Pro Ala Trp Gln Pro Phe Leu
Lys Asp 1 5 10 15 His Arg Ile Ser Thr Phe Lys Asn Trp Pro Phe Leu
Glu Gly Cys Ala 20 25 30 Cys Thr Pro Glu Arg Met Ala Glu Ala Gly
Phe Ile His Cys Pro Thr 35 40 45 Glu Asn Glu Pro Asp Leu Ala Gln
Cys Phe Phe Cys Phe Lys Glu Leu 50 55 60 Glu Gly Trp Glu Pro Asp
Asp Asp Pro Ile Gly Pro Gly Thr Val Ala 65 70 75 80 Tyr Ala Cys Asn
Thr Ser Thr Leu Gly Gly Arg Gly Gly Arg Ile Thr 85 90 95 Arg Glu
Glu His Lys Lys His Ser Ser Gly Cys Ala Phe Leu Ser Val 100 105 110
Lys Lys Gln Phe Glu Glu Leu Thr Leu Gly Glu Phe Leu Lys Leu Asp 115
120 125 Arg Glu Arg Ala Lys Asn Lys Ile Ala Lys Glu Thr Asn Asn Lys
Lys 130 135 140 Lys Glu Phe Glu Glu Thr Ala Lys Lys Val Arg Arg Ala
Ile Glu Gln 145 150 155 160 Leu Ala Ala Met Asp 165 20 137 PRT Homo
sapien 20 Met Gly Ala Pro Thr Leu Pro Pro Ala Trp Gln Pro Phe Leu
Lys Asp 1 5 10 15 His Arg Ile Ser Thr Phe Lys Asn Trp Pro Phe Leu
Glu Gly Cys Ala 20 25 30 Cys Thr Pro Glu Arg Met Ala Glu Ala Gly
Phe Ile His Cys Pro Thr 35 40 45 Glu Asn Glu Pro Asp Leu Ala Gln
Cys Phe Phe Cys Phe Lys Glu Leu 50 55 60 Glu Gly Trp Glu Pro Asp
Asp Asp Pro Met Gln Arg Lys Pro Thr Ile 65 70 75 80 Arg Arg Lys Asn
Leu Arg Lys Leu Arg Arg Lys Cys Ala Val Pro Ser 85 90 95 Ser Ser
Trp Leu Pro Trp Ile Glu Ala Ser Gly Arg Ser Cys Leu Val 100 105 110
Pro Glu Trp Leu His His Phe Gln Gly Leu Phe Pro Gly Ala Thr Ser 115
120 125 Leu Pro Val Gly Pro Leu Ala Met Ser 130 135 21 142 PRT Homo
sapien 21 Met Gly Ala Pro Thr Leu Pro Pro Ala Trp Gln Pro Phe Leu
Lys Asp 1 5 10 15 His Arg Ile Ser Thr Phe Lys Asn Trp Pro Phe Leu
Glu Gly Cys Ala 20 25 30 Cys Thr Pro Glu Arg Met Ala Glu Ala Gly
Phe Ile His Cys Pro Thr 35 40 45 Glu Asn Glu Pro Asp Leu Ala Gln
Cys Phe Phe Cys Phe Lys Glu Leu 50 55 60 Glu Gly Trp Glu Pro Asp
Asp Asp Pro Ile Glu Glu His Lys Lys His 65 70 75 80 Ser Ser Gly Cys
Ala Phe Leu Ser Val Lys Lys Gln Phe Glu Glu Leu 85 90 95 Thr Leu
Gly Glu Phe Leu Lys Leu Asp Arg Glu Arg Ala Lys Asn Lys 100 105 110
Ile Ala Lys Glu Thr Asn Asn Lys Lys Lys Glu Phe Glu Glu Thr Ala 115
120 125 Lys Lys Val Arg Arg Ala Ile Glu Gln Leu Ala Ala Met Asp 130
135 140 22 165 PRT Homo sapien 22 Met Gly Ala Pro Thr Leu Pro Pro
Ala Trp Gln Pro Phe Leu Lys Asp 1 5 10 15 His Arg Ile Ser Thr Phe
Lys Asn Trp Pro Phe Leu Glu Gly Cys Ala 20 25 30 Cys Thr Pro Glu
Arg Met Ala Glu Ala Gly Phe Ile His Cys Pro Thr 35 40 45 Glu Asn
Glu Pro Asp Leu Ala Gln Cys Phe Phe Cys Phe Lys Glu Leu 50 55 60
Glu Gly Trp Glu Pro Asp Asp Asp Pro Ile Gly Pro Gly Thr Val Ala 65
70 75 80 Tyr Ala Cys Asn Thr Ser Thr Leu Gly Gly Arg Gly Gly Arg
Ile Thr 85 90 95 Arg Glu Glu His Lys Lys His Ser Ser Gly Cys Ala
Phe Leu Ser Val 100 105 110 Lys Lys Gln Phe Glu Glu Leu Thr Leu Gly
Glu Phe Leu Lys Leu Asp 115 120 125 Arg Glu Arg Ala Lys Asn Lys Ile
Ala Lys Glu Thr Asn Asn Lys Lys 130 135 140 Lys Glu Phe Glu Glu Thr
Ala Lys Lys Val Arg Arg Ala Ile Glu Gln 145 150 155 160 Leu Ala Ala
Met Asp 165 23 137 PRT Homo sapien 23 Met Gly Ala Pro Thr Leu Pro
Pro Ala Trp Gln Pro Phe Leu Lys Asp 1 5 10 15 His Arg Ile Ser Thr
Phe Lys Asn Trp Pro Phe Leu Glu Gly Cys Ala 20 25 30 Cys Thr Pro
Glu Arg Met Ala Glu Ala Gly Phe Ile His Cys Pro Thr 35 40 45 Glu
Asn Glu Pro Asp Leu Ala Gln Cys Phe Phe Cys Phe Lys Glu Leu 50 55
60 Glu Gly Trp Glu Pro Asp Asp Asp Pro Met Gln Arg Lys Pro Thr Ile
65 70 75 80 Arg Arg Lys Asn Leu Arg Lys Leu Arg Arg Lys Cys Ala Val
Pro Ser 85 90 95 Ser Ser Trp Leu Pro Trp Ile Glu Ala Ser Gly Arg
Ser Cys Leu Val 100 105 110 Pro Glu Trp Leu His His Phe Gln Gly Leu
Phe Pro Gly Ala Thr Ser 115 120 125 Leu Pro Val Gly Pro Leu Ala Met
Ser 130 135 24 132 PRT Homo sapien 24 Met Ala Phe Asp Ser Thr Trp
Lys Val Asp Arg Ser Glu Asn Tyr Asp 1 5 10 15 Lys Phe Met Glu Lys
Met Gly Val Asn Ile Val Lys Arg Lys Leu Ala 20 25 30 Ala His Asp
Asn Leu Lys Leu Thr Ile Thr Gln Glu Gly Asn Lys Phe 35 40 45 Thr
Val Lys Glu Ser Ser Ala Phe Arg Asn Ile Glu Val Val Phe Glu 50 55
60 Leu Gly Val Thr Phe Asn Tyr Asn Leu Ala Asp Gly Thr Glu Leu Arg
65 70 75 80 Gly Thr Trp Ser Leu Glu Gly Asn Lys Leu Ile Gly Lys Phe
Lys Arg 85 90 95 Thr Asp Asn Gly Asn Glu Leu Asn Thr Val Arg Glu
Ile Ile Gly Asp 100 105 110 Glu Leu Val
Gln Thr Tyr Val Tyr Glu Gly Val Glu Ala Lys Arg Ile 115 120 125 Phe
Lys Lys Asp 130 25 201 DNA Homo sapien 25 ggctgtgcag acaaagggaa
aatcaaccct gcatctttag acaaggccat gtccattccc 60 atgtgggtgg
acaatgttca gtgtccaaaa ggacctgaca ygctgtggca gtgcccatca 120
tctccatggg agaagagact ggccagcccc tcggaggaga cctggatcac atgtgacaac
180 aagataagac ttcaggaagg a 201 26 201 DNA Homo sapien 26
ggctgtgcag acaaagggaa aatcaaccct gcatctttag acaaggccat gtccattccc
60 atgtgggtgg acaatgttca gtgtccaaaa ggacctgaca ygctgtggca
gtgcccatca 120 tctccatggg agaagagact ggccagcccc tcggaggaga
cctggatcac atgtgacaac 180 aagataagac ttcaggaagg a 201 27 201 DNA
Homo sapien 27 ggctgtgcag acaaagggaa aatcaaccct gcatctttag
acaaggccat gtccattccc 60 atgtgggtgg acaatgttca gtgtccaaaa
ggacctgaca ygctgtggca gtgcccatca 120 tctccatggg agaagagact
ggccagcccc tcggaggaga cctggatcac atgtgacaac 180 aagataagac
ttcaggaagg a 201 28 201 DNA Homo sapien 28 ggctgtgcag acaaagggaa
aatcaaccct gcatctttag acaaggccat gtccattccc 60 atgtgggtgg
acaatgttca gtgtccaaaa ggacctgaca ygctgtggca gtgcccatca 120
tctccatggg agaagagact ggccagcccc tcggaggaga cctggatcac atgtgacaac
180 aagataagac ttcaggaagg a 201 29 201 DNA Homo sapien 29
taagaagaag gatccaaagt gtatatttgc atgtgaagag atgtcaggag aagttcggtt
60 tagtagccat ttacctcaac caaatagcct ttgtagttta rtagtagaac
ccatggaaaa 120 ctggctacag ttgatgttga attgggaccc tcagcagaga
ggaggacctg ttgaccttac 180 tttgaagcag ccaagatgtt t 201 30 201 DNA
Homo sapien 30 attaaccctt ggtgaatttt tgaaactgga cagagaaaga
gccaagaaca aaattgcaaa 60 ggaaaccaac aataagaaga aagaatttga
ggaaactgcg ragaaagtgc gccgtgccat 120 cgagcagctg gctgccatgg
attgaggcct ctggccggag ctgcctggtc ccagagtggc 180 tgcaccactt
ccagggttta t 201 31 201 DNA Homo sapien 31 attaaccctt ggtgaatttt
tgaaactgga cagagaaaga gccaagaaca aaattgcaaa 60 ggaaaccaac
aataagaaga aagaatttga ggaaactgcg ragaaagtgc gccgtgccat 120
cgagcagctg gctgccatgg attgaggcct ctggccggag ctgcctggtc ccagagtggc
180 tgcaccactt ccagggttta t 201 32 201 DNA Homo sapien 32
agtgtttctt ctgcttcaag gagctggaag gctgggagcc agatgacgac cccatgcaaa
60 ggaaaccaac aataagaaga aagaatttga ggaaactgcg ragaaagtgc
gccgtgccat 120 cgagcagctg gctgccatgg attgaggcct ctggccggag
ctgcctggtc ccagagtggc 180 tgcaccactt ccagggttta t 201 33 201 DNA
Homo sapien 33 attaaccctt ggtgaatttt tgaaactgga cagagaaaga
gccaagaaca aaattgcaaa 60 ggaaaccaac aataagaaga aagaatttga
ggaaactgcg ragaaagtgc gccgtgccat 120 cgagcagctg gctgccatgg
attgaggcct ctggccggag ctgcctggtc ccagagtggc 180 tgcaccactt
ccagggttta t 201 34 201 DNA Homo sapien 34 attaaccctt ggtgaatttt
tgaaactgga cagagaaaga gccaagaaca aaattgcaaa 60 ggaaaccaac
aataagaaga aagaatttga ggaaactgcg ragaaagtgc gccgtgccat 120
cgagcagctg gctgccatgg attgaggcct ctggccggag ctgcctggtc ccagagtggc
180 tgcaccactt ccagggttta t 201 35 201 DNA Homo sapien 35
agtgtttctt ctgcttcaag gagctggaag gctgggagcc agatgacgac cccatgcaaa
60 ggaaaccaac aataagaaga aagaatttga ggaaactgcg ragaaagtgc
gccgtgccat 120 cgagcagctg gctgccatgg attgaggcct ctggccggag
ctgcctggtc ccagagtggc 180 tgcaccactt ccagggttta t 201 36 201 DNA
Homo sapien 36 aatgggtgtt aatatagtga aaaggaagct tgcagctcat
gacaatttga agctgacaat 60 tacacaagaa ggaaataaat tcacagtcaa
agaatcaagc rcttttcgaa acattgaagt 120 tgtttttgaa cttggtgtca
cctttaatta caatctagca gacggaactg aactcagggg 180 gacctggagc
cttgagggaa a 201 37 52764 DNA Homo sapien misc_feature 6568, 6569,
6570, 6571, 6572, 6573, 6574, 6575, 6576, 6577, 6578, 6579, 6580,
6581, 6582, 6583, 6584, 6585, 6586, 6587, 6588, 6589, 6590, 6591,
6592, 6593, 6594, 6595, 6596, 6597, 6598, 6599, 6600, 6601, 6602,
6603, 6604, 6605, 6606 n = A,T,C or G misc_feature 6607, 6608,
6609, 6610, 6611, 6612, 6613, 6614, 6615, 6616, 6617, 6618, 6619,
6620, 6621, 6622, 6623, 6624, 6625, 6626, 6627, 6628, 6629, 6630,
6631, 6632, 6633, 6634, 6635, 6636, 6637, 6638, 6639, 6640, 6641,
6642, 6643, 6644, 6645 n = A,T,C or G misc_feature 6646, 6647,
6648, 6649, 6650, 6651, 6652, 6653, 6654, 6655, 6656, 6657, 6658,
6659, 6660, 6661, 6662, 6663, 6664, 6665, 6666, 6667, 6668, 6669,
6670, 6671, 6672, 6673, 6674, 6675, 6676, 6677, 6678, 6679, 6680,
6681, 6682, 6683, 6684 n = A,T,C or G misc_feature 6685, 6686,
6687, 6688, 6689, 6690, 6691, 6692, 6693, 6694, 6695, 6696, 6697,
6698, 6699, 6700, 6701, 6702, 6703, 6704, 6705, 6706, 6707, 6708,
6709, 6710, 6711, 6712, 6713, 6714, 6715, 6716, 6717, 6718, 6719,
6720, 6721, 6722, 6723 n = A,T,C or G misc_feature 6724, 6725,
6726, 6727, 6728, 6729, 6730, 6731, 6732, 6733, 6734, 6735, 6736,
6737, 6738, 6739, 6740, 6741, 6742, 6743, 6744, 6745, 6746, 6747,
6748, 6749, 6750, 6751, 6752, 6753, 6754, 6755, 6756, 6757, 6758,
13921, 13922, 13923 n = A,T,C or G misc_feature 13924, 13925,
13926, 13927, 13928, 13929, 13930, 13931, 13932, 13933, 13934,
13935, 13936, 13937, 13938, 13939, 13940, 17414, 17415, 17416,
17417, 17418, 17419, 17420, 17421, 17422, 17423, 17424, 17425,
17426, 17427, 17428, 17429 n = A,T,C or G 37 ctgactttga caaaattagt
gactttattc cgaaaaggga cacaatgaga ttactaacaa 60 gcctcaaatc
tattccaaaa ttcagaacaa gtttcaaaat ctgataccat tccctttatt 120
tattcctctt ctttttccca ttccccatga atatgcccac ataaaaatgt cacaaatttc
180 tgagtaaatc atctgctaac ctacaggtcc actcaacatt tttgtgtctt
cagcttcccc 240 caagcctggc cctttttgga gggtttgact tcacaattgt
tgtaaatctt aaagattgag 300 ctgctggtgg acttctctct ttcttttgtc
ctccaccaat aatagctcac aaagattttt 360 aaaaacatct ttattggagc
atagttaata tacaataagc tgcacatatt taagtgttcc 420 attagtttag
ttttgacgta tgtgtatacc tgtgaaatta tcaacacaat aaagacaatg 480
aacatatcaa ttactccaaa aaacttcatc atgattaccc ctcattgtga ttctttcctt
540 gcactccttc ccatccatct ctaagcaacc actgcatcta gtcacagtcg
attaattttc 600 actttccaga attctatata aatagaatca cacggtataa
gatttgtaag aatatactgt 660 ccaacactgc tattgaaatt aacctatttt
ctgtttttac agcaccaata acaattagct 720 tttaaacttt atcttttgcc
attatctaga cttgcacact taatcacaac attttacaag 780 agtaaaattt
cacagtctct ttcactgtgt cataaaatac cgtaagaagg aatcaatttg 840
gcttccccaa taccaaatct gtattagtca gggtccttca gagaggcgga tccagtaaga
900 catatatata tatagacatt gagagagaat ttattagggg aattggcttg
cataattttg 960 gtggctgaaa agtcccatga taggctttct gtaagctgaa
gaccatggga tgactgtagc 1020 atgactcagc ccaagtccaa aggcctgagc
accaggggag cccatggtgt aactctcagt 1080 ctgagaccaa aagtcttagc
attctggagc cactccagat gtaagtgctg aagtcgaaag 1140 gccagtgagc
ctgcagttct gctgtccaaa gtagcacaag aaaagtctgg cccagctctc 1200
aaagagagac caattttcct tctgtatttg ttctctctgg atccctgatg caggccagcg
1260 ctgagggaag gtcttcccca cctagtccac tcagactcac acactaatct
tatccattat 1320 tttggatgtg tctggaagca ctcttacaga catgaaaaat
aatgctttat caagtgtcta 1380 gaatccttaa tccaatcaag ttgacaccta
aaattatgtc tataaatcca ccccttgtca 1440 acttggcacc catacttaat
tgcatctcct taacctatat ttaatttcca aataaagaca 1500 ataccaaggg
aatagttccg cctaacgtgt tgtaaacaac agaatgcaac tatcctgtat 1560
gcaaccaaaa atgtactgat ctgttccaca gaattcaact ttcagaattt caacattctg
1620 aattgtaatt tttgcaattt ttgatgttag gaatttttag atttcagaga
tgttgatctt 1680 tagggatttt gaactttggg attttaacat ttaggattat
ggcatttgta attgcatgtt 1740 ttgagattat aatcagcact ctccccaact
ccccaaaaaa gaaactacac aacatagact 1800 ccgagtttta taaacagcaa
tttatttctt tgttttggag gctggaagtt caagatcaag 1860 atgctgtcag
attcagtgtc tggtgagagt ccacttccag ggttatagat taccattttt 1920
tagctgtact tcacacggca aaagggatca gtcttctctc agggatctct ttcatgaggg
1980 caataacccc attcatgaag gcagagccct catgacctaa ttatctccca
aaggccctac 2040 ctactaatac ctttgaggtt aggatttcaa catatgaatt
ttgaggagac acaaacattt 2100 agaccatagc actaagtaat attttgttta
attccagaca ttgtaaacta aacttcgctg 2160 agttttaaat tatttttatt
gctttagata ttctaacgtt tagtctagaa tgttgttata 2220 ttcttggaaa
caatttgatt ctttgggtct tgtctttaaa atttgttagg tgggatctaa 2280
tcagcaagta gctagggcta cttatttccc accactgtgg taagactcat ctgtgaactc
2340 tacccaatac cccatgactt gtgagttttc catgtgaaca gtcacttttc
ctggccctgt 2400 gtgtaactct aatcctttca ttattctttc cccaggctca
ggtgatttgc tcacatgcat 2460 gcacttaaca atactcagct gaatactcaa
aggggactct acagatattc agctctctct 2520 ccttgcctcc tccctactct
ctctctctct ctctttctct ctctctttct tcttgcaagt 2580 ttctcctctg
tgagactatt tgctgtgaat tctagccacc ttagtctccc agattctcag 2640
ctgcttcttc ccaactctga gactcctctg atcattctct gggtttccta ttccatggta
2700 gcctggaaac tatctcaagg cagtaaactc agcttagctt ctcatctctc
atggatcact 2760 atctttgaga tccaggcaac aactcttctt gttgcctcat
tttcagtgtc ttgaaaatca 2820 ttgttacaca tgtttggctt tgtttttagt
tgtttctggt gggacagtaa acacacttcc 2880 tgtttcttca ttttgaataa
aagcaaaaat gtcttgttca cctagctttt taattttgca 2940 ccaatctatt
tcttccattt ttctactgta tcttttcaat cgggctctta ttctattaga 3000
tttttgctat aatattcact tgcactgcat ttaatcttgc tcttatttcc atttttaccc
3060 tggccaccaa tcatcctcaa agttgctacc aagatgagat atgtaaaaga
caaagcaatt 3120 taatactcat gtttcaaact atttaatgtc attacttgcc
tacagtgtaa aattataatt 3180 ctttaacatg gtatataaaa ctcctcttca
tgtagtcctt gtttacctca gcaataataa 3240 caacattgtc aacaacaaca
acaaaatagg taatattgag cctagcatct ggcagggact 3300 ggtagatatc
tttttgaaaa aaacacagtt cctatcctgg tatcttaaga ctagcaagta 3360
agataaacaa taaatataaa aatataattg ggaggggtgg agccaagatg gccaaatagg
3420 aacagctccg gtctacagct cccagcgtga acaacgcaga agacgtgtga
tttctgcatt 3480 tccatctgag gtactgggtt catctcacta gggagtgcca
gacagtgggt gcaggatagt 3540 gggtgcagcg cactgtgcac aaaccgaagc
agggcgaggc attgcctcac tagagaagcg 3600 caagaggtca gggagttccc
tttcccagtc aaagaaaggg gtgacagagg gcacctggaa 3660 aatcaggtca
ctcccaccct aatactgcgc ttttccaacg ggcttaaaaa acagcacacc 3720
aggagattat atcccgcaca tggctcggag ggtcctacac ccatggagtc tcactgattg
3780 ctagcacagc agtctgagat caaactgcaa ggcagcagcg aagctggggg
aggggcgcct 3840 gccattgccc aggctcgatt aggtaaacaa agcagctggg
aacctcgaac tgggtggagc 3900 ccaccacagc tcaaggaggc ctgcctgcct
ctgtaggctc cacctctggg ggcagggtac 3960 agacaaacaa aaagacagca
gtaacctctg cagacttaaa tgtccctgtc tgacaacttt 4020 gaagagagta
gtggttctcc cagcacgcag ctggagatct gcgaatgtgc agactgcctc 4080
ctaaagtggg tccatgaccc ccgagcagcc taactgggag gcatccccca gtaggagcag
4140 actgacacct cacacggccg ggtgctcctc tgagacaaaa cttccagagg
aacaatcagg 4200 cagcagcatt tgcagttcac caagatccgc tcttctacag
ccactgctgt tctgcagcca 4260 ctgctgctga tacacaggca aacagggtct
ggagtggacc tccagcaaac tccaacagac 4320 ctgcagctga gggtcctgac
tgttagaagg aaaactaaca aacagaaagg acatctgcac 4380 caaaaaccct
tctgtacatc atcatcatca aagaccaaaa gtagataaaa ccacaaagat 4440
ggggaagaaa cagagcagaa aaactggaaa ctctaaaaag cagagtgcct ctgctactcc
4500 gaaggaatgc agctcctcac cagcaatgga acaaagctgg atggagaatg
actttgacga 4560 gctgagagaa gaagtcttca gatgatcaaa ctactccgag
ctacaggagg aaattcaaat 4620 caatggcaaa gaagttaaaa actgtgaaaa
aaaaatagat gaatggataa ctagaataac 4680 caacgcagag aagtccttaa
aggagctgat ggagctgaaa gccaaggctc aagaactacg 4740 tgaagaatgc
agaaggctca ggagccaatg cgatcaactg gaagaaaggt tatcagtgat 4800
ggaagacaaa atgaatgaaa tgaagtgaga agggaagttt agagaaaaaa gaataaaaag
4860 aaacaaagcc tccaagaaat atgggactat gtgaaaagac caaatctacg
tctgattggt 4920 gtacctgaaa gtgatgggga gaatggaacc aagttggaaa
acactctgca ggatattatc 4980 catgagaact tccccaatct agcaaggcag
gccaacatac agattcagga aatacagaga 5040 acaccacaaa gatactcctc
gagaagagca actccaagac acataattgt cagattcacc 5100 aaagttgaaa
tgaaggaaaa aatgttaagg gcagccagag agaaaggtcg ggttacccac 5160
aaagggaagc ccatcagact aacagcggat ctctcggcag aaactctaca agccagaaga
5220 gagtgggggc caatattcaa cattcttaaa gaaaagaatt ttcaacccag
aatttcatat 5280 ccagccaaac taagcttcat aagtgaagga gaaataactt
tacagacaag caaatgctga 5340 gagattttgt caccaccagg cctgccctaa
aagagctcct gaaggaagca ctaaacatag 5400 aaaggaacaa ctggtaccag
ccactgccaa aacatgacaa aatgtaaaga ctatcaaggc 5460 taggaagaaa
ctgcatcaac agaaatcata gtaaactgtc tctcagacca cagtgcaatc 5520
aaactagaac tcaggattaa gaaattcact caaaactgct caaccacatg gaaactgaac
5580 aacctgctcc tgaatgacta ctgggtaaat aatgaaatga aggcagaaat
aaagatgttc 5640 tttgaaacca acgagaacaa agacacaaca taccagaatc
tttgggacac attcaaagca 5700 gtgtgtagag ggaaatttat agcactaaat
acccacaaga gaaagcagga aagatccaaa 5760 attgacaccc taacgtcaca
attaaaagaa ctagaaaagc aagagcaaac acattcaaaa 5820 gctagcagaa
ggcaagaaat aactaaaatc agagcagaac tgaaggaaat agagacacaa 5880
aaaacctttc aaaaaattaa tgaatctggg agctggtttt ttgaaaacgt caacaaaatc
5940 gatagactgc tagcaagact aataaagaag acaagaaaga agaatcaaat
agacacaata 6000 aaaattgata aaggggatat caccaccgat cccacagaaa
tacaaactac catcagagaa 6060 tactacaaac acctctacgc aaataaacta
gaaaatctag aagaaatgga taaattcctc 6120 gacacataca ccctcccaag
actaagcgag gaagaaattc aatctctgaa tagaccaata 6180 acaggctctg
aaattgtggc aataatcaat agcttaccaa ccaaaaaaag tccaggagca 6240
ggtggattca cagccaaatt ctaccagagg tacaaggagg agttggtacc attccttctg
6300 aaactattcc aatcaataga aaaagaggga atcctcccta actcattatt
ttatgaggcc 6360 agcatcatcc tgataccaaa gcctggcaga gacacaacca
aaaaagagaa ttttagacca 6420 atatccttga tgaacattga tgcaaaaatc
ctcgataaaa tactggcaaa ccaaatccag 6480 cagcacatca aaaagcttat
ccaccatgat caagtgggct tcttcgctgg gatgcaaggg 6540 tggttcaaca
tactcaaatc aataaatnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 6600
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
6660 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
nnnnnnnnnn 6720 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnca
ccactcctat tcaagatagt 6780 gttagaagtt ctggccatgg caattaggca
ggagaagaaa ataaagggta ttcaattagg 6840 aaaagaggaa gtcaaaattg
tccctgtttg cagatgacat gattgtatat ttagaaaacc 6900 cccatcgtct
cagcccaaaa tctccttaag ctgataagca acttcagcaa agtctcagga 6960
tacaaaatca atgtacaaaa atcacaagca ttcttataca ccaataacag acaaacagag
7020 agccaaatca tgagtgaact cccattcaca attgcttcaa agagaataaa
atacctagga 7080 atccaactta caagggatgt gaaggacctc ttcaaggaga
actacaaacc actgctcaat 7140 gaaataaaag aggataccaa caaatggaag
aacattccat gctcatgggt aggaagaatc 7200 aatatcatga aaatggccat
actgcccaag gtaatttata gattcaatgc catccccatc 7260 aagctaccaa
tgactttctt cacagaattg gaaaaaacta ctttaaagtt catatggaac 7320
caaaaaagag cccacatccc caagtcaatc ctaagccaaa agaacaaagc tggaggcatc
7380 acgctacctg acttcaaact atactacaag gctacagtaa ccaaaacagc
atggtactgg 7440 taccaaaaca gagatataga tcaatggaac agaacagagc
cctcagaaat aatgccgcat 7500 atctacaacc atctgatctt tgacaaacct
gacaaaaaca agcaatgggg aaaggattcc 7560 ctatttaata aatgttgctg
ggaaaactgg ctagccatat gtagaaagct gaaactggat 7620 cccttcctta
caccttatac aaaaattaat tcaagatgga ttaaagactt acatgttaga 7680
cctaaaacca taaaaaccct agaagaaaac ctaggcaata ccattcagga cataggcatg
7740 ggcaaggact tcatatctaa aacaccaaaa gcaatggcaa caaaagccaa
aatagacaaa 7800 tgggatctaa ttaaactaaa gagcttctgc acagcaaaag
aaactaccat cagagtgaac 7860 aggaaatcta caaaatggga gaaaattttc
gcaacctact catctgacaa agggttaata 7920 tccagaatct acaatgacct
caaacaaatt tacaagaaaa aaacaaacaa ccccatcaaa 7980 aattggcgaa
ggatatgaac agatacttct caaaagaaga catttatgca gccaaaaaac 8040
acatgaaaaa atgctcatca tcactggcca tcagagaaac gcaaatcaaa accacaagga
8100 gataccatct cacaccagtt agaatggtga tcattaaaaa gtcaggaaac
aataggtgct 8160 ggagaggatg tggagaaata ggcatgcttt tacactgttg
gtgggactgt aaactaattc 8220 aaccattgtg gaagtcagtg tggcaattcc
tcagggatct agaactagaa ataccatttg 8280 acccagccat cccatttctg
ggtatatacc caaaggatta taaatcatgc tgctataaag 8340 acacatgcac
acgtacgttt atagcggcac tactcgcaat agcaaagact tggaaccaac 8400
ccaaatgtcc atcaatgata gactggatta agaaaatgtg gcacatatat accatggaat
8460 actatgcagc cataaaaaat gatgagttca tgtcctttgt agggacatgg
atgaaactgg 8520 aaaccatcat tctcagcaaa ccatcgcaag gacaaaaaac
caaacaccac atgttctcac 8580 tcataggtgg gaattgaaca atgagaacac
atggacacag gaaggggaac atcccactcc 8640 agggacggtt gtggggtcag
gggagggggg agggataaca ttaggagata tacctaatgc 8700 taaatgacga
gttaatgggt gcagcacacc agcattgcac atgtatacat acgtaacaaa 8760
cctgcatgtt gtgcacatgt atcctaaaac ttaaagtata ataataattt aaaaaaagga
8820 aaaaaatata tgtataattg acaaccctca taagtacagg ctatcatgag
tgcatatttt 8880 ctggggtctt tcagcgttag agttatttct cctagtgaag
ggaggtttaa ctgagaaata 8940 aagtatatgt gagtgagagt tatcttagag
aaaggtacgg gtagggaggc agtaaacaca 9000 gaaggaacaa tatgtatgac
ataccatgag gcagggagag atttggcatg cccgagaagt 9060 ggaagaaaac
tggggtgcct ttgttgactg ggttctagtg aatgtctctc tggaaggctg 9120
gagttgctct ttaattcccc atttcaggcc catcactaac acctgacaat atgtatgcca
9180 tgatgaacta ttagtagttc ttttgagctt atatgttttt atatacaaat
attcatttgt 9240 atgtgctatt tttatgcctt aaactccttt atcatttgtc
cctttggcaa tgtttttctc 9300 atcttttagg acacaattta agcctgtcct
cagagaaaat agttttctga ctgttcattc 9360 ctttctctca atagatagga
ctactttgtc cattattttt aacactgcac tttagcttta 9420 tgttcattgt
tgttatcata tgtgtttctc ttacttgaat ctgagttata tggagctaaa 9480
atcatgtgtt gctaattttt gtttcaccat ttgtaatatc agtattagtc atggctaatt
9540 ctcttggtgt acttcatccc attagaaaag aaatgacaaa tgctgtgtct
caacaactta 9600 cacaaaatta ctcattagac acatttgatt atggaaataa
aattaaaagt gcatatgata 9660 aaatgttatt taattatgtt ttgcctgttt
tgctttagtt ttttacataa tttttctaca 9720 tgacaattag taattttttg
tgtcttatat atttgtccaa aatgaagttc aaaaatgtaa 9780 aatatttaat
tcagcaacag cagcatatga gttagtattt cctctaattt ttcgaaatct 9840
gtgggaagtg tttcccaatt tcctttggtt gtttcatgtg ctatattgaa gaaaacatga
9900 gtatgaaatg gaacctcagc tctttcaatg acttcccttt ttgagttgac
tccgcctcca 9960 tatgtagcct tttcattttc atgaaagtga agtgattttt
agaattctta gttgttttct 10020 ttagaagaac atttctaggg aataatacaa
gaagatttag gaatcattga agttataaat 10080 ctttggaatg agcaaactca
gaatggtgct acttgaagac tctggatctg ctggtaaaag 10140 cttctcattt
attctacatt tcccctttaa tggggtatgt aattattatg acagtcaatg 10200
gatgtgattt aaaagtgatt ggcatcagga gagtaaggag tggaaaaagg acataggctt
10260 atatggcagc acctttgatc tgccatagat tcaaaattga agatgtgata
tgggaatcag 10320 agtcggcatt
aattttgtaa aactgctgag gtgaaattaa gtcaggaact aaagttaact 10380
gaaatgtact gagtatgaaa aaatagttgt tagaaaagat agaatatgca gaaaggaata
10440 ttgatagaga gttaattaca taaagaaagt tatctactct gataagtgag
ttatctattt 10500 taagacttat ttggacatgt taaaatcatt tagcattttt
agttttaaat tagtgttggg 10560 ttagagtaac gagaagatat gagagtaata
acagcattaa agagtgtatt atgtgataag 10620 gatacaggta gagatgacaa
gatttaatat ttgagttttt cagatatgga tatcatcact 10680 taggtattgt
tctgctagct actatttggc aagcatcaag ccaggtactg gtgaaattat 10740
ggtgaacatg atagatataa ttcgtttact catggaactt atcctctatc agtgcggcta
10800 ggtagtctta aagatggggg catccataat tgaatatcaa ggtggttagg
gaatagagaa 10860 atctcaggat accttccctg tgcctgctgg gagaattgtc
cactactaag tttcattgat 10920 gttgtttcca ttttccaggt tttaagggtt
gaccttgcaa ttctcgtttc caccaccaac 10980 cttctgctaa tccgtcacct
ctaggttgcc tataaacatg aacattttca aaaccatttc 11040 actgaaacta
tatgagcctg ggtatgaatg ttttctgcct gccttgcaat ttagattgaa 11100
gagttttctg tcttggtttg agcttctttg ttaattaacc tctgaaacca tctatcttct
11160 ccaaagtctc tttttgggaa gatgctactt ttgccctctt ttctttttca
cagacttcag 11220 aagacatttt gtcaacctga gtcccttcac cattactgtg
gtcttacttc tcagtgcctg 11280 ttttgtcacc agttctcttg gtgagtactt
tgtcaaattt acttacagcc ttagcccact 11340 ctgacaagaa cagttaaaag
gcaaacaatt tttctgaagg ccaagtagca tctaaaaact 11400 aggtgacatg
ctccatgtat tcaaaggatc agatattatt aaaagcacac aataaaaatc 11460
taaccaaaat attagcatgg ttcctcacca aatgaaataa actcacagga gagaaatgaa
11520 ttttacttaa agattagaaa taatgttgaa ctgaaaaata ggaaagaata
caaaggagga 11580 gaaacttcag aatcattaaa acaaaagtga atttgggaaa
cacattagta aattcaaatg 11640 ccaaaattta tctaaaaatg tattgagaga
tgccaggttg gtgggaccag attcctacat 11700 aaagctaaaa taatgccaag
gaggcaactg agatcatcta tcatctgggg atgggaagag 11760 agtacaaaga
ggtttatttt atggtagaaa atagcagaaa atatctccat cttaatttta 11820
gataagtttc agtctagcgt ttttaagatt tatgctattt tacatatacg tatgaaataa
11880 gagttaaatt tcttacaact agtcttttca tcttcataaa ttctctatca
gccttctttc 11940 tatcgacctt aggattttgg agaaatgtaa aaagttcatc
cccataacgt tttgtagcat 12000 cttatttaac tcgaaaatgc ttcacaggaa
ttatataaat tgccactatt aaggctttcc 12060 agccataaaa taaagctatt
aaacactatt gtgttactta cctttactgg tcatatagca 12120 ctgcttaatt
tgaaaccagt gatactgata cttatatttg aatgactgtg aaataacatt 12180
cttctgtttt accatggata ggactaggat ggttctcaaa tctgagtttg aaggataaat
12240 aagtaatgta aaaaaataat tttctacttg ctctttgcga ccttggccaa
ttactgtctt 12300 aatccattgt tgcaggagga acagacaagg agctgaggct
agtggatggt gaaaacaagt 12360 gtagcgggag agtggaagtg aaagtccagg
aggagtgggg aacggtgtgt aataatggct 12420 ggagcatgga agcggtctct
gtgatttgta accagctggg atgtccaact gctatcaaag 12480 cccctggatg
ggctaattcc agtgcaggtt ctggacgcat ttggatggat catgtttctt 12540
gtcgtgggaa tgagtcagct ctttgggatt gcaaacatga tggatgggga aagcatagta
12600 actgtactca ccaacaagat gctggagtga cctgctcagg taagacttgc
attagtcaaa 12660 gcctcaacac aaaatccttg gtgggaaaaa aatatgtaga
tgggttaaaa cctagaataa 12720 gccactttcc tgtaagcaat ctagttcatg
tataaaagta ctccatccat tgctagaaaa 12780 ccacaaaaca cgaggtcatt
tttttttaat aaaaaaaatt ctgaacactg tgattaatga 12840 ggaggtgact
ggcttcaatt ttatacatga tcttagccaa aaagccaaga agtggtggtt 12900
taaatttgat attcagtaag cttactgtaa cctactattc ctatatcata aaaatatcat
12960 gacatctaag ttatttcctt gttctttccc agtgacttgt tttgatatgt
tgacccacaa 13020 attattacca ttttgtccat taactgataa tcaacttagt
gaatcaaaaa acaactattt 13080 ctagcattaa tacatcattt cttgttgtag
gaaggattct ttgaaagtat aattgcttga 13140 ccagtgtgca atgctagtca
tttcatttac attgtgcttt taatttacaa aacatctcca 13200 tatgttttat
tgcaattgac cccacaataa ctgagaaagt catgtgtcag gcctttctgc 13260
tgacaaatcc caagggcaca tttctgatcc aactcagatt cctgggatcc ccatttcttt
13320 ttgtttgttt cattcctttt tttttttttt tttttttttt tgggacgaat
tgtcgctctt 13380 gtcccccagg ctggagtgcg atggcgctat ctcggctcac
tgcaacctcc acctcccggg 13440 ttcaagcaat tctcctgcct cagtctcccg
agtagctggg attataggca cctgccacca 13500 cacctggctg atttttgtat
ttttagtaga gacagggttt caccatattg gccaggctgg 13560 tctcgaactc
ctgagctcag gtgatccacc tgcctcggcc tcccaaaatc ctgggattac 13620
aggtgtgagc caccgtgact ggcctgtttc attaattttt atacaatcct ctcacctccc
13680 agcctctctc tctcccatag gactgactat agaaaggact tatctctgtc
caggcatcat 13740 gatgtgataa gagaaacaca cagtagcatc tagaaaaatg
ctgaaataaa ttttcacata 13800 gctaatatcc tgtgattttg tgcaagttgt
caaatctctc taaacttcag tttctgagga 13860 ttcaatgata taatacatgt
aaaactactg gtacactgtt tggtatgtaa caggtgctga 13920 nnnnnnnnnn
nnnnnnnnnn gtgactcagt tctctttctt ctttctccgc ttcctcttcc 13980
tcttccttct cctccttctc ctccttcttt cctcttattt ttaaccacta tgggaggtgt
14040 gaaatgggga ataacaaaag taacatctac tgcaaagttt aaaattatca
taaattttag 14100 caggctctta tcataattat tgctgtgtat agaactggct
gtttagctaa aacagtgtag 14160 tacttttagc tctttttttc tgttgtttta
taactaccag tggagttaat caatttgctt 14220 tttgaaattt atacacttaa
agctttaacc tgagtcaaat ttaaataact tgagtcgaat 14280 ttatctattt
ctgtacaaaa aagagtattt acatctgtcc tagtaatagg tggaaacaaa 14340
acaaaacaga aagtttaaaa actttaacct gtaaacattt tcctttgtaa gtagatatag
14400 ataaaaatcc taattgcttc ctagatttta acatattaaa attctctact
gtgtggagct 14460 gggcaatgtc caaaatcagg caccatttta aggacaacat
aaaatcccaa gtgtccagag 14520 gaaacttctt ttctaaaatc acttctaaat
gaatgtaact tctgaactct ggtcttttct 14580 ctgctccaga tggatccaat
ttggaaatga ggctgacgcg tggagggaat atgtgttctg 14640 gaagaataga
gatcaaattc caaggacggt ggggaacagt gtgtgatgat aacttcaaca 14700
tagatcatgc atctgtcatt tgtagacaac ttgaatgtgg aagtgctgtc agtttctctg
14760 gttcatctaa ttttggagaa ggctctggac caatctggtt tgatgatctt
atatgcaacg 14820 gaaatgagtc agctctctgg aactgcaaac atcaaggatg
gggaaagcat aactgtgatc 14880 atgctgagga tgctggagtg atttgctcaa
gtaaggactg atcttggctc attctattct 14940 ccaggagagg acaaaagaaa
ggggtaataa atcttaagag cctgaactct taagaacata 15000 atgaagtctg
cttcttttgt caccgcattt taacctaact tcaggttcca gttctgatac 15060
ctgtggactt tttaccttaa ctgaaattag gagaaatagg ttagggaagt gcatagtgaa
15120 actgagatcc aggtcataag aaaagaagaa agtgtttagg aaaagcccat
aaagaaaggg 15180 agagaccgaa aagaaaaaga ggggaaaaag gaaagaagga
agggctttac taggaaattt 15240 tgcactatga agttttatga ccaaaccact
ccagttatca tctaatccac aaaagcctcc 15300 tcattagcct cattaaccca
ctccttccca aatcattttt caagttcaca gaatcgaatc 15360 acttgttaaa
aatcaatatt ggtttctcat taaaacttta tttatactag atatgcattg 15420
tatctgtcta gagactttca cgttgtatac tcagactggg ggcggggaaa atcgtttgtg
15480 tgtgcagaga taccactctt cgtttttctg ttgaattctt agtataaaag
gagattgagt 15540 gtgtcttgaa ccccagagaa cacttaggaa aataagagta
aaaaatccct atcttcacca 15600 gatccctttc tctctctctc tctctctctc
tgtgtgtgtg tgtgtgtgtg tgtaaataaa 15660 ggaacttcca ttcaaacttg
tttaaaaaga agagaatact ggctcacaaa actgggaaat 15720 ccagagggta
tagttagcct catgcatggt tggattcagg ttcttgaaca atgtcatctg 15780
ggttctatct ctttctgcat gtcttgaaca agctttttct tgttgaacaa tgtgatcagc
15840 tatagcctga accacaatga atggaactcc tacaagaaag atagattttg
ttaggttaag 15900 aatgtgaaga atttggagta gaaaaaacac agactgtacc
ttccattctc ttttacccat 15960 caacatttct ctaggccccc agttagcaca
tgctgacgaa tcacaatccc acaatgaata 16020 acattgtggg aatgaaaacg
gaaactgcca gaggaaacaa ccagtttaat ttattacaga 16080 atattcttca
gaatcaacgt ctgttctttg aagaatattg atttggggaa ggtgaaaatt 16140
gtagtactga agcctccttt ctctttgctc tgcttttaaa ctaaggaagt ggtcacaaca
16200 ggttcaggtg tacaacacag gctaggtagt atcaatggag aatgcagtca
gaggcaaaag 16260 ggattgctaa gtatggaggc aaactgaagg atgagggagt
cacctaaaca catgcaaata 16320 tattttaacc agaaaaagta aacagttcta
acaagaactc tgtggtggtc aaatagcagg 16380 tctctagatg aaattcagtc
cctgtatctc ctgttagaaa cacttcctgt agtatatgct 16440 gtctatactg
agatgtccca tcttcttgcc tgatccactc ctccttccat cttgaatgat 16500
ttccagtggt agtacagtac atcacagttt tctctgtctc cacttatatc tctccccttt
16560 tccttaactc atctcaggta tagctattta tagtctcatg ttcttgcttc
cttatcgcag 16620 acctgttgtg tgtctctgtt acagagggag cagatctgag
cctgagactg gtagatggag 16680 tcactgaatg ttcaggaaga ttagaagtga
gattccaagg ggaatggggg acaatatgtg 16740 atgacggctg ggacagttac
gatgctgctg tggcatgcaa gcaactggga tgtccaactg 16800 ccgtcacagc
cattggtcga gttaacgcca gtaagggatt tggacacatc tggcttgaca 16860
gcgtttcttg ccagggacat gaacctgctg tctggcaatg taaacaccat gaatggggaa
16920 agcattattg caatcacaat gaagatgctg gcgtgacatg ttctggtaag
ttaaaaccaa 16980 accaaaacca aaacactaaa caaaaagaaa agcagaaacg
gaaagtcctg tgctccttaa 17040 gagtaggaac gtgctcctta agaagtagaa
atgcctagta aattccttgc tgggaattcc 17100 tttatacctg taatctttga
cacattcttt attcaactag tttggaaggt tgtctgacgg 17160 cattctctgt
aataattact cagactgctt ttatggagca cacttcaaac tcagaatcta 17220
cctggagctc agagagacaa aatttcctag gaatctgttt ttaaattttt ttaatttact
17280 tattttttta ctttaagttc tgggatagat gtgttgaatg tgcaggtttg
ttacatgggt 17340 aaacatgttc catggtggtc tgcttcacgt atcaacctgt
catactaggt tttaaccccc 17400 catgcattag gtannnnnnn nnnnnnnnnn
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 17460 nnnnnnnnnn nnnnnnnnnn
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 17520 nnnnnnnnnn
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnatggg catttgggtt 17580
ggttccaagt ctttgttatt gtagatagtg ctgcaataaa catacatgtg catgtgtctt
17640 tatagtagaa tgatttataa tcctttgggt atatacccag taatgggatc
gctgggtcaa 17700 atggtatttc tagttctaga tccttgagga atcaccacac
tgtcttccat aatggttgaa 17760 ctaatttaca ctcccaccaa cagtgtaaaa
gctttcctat ttctctacat ccttgccaac 17820 atctgttgtt tccagacttt
ttaatgatcg ccattctaac tgggatgaga tggtatctca 17880 ttgtgatttt
gatttgcatt tctctaatga ccattgttga taagctattt ttcatatgtt 17940
tgttgtccgc ataaatgcct tcttttcaga agtgtctgtt catacccttc acccactttt
18000 tgaggaatct cttttaaagc tgaattgctc atatctttct gccattgaca
atttttatca 18060 acccttgaaa ctacagtccc atttctaaca aaaatttatg
gtctatatta ttgtttctta 18120 aaaagtcagg aaataagtta cctataattt
tcaatgcatc agtatcagag aagaaaaaag 18180 acaaaaaaaa atcctctgga
agctaacctc catatgtatc gatggtttaa gtttatgttt 18240 cttttgcaga
tggatcagat ctggagctaa gacttagagg tggaggcagc cgctgtgctg 18300
ggacagttga ggtggagatt cagagactgt tagggaaggt gtgtgacaga ggctggggac
18360 tgaaagaagc tgatgtggtt tgcaggcagc tgggatgtgg atctgcactc
aaaacatctt 18420 atcaagtgta ctccaaaatc caggcaacaa acacatggct
gtttctaagt agctgtaacg 18480 gaaatgaaac ttctctttgg gactgcaaga
actggcaatg gggtggactt acctgtgatc 18540 actatgaaga agccaaaatt
acctgctcag gtatgacttt caatcaactt gttaagaaga 18600 agggtgtaaa
tttccagtac tacctttgaa attgaaaaga aacattggtc cttatgacta 18660
gatcactctc aagaaaccag gaaatatcta cacaaattcc actgggagag tcagataagt
18720 ttacaattca tgttgcactt tagatagaag tgtttgggtt ttgctcactt
tggtttttga 18780 gtaattttta gatcaataga ggtataagat taaggaggat
taaaatttaa aaaatagaat 18840 tcaatatttc tttcacagga tgtcaccaaa
agatttaaaa aataaccatg ttagataaac 18900 tgacataacc ttcagctccc
tctattgtac tgtgaaagaa ttttcttttt ttttggaaaa 18960 cctgtgaata
tttcactgca acctctttaa aaagctcaaa aatgagttgt aaactggtta 19020
ggtttgtgga taatctttag tattttatgc taattatatc tatacaaatg ttaactttgt
19080 tgaaaagtga tgctttttaa ataactctaa agtaaatgta acttttactc
atgcaacaaa 19140 aaggcttcag aaaacacagt gtacaagaaa accattaaac
atgtagagac ctcacagaat 19200 ggaaacttgt acgtgctgca tgtcctctaa
ccaacaatat ggctccagat acctcccaac 19260 ctaggcatgc agtgaggggc
atttccctga tgttacaaat ggtgcttacc tctagttaag 19320 tagaaattat
cctccgattg aaaaaaggca atcatttaag atataaattt gaatttctat 19380
attacttttt aaaaaatatt ctggaccagg caatataatc acatggttta aacttcaaaa
19440 taaaaaagtg taaactcttc ccccactact atccaattct tttcctcaca
gacaacttat 19500 gtggttggaa tatttatata tcatttgaga tatttcatgt
aaatacaaga aaatacaaac 19560 aaatctatat atagctatca tttgctttca
ttatccaaat ggtggcatgt tgtataaatt 19620 gttttgtacc tgaagtattc
acttagtaaa atttcttgat caaatttcac atcagtattt 19680 aaagagctgt
ctgaaattac ttccatatct ataacatttc ttataattta ttaccgtgtg 19740
tattcattta ttcttttaat tttttacctg acaaggttag ccattctttt acctaaattt
19800 ttttttaaaa aaccattatt tggattttaa aaaattaatg ttacttttat
ttcctttcta 19860 attcattaaa atttactttt ctttctaata attctcttca
gatttcctaa agtttgattt 19920 ctttttggcc cccaacttct tgagttgaat
cttaagtttt atttcttctg atttactaat 19980 ataaatattt taggttatag
attttccttc taaatttata atggtttgta tattttctta 20040 ttaattgtat
tttacattga ttagcattaa gttctgctgt atctaaaaga gaaacaaata 20100
atagtgactg aagaaagaca gtgttcactt tttctcattt aaaattagtc aaatagataa
20160 gcagtttaag gctgttttgg tgtctctgtt gtcaccaaga aacatccaga
atagatttgc 20220 tgcagtaagt atcaaatcct accggacagg aaggataagc
caccccgcag ctcagtgagt 20280 tccttggaaa gccaatctgg tagacgaagt
gctggcctct gagaaggtgt cttctgtccg 20340 ctgtccttca tgggccccag
tgttgctctc agggcagttc ttcctggcac attattcttc 20400 acagttacat
ccaagtttgg tttggagagt atctctatct aaggacagca cacctttcac 20460
aacccacttc ctgctaatat tggtttaggg acctttggat taaagattga ctaatcaagg
20520 gctgatgtca gctgccctca aattcaacaa aaaagttctt tccaaggtta
ctaagcttct 20580 agtctacttg ctttataacc caatgccatc tgaaagtaac
tatagtccaa gagcatataa 20640 atatactttg atctatgctg tgaccagccc
ttctctttaa ttattgacca ctgccttaag 20700 gccgtttgaa actgtgggtt
tgggaggaag gacaagacct aaattccgtc tttgccggta 20760 gactcattgc
ctttgtatct ggagtatcaa ttttatcttc tgacaaggct acagttttac 20820
caccacctca atttgggggg acaggatact tagagttttt tcttcaattc ttcaattcca
20880 aattatagga ctcttgatta acaatataca acacctgcag gcagaccata
cccccttata 20940 aaattgatat ttctcctccc atttctgctt gcattctgac
tgctggtagt ctgaattcat 21000 ctttcttgta aatgctaaaa gtggcaagga
gaatccacgt catgcaaata ttctgatttt 21060 ttgtagtttt gtttcttgta
ctttgcccaa taaattctgc attttttaag catcagtgag 21120 cacacggttt
tcttttcaaa gtgaggctta cccaaatatt ttaccatgtg taagactcct 21180
taggttttta gtctctcaca cctgagtcat taatgcattc tgcccatcaa ctccattcaa
21240 ccacacccat attttaggtt gcattactag aataccttac ttatggtaac
atgttctaaa 21300 ttattcagga ttctattcag atgtgtataa tagaaatcat
gaataacata aatttgcaac 21360 ttaaggtggg ctccgtactt cacagagctg
tatttttgtt tgctatttgc catgtgcact 21420 ttttgtttcc ttttacaaac
tcttttgtta tattgctgta ggttttggaa gttcttctac 21480 ttatgttggg
ggccttggtt tttagctttt tacatgtttt gttagtaata tagtttgtac 21540
ttttgtttct ttatttactt gatactttca atgatgacat actgtctctg tcaatgatga
21600 tatgctgtct ctgtgcctgc caaattcaag ctacgaattt gatcatctgt
ttaaattgtt 21660 taacttttat atagtacagg gctatacagc ttaaaagaac
caggagaatt caactaagca 21720 ggagaattgg aaacaattta agataaattg
ttgacagaaa aaggtctagg aaattttggc 21780 cacaaaatta ctttgaatct
cagacatttt gtttgtgatg tctgtgtatg tactgtagag 21840 gacatctgac
cttaagaaac agttcatttg agctatgctg tcttttaact taggatggag 21900
aaataattct gagattatga tgatctatac ttcttcagag tctgaaaagc attaaatgtt
21960 gaaaatacat taaagcttct ttaaatcata attgtatatt ttatttatca
ctaaacaaaa 22020 ggaagtaatt ctagttaatt atgtctctaa ttatgttaaa
gctctttgta gatctattta 22080 tttttctaga aaaagaaaga aaaacagatt
cagttactta tgccagggga aaactgtagt 22140 tgttgatcaa tggtgctaaa
tccaggtgca aaaatttact taatggtttt tatgtaaaaa 22200 cacagtcaga
gacttagcag aaaaaaatcc attaaaggga tacatttcca atattctatc 22260
catgtctaaa gagtaacgat ataagcaaag cgacccatag gatagattaa catgttacta
22320 gcagattcac gaaataaagg ctgaggatgg agtctccaag tcactttctg
agttggagaa 22380 tttgtcctct gccttgactc aaatatgtta gtacaaagaa
aacaagagga tacttataga 22440 agggactttt tctcaattgc ttaatgagtt
ttcacaatgg aaaagaaaat atttttgttt 22500 gtttgtttgt ttgtttttca
ctggttttct tttggtttat tttgttcaag atgggtccaa 22560 atgaaagagt
aagaagttac aagaaaccag aatttggctt gatctaggag actgccttta 22620
cttaaagatt tagggctgtc caaaagaggt tgggatcata tgaatactag ttaatgatat
22680 cttagagaaa tttcatgctt gaataaagat tcaaaaagaa gaaatgaaag
accatcccac 22740 tcaagggtca ttaattattt gtattgtaga ggtgttattt
gctttaaact ctgagctatg 22800 tttagaaagc aacatgtttt tcatttcaaa
tttggatagg aatttgcttt gcgcagattt 22860 aatggataat gggtgaaatt
ttgacatgtt gaggtattct atattgctgt agtacagaat 22920 atggtcatat
tcgaaacatt ttctaagaca tttttgtatc aggtgaaagg tttatgagag 22980
atgaaaggaa atcagagtat gtagattggc aagggctgat ggtgtatcta tatcattgtg
23040 atagtaacat caagagatta caaaaattgt tgatatttga tagcaggtat
ctataaagga 23100 cttaagacct tcagaaggtc taattttcaa ctgcattttc
tttatttctc ttaaagaaca 23160 agttaaaaga aatagtttgt atctcttaag
tactttatat tgcctctgag tattttcaaa 23220 gaaactcact tatcttgatg
attgtgtttc ttaacttgac aaagagcatc ttcataatat 23280 aaattccatt
tattatttct ggtgtgtctt ttctttatat tttaaatttc agctacagta 23340
agttacttat aattcttcaa tgacaccatg catttcgtga ccatattcct taaagattgt
23400 ttcacacact gtgtatttgg caaactcctc atcttttttc aagatccagt
ttagatctat 23460 ggtattctct cccccatagt ctaaaacaga gtgaatgact
cctttatttg ttagttttat 23520 gtctaataca tatttctatt tttgggttca
tcactattaa aatatgtata tcttgtagtg 23580 aactgtgttc atggaggaga
aggctattag tgagaaattg aacagccttg aaaagaagta 23640 gccttttcct
aggtatataa ggtagggaaa taaattttaa gtagagagaa aagtttctac 23700
agaggcacag aagcatgaaa attacggcat gcccaagcat ctttaggctg gagcatcaaa
23760 ttaggttgat aaagtaatag atattaaagc ttttaaggta acagcatact
attttaaagt 23820 gtactaaaga attagtgttt atcacctctc cttaatttta
atagctggac gaaattaggg 23880 aatctatgaa aactgtttgt gctgcagttt
gctcagaaga aaaatgggag tggtaatagg 23940 gcctacctca caggaattac
acagtgagct ataaatacat gttagctaat atgattatga 24000 ttattatttt
tagtattatc tatcttcact gacagactga attctgtggc ggccttttta 24060
acccacccat catcaaatgc ctaaaataga gtcagagaac attacaggtt gttacattat
24120 aggtcattac attgatactt ttggattaat tattgcatac atgattatta
cagaaattat 24180 aagaaattat gtaagggatc attgcataca ttttaaggaa
acgttaagtc atctaaagga 24240 aaaacatcac atgaaaatat ttgaaattta
ttaatatatt tataatcatg gcataaattt 24300 cttggaaaaa agactttgaa
gcaaggaaac aatttacgaa gctattgtag aggtctgcat 24360 gacagatgat
gacaaatcac tttatggcaa tagcaacaaa tatgcagaag aagaaatatg 24420
ctggaggacc aacagatatg gtggttttct acatgttcaa tgtatcatga aagtcaacat
24480 tctaggctct ggaatatact gtctggcttc aaattctgtt ctcttactaa
ttctgtagta 24540 atttcttgtg tgaccttagc aaattactac acctcacctc
tctatgtttc agattccctg 24600 tctgtaaaat cagaatgata ttaacaatta
tctcctaagg caattttaag gattgaatga 24660 aataatgttt gttaaatgct
tattggaatg ccatatatgt aatataatag acgctcactt 24720 aatattcact
gctatcatta ttaccattat ttgtggtgag ataaagtggg tcaattttag 24780
gtggttctaa tttggcctgt atatcattaa ttaagtaaag aatatagtgg gcttgtcata
24840 caataattca ttcaactttg aatactttaa gctccacatg tctaagcaga
tacgtttgtg 24900 gatctaaaat taaagaaaaa agtatggatg aaaatatatt
tattagcagt tgagacatgt 24960 ccattgagca gatgagaagg ttacggacag
aactttgact caaggaatac acgaaatgtt 25020 ttgataaagc actgtcagaa
agaatggagg agaaaatggg acaagaaggg tcacgtatac 25080 caaacgtaac
agacgtcaaa aggaagaagt ctgatgttca ttctgaaaat tagaagtata 25140
ttgaaaagct ttacatagaa caattcaaaa aggttgagca gaccccagat tgcagtacat
25200 ggaggaatga gtgcaagact aagagatagc gatagtagat acacattgtt
tgtggtctct 25260 aatgttatta tgaataaagt aagcaaaatt ttggaagaac
agagttatca ttgcctttgg 25320 ggatggttgg ctgcacaagg tgaagtccca
gaattctctc agatggatga gttcagctag 25380 gaatggggag
gtctagattt atccataccc aactcgaggc agaaaagcca agacaaacaa 25440
gtgtaggaaa tcactgagca ccctggtctc catttcccac tccacaccac cttcttcacc
25500 ctcatcatga atggaaatct ggaaatcata ttacctctca ctgaaataat
atttattttt 25560 cagcccacag ggaacccaga ctggttggag gggacattcc
ctgttctgga cgtgttgaag 25620 tgaagcatgg tgacacgtgg ggctccatct
gtgattcgga cttctctctg gaagctgcca 25680 gcgttctatg cagggaatta
cagtgtggca cagttgtctc tatcctgggg ggagctcact 25740 ttggagaggg
aaatggacag atctgggctg aagaattcca gtgtgaggga catgagtccc 25800
atctttcact ctgcccagta gcaccccgcc cagaaggaac ttgtagccac agcagggatg
25860 ttggagtagt ctgctcaagt aagacccaga aaacatcttt aattggttct
catactgtga 25920 aagggacagg gttagggagt catagctgtc tttttctaaa
gccctgtctc cttccaggat 25980 acacagaaat tcgcttggtg aatggcaaga
ccccgtgtga gggcagagtg gagctcaaaa 26040 cgcttggtgc ctggggatcc
ctctgtaact ctcactggga catagaagat gcccatgttc 26100 tttgccagca
gcttaaatgt ggagttgccc tttctacccc aggaggagca cgttttggaa 26160
aaggaaatgg tcagatctgg aggcatatgt ttcactgcac tgggactgag cagcacatgg
26220 gagattgtcc tgtaactgct ctaggtgctt cattatgtcc ttcagagcaa
gtggcctctg 26280 taatctgctc aggtaagagg atgacgggca gccagtgatg
ggtcttaaga gaaggtgtac 26340 atagggatga agaacaaaag taaaacgcag
tgatccattt tgaagggtca tgagaagaat 26400 gaatactaat tcatgcttca
gaatattaac ctcatttgtc ttttcagagg gtgagcaaac 26460 ctaattttga
tggacttctt agttatccag tgcttactat gtgccaagcc catctcaaag 26520
cacctgattt atttcaaact tgttgaccat ccaccttccc ctctctctct aagctatgtt
26580 tgtcctcaac tcatccatat aacatttcat ctgactcccc taaccttttg
tgtcttgtat 26640 gactgagtac cttggtgttt gcaggaaacc agtcccaaac
actgtcctcg tgcaattcat 26700 cgtctttggg cccaacaagg cctaccattc
cagaagaaag tgctgtggcc tgcataggta 26760 agactctgtg acaaatctat
gaaaatatta ataacaagtg gaatgctcat tattaatagt 26820 ttcatttctc
tttgcagaga gtggtcaact tcgcctggta aatggaggag gtcgctgtgc 26880
tgggagagta gagatctatc atgagggctc ctggggcacc atctgtgatg acagctggga
26940 cctgagtgat gcccacgtgg tttgcagaca gctgggctgt ggagaggcca
ttaatgccac 27000 tggttctgct cattttgggg aaggaacagg gcccatctgg
ctggatgaga tgaaatgcaa 27060 tggaaaagaa tcccgcattt ggcagtgcca
ttcacacggc tgggggcagc aaaattgcag 27120 gcacaaggag gatgcgggag
ttatctgctc aggtaaatta tgcatatgac ctttggtcat 27180 aattacatag
agaaaggact gaggaagggt agtgaggggt attaatctta aagattttcc 27240
tgaaaagttg atccaaaagg aactattgtg ggagtcagga caacagaagt ctgggtttga
27300 tgtgctagtg gtacttcatg agcaaggtag tgcaacattg taaagagaaa
agaccagaaa 27360 tggtcttgta tatcatgaaa atgacaacaa agttaataat
gaaggttttt aataaatata 27420 tttggggcca gatattagat agcgatccaa
tagcaacact tgataattac agtattagta 27480 aagaagtaga tttaaaggga
caagaagaaa tatatatgtt gcatcattct acagaaaaaa 27540 agataattac
tatgcccaag caatttttca tggcagataa atgagactta cttgtcatag 27600
aaacaaaaag tagcatgagg ttactgccct catactgctt tgtatacatg gtgagtttta
27660 gaaaacattc tgaaatttaa aaatgtacta tttcatatgt attatgggga
acagaactat 27720 gattcagtaa cttaaggaag tgaaaaactc tgtatctcag
aagtgattta ttcctatttt 27780 aatgacataa atctggtaga agacatgcac
tgcaaaactg tgaagtgtca gcaaatgaac 27840 tgtcatttta agagatgtac
acaaaacatt gattttagtg cctgagtgtc taacaatttt 27900 aactatcaaa
gacagttgca aaaaaaggag aatatttttg ggaaaaaatc caaactatgt 27960
aactgaaaaa agcctattag aatgattgag ggtaatctaa agaggactct gagtccaaaa
28020 ccaaagacca aaaaactata atgtatttat agaataaata aaatatttta
ataatatcta 28080 tgaaatcatc cagaagttac taacgaatta ttagtaactg
acatattgta ctattgtacc 28140 tatttttaaa gccattctta catcaaggta
aagggaagga gtctctgaat cttcagaaaa 28200 gatgcaaaac tactaattct
atgttttgct gcagaattca tgtctctgag actgaccagt 28260 gaagccagca
gagaggcctg tgcagggcgt ctggaagttt tttacaatgg agcttggggc 28320
actgttggca agagtagcat gtctgaaacc actgtgggtg tggtgtgcag gcagctgggc
28380 tgtgcagaca aagggaaaat caaccctgca tctttagaca aggccatgtc
cattcccatg 28440 tgggtggaca atgttcagtg tccaaaagga cctgacacgc
tgtggcagtg cccatcatct 28500 ccatgggaga agagactggc cagcccctcg
gaggagacct ggatcacatg tgacagtgag 28560 tatccatcga cctatatgaa
aattccattc tgcagccccg ctatcatctt ctgataatgg 28620 acactggatt
ttattttcct attctcacac tggcttaaat tgttagattg aattaagcaa 28680
gaatgggtaa cttgtgggta atatctattc tacctggata gtgtaactgg ttttatttaa
28740 tttggcctgt agatattcag gatcagctat tattattcta aagaatatca
tactccccat 28800 ggtctgcttg aaagatttat tcaattgctt tctctgagga
ggagagaata tgtggtttaa 28860 tatttgactg ttgaatggtc taccttacct
tcccattttt ctttcaaata ttatcttgtt 28920 acactaatat aaggagaaaa
gcagtcagaa atcagaaaga tcaattctga agtatttcca 28980 cattctacag
gttttttttt tttttttttt tttgagacag agttttgctc ttgttcccca 29040
ggctggagtg caatggcgtg atcttggctc actgcaacct ctgccttctt ggttcaagca
29100 attctcctgc tccagcctcc tgagtagctg ggattacagg catgagccac
catgcccagc 29160 taattttgta tttttagttg agatggggtt tccccatgtt
ggccaggctg gtcttgaact 29220 tctgacctca ggtgatccac tccccttggc
ctcccaaatt gctgggatta caggtgtgag 29280 ccactgtgcc tggcctctac
atcttcttaa actgtgagat ttgtgacaac ctgccagttt 29340 cccaagaatc
ttgcctattt gaaagaattt gacatcacat atttataatg ttaactttaa 29400
tcatttaaca tttactgggt cacattatgt atcttatgtg caggatggaa tatacctagt
29460 tggctgtaat gacaagtcct cttataaaag ataaattttg taaaaatata
aattttaggc 29520 caggcacggt ggctcacgcc tgtaatcgga ctttgggagt
ccgaggtggg tggatcacga 29580 ggtcaggaga tcgagaccat cctggctaac
atggtgaaac cccgtctcta ctaaaaatac 29640 aaaaaattag ccgggtgtgg
tggtgggcgc ctgtagtccc agctacttgg gaggctgagg 29700 caggagaatg
gtgtgaacct gggaggcgga gcttgcagtg agccgagatt gtgccactgc 29760
actccagcct gggtgacaga gcgagactct gtctcaaaaa aaaaaaaaaa atatatatat
29820 atatatatat atacatatat atatataaat tttaatatgc acattcagta
aaattttaaa 29880 tagttttttt ttttttttaa gtttcaaatc tctcttttca
cctgtgaact cctggaaact 29940 tttagaagtc ttagaaatag gctacatgtc
tctgattttt cagacaagat aagacttcag 30000 gaaggaccca cttcctgttc
tggacgtgtg gagatctggc atggaggttc ctgggggaca 30060 gtgtgtgatg
actcttggga cttggacgat gctcaggtgg tgtgtcaaca acttggctgt 30120
ggtccagctt tgaaagcatt caaagaagca gagtttggtc aggggactgg acccgatatg
30180 gctcaatgaa gtgaagtgca aagggaatga gtcttccttg tgggattgtc
ctgccagacg 30240 ctggggccat agtgagtgtg ggcacaagga agacgctgca
gtgaattgca caggtaagtg 30300 ccagaagcct ggattgagct tggaatctct
ggcagcaaag aggggttggg cagtgatgtg 30360 tgttgtgtaa aaaccggatt
tatcagcaca gggatctggt ggggattctc agagaatcat 30420 aatgtctcta
ctgaaagaat gatgaaaaca attgttatac aatggccttc accaaggaga 30480
cacatacaac ctatattccc tgtgcagaga aagaaagact agaggactct agttttaggt
30540 gggaaagagt catatacttg gttgagaggt actaaggggt cagacaagga
aatcagaagg 30600 cttgttctat atatcatgca aagaattaac cctctctctt
tcttttctcc tacagatatt 30660 tcagtgcaga aaaccccaca aaaagccaca
acaggtatat catggagttt atgatgatga 30720 gaccaattat ctgcatacat
agaatttttc ttgtttagaa attttaggtc agacgttctg 30780 aggatttgat
atttacctca gggatcaagt atttgaccta agagagaatc agtaactagg 30840
gaccttgatc ttttcgataa gacatgatca tatctcttga tatttacttt tacttaggtc
30900 gctcatcccg tcagtcatcc tttattgcag tcgggatcct tggggttgtt
ctgttggcca 30960 ttttcgtcgc attattcttc ttgactaaaa agcgaagaca
gagacagcgg cttgcaggtt 31020 tgagccaaat ttggtttatc taatatgtct
acaggataag ggggttcttg taagacgaaa 31080 atattagact caagtctaat
ggccagaaag atggactcta aaagaggttt gagcagtttc 31140 tcaaaacaga
gggaagggcc aggcacagtg gctcacactt gtaatcccta cactttggga 31200
ggctgaggca ggagaatcgc ttgagtccag gagttcaaga ccagcctggg ccacacaggg
31260 agctcccatt tctacagatt tttttcattt tttaaataga gggaataaga
tttcagagga 31320 gaaaaggggt ttaggttgca ttgatgattc cctgtgttct
atctacatgc ttaactggaa 31380 cattacagaa ttgaaagttc agaatgaata
tgttagatat tgtttagnnn nnnnnnnnnn 31440 nnnnnnncat gggacaataa
atcttggtgg aacttcaagt ctagtggaag agatatatat 31500 gtaaacaaat
acactacaat gtagccattt tacattaata ggaaaaagga acaaaagtgg 31560
cacagagaag ggagtaatta actgaaaggg gagggggatt actggaggct tcaaagaggc
31620 ttggggacaa atttcgaaca tagaggctca gagaattggg tgaggaacat
tctagagaag 31680 aatatgtaca agtctagaga agcctaaagg atgaagcatt
tcttgttttt ttcctgaggg 31740 gcggggggag aaaagttgag tgtgaggaga
gaggctgcta agtctgaaga agagaaaagg 31800 cacaaattat gaagttctta
tgtttcacat aagattcata gtcattaaag agtatcatgt 31860 aggggagtga
aatggtcaag atatctttta ggaagatctt ttggcaacgg gatctaggag 31920
gattgaagga ggaccacttg actgacagga atggcatttt gtaacagttg aggtgaaaaa
31980 tgaaaagtac tataacatat atagtgagaa tggaggggag aggaaagact
tttagagata 32040 cttaagaaaa gctccagtga gttatggaag actgctggct
tcccttgggt aaaaacagtg 32100 ggttgtttat attaatacaa tggcagtggg
gaagatttag ccttgtgcct atttcatttg 32160 agtgtgcgga attgtaaggg
aagagcttgg tagtagaatg ggatagaaac aatagcaggg 32220 ataaagacga
ggtatgtttg cagcgcttgg aaaaaacaca cttacagcaa gggaaatttt 32280
attatgatag ctgatcccta acctagggat ctctgttttc agtttcctca agaggagaga
32340 acttagtcca ccaaattcaa taccgggaga tgaattcttg cctgaatgca
gatgatctgg 32400 acctaatgaa ttcctcaggt ctgtgggttc ttggagggtc
tattgcccag gggttcagat 32460 cagtggctgc agttgaggca cagacattct
actttgataa acagttaaaa aagtctaaaa 32520 atgtaatagg aagcttagat
gcatataatg gacaagaatg actgaaaatt attcttggag 32580 aatatcaaaa
ttgcaatcat agggaggcct ttagcttaag aggcctgtga ttattcctga 32640
tagaggtatg gaaagaacca tgcagaggaa tattatgact tggacctcat tttattaaaa
32700 cagaaattaa tcttacaaaa gattgtcata agtgacagtt taactttttt
ctttaaattt 32760 tgttgtgtat atttaaggta tacaacatga ttttatggga
tgtatataga tagtaaaaag 32820 cttactaaag caaagcaaat gaacacaccc
atcatctgac atagttaccc ttttttgtgt 32880 tgttcttgtg gcaagagcag
ctaaaaccta ctcacttagc atgaatccta catacagcac 32940 aatgttatta
cctataatcc tcatgttgta cattagacct ctagactggt tcattctacg 33000
tatctgctac tttgtatcct ctgacctaca tacgtctttc acagtttctt ccattcccat
33060 ttcctgtcat tttttttctc tagcttgata tttattatat ttttccctaa
aagtctaaaa 33120 ccttaaactt tcaatatctt tattgcatga gaagccatac
aaatccacag aactagcctt 33180 atttctcatc acatcatgct gttttatcct
tgaacttcta tttagcacca gtgcactaat 33240 tctgcatctg ggcaggatga
ctttactggg ttggaagaaa tatcccaaaa cccattgtct 33300 ttactccatg
aagggtccct gaccttctga gaggggcctg cctcacttct tccatccaaa 33360
gaattatgca tctgctactg tgtcagggaa catatttaag gaacatgtac tgttactgtg
33420 tcaggaaaca tatttaagaa ataggaaaga ctttctctgc cccttaaatc
acacatgctt 33480 ttcttcctag ttatgggtgg tgtttttagt tgctcaaaga
gcctcacagt tacgtgagaa 33540 gaggtctggt ttatttccca gtaattattt
tcttcctttc agaaaattcc catgagtcag 33600 ctgatttcag tgctgctgaa
ctaatttctg tgtctaaatt tcttcctatt tctggaatgg 33660 aaaaggaggc
cattctgagc cacactgaaa aggaaaatgg gaatttataa cccagtgagg 33720
tgagtgatga gaatttatta gtcattgttc aaaacagctg cattcctttt gaggactgag
33780 agctcttctg gcattagaaa gaaagaatgt atataaacat attttatttg
cttatgtgca 33840 tatgtttgta tatgtatata tctatattga gaatatataa
aagcttataa atataactga 33900 ttcatctctt aattatacac acaaatagta
ataacacatc catggagcta aggagaaact 33960 gagtaaacaa atacaaatta
atcttttgaa gacattacag gtatcatttc aggaaaaaga 34020 taagacatat
tgtacatcta catcgtataa tacatgctat ggtaaagact gtacaaggca 34080
ctacagaaag tttgcctaag aaactttggg gttgttgtaa gatgatatga tatttaaggt
34140 aaactttaaa aataactgaa aaatagacag ttccaaagaa aggacacagt
atgtgcaaac 34200 tagaggcaga aaaaagaacc atgcctttag ggaactataa
gaagtttttt attcaaaaac 34260 atatattgaa cgtatttatg gtaacaagtg
gtaccgtgag cactgaagat agagataaaa 34320 tctactattg gcaaagacat
gattgtttaa taaaagatat catagtctat aggtggtaat 34380 atgaagaggc
catgtggaga gaagagaggt ggttagaatc taagctgaaa aggctgcata 34440
gagccctgta atgaatggtc ttacctgcta tactttatgt tctctttcac tgggggtctg
34500 ttgaagaggt gatgatgagc tttgaaagtt tcagatgaga caagatgatt
agaattgcat 34560 tttacaactt gctttcatga gaaaaacata aaaaaaaatt
caaatgtgaa caagacgtat 34620 agttaagcta tacatcaaga gaaaaagatt
gtgaaatgag ttactagata cattttctgt 34680 ttttaaaata ataatttggg
gaagactaca tctagaaatg aaatgtagaa aaagtatttt 34740 ttatagaaat
attatatatt cctataattc atctacatat tatatatcta cattgtataa 34800
tacatgctat ggtaaagact gtacaagaca ctacagaata tttgtatata cgaaatatag
34860 attaagacaa aggataagag ttggaactct taaagtacat gaaatataaa
ttgtgaatat 34920 ctataaaatg cagagctaat tttacatact atatatatac
acacatagat acatagagtt 34980 cttagaaact gtacagagca caaaagggtt
aatatatgtg aatgtaaatt gtatgtatgt 35040 gtatgtgtaa catctaatag
ataattgggc aaagctatga aaagacaatt gacacaaaat 35100 gaatttcaaa
tgacagacat ttttaaatgt ctggaggatt gaaattataa ttaacaatga 35160
gctatcactt tatacaaatc agactaacaa aaatttgaaa taccatcaat atttattact
35220 ggcgggaata cagagaaaaa ggccatttca aacactgctt ttggaagtgt
gagatatggt 35280 aggtagcctt tggagaaagc aatctggcag tttctattaa
aaattatttt caaagattca 35340 cagacccttt gacccaataa ttctatccct
gagaatttct tcttaccaat aaagtcacta 35400 gtatgtaatg acaaatatac
aaagattcct ttctgccatg ttgttgaaag tggcaaaaac 35460 tagaaacaaa
gtgaatgcca attgatggag taatgactgg aaaaatgacg gtttaggcaa 35520
ctaaaaaaga aaaaatcatt ataaaatata ataatattcc atttatgtat tggctataat
35580 taaatacaat gcttacatct ataaaattat gcattcatat gtgcataaag
aagctaaaaa 35640 ttctagtgtt gtaaactgag aaccatgaat aatatttgat
ttataatctg aagcatatga 35700 atgttttctg atgtgataaa tgataattga
actacttttc cctgaaagag taatcactgc 35760 tgaataactg cttcagaatc
acatgagggt gcttttaaaa aatgacgatt ctttttttgt 35820 ttttaatctt
taagttctgg gatacatgtg ttgaatgtgc aggtttgtta cataggtata 35880
cacgtgtcat ggtggtttgt tgtgcctatc aacccgccat ctaggtttta agccctgcat
35940 gcattaggag cttaaatttg tcttaatgct ctccttcccc ttgcccccac
cccgcagcag 36000 gccccatgtg tgatgttccc ctccctgtgt ccatgtgttc
tcattgttca actccaactt 36060 atgagtgaga acatgcggtg tttggttttc
cgttcctgtg ttagtttgct gagaatgatg 36120 gtttccagct tcatccatat
ccctgcaaag gacatgaact cattcttttt tatggctcta 36180 tagtattcca
tggtgtgtat gtgccacatt ttctttatcc agtctatcat tgatgggcat 36240
ttaagttggt tccatgtctt tgctattgtg aatagtgctg caataaacat gtgtgcctgt
36300 gtctttatag tagaatgatt tataatcctt tgggtgtata cccagtaatg
ggattggtgg 36360 gtcaaatggt atttctggtt ctagatcctt gaggaattgc
tacattgtct tccacaatgg 36420 ttgaactaat ttacactccc accaacactg
taaaagcatt tctatttctc cacatcctct 36480 ccagcatccg ttgtttcctg
actttttaat gatcaccatt ctaactggtg tgagatggta 36540 tctcattgtg
gttttgattt gcatttctca gatgaccaat gatgatgagc ctttcttcac 36600
atgtttattg gccacataaa tgtcttcttt tgagaagtgt ctgttcatat cctttgccca
36660 ctttttgatg ggttgtttgt gtttctcttg tagatttgtt aaagttcctt
gtagattctg 36720 gttattagct ctttgtcaga tgcctagatt gcaaaaattt
tctcccattc tgtaggttgc 36780 ctgttcaccc tgatgatagt ttcttttgct
gtgtagaagc tatttagttt aattagatcc 36840 catttgtcaa tttttggctt
ttgttgccat tgcttttggt gttttagtta tgaagtcttt 36900 gcccatgcct
atgtactgaa tggtactgcc taggttttct tctagggttt ttatggtttt 36960
aggtcttatg tttaagtctt taatccatct tgagttaatt tttgtatacg gtgtaaggaa
37020 gtggtccagt ttctgttttc tgcatatggt tagccaattt tcccagcgcc
atttattgaa 37080 tagggaatcc tttcctcatt gcttgttttt ggcaggtttg
ttgaagatca gatggttgta 37140 gaggtgtggt gttacttctg aggcctctgt
cctgtgccat tggtctatat atctgttttg 37200 gtaccagtac catgctgttt
tgataactgt agacttgcag taagtttgaa gtcagatagc 37260 gtgatgcttc
cagctctgtt ctttttgctt aggattgtct tggctataca ggctctattt 37320
ttgttccata tcaaatttaa agtagatttt tccaattctg taaagaaagt cgatggtagc
37380 ttgatgagaa tagcattgaa tctataaata actttgggca gtatggccat
tttcacaata 37440 ttgattcttc ctatctatga gcatggaatg tttttccatt
tgtttgtgtc ctcttttctt 37500 tccttgagca ctggttggta gttctccttt
aagaggtcct tcacatccct tgtaagttgt 37560 attcctaggt attttattct
ctttgtagga attgtgaata ggagttcact aatgatttgg 37620 ctctctgctt
gtctattatt ggtgtatagg aatgcttgtg atttttgcac attgattttg 37680
tatcctgaga ctttgctgaa gttgcttatc tgcttaagga gtttttcggc tgagacaatg
37740 gggttttcta aatatacaat tatgtcatct gcaacaagag ataatttgag
ttcctattcc 37800 tgtttgaata ccctttattt ctttctcatg cctgattgcc
ctggccagaa cttccaatac 37860 tctgttgaat aggagtggtg agagaagaca
tccttgtctt gtgctggttt tcaaaaggaa 37920 tgcttctagg ttttgcccat
tcaatattat attggctatg ggtttgtcat aattagctct 37980 tattattttg
agatacgttt catcaatatc tagtttattg agagttttta gcatgaagca 38040
ctgttgaatt ttatcgaagg cgttttctgc atctgttgag acaatcattt ggtttttgtg
38100 atttgttctg tttatatgat ggattacgtt tattgattta catatgttga
accagcctta 38160 catccaggga tgatgccaac ttgattgtgg tggataagct
ttttaatgta ctgctggatt 38220 tggtttgcca gtattttatt gaggattttc
gcatcaatgt tcatcaggga tattggcctg 38280 gaattttctt ttctttgttg
tgtctctgcc aggttttggt gtcaggatga tgctgacctc 38340 ataaaatgag
ttagggagga gtctctcttt ttctattgtt tgggatagtt tcagaaggaa 38400
tggtatttgc tcttctttgt acctctggta gaatttggct gtgaattcat ctggtcatgg
38460 tttttttttt tttttttttt ttggttggta ggctattact tactgcctca
atttcataac 38520 ttgttattgg tctagtcaga aattcgactt cttcctcgtt
tagtcttggg aggatgtatg 38580 tgtccaggaa tttatccatt tcttttagat
tttctagttt atttgtgtag aggtgtttat 38640 agcattctct gatggtagtt
tgcatttctg tgggatcagc ggtgatatcc cctctatcac 38700 tttttattat
atctatttga ttcttctctc ttttcttctt tattagtctg gctagtggtc 38760
tgtcgattct gtttatcttt tcaaaaaacc agctcctggg ttgattgatt tttgaagggt
38820 ttttcatgtc tccatctcct ttggttttgc tctgatctta gttatttctt
gtcttctgct 38880 agcttttgaa tttgtttgct cttacttctc tagttctttt
aattgtgatg ttagggtgtt 38940 gattctaaat atttcccact ttctgatgtg
ggcatttagt gctatacatt tccctcttaa 39000 cactgcttta gctgtgtccc
agagattttt gtatgttgtg tctttattct cattggtttc 39060 aaagaacttc
attatttctg ccataatttt gttacttatt cagtagtcat tctggagcag 39120
gttgttcagt ttccacgtag ttgtgaggtt tcgagtgagt ttcttaatct tgagttctaa
39180 tttgattgca ctgtggtttg agagactgtt atgatttctg ttcttttgca
tttgctgagg 39240 agtgttttat ttccaattat gaggtccatt ttagaataag
tgctatgtgg tgctgagaag 39300 aatgtatatt ctgttgattt ggagtggaga
gttctgtaga tgtctattag gtccatttgg 39360 tccagagctg tgttcaggtc
ctgaatatcc ttgttaattt tctgtctcat tgatctgtct 39420 aatattgaca
atggggtttt aaagtctccc actactattg tgtgggagtc taagtctatt 39480
tgtaggtctc taagaacttg ctttatgaat ctgggtgctc ctgtattggg tgcatatgta
39540 tttagtatag ttagctcttc ttgttgcatt gattccttta ccattatgta
atgcccttct 39600 gtgtcttttt ttaatcttta ttggcttaag gtctatttta
tcatagacta gaattgcaac 39660 ccctgttttc ttttttttgt tttccatttg
cttgttatgt tttcctccat ccttttattt 39720 tgagcctgtg tgtgtcttga
acatgaggtg ggtctcctga atacagcacg ccgatggatc 39780 ttgactcttt
atccaatttg ccagtctgtg tcttttaatt gggggcattt cacccattta 39840
catttaaggt taatattgtt gtgtgtgaat ttgatcctgt catcatgatg ctaactggtt
39900 attttgcaca ttagttgatg tggtttcttc atagtatcat tggtctttat
attttggtgt 39960 gtttttgcag tggctggtac tagtttttct tttccatatt
tagtgcttcc ttcaggagct 40020 cttgtaaggc aggcctgatg gtgacaaaat
ccctcagcat ttgcttttct gtaaaggatc 40080 ttatttatcc ttcactcatg
aagcttagct tggctgaata tgaaattctg ggttgaaaat 40140 tgtttccttt
atgaatatta aatattggcc cccactctct tctggcttgt atggtgtctg 40200
gagaaagttc tgctgttagt cagatgggct gccctttgta ggtaacctga cgtttctctc
40260 tggtgccctt aacatttttt cattcatttc agccttggag aaacggatga
ttatgtgtct 40320 tcgggttgct cttcttgggg agtatcttag tggtgttccc
tgtatttcct gaatttgaat 40380 gttggcttgt cttgctaggt tgtggaagtt
ctcctggata atatcctgaa gagtgttttc 40440 caacttggtt
ccattctccc catcactttc aggtacacca atcaatcata ggtttgttct 40500
tttcacatag tcccatattt cttggaggct ttgtttgttc cttttcattc ttttttctcc
40560 aatcttgtct tcattccttt tttcagtaag ttgatcttca attactgtta
tcctttcttc 40620 cacttgatcg atttggctat tgatacttgt gtatacttca
cgaagttctc atgctgtgtt 40680 tttcagctcc atcaggtcat ttgtgttctt
ctctaaactg gttattctag ttagcagttc 40740 ctgtaacctt taatcaggtt
cttagcctcc ttgcattggg ttagaacatg ctcctttagc 40800 tcagacaagt
ttgttagtat ccaccttctg aagcccactt ctgtcaattc atcaacctca 40860
ttatctgtcc agttttgtgc ccttgctgga caggagttgt gatcaattaa aggagaagag
40920 ccattctggt ttttggaatt ttcagcattt atgcactggt ttttcctcat
cttcatcgat 40980 ttatctacct ttgatatttg gagctgatga cctttgttgg
ggtttttgtg taggggtcct 41040 ttttgttgat gttaatatta ttgatttctg
tttgttagtt tttcttataa cagtcaggcc 41100 cctcttctgt aattctgctg
cagtttgatg gaggtccact ccagacccta tttgcctggg 41160 tattaccagc
gaaggctgca gaacagcaaa gattgctgcc ttctcattcc tctggaagtt 41220
tcgtcccaga ggggtactgg cctgatgcca gccagagctc tcttgcatga ggtgtctgtt
41280 gacccctgct tggaggtctc tcctagtcac taggcatggg ggtcagggac
ccacttgagg 41340 aggcagtctg tcccttagca gagctcgagc gctgtgctgg
gagaattctc cttgtcagga 41400 tctgctgctc tcttcagagc cagaagacag
gaatgtttaa gtgcgctgaa gctgcaccca 41460 cagccgcccc attcccccag
gtgctctgtc ccagggagat gggagtttta tctataagcc 41520 cctgaggggg
gctgctgcct ttctttcaga gatgccctgc ccagagagga ggaatctaga 41580
gaggcaatct ggccaacggc ctccttggcc tccttagcgc tgtcacggga aaaccgtcta
41640 ctcaagtctc agtgatggca gatgtccctc ccccgaccaa gcttgattgt
ccgagttcga 41700 cttcagactg cagtgctagc atcgagaatt tcaagtcagt
agttcgtgac ttgctgcgct 41760 ttgtgggagt gggacccgct gagtgagacc
acttggctcc cttgcttcag ccccctttcc 41820 aggggagtga aaggttctgt
ctccctgggg ttctaggcgc cactcagttg gaaatgcaga 41880 aatgactcag
ttggaaatgc agagacttct gcgttggtct cactgggagc tgcagactgg 41940
agctgttcct attcagccat cttgccagct cctataaatg gcatttctta atcttactgt
42000 atttccatgg aataaaagta attcttatgc acactgaagt taaaaaaaaa
tagcatttaa 42060 aatttctgct caggaaagta gtataatttt taacataagt
gagtttctct ttgtgttata 42120 atgtaatgaa ttcttatacg catatggaga
ggaaaatgac atttttctat ttatggtttt 42180 agttcagcct ttaagatacc
ttgatgaaga cctggactat tgaatggagc agaaattcac 42240 ctctctcact
gactattaca gttgcatttt tatggagttc ttcttctcct aggattccta 42300
agactgctgc tgaatttata aaaattaagt ttgtgaatgt gactacttag tggtgtatat
42360 gagactttca agggaattaa ataaataaat aagaatgtta ttgatttgag
tttgctttaa 42420 ttacttgtcc ttaattctat taatttctaa acgggcttcc
taattttttg tagagtttcc 42480 tagatgtatt ataatgtgtt ttatttgaca
gtgtttcaat ttgcatatac agtactgtat 42540 attttttctt atttggtttg
aataattttc ctattaccaa ataaaaataa atttattttt 42600 actttagttt
ttctaagaca ggaaaagtta atgatattga agggtctgta aataatatat 42660
ggctaacttt ataaggcatg actcacaacg attctttaac tgctttttgt tactgtaatt
42720 ctgttcacta gaataaaatg cagagccaca cctggtgagg gcacaaagac
tcagtggttt 42780 cgtttctcat tcctcaggac attataagta aagcttaaat
acatacataa actatgtctt 42840 ttctatgtaa atataatggt tataaagtaa
gtttgtatat tactaaaagt tacttatgat 42900 acatattcac ataaagaaaa
gatagcattt aaagtagtac aacactaaac ttagggccta 42960 ctaacttgct
aaatatccta gttttctatg caattgattt ttttttaatt ttagtgggtc 43020
cactgtaagt gtatatatat atggggtata tgagatattt tgataaaggc acacaatgtg
43080 taatcatcac atcagggtaa attaggtatt catcacctca agcatttatc
ctttctttgt 43140 gttatttatt ttttaaatga caaaatttta catatttatc
atgttcaata tgatattttg 43200 aaatatgtat acattgtgga atggctagat
caagctaatt gacatatgca ttacctaaca 43260 tacttacttt tttctgtgag
aacacttaaa acctcctctg agtgattttc gagaatacaa 43320 taccttgtta
ttaactgtag tcaccatgtt gtacaataga tctcttgaac tttctcctcc 43380
tacccaatga aaattttgta tcctttgacc aacatcctct agacaccttc ctccagctac
43440 tggtaaccac cactctactc tctgtttcta taagttcaac tttttcacat
tccacttata 43500 aatgaggtaa tatgacattt gtctttctgt gcctggctta
tttcacttaa cataatgtcc 43560 tctaggatca tttgttttgt tgcaaaggac
aggatttccc tctttttaaa ggctaaatag 43620 tattccattg tgtatatgta
ccacattttc ttcattcatc catcagcaga cacttaggtt 43680 gattccatat
gtgggttatt gtgactagtg ctacaataaa catgagcatg cagatgtctc 43740
ttcaacatgc tgatttcctt tactttggaa atatagacag aagtggaatt gctgaatcat
43800 atggtaattc tatttttaat ttttaaggct gtcctaattt gcattctcac
cactagttta 43860 caagggatcc cttttcccca caacctcatc aatcctcatc
tttcatgttt ttgattatag 43920 ctatctaata ggtataaggt gaggtgatac
tgtggtttta atttgccttt tcctgatgat 43980 tagtgatgtt aaggattttt
tttcatatac ctgtttgaca gttgtatgtt ttcctttgac 44040 aaatatctgt
tgttctgccc atttttaatg aggttgtttt ctcaatattg ggttgtttga 44100
gttctctcta tattttggat attaacccat tatcagtatg atgtaaactc attcgtttac
44160 gttcatcttt gttgtctgtg ttttatttcc aaagaatcat tgtcatgaaa
tttcccgtgc 44220 cccacccttt attattatta ttgttgtact ttaagttctg
ggatacacgt gcagaacatg 44280 cagttttgtt gcataggtat acatgtgcca
tggtagtttg ctgcacctat caacccatca 44340 tccaggtttt aagccctgca
ggttttaagg tatttgtcct aatgctctcc ctctccttgc 44400 acccaactcc
ccgacaggcc ccagtgtgtg atgttgctct ccctgtgtcc atgtgttctc 44460
attgttcaac tcccacttat aagtgagaac atgtggtgtt tggttttctg ttcctgtgtt
44520 agtttgctaa taatgatggt ttccagcttc atccatgtcc ctgccaagaa
cataaactca 44580 ttctttttta tggctgcata gtattccatg gtgtacatgt
gccacatttt ctttatccag 44640 tccatcactg atggacattt gggttggttc
caagtctttg ctattgtgaa tagtgctgca 44700 ataaatatac gtgtgcatgt
gtgtttatag tggaatgatt tataatcatt tgggtatata 44760 cccagtaatg
ggattactgg gtcaaatggt atttctagtt ctagatcctt gaggaatcgc 44820
cacactgtct tccacaatgg ttgaactaat ttacacttat acaatgagtt tggaagtatt
44880 cactcctcct ctattttttg gaatagtttg agtaagattg gtattagttc
tcttctttaa 44940 atgcttggca gaatcagcat tgaacccatt ggatcccaga
atttttattt tttaatggga 45000 gattttttac tatggcttca atcttattat
tgttattgat ctgttcaggt tttggatttc 45060 tttattgttc aatcttcgta
ggttgtatgt gtctaggagt ttgtccatgt cttctagatt 45120 ttccaattta
ttagcttata gttagtcata gttgccacta atgatccttc gaatttctgc 45180
agtaataatt ttcatgtctt ctttttcatc tctgacttta tttattcgga ttctccctct
45240 ttttcttagt ctagcttact aaactaaagg tttgttaatt ttgtttaatt
ttcaaaaaac 45300 caactgtttg atttgttgat cttttgtatt gttttattca
ttttaatttt atttatttct 45360 gctctgattt ttattatttt gtttcatctt
ctaatttcgg ctttgatttg ctcttgtttt 45420 tgctccagtt ttttaagatg
catgattagg ttgcttattt gaagtctttc tacctttttg 45480 atggatgtgt
ttattgatat atattttctt cttaatattg cttttgatgt atccaataag 45540
ttttggtatg ttttgtttcc attttcattt gttcacaaaa tttttaaatt ttatttttaa
45600 tttcttcatt gaccaatgtt cattcaagag catatttttt aatatctgtg
tatttatagt 45660 ttccaatgtt acttttgtta ctgatttcta gtttttttct
actgtggtca gaaaataaaa 45720 ttgatatgat ctcaatttct ttgaacttct
tgaggcttgt tttgtcctct aacatgtggt 45780 ctatcttgaa gaatattcta
agtgctgaga agaatgtgta ttctgaagct cctgaatgaa 45840 atgttctata
aatgtctctt aagtctattt agtctatagt gcagattaac tctgatgttt 45900
ctttgttgat tttctgttgt gatgatctat ccagtgctga aagtggaata ttgaagttcc
45960 caacttcagt tgtattgggt ttatttctct ttaacttaat aacatttaat
aacatttgct 46020 ttatgtattt ggggactcca gtactgggtg catatatatt
tacaattgtt atatcctctt 46080 gctaagtcga ctcctttatt attatataat
gatcttcctt gtttcttttt atagatttgt 46140 cctgaaatct attttgtctg
atataagagt gactattccc ttttcttttt tggtttaaat 46200 ttgcatggaa
tatcttttct atcctttcat ttttagtcta tgtgtgttta tataggcaaa 46260
gtgaatttct tgtaggaagc atgcagttgg gtctttttgt tattattcat tcagccactc
46320 tatgtgttct aattggagaa tttagtccat taaccgtcaa tgtattattg
acaggaaagg 46380 acttactact gccattttgt tatttgtttt ctagttgttt
cattggtcct gtcttcattt 46440 cttactgtct tcctttgtgt acaagtgatt
ttctctggta gtatgtttta gttttttgct 46500 ttctagtttt tatgtatcta
ttttagtttt tcttctcatg gttaccatga ggcttgtaaa 46560 taacatctta
taacagattt ttttaagctg atgaaaactt aattatactc ataatagaaa 46620
cggaaacaag caaagagaaa actaaaaaat ctacacttta actttattct ccccttatca
46680 ttttttgagt tgttgtctat atatcttttt atattgcctg tatttaacaa
atcattgtag 46740 ttgttattat tcttgatagg cttgtctttt agtcttcata
ctaaatatat gcatggttta 46800 cacactgcaa ttacagtgtc agaggtgttc
tatatttgtt tgtgtactta cttttactgg 46860 tgagttttat actttcaggt
gatttcttgt tgtttgttag caccttatct ttcagactga 46920 aaaattccct
tcagcaaatt tttgcaaagc agggctgatg ttaatgaaat cccttaggtt 46980
tttgtttgtc ttggaaagtc tttatttctc cttcatgttt gaaggataat tttgctggat
47040 ataacattct aagttgaaga ggtttttttc ttcactactt tgaatatatt
atccccatcc 47100 cacttggcct gtaaggtttc cactaaaaag tcaactgcca
gatgtatcag agttccttta 47160 catgttattt gcttcttttc tcttgctgct
tttaggatac tttgtccttg aactttgaga 47220 gtttgattat tttgtgtctt
gaggtagtct catttgaatt gaatctcctt ggtcttcttt 47280 gagctttatg
tacctgggta ttcaaatctt cctctaggtt tgaaaagttg ctatttcttt 47340
gaataaaact tcttcactga tctcttattt tacatccttt ttaaggccaa taactcttag
47400 atttgccctt ttgagggtat tttctagaac ttgtaggtgt tttcactcct
tttaactctt 47460 tttttctcct tggactctgt ctttttaata tagctatttt
caagctcact aattcttttt 47520 tttctttacg tcttctaaaa aaatgggata
catgggcaga atgtgcaggt tggttacata 47580 ggtatatgtg tgccatggtg
gtttgctgta actattgacc catcctctaa gttccctccc 47640 ctgactcccc
acctcccaat aggctctgat gtgtgttgtt cccctccctg tgtccatgtg 47700
ttctcaatgt tcaactgcaa cttatgagtg agaacatgta ttgtttggtt tactgttcct
47760 gtgttagttt gctgaggatg atggcttcca gcttcatcca tgtccctgca
aaggacatga 47820 actcattcct ttttatggct tcatggtatt caatggggta
aatgtgccac attttcttta 47880 tccagtgtat cattgatgag catttgggtt
ggctccaagt ctttgctatt gcaaatagtg 47940 ctgcaataaa catacctgtg
catgtgtcct tatagtagaa tgatttatat tcgtttgggt 48000 atatatccag
taatgggatt gccgggtcaa atggtatttc tggttctaga tccttgaaga 48060
atcaccatac tgtcttccac aatggttgaa ctaatttaca ttcccaccaa cagtgtaaaa
48120 gtgtccctca ccagcatcta ttatttcctg actttttaat aattgccatt
ctgactggca 48180 tgagatggta tcttattgtg gttttgattt gcatttttct
gatgaacagt gatgttgagc 48240 tttttttatg tttgttggca gtgtaaatgt
cttcttttgt aagctcatta attctttatt 48300 ctgcttgatc aatactgctg
agagacactg ttgtattttc cagcttgtca attgaatttt 48360 tctgctccag
aatatctgct ttgtttttta aattatttaa tatctttgtt aaatttctct 48420
gatagaattc tgtacttctt ttctgtgtta tcttgaagtt tgttgagctt ctgaaagaca
48480 gctattttta attccatgtc tgagaggttg catgtctcca tcactctaga
attaatcact 48540 ggtgccttat ttaatctact taatgaaatc acattttcct
caatgttcct gatgcttgca 48600 aacattcact aatgtctaga cattgcagag
tcgattgttt atttcaatct tcacaatctg 48660 ggcttgtttt tattcatcct
ttttgaaagg gcttttaaat aattcaaatt agaaattaag 48720 acagagaaat
tcacccaaag ccatacagtt acatggaaat tgaataacat gatcctgaat 48780
ggcttttggg taatgaaatt aaggcagaaa tcaagaagtt ctttgaaact aatgagaaca
48840 aagataaata taccagaatc tctgggacac agctaaagca gtgttaagag
ggaaactcat 48900 aagactaaat tcccatatca aaaacttaga ccgatctcgt
taacaaccta atatcacaac 48960 taaaagaact agagaaccaa atgcaaacaa
actccaaaag tagcagaaca caagaaatta 49020 ccagtcagtg ctgaactgaa
aaaaatagag acatgaaaga acatccaaaa aatccagaaa 49080 ttcaggggtt
ggcgttttga aaaaattaat aaaatagacc actagctaga ctaataaaga 49140
agaaaagaga gaggattcaa acatacacag tcagaaatga taagggggat attaccactg
49200 accccacaga aatacaaaca accatcagag aatattatga acacctctat
gcacataaac 49260 tagaaaatct agaagaaatg gataaattcc tgaacacaca
acaccatccc aagactgaac 49320 caggaagaaa ttgatttcct aagcagacca
ataatgagct ctgaaattaa ggtagtaata 49380 aatagcctac caaacaaaaa
aaaaaaaaaa agcccagcac cagagggata gagggattca 49440 cagctgaatt
ctaccagctg tgcaaagaaa agatggtact atttatattg aaactattac 49500
aaaaaattga ggagaaggaa ctccttccta actcattcta tgaagttagc atcatcctga
49560 taccaaaaac tggcagagat acaacaaaaa gagaaaactt caggccaata
tccttgatga 49620 acatcgatgc aaaaatcctc aacaagatac tggcaaactg
aatccagcag cacatcaaaa 49680 agcttatcca ctacgatcaa gtaggcctca
tccttgtgat gcaagattgc ttcaacatat 49740 gcaaatcaat aaatgtgatt
catcacatac acagaattaa agacaaaaac cacatgatta 49800 tatcaatagg
tgcagaaagg actttcaata aaattcatca tcgatttatg ctaaaaactg 49860
tcaataaact agttgttgaa ggaacatacc tcaaaataat aagagccatc tatgacaaac
49920 ccacagccaa catcatactg aatatgcaaa agctagaagc attcaccttg
aaagccagca 49980 caagacaagg atgccctctc tcacactcct attcaacata
gtattggaag ttctggccag 50040 gccaatcagg caagagaaag aaataaagcg
catccaaaca ggaagagagg aagccgaact 50100 atctctgttt gcaggcgaca
taatcctata tctacaaaac cccatagtct cagcccaaaa 50160 gcttcttaca
ctcataaata tcttcagcaa agtctcagga tacaaaatgt gcaaacatca 50220
ttagcattct tatacaccaa caacagtcaa gcagagagca aaatcaggaa cacaattatc
50280 attcagaatt gccacaaaaa gaatataatg cctaggaatt cagctaacca
gggaggcgaa 50340 agatctctac aaggagaact acaaaccact gcttaaagaa
atcagagatg aaagaaacaa 50400 attgaaaaac attccatgct catggataga
aagaatcaat attattaaaa tggctatagt 50460 gcctgatatg gtttgactct
ctgtccccac ccaaatctca ccttggattg taataatccc 50520 cacgtgtcaa
gggtgggacc aggtggagat cattgaatca tgggggcagt ttccaccatg 50580
ccgttctgat aatgagtgag tctcttgaga gctgatggtt ttataagaag cttccccctt
50640 cactctgctc tcattctctc ctgccatcct gtgaagaagg atgtatttgc
ttccccttcc 50700 accatgattg taagtttcct gaggcctccc cagccatgca
gaactgtgag tcaattaaac 50760 ctcttttctt tataacttac ccagtcatgg
gcagttcttt atagcagtgt gagaatgaac 50820 taaaacaatg cccaaagcaa
tttatagttc aatgctattt ctgttgaacc accattgaca 50880 ttcttcacag
aactagaaaa aaactatttt gaaattcata tggaaccaaa aaggagtctg 50940
aatagccaaa gcaatcctaa gtaaaaagaa tgaagctgga gttatcacgc tacttggctt
51000 taaattatac tacagggata cagtaaccaa aacagcatga tactggtatg
agaacagaca 51060 catagactga tgaaacagaa tagagaaccc agaaataaga
ccacacacct acaactatct 51120 gatctctgac aaacctgata aaaacaggca
atggagaaag gattccctat tcaataaatg 51180 gtgctgtggc caggtgtggt
ggctcaggcc tctaatccca gcactttggg agaccgaggt 51240 gggtggatca
cttgaggtca ggaattcgag gccggcctgg ccaatatggc gaaaccttgt 51300
ctctacaaaa gatacaaaaa ttagccaggt atggtggcag gtgcctgtaa tcccagctgc
51360 ttgggaggct gaggcacgag aattgcttga ccccaggagg cagacattgc
agtgagccaa 51420 gatcgcacta ctgcactcca acctgggcaa cagagtgaat
ctctgtctca aaaaaaaaaa 51480 aaaatggtgc tgggataact ggctagtcat
atgcagaaga ttgaaactag actccttcct 51540 tacactatac acaaaaatta
acacaaggtg gattaaagat ttaaatgcaa aacccaaaac 51600 tgtaaaaacc
ctgaaagaca acctaggcaa tagagttcag gacatgggca gggcaaagct 51660
ttcatgatga agatgccaaa agcaattgca acaaaaacaa aaacagacaa attgaatcta
51720 attaaagagc ttctgcacag caaaataaac tatcaagaga gtaaacagac
aacctacaga 51780 atgggagaaa aattttgcaa actatgcatc caacaaaggt
ctaatatcca gcatctataa 51840 ggaacttaaa tttacaagaa aaaaacaacc
ccattaaaaa gtggacaaag gacatgaaca 51900 gagacttctt agtagaagat
gtacatgcag ccaacaggca tatgaaaaaa gcttgatatc 51960 gctgatcatt
agagaaatgc aaattaaagc cacaatgaga taccatctca caccagtcag 52020
aatgactatt attaaaaagt caaaaaataa cggatgctgg caaggttgtg gaaggaaaag
52080 aatgctgttg gtgagggtgt aaattagttt agccattatg gaagacagtg
tggcgattcc 52140 tcaaagacct taaagacaga aataccattt gactcagcaa
tgccgttact tcgtatatac 52200 ccaaggaata taagtcattc tgttataaag
acacattcac gtgtatgttc actgcaacac 52260 attcacaacg gcaaagacac
gtaatcaacc taaatgctca tcaatggaag actggataaa 52320 gaaaatgtga
tacgtataca ccaaggaatt ctatgcagtg agactaagtc ctttgcaggg 52380
acattattct tggcaaacta agtcaggaac agaaaaccaa atgccacgtg ttctcactta
52440 caagttggag ctaaatgatg aaaacacacg aacacataaa ggggagccac
acacactggg 52500 tttttcggaa ggtagaaggt gggaagaggg agaggatcag
gaaaaataac taatgggtac 52560 taggcttaat acctgggtgt taagggtagg
ctattgggtg ataaaataag ctgtacaaca 52620 agcccccatg acacagcttt
atctatgtaa caaacctgca catgaacccc tgaacttaaa 52680 ataaaagtta
aacaaaaaaa agagacagag aattcaaatg ggtttgagtg ttgttaccta 52740
agcctgtggt catggcagcc tctc 52764 38 61329 DNA Homo sapien 38
gagtgcagtg gcacgatctc ggctcactgt aacctccgcc ttccaggttc aagcaattca
60 cctgcttcag tctcccaagt agctgggatt ataggcacgt gccaccatgc
ccagctaatt 120 tttgtatttt tagtagagac ggggtttcac cttgttggcc
aggctggtct tgaactcctg 180 acctcgcgat ccatttgcct tggcctccca
aagtgctggg attacagtca tgagccaacg 240 cacccggtct ggataattgt
tttctaacgt atctatattt gcctgtactg gtactcattg 300 ggcccttctg
tgctcactca tggcatcttc ccatcagaga atatctgagt ggtagaccat 360
tcttgcatgt tttggttttt acagaaatat ttttagttgc cattaaaggc tgacatccat
420 tgttggaaat cattcttttc tgcctctttg gatgcatctg agtttgaggt
tttactttcc 480 ttctacccca ttaggttgta tgtttgcttt tgttttgtag
aatttagtca gcagcttaag 540 gcttaggaac attagatgct aatcatgcca
tcttagcttt ttttgtgagc ccaaggagta 600 tattcggggg tgggggtggg
aatgaaagtt actggggagg cagaaagggg tcattttttt 660 tttttttttt
tttttggaga cggagtctcg ctctgtcacc caggctggag tgtagtggcg 720
cgatctcggc tcactgcaac ctctgcctcc cgggttcaag caattctcct gtctcagcct
780 cccgagtagc tgggactaca ggcacatgcc atcacaggcc tggctaattt
ttgtattttt 840 agtagaaatg aggtttcacc atattggtca gcctagtctg
gaacttctga cctcaggcgg 900 tccacctgcc tcggcctctc aaagttttgg
gattacaagc gtgagccact atcccagccg 960 aaagtggtca ttttgaaatc
agaagttgta tctttttgtg gttgagggtg ataaaaactg 1020 aaaaactgtt
ttcaagtttg gagcatggga gggaaaaccc agcgatttgt tctggtctcc 1080
ctcagctcag tgtgtttgaa gccaaaagcc tgcttgtgtg cctctcccaa ttcatttgct
1140 cctagttcat ttgctacttg catagaaatt ttcatgccga atctcactct
tcagcggata 1200 cctgagaaca ttgttgctac aatctctaaa atcagatttt
ttcaagaaag aatttttatt 1260 aataacctca agtacttttc gcctacaccc
tcaggctcac tttttgtata ggtttttctc 1320 cctggagaat cctacatgct
gtattgatgt ctcaattctg tgtgctgctt cttcagtgct 1380 accttgccct
ggccaaagga ggcttatctg atgaccatgt cctcatcctg cctattggac 1440
actaccagtc agtggtggag ctttcagcag aggtggtaga agaggtggag aagtataagg
1500 ccactctgag acggttcttt aagagtcgag ggaaatggtg tgttgtattt
gagagaaatt 1560 ataagagcca tcacctccag ctacaggtag gtggtctgtt
catagaaact tggaagggcg 1620 ctatggggac tttgatattg gtaaataagt
atttaaatag gctgggcgag gtggctcacg 1680 cctgtaatcc tagcactttg
ggaggccgaa gcggttggat tgcctgagct caggagttca 1740 agaccagcct
gggcaacacg gtgaaaccct gtctctacta aaagtacaaa aaaattagct 1800
gggcgtggtg gtgcatgcct gtaatcccag ctactcggga ggctgagaca ggagaatagc
1860 ttgaacctgg gaggcagagg ttgcagtgag ccgagattgt gccactgcac
tccagcctgg 1920 gcaacagagc gagactccat ctttaaaaaa aaaattatat
aaagtttagc tttaatatgt 1980 aaaatattta atatttttat attaatattt
aaataacatt catttaaaaa gtttttaaag 2040 agtatctggt ggtaacccca
gagacttcac agggaagtgt gtggtcagta catgtaaaca 2100 gcatagatag
taagttcctt gtttagaaaa ggaatgggta gttgaggaag ttccaaggaa 2160
gatgtggggc ttgctggata agaatttgga agaaagtgag tattttgtgt ggaggcccag
2220 catgagggaa agcttagtgg taggaaatag tctcaaagcc tgcatggggc
tagcagctca 2280 gaaaatcaga ttgtaaatca tttgtagttg tggtggtttt
ctttttggca catttgttaa 2340 gttttcaaat gggcctaact ctgccacata
tataatatcg gagatggcaa aggcttgtga 2400 cggagatatc tctcttaagc
ctttcctgca tcagagaatg gctcccacat gtgtcaggct 2460 atctcataag
tttaagtcct tacagacagg ctgacacagg cttgtctggc ctgaggatat 2520
tctgtgctga ggggtgtttc tccacccctg tgatggattt gctacttctc agagactgtg
2580 caggcacaac ttctctggga cctgcacaag gaatttctcc tgtctcccca
acccccaggt 2640 cattcctgtc ccaatcagct gctctactac tgatgacatt
aaagatgcct tcattaccca 2700 ggcacaggag cagcagatag agctgttgga
aatcccagag cactctgaca tcaagcaggt 2760 gaaacagggg atggtgatat
tcagggaaga gtaaatatct gttttttttg ttgttgtttg 2820 ttgtttgttt
gtttgtttga gacagagtct tgctctgttg cctaggttgg agtgcagtgg 2880
catgatcttg gctcactgca acctccgcct tccgggttca agtgattctc ctgcctcagc
2940 ctccccagta gctgggacta caggcatgcg ccaccatacc tggctacttt
ttgtattttt 3000 agtcgagacg gggtttcacc atattggcca ggctggtctc
gaactcctga ccttgtgatc 3060 tgcctgcctt ggcctcccga aggtattaca
ggcatgagcc acccacgcct ggcaagatct 3120 ggtttttgac atgttgagcc
taaggttttt gacatgttga gctatcaagg cattccaaaa 3180 atgttattaa
ataggcaatg ggttgggcat ggtagctcac acctgtaatc caacactttg 3240
ggaggctgag gcgggaggag cacttgaagc caggagtttg ataccagcct gggcaacaaa
3300 gtgagacacc ttgtctctac aaaaaataaa aaaataagct tggcatggtg
gcatgtgcct 3360 gtagtcttag ctactcagga ggctgagagt caggaggatc
acttaagcct aggagtttga 3420 ggatgcagtg aggtatgatt gcaccactgt
acttcagcct gggcaacatc gcaagaccct 3480 gtctctaaaa aaaaaaaaaa
aaaaaagagg cagtggacaa tttggaacta gagcactggg 3540 tattgactag
actggagatg aacagggcat tactccaaag tttagtgctt tcctgggaag 3600
agcaagaaat tagaatttcc taaggtagga acttagaagg atagacgcca ctgggaggat
3660 attgttagta tcttccagtt agacttttta agccctagtt ccactaagtg
gcttgggatc 3720 tatagcaaca tacatatctt tttttcttgt gcatttagac
tgaattattt gcctagagat 3780 gaactcagat tatagatttt taaagcatta
ttggacatcc tcatacaact ctgtttactt 3840 tagattgcac agccaggagc
agcatatttt tatgttgaac ttgacacagg agaaaagctt 3900 ttccacagaa
ttaaaaagaa ttttcctttg cagtttggaa ggtatgtttt tcttcttttc 3960
aatgtatttt cactagacct gctatgaaca gagtggtggg ctacatggga caattgataa
4020 aaagctaaat tgtaaacagt ccttgcccca gaggacccat ggtcttaggg
cagatgttta 4080 catatatgac agtaatgtaa tgtagaatgt gaagagtacc
atgacagagt tttggagaac 4140 aatgtgctgt agaaattcag aaaagggaca
gctcatttct agttggtgaa agccttcatg 4200 aaagtggcaa agttgaacct
gagaggcata tttagagggc aaagattgag gtgagggcat 4260 tctagccaga
agtgctaggg aactttaatt gagcacatac tctgtgccaa gaactgtacc 4320
aagaacttta cacatacatg actcatttaa tcttcataac tctgtaaggt aggtactgtc
4380 atcccgattt accagtgtgg aaaatgaggc tctagagcag tgcccggtag
acttttcaca 4440 acgatggaaa tgctcaaata tctgcactgt ccaatacagt
tgtgactagc cacttgtggc 4500 cactgagtgc tgaaatgtgg ctaatgtgac
tgaggaacta aatttttgat tttatttaat 4560 taattagaat ttaaatttaa
atagccatac ctggctaatg gttaccccac tggatagcag 4620 gctgtagaga
gcttaggtag cttgtcctag tcacacagtg gtaggttcaa acccaggcct 4680
gttggacctc aaaacttgag gtcttaacct cagtgttcta cagtatggta aaggcctaga
4740 ggaggcttca gttctgttcc tcgtatcagg caaaacatga ctgcaaaacc
atagagggag 4800 taacataact taagcaaaga ttgaagatgg gaagttgctg
gcaagaagat acagtttgaa 4860 ggggagtgga aaatgtgtag aacagagtat
ctgatgctgg tgtctgtgat ttgcagctgt 4920 tttttctccc ctagttaaaa
atgggctttg agtgagcctt gaacctgtct tggactaatt 4980 cagaaagcct
ccaagagatt gcccccttag ataagtccag taaatgtagt tgtctacatt 5040
tttcaaaaag ataaatggga acaagtgtgc tacgagattc agacatctga cgtgatgttt
5100 tcgttgttgc ctcagcaaca gatgcggtta tgaacattga caacataagc
acacgattgc 5160 agatacacat aggcgtataa agcacacaaa aagaagattc
tctgaatggt tttgtgctta 5220 tgatgtattt tcaatattga gtaattatca
agatactaat ttatagttat atagcatttt 5280 atagttaaca aaatatttga
aatggcatat ttttttcagc tgtacaagaa aggtagttct 5340 attttccttt
tatatttgaa ataggctcaa agtttgccca gggagcttag gttctttgac 5400
ttatagagaa ttcatgattt gtttctaact cttctattta ctgtaacttc aaagtggaaa
5460 atgaaccata taaatgtgta atagaattat ttctaatata tcaaaataag
taaaaataaa 5520 aatataaagc aatggcgatt agaaggcagc ttgtgtgtga
gtgtgtctgt atgtagttat 5580 tctttagtcc taagagccaa gatagttgtt
ttgttgacat tgggcttgtg ctgctgttac 5640 gtgggcctga gtcccctaga
aggagaggca gaggaatttt aaatcagagt gggtatgact 5700 aggctcctga
ctctgcaaac tcatgtctag ttgaggtgac aaaactacag tttaaggcag 5760
gttgtaatca cagctaatat ggcactttga ctgtgttatc tctgtacctc acaaaaaccc
5820 tgtatggtgg gtgttcttat ttgcatttca taggtgagga aatcaagact
cagagatgaa 5880 atcatctgtc tagatcatag atagcaagtg gcaaagctgg
aaatgggatc cagtctctga 5940 tctttctcac accagggcct gccctttttc
tatgtggtgt tgcttctcta agtaccaata 6000 atatagttcc aagtttttta
agagagggag aagcaagtgg ggtttgaagg atcaggaaat 6060 atatagtacc
tgagctggtc atttaaaacg ggtatgattt tgatcaatag tatggcagag 6120
atggcactca gggcaggaga gcaaagtcag agaaatttgc atacctcctt tgggtgatgt
6180 tgaggagagc tcaggctgat agtttccatt ttgacttctt tgtgctattt
ctgcagggag 6240 gtcctggcca gtgaagccat ccttaatgtt cctgataagt
ctgactggag gcagtgtcag 6300 atcagcaagg aagacgagga gaccctggct
cgccgcttcc ggaaagactt tgagccctat 6360 gactttactc tggatgacta
aaacaaaggg aagaactttt tatgaactcc acaggaagta 6420 gtaaagcttt
tttttttttt taattaaaag aatttttttt gagacaaagt ctcgctctgt 6480
cacccaagca ggattgcagt ggcataactg tggctcactg tagcctcaac ctcctgggct
6540 ctagagttcc tcccacctca gcctcatgag tagctgggac cacaggcgca
tgctaccatg 6600 cctggcaaac ttttttgatt ttttatagag acaggagggt
ctccctgtgt tgcccaggct 6660 ggtctgtaat gcctaggctc aagggatcct
ctgccttggc ttcttaacct gctgggatta 6720 caagcatgag acaccattcc
tggcctagaa gcctattttt aaagaaacta caatctccca 6780 tggggactgt
ttccctgcct cttttgtgca gtcccatgga acttgcctac agcaagaggc 6840
ctaagattga atctttttgg ggaaaagtca ttctaggatg aaaatcctat gttaaggccg
6900 ggcgcagtgg ctcacgcctg taatcccagt actttgggaa gccgaggcag
gtggatcacc 6960 tgaggtgagg agtttgagac cagcctggcc aacatggtga
aaccccgtct ttactaaagc 7020 tacaaaaatt agctgggcgt ggtgccaggc
acttgtaatc ccagctactc aggaggctga 7080 ggcaggagaa ttgcttgagc
ctgggaggtg gaggttgcag tgagccaaga tcgctccatt 7140 gcactccagc
ctgggtgaca gtgaaactcc atctcaaaaa taaaagaata aaagtatgtc 7200
tgtcatccag ctcctatgtc tgttatccag ctccaagtac agcttgtgta tatcaacatt
7260 ttcaaaaacc tttaaactac caaatgaaat ccagatgttt ttccatggtt
gagaagtgtc 7320 ttgtgttcat gttgcctgtg accgttgaaa tgaagtagta
tgtgtaatag caggggctag 7380 agctttaggc ccagctgtta ctccaaaatc
acacagggca gtttctccct gaacaaccca 7440 aagtggaaaa gggatccaag
gcttttgcta tttttctctt aaagcattca tcttattcat 7500 agataagagc
ccatttagtc cttcggatat ttaactccta tgtgccaggc atttttctaa 7560
gctccaggac catagtcatg tcccaatatg gtccctgttg tcccagagat tacagtttaa
7620 tggggaatag aagggacaga tttaagaaaa aaaagataat tctcctttgg
atattgtttt 7680 aggtgcttga tatatctaat atgtgtctgt tgtgctgctg
ggcagttcta aactcatcag 7740 tacctgggtt ggttacaggg ctttggggat
catcagggta taggccatag gtgaaattaa 7800 gatagggtaa atacagtcct
gagctgaaga tgattacctg agaaatatca gtatttaagg 7860 agtgcttgaa
aatagtgatc taacatttac atagagaagg agcaaagaga atgctacata 7920
ttaggcgcaa aaagaaaaat atttttacct ttatattggg cacttatggt ccagacactg
7980 tttcatttaa tcttcacaaa atctctgtga ggtaggtatt acctctattt
tactcatgaa 8040 aaaaaaactg agactcggaa aattgaagat tgtggagcct
ggggtcaaca ggcttaggct 8100 ttttaaaaaa caggccaggt ggctcatgcc
tgtgattgca gcactttggg aggctgaggc 8160 aggcagatga cctgagatca
ggagtttgag accagcctag ccaacacggt gaaaccccgt 8220 ctctactaaa
aatgcaaaaa attagctggg cgtggttgca tgcgcctgta atgccagcta 8280
ctcgggaggc tgaggcagta gaattgcttg aaccctggag gtggaggttg cagtgagctg
8340 agatcacacc attgcactcc agcctgggtg acaagagtga aactccatct
caaacaataa 8400 caacaacaac aacaaaaaac aactcaaatt actcatggcc
ctgggaatgg gggcatttaa 8460 aaagttaacc aagggaacca acctggtttc
cacaaagatg gccacttctg ggtcagccca 8520 atcatcagta tccctgggtc
ctcagcatcc cttcctacct cttttgactg ggaattacga 8580 gattacagcc
ccacctacgg aaattgagca cattagcact ggccatgctg tacatacagc 8640
aaatgagagg gcttcatact gttacaacaa tctgtaccac aattctctgt gcggtctgcc
8700 cagctctaaa tgacatgtcc tcagaggcat catcatctcc aagtccatgg
aaaccatgca 8760 cttacagaaa ccagaacctc ccataccaca tgttactaat
gacctacaac atcgacttcg 8820 tctgtatcat gcagtgctca acattctggt
tgcagcatca catttatcaa taatttgatt 8880 ccatgctgat tcctttactt
ggagtattga gatcccccag gtacagtgaa ggtatgaccc 8940 tcttaaatgg
aagaccagga gttgtttccc ctatgctatg gcaatccgcc acccggaagc 9000
acttgctgca ccaacagcat tcccaaagct tgcctctgca gccctgtagc acccattaga
9060 aacacttctc acaatgaact actgtgctgg caaacccata actggactgc
cggacagcaa 9120 gctctgtgag gcatctagga aaaaagctcg cgaaatcccg
agaaacactc ttcactccgg 9180 aatagcgtca cattcttgag tcatttggga
tgacagggga tatcctggag caggtgtatt 9240 gggaagtcag tgaacaagaa
ggctaattca ttagtgggcc tgctagcgta gtattttaac 9300 gatttcgcag
cctttaaatc attggtggtc tttagcgtgc cggatcagct ggtggtggag 9360
ttggaggata gatggcaggg atctgaagaa aggtgaagtg tagataggag tagagacaat
9420 atgaagttta gaaaagacta attgctggac gcggtggctc atgtctataa
tcccagcact 9480 ctgggatgcc aaggtgggag gactacttga gaccaggagt
tcgggaccag cctgggcaac 9540 gtagcaagac catgtctcta caaaaaataa
aaaattagcc aagcgaggtg gtgcgcgcct 9600 gcaatcccag ctactcggga
ggtgagaaga tcgcttaatt ccgggaggtc gaggctgcag 9660 tgagccgtga
tcgtgccatt gcactccagc ctgggcaacg gagcgagacc ccatcttaat 9720
taaaaagaaa aaaatacagg agagactggg ctgctttgaa aagtggattt gacgggaatg
9780 tggggtttgg agagatctta tgttttccaa gactagaaaa acattgtggt
tccgttcagc 9840 cctcccggaa aatacgtagg ggacttaaag agcggaccga
gtacctgttt gtttccccat 9900 cccttagccg gcagggctct ggcagcgccc
tgggagaact tagggaaggg ccttctcgca 9960 gcggttaccg gaagtgacgc
attcattctc gcgagaacaa agacgcgcga gcatcggcgg 10020 cccggaaccg
gccttggaac aactgtggaa cctgaggccg cttgccctcc cgccccatgg 10080
agcggccccc ggggctgcgg ccgggcgcgg gcgggccctg ggagatgcgg gagcggctgg
10140 gcaccggcgg cttcgggaac gtctgtctgt accagcatcg ggtgaggcgg
ggcgtgagag 10200 ggagcggcgg tgggttggag cggcggtggg tttgagcgtc
tgtggaacac agggctgaca 10260 gtgtgggata gatggttggt gggaccttgg
gcagtatttg ggggccagtg ggcttgtgtg 10320 tgtgtattag agagaaagat
actatgcctg tgtgtccaaa gttgtatttg aggtactggg 10380 catagataga
gctgttgcgg ggaacttggg tatgactgtc aggctgtgac caggtgttta 10440
gtgagtgtga cagtgcgtgt gtgtctctgt gaatgtggag tgtcagggtg tgtgaggcag
10500 atgcacgtca cagtgtgtat gtaggagcac gaatgtgtga gaagagggct
gaaggggact 10560 gcctttggga atgtgtgcgt ccgtgtgtgt gtgtgtgtgt
gtgtgtgtgt gtgtgcgcgc 10620 gcgcgcgcgc gcttggaggc catatgatat
cagtggggac tggagtagac aaggaaggct 10680 tcatgatggg tccagagtta
agcctttgga agcatggatg ggctttggaa ttggggatgt 10740 attccaggtc
gggacagtat gagggtgagt tggtgcagag catggcttca aacgaaaggc 10800
tatattagta gatggagatg gggggcgctc tagtaggtgg aggacagtta acatcagact
10860 caagaatttt actttgattt aatagagaat gtttaaccat tgtagtagca
gttaaaaaat 10920 catatttggg tcttgtttgt ttgtttttgt ttttgagacg
gagtctcact ctgtcgccca 10980 ggctggaatg cagggcgcga tctcggctca
ctgcagcctc ctcctcccga gttcaagcga 11040 ttctcctgcc acagcctcct
gagtagctgg gattacaggc gcccgccacc gtgcctggct 11100 aatttttttg
tatttttagt agagacgggg tttcaccatg ttggccaggc tggtctcgaa 11160
ctcctgacct caggcgatcc acctgtctca ggctcccaaa gtgcagagat tataggcgtg
11220 agccacggca ccaggcctgg gacatgtatt ttttgatata ccgataaaag
ctgtgaaccc 11280 tttccccaga aaaatgcaca gatgcacatt cacacaacca
tgcattgcca tttgagaggt 11340 tcgtgccttc ctaaaggaca ttttaggtaa
atagaccctg gactaataat tcctgtaata 11400 taggctttgt tttgttgcct
gtaaagtaag tatagtataa taattacatg catcatggat 11460 ttgagagtta
aataaaatag taatgtgtgt aaagctaaat acggtacctg acccatgaaa 11520
gccctcaact gttagcacct gctgctactt tgttgttgtt gctgctatga tcataatgct
11580 cctcattaat tggccttaag gaaatgtcaa gataaaaagt gttgttctag
gaaaatggtt 11640 ttgtaaagga taaatttgag tggaacaaca tatgggacaa
gatgaacagg taagatctta 11700 ctaaaatgcc tgggcttggc ttgtgaatag
agaagttaat tcaagaggaa aagtaataat 11760 ttgttatcta aaggaacatg
ggtgtagtga gaaagctcgt aatttggggt gggagaaagg 11820 ggagatagga
gtctaccttg ggaaaatagg catatttagg gtaattagaa tgacaactga 11880
aactgttaga actgaggaag ttttgtgaac caggtttcta gatagggcat aggagaaaat
11940 cttggagaaa gagaatggaa ggtgaaggac taaggcatta agtcctgtga
tagcaggata 12000 agtagtaaat ttaataattc tcaccatcag agggaacata
acacactaaa aaaaaaaaaa 12060 agtgggagcc aagtgcttcc agtggcatgt
tgcctgtgtt acctaagcct gcctatccgc 12120 ctggtcataa ggaaaaagga
tgcatgctgg cagcttttgt gacttctttt cttggtccct 12180 gggtggggat
ggatgaatta atcacttttt gttttattat ggtaaagtat acataacaca 12240
acatttatca ctttattttt aagtgtacag ttaggtggca ttaagtacat tgacgttgtt
12300 gtacagccat aaccctatct ccagaatatt ccgacttttt tgttgttgtt
gtacatctat 12360 aaacttccca tagaatattc tttgtgaaaa ggtagaattt
cacagagttg atgaaatata 12420 aattaaggag ttgaaggatc ttacaaagtg
ggatttttga aatagataac ttattgctcc 12480 attaacatga gcattcaggg
ctcagtgttg ttcatggttg tgcctgattc tcatttccat 12540 ggtggtaaag
gcctgcatgg gacatagata taactggtca cctgctcata acaaaaggaa 12600
gattgaaggg agtgagaact gtgggtaggc tagcatgcag tctatcctag tttccttctt
12660 ctctatgcac caggtatcat cttatctcta gcttcttttt tcctctcacc
ctccaattca 12720 cactacccta tccaagctat gtcttgcttc cccacctata
atatattttt tctgtcctgg 12780 cccctgggct cttctctctg tattcataca
cagcctttat tacaactctg gtttgtgtca 12840 gagattacat atgttggaaa
ttcttttgaa ctctccttgt gatatcagaa acatagtgta 12900 gcctgtttag
ccctgactgc tatttctgag ctgcattatt aatgttgcca gaatatgatg 12960
gggtctggct atttcctgta atatgcatca gaatgggttt aatttcagtc agaattggct
13020 aaattactag tcacaggcaa gagatgttag gttcattatg tagctatgga
agcagaagta 13080 cactcattgg taggattttt tttcttcatg tcatttgaaa
tgcatacatt aggattggag 13140 ttttttgctt taggcttggt tatattaaaa
ctttctcttg ctgtttggag tccaggaagc 13200 cctgaattac cttcttcatt
ttagttatgc tagaagctgg ggcacatatt agttgtcttg 13260 attaagctct
gaaaggggtt tgctgaggag acaggatctt tttttttcta acattatata 13320
tgagagagag ttctgaaaag acctgttata aaatcagttt caggcacttt tcccttgtat
13380 gctgttgtgt gtttgaaggt cagttttaaa gcattggtgt aatagtctca
tggtttatct 13440 ttttaaaaag cattgcctca agagagctgt ggataaatat
atttaattag taaagctaaa 13500 aagttttgtt gtcttaagtt atttctttgg
tagcaaataa atcttcacaa gaacctaatt 13560 ttttttttct gttgattttc
tattttggca ggaacttgat ctcaaaatag caattaagtc 13620 ttgtcgccta
gagctaagta ccaaaaacag agaacgatgg tgccatgaaa tccagattat 13680
gaagaagtaa gtgtattcct tgacttacat ttccctggtt ctacagccca accagatgtc
13740 atcatgggga aagaggccag gaatccactc ttctcaggat ctttttggga
acatatattt 13800 ttttctctca ttaaatatat tctatttcaa ataataatat
cacattaaga ttaagaaggt 13860 ttagttaaaa tcagctagga cacttacaca
ctgttggtgg gaatgtaaaa ttgtgcaact 13920 gttacggaaa acagtacggt
gattcctaaa aaattaaaaa tagaattacc atatgatcca 13980 gcaattccat
ttctgggtat atacaaaaat gaattgaaag cagggactta aagagtaatt 14040
tgtacactca tattcataac attattattc acaatagctg aaaggtaaaa gcaacccagt
14100 atctaccagc agagacatgt ataaacaaaa tgtggtatat tcatgaaacg
gaatatcatg 14160 cagccttaaa aaggaatgaa actctgacac atgccacagc
atggatgaat cttgtggaca 14220 ttatgctaaa tgaaataagc cagtcacaaa
aagacaaata ctgtgtcatt ccacttatgt 14280 gtggtaccta gagtagtcaa
actcacagtg aaagaaagta gaatggtgtt tgccagaggg 14340 tgaagggacc
agcaaataga gaataatttt taataggtgc ggagtttcag ctttgcagaa 14400
ttaaacagtt ttgtgcagta tagtgatagt agcaaaacat tgtgaaagta tttgatgcca
14460 ggctgggcat gttggctcac actgtaattc caacactttg agaggctgag
gtgggtggat 14520 tgcttgagct caggagttcg agaccagcct gggcaacatg
gcaaaacccc atctctacaa 14580 aaaatataaa aactagctat gtgtggtggt
gcatgcccgt agtcccagtt acttgggagg 14640 ctgaggtggg aagattgctt
gagcccagga ggtcaaggct acagtgagct gagatcatac 14700 cactgtactc
cagcctgggt gacaagagca agaccctgtc tgaaaaaaaa ggaaagcatc 14760
ttgatgccac taaactgtac acttaaaaat ggttaaggtg gtaaatttta tgctatgtgt
14820 gttttaccac aatcaaaaaa ctatttcatt tcagttaaca aaagtcagct
agggccgggc 14880 gtggtggctc acacctataa tcccagcact ttgggaggcc
aaggcggatg gattacctga 14940 ggtcaggagt ttgagaccag cccggtcaac
atggcaaaac cccatctcta ctaaaaatac 15000 aaaaattagc caggtgtggt
ggcgggtgcc tgtaatccca gctacttggg aggccgaggg 15060 aggagaattg
cttgaacccg ggaggcggag gttgcggtaa gctgagatca taccactgca 15120
ctccagcctg ggcgactccg tctctaaata aataaataaa taataaaaaa taaattaccc
15180 caacatttag tggcttaaaa tagcaaacat ttagtatctt atagtttctg
tgacttagga 15240 atttgggagt ggcttagctg ggtgattcta gcttagagtc
tttcctgaga catctcagga 15300 tattggtcgg agctgcagta atccaaaggc
ttgactgggg ctggagtatc tgcttccatg 15360 ctcacttata tgaatgttgg
ccaaaagcat cagttcttcg ccatgtgggc ctcttcatgg 15420 ggaagcttga
atgttttcat gacatggaag ctagctttcc ccagaaagat cttttcctca 15480
gaaagagtga gtgatctgag agagagagag agagagagag agtgaacgag cgagcaagat
15540 gaaaaccgac gtatctttta tgatctttac ttggaagtga gactccatcc
ttttccacca 15600 cattctgttt gttacagtga gtcactaagt acagcctaca
ttcaagggga ggagagttaa 15660 gctcttaaaa agagaagtag acatatttta
aaacttccac aggtagtgtg ctaagtactt 15720 tacagatcat tcttccagtt
atcacagcaa tcttatattt aggtattatt ttgcaaatga 15780 ggaagttgaa
gcttatagaa tcacatagct ggtggttggt aaagtgtgga tttgaaataa 15840
atactttctg actctaatat ctgtgcttaa ttccattttg attctcaaac ttggttatgt
15900 ggcagaatca ccaggaaagc ttaatagtat aaattatcag gtcctacatg
tacatggaaa 15960 ttctgtttgc ttctttttct ttttttcttt ttttttttaa
aagagatgtg gtcttgctat 16020 gttacccagg cttgacttga actcctgggc
tcaagcagtc ctcccacctc agcctctgga 16080 gtagctggga ctataggcat
gcgccaccac acccggctaa tttttaattt tttgtagaga 16140 tgaggtctca
ctgtgttacc catgctgatc tcgaactcct ggcctcaagc aatcctccca 16200
cctcagcctc ccaaattgtt gggattacag ttacaactac acccagccta gaacgccaga
16260 tttgacttac tcactgtgtg atcttgttac ttagccaatt tgtgcttcaa
ttttctcctg 16320 tgtggtgaaa gtattaataa atctacctat aataggatta
ttgtgacaag taactgagtt 16380 aatatattta aaatgtttac tgcttggcac
ttagaaatgt aatagtagta atagtgatgg 16440 tatttttatg tgtgttttct
tttaaaaaat tactaatttt taattataca aatcagaaat 16500 tagaatatta
taaataatac tgatctttcc ccctctgttc atccacccac aattaagtag 16560
catttcaaat ccagtccctt tttattggaa ataatttttt ttattgtaat gtttttcttt
16620 cttattaggt tgaaccatgc caatgttgta aaggcctgtg atgttcctga
agaattgaat 16680 attttgattc atgatgtgcc tcttctagca atggaatact
gttctggagg agatctccga 16740 aaggtagtgt ggatgttttg agatgagaga
gagtaaatca caaactttag caatgtttga 16800 atcatgttaa atataataaa
ctagaataag aatcggaaaa actatggtct gcttcctgtt 16860 ttgtaaataa
agttttattg gaacacagcc acattcattt ctttacacat tgtctatggc 16920
tgctttctta tggctgcaga agtgagtagt tttaacagag actatgtgac ttgcaaaacc
16980 taaagtattt accatatggt tccttacaga aaatgttgac tcctgattta
gaaaatgcat 17040 ttttgttgtt gctgtttgat gggactatag caatggtgtt
agatgaaaat ttcagtggtt 17100 aaggtgctta taaaaagttg ttttctgtat
tgcaatcttt tttttctacc taattttaaa 17160 tgtgcccata tgaccatttt
tatatatatc catgagattt gtcagccttt tccagtttca 17220 aatgcttttt
gcctcttttt acttttatga gccttatcat gaaatttaca tgccctcagt 17280
atggatttgg taattttatt cccttgtatt ttattatgac ttattttctc atttgaatat
17340 tcttttagag tatagtaatt aatgtgtggt cactattgca gcccagcaat
ttgcagaaca 17400 gatttttaga tttctttctt tttttagctg ctcaacaaac
cagaaaattg ttgtggactt 17460 aaagaaagcc agatactttc tttactaagt
gatataggta cgtatcagta ttaaaatacc 17520 attgtaatgt aatgtccctt
tggcccataa aagatctttt tgggaattca ggagaattgt 17580 tgatacacta
tacaaagtct atagaatttc aacatctata agaagttata aaaatctatt 17640
actattgatg ctaaattaaa actaggatat tttggctagg tgcggtggct cacgcctgta
17700 atcccagcac tttgggaggc caaggtaggc ggatcacaag
gtcaggagtt caagaccagc 17760 ctggccaaca tagtgaaacc ccatctctac
taaaaataca aaaattagct gggcacaatg 17820 ccgcatgcct gtagtcccag
ctactcggga ggctgaggca ggagaatcgc ttgaacccag 17880 gagatagagg
ttgcagtgag ccgagattgc accactgcac tccagcttgg acaacagagt 17940
gagactttgt cacacacaca caaaaagaaa ctaggatatt ttattattta tttgtggcta
18000 agagtccttt ttttgtctaa tagatgttcc aatagctgac aaaaaaatca
tgtgaaatga 18060 ccttactcaa tttctctccc cccctaccat tgacatagta
attttgttta cagtggaatt 18120 acatctcctt gagggatggg tcctcaagtc
aactcttttt tgtttgtttg ttttgagacg 18180 gagtctagct ctgttgccca
ggctggagtg cagtggtgcg atctcagctt actgtaacct 18240 ctgcctccca
ggttcaagcg attctcctgc ctcagcctcc cgagtagctg ggattacagg 18300
tgcccgccat cacacctggc taatttttgt atttttaaca gagacggggt ttcactgtgt
18360 tggccaggct ggtctcaaac tcctgacctc gtgatctgcc cgcctcggcc
tcccaaagtg 18420 ctgggattac aagcgtgagc caccgcaccc agccaagtca
actcttcagg gtgaaaaatc 18480 tctgcttaat cagaatgact ccagaaaaat
tcctaaaata accagtacct aaccttttgg 18540 aggaacctca aaaaccaaaa
atttgtttcc ccatagattt tggttttgcc tcttttccaa 18600 ggtcatgagg
gctaaagtcc aattgaaatg agtgatttag gtgggagaaa gaggcttata 18660
agagtttcag ggattttccc aattgcttgt tgcatttcta tttggtgtta attctaatga
18720 gttaaatgaa catatagtat aattttgtag tatttgttta atgttacttt
tttttctgtg 18780 aagaaaacct ggacactgta gtattagttg tgtagttacg
atgatccata taaacttgag 18840 catcagagta gatttgtaca aaatgtgagg
tgaatttttc ttttaactca atgatttctg 18900 ttacttcaaa gtatatttta
aactttattt agggtctggg attcgatatt tgcatgaaaa 18960 caaaattata
catcgagatc taaaacctga aaacatagtt cttcaggatg ttggtggaaa 19020
ggtaggtgaa acctgaagta atgtttcatg cctattgaat atagtctctc tgatatagtt
19080 agtccttgag ctttggtgtc tttgctgtat gcccataaca gtcttacttt
ttccatttca 19140 gacctcacca cactgaactg tagttttgtg tttagttttt
ttgtcccttc tgctaaacag 19200 tggttccatc agagaaggaa cagcacctgc
tttttttttc tttttctttt ttttcatttt 19260 tcatacttaa cccagctcct
agacttcacc tgtcttgttc aagtctgttt ttccatcata 19320 gtatctagca
aagatacagg ctgtgaatac ataatatgtt aacacttatt attttattct 19380
tatgaagcac atccaaaaca catacatgca catcctgggt ccactttaag aaaaaaaaga
19440 aagtatatat atttttaaaa cttattttcc ttcccccaat ttaaatgagt
catctggaga 19500 agaaccagaa attgtgaatt gactcttaaa atgttcattt
cattggtaac ttaagcagtt 19560 ttcttgtttc cagcatccaa atttcattcc
aataaagact taaaggccaa gagtctgaac 19620 cttgtgttga gttgtgtgtg
ccgtctcttt agttagtgaa aatagttgtt ggcctcttgg 19680 aggtgttgga
accttggtga tgcctcttct cttaggaacc tttcacgtat gcttggcacc 19740
aaatatctct gctttttttt tttttaatcc aaagacttca ggacattcct gaagttagat
19800 ctaagtattt actcacattt aatctactat aaggaggtat gatgcttctt
caagtttgga 19860 agtcaccctt gcctttactt tccagtttaa tggattgagc
ttattttgaa tatagtaaaa 19920 taaaaattaa tagcccacct gtattcaatt
tttaccatgt accaagcact gtgttacctt 19980 ctttactttc gttatttcat
tggatcctaa taacaattct gtgaagcagg gactgttagt 20040 catccaatta
gattactgag agaggttaaa taacttggta gatcacatgc ctagttaagt 20100
ggtggcactc agacttgaaa tatttttgtg ttattacaga gcccaaggtc ctcaccacta
20160 ccctgtctac agcctcgtaa ggatattgtt gtctttcctt tttaaatttc
tctaccattt 20220 aatacttaat ctgtcttatt attttgcaga taatacataa
aataattgat ctgggatatg 20280 ccaaagatgt tgatcaagga agtctgtgta
catcttttgt gggaacactg cagtatctgg 20340 tgagacctgt tttgtttctt
tgaatttaag tgtgtatagg attctcttat ttctgaagat 20400 catgaatata
gtaggtcaaa aataaagtag cttttcccat acagtgttgg tgtttaaagc 20460
attttctctt ttgaaggccc cagagctctt tgagaataag ccttacacag ccactgttga
20520 ttattggagc tttgggacca tggtatttga atgtattgct ggatataggc
cttttttgca 20580 tcatctgcag ccatttacct ggtaagaaat ggatgagaac
ttgggcatgc ttaatgggaa 20640 tggaaatttt ttcatttgtc tttttttttc
tttctttaat tgtttaattg cctttgaaat 20700 atacaattaa atataattaa
atggcaagaa atcttaattt gtgagaatca ctttgtcaac 20760 tgaggtagct
tttgatttta taggcatgag aagattaaga agaaggatcc aaagtgtata 20820
tttgcatgtg aagagatgtc aggagaagtt cggtttagta gccatttacc tcaaccaaat
20880 agcctttgta ggtaagtata ttttagttaa ggaagtttgc ctcagtttct
ccagagttgt 20940 aaaggagcag gagagtagtt gtctgtttca agttgcatta
tccagtagaa ctttctgtga 21000 tgatgtagaa accttctata tttctgttgg
ccacatgagg ttattgacta cttgacatgt 21060 agctagtttg gctagagaac
gaattttgaa taaatagtaa atataaatag ccacttggga 21120 ctagtggcta
ccattttgga cagcacaggt ctggtggaaa caagaactgc catttaactg 21180
tttgacatat gtcctaagaa ggctcttagg ataatgaagt aaatgggtta tttgcagaat
21240 ggatttggac tcatccgtca gagttgtcag taactaacat atgtggagtt
ttgagtagtt 21300 tttgatggaa aaaagtttgt cccctgtcat ttcatgaatt
tgaaagtaat ttatgaaatg 21360 gtacagacaa gtgatgtggg aaagaaaatg
gacaacctgc aaaattacct ttctaacagt 21420 ataaagggaa attggaacat
aatgatttga ttttcaccct attttgtatg ttttctagtt 21480 tagtagtaga
acccatggaa aactggctac agttgatgtt gaattgggac cctcagcaga 21540
gaggaggacc tgttgacctt actttgaagc agccaagatg ttttgtatta atggatcaca
21600 ttttgaattt gaaggttagt cttaatgggt ctactgcata agaccagcat
tgaaagtaac 21660 aagtttattt ggaaaattag cataaataca gtttaataat
cttttcctgt tgcttattta 21720 tttatttatt ttgagacagg gtctcactct
gtcacctagg ctggagtgca gtggcatgtt 21780 tttggctcac tgcaacctgc
gcctcccagg ttcaagcgat tctcttgcct cagcctcccg 21840 agtagttggg
actaaaggtg tgcaccacca tgcccagcta atttttgtat ttttagtaga 21900
ggtagggttt cgctatgttg tccaggctgg tctcgcactc ctgacctcag gtgatctgcc
21960 cgcctccacc tcccaaagtg ggattacagg cgtgagccat tgcgtccagc
cctgatgctt 22020 atttgaaaaa gaaaattata ttcttagagc tgatttttta
aaaagtttat ttagcaagct 22080 gtctcagttt gcttctctgt ggtctttgtt
aattactcct ttgaagcaac ttttttattt 22140 accttttccc atgttgttgc
cattatcctc gattcattta tgtctagtca tactggattt 22200 gatttggtaa
ggaaggttcc tatctgcctt tcatggctct gcacttactc cttaacctat 22260
agttcattca atatgtacta cctatctcat cttctgaaaa tgctttttta ttcttctgcc
22320 caagaaacta acatagttct tttttttttt ttttttaaca agtcaagtgt
aaactgatct 22380 gcttggtttt ttgggggggc tttccctaac caagttttgc
cttgtacctg cttctcttaa 22440 cctttacctc atacaagacc agcttgtcac
tgctccttat gcctcaaacc cgttatctac 22500 ctttgtgctt tgacagatgc
tatccttctg cctggatatc catttctgtg tatcttctca 22560 cctacactgg
catatcttta agagcttttc tttaaatggt ctctgccaca aaatttgtca 22620
tttaaaggct atattctctt tctcgtcttc cattcatgca tcttcatata gactgttgtc
22680 ttcttcaatg ttgggattgt tttacttctt ttattaacaa tcacccaccc
tacccttcct 22740 aaccatggca ttctgtacag agcagggcat gtatcaggtt
cttagtaaat tatttattga 22800 tatgaatgag agctatagat agttctatta
tgcagtgtac ccagaaacaa ctctcatcaa 22860 tagtctttac atttgtattg
attgtttgta gtatacaggt ttctaaaatt gaaatgagaa 22920 attataatca
tgtaatgttg gagtacatgg cgagatagct cactttagtc tgcaacccct 22980
gagctcaagt gattctcctg cttcagcctc cagagtaaat gggactacag gtgtgcgcca
23040 cctctcctgc ctaattttta aatttttggt agaaatgggg tctcactgtg
ttgagcaggt 23100 tggttttgaa ctcctggcct caagtaatcc tcctacctcg
acttcccaaa gtgctgagat 23160 tacagatgtg ggctactgtg cccgaccttg
gcatacttta ttaaatagaa acttgaggcc 23220 atgtggatgt cattcttggc
taacactgaa aagatacctg gcttcttctt gatttggttt 23280 ccttctaatg
tgccaaagaa taaattatcc ctcagtgtgt taggaaagca gggctttagt 23340
tctgtgttgc ctctgaaaca tttcagctgt gaatgaacca catctcgcat ttgtttcata
23400 cttgtcatag gaatcatagc atagtttctg ctctgaaggg catctgtgag
cctcttgtta 23460 tagggcacgg ggaaagagtc cacattatgg agtttatagt
ttacagactt taattcagtt 23520 tgcagcacca ctatttacct atctgtgaac
ctgagcaaat gtctctctat ctcaaggaca 23580 gtaatatctt cctcaaagca
tcaatattat tatgaggtta tgcagaataa ggtatgtata 23640 aggtactagt
tatagtagtt caagaagctt caaggatgct tgttagacat gataatagga 23700
ctgaattttg aagaatgaga gtttttaggg aaagggtgag ggcgggagag atttataggc
23760 agcagagcat catgagcaaa agtacagaag catgaactgc aaaacagctt
ggtgttactg 23820 taggacattt tcccaaaatg tttatatata ttgtacaaga
aaggaatgat gtctgtttga 23880 ttttcactta aaaaaggaag caacaaaaca
gttactgaaa tatttccctt ttgaaaaaat 23940 agcttagcaa gtctgattgt
atgcatattt tccaactgtt tattatgaac atctttagat 24000 atgtagaaga
gttgaaagaa gtgtactgaa cctaccacta aaaataatga tgaaacagta 24060
gcatttaatt attatttttt tttgcctcct ctttcacgcc cctaaatgta actatattct
24120 gagtctcttc ttggttctct ctttttccat tacatttcct ttcttttatt
aggcagcctc 24180 ttcaactgta tagctttact tataaactac atatggctaa
ctcctaaagt tatttatctc 24240 tgtagctgtg actcttctgc caaggactta
taactgctgt ataactgtat aactacctgg 24300 ctgttccact ggcacttaca
acttaattta ttaaaaactg aacatatttg ttctatccat 24360 taaatctact
tctttttttt tttttttttt ttttgatatg gagcctcact ctgtctcacg 24420
atctcaactc actgcaacct acacctcctg ggttcaagtg attctcgtgc ctcagcctcc
24480 cgagtagctg ggattacagg tgcctgccac cgtgtccagc taatttttat
atttttggta 24540 gaaatggggt ttcaccatgt tggccaggct ggtcttgaca
ccctgacctc aggtgatctg 24600 ctcacctcgg ccttccaaag tgctgggatt
acaggcatga gccactgcac ctggcccatt 24660 aaatctacct ctatagattt
cctacacttc ttatttgcac tgtcaccttt ccagttaccc 24720 aggtgggaaa
ctttggatgt gttgactctt cctttctctt caatcgatat atccaattat 24780
tttatcaaat ttatcttttg aatatcttaa caattttgag cttgccaggt tctcaattta
24840 gaaaggcttg catagttttc tttcacttat tgaataaagt ccagaccctt
tcattggtac 24900 tcaaaaccta tcttggtttc atatcaactt acactcctgg
acttatctct tactaatgcc 24960 ctgagaggcc atcattttag gtacatttga
actggattaa ttgtattctc tgattatgcc 25020 catccctttc tacttttctg
cttctgctca ccttgttttc tgcctaaagg aactcttttc 25080 agtcttttcc
catttcagtc tttcatttca aagtttagct caagtagtgc ctcctttatg 25140
aagtgcctgc ttcctactgc caagtgtatg atactttaca ctgattataa cacctatcac
25200 attttgcctg gtaccttagt tctcttagta tatcttaata tcttctgttt
gactataact 25260 gtatggaagg caaggatcat aaggtactca gtttgtatac
ccactactac caccacaaca 25320 cacttaatag tatgtaccaa tagtatgtac
ctaccaatag taggtactca ttaaatatta 25380 tagtgagcaa aaagataaat
ttaggttgta tttcctcttc tttcttttgc ttccttctgg 25440 cctggagctt
tggtagtgag atgccatgat gacatactct aatatcctgt gtaaaagtag 25500
atgtccatta taggattttc tattgctata agtggcagct gtcatagtgt gaaatctgcc
25560 taaattggga aggcatggaa agtctcaggt agtataattg tattgcaaat
ccgtatcagt 25620 tttgaagatt tccctactcc ccttttaaac ataaagattt
atattaaaga aaagtctctt 25680 aataagagaa atactggcaa ttgtgttatg
acatttggaa ttcacatgaa aaggatcagc 25740 ttcaggttat aaaaacagtt
tggtatttct tttttaaaat tttgttgata caggtgtggt 25800 ggctcatgcc
tgtaatccca gcactttggg aggtggaggc aggcagatca cttgaggtct 25860
ggagttcgag accagcctgg ccaacatggt gaaaccccat ctctactaag aatacaaaaa
25920 ttagccaggg atggtggtgc atgcctgtag tcccagctac acgggaggct
gaggcagaag 25980 agttgcttga acctgggagg tggaggctgc attgaaccaa
gatcgcacca ctgcactcca 26040 gcctgggtga cagagagaga ctctgtctca
aaaacaaaca aacaaaattg ttaatacgct 26100 tttaattttt tttttttttt
ttgagataag agtcttgttc tatcacccag gctagagtac 26160 catggtgcaa
tctccactca ctgcaacttt cacctcccag actcaagtga ttctctcgtg 26220
cctcagcctc ccgcgtagtt ggaattacag gtgtgcgcca tcatggccaa ctaatttttt
26280 gtgtgttttt agtagagaca gggttttgcc atgttggcca ggatgttctc
aaactcctgt 26340 cctcaagtga tctacctgtc ttggcctccc aaagggctgg
gattacaggt gtgagccacc 26400 gcgcccagcc taatttttta aattgacaaa
taattataca aattaatggg gtaaatagtg 26460 atgtttcaat acatgcactg
tgtagtgatc agattagggt aattagcata tccatcagct 26520 aaaacataat
ttctctgtgt tgggaatatt cagtatccct ttcttttttc tttttttttc 26580
tttttcttct ttcttttttt ttttttttga gacaaagtct cactctgtcg cctggagtgc
26640 agtggcgcga tcttggctca ctgcagcctt gatctcccag gttcaggcga
ttctcctgcc 26700 tcagccttct gagtagctgg gattataggc gttgtgccac
caagcccagc taatttttgt 26760 gtttttagta gagacaaggt ttcaccatgt
tggccaggct ggtcttgaac tcctgacctt 26820 aagtgatcta cttgtcttgt
aatcccaaag tgctgggatt acaggcgtga gccaccacgc 26880 ccagcctcag
tttagtattt ctgtagcacc acttatttgg gcatctgtgt taggatttgt 26940
aagtggaaat aatagtgggc cttcaaacta aaaagtgtac agtagaaaaa ataatcaaca
27000 aaacgaaaaa gcaacccaac ccatggattg ggaaaagata tttgcatacc
atacatccaa 27060 taagaaatta atatcctaaa tttataagaa actcatgcaa
ctcaatacca aaaatgtaaa 27120 taacccaatt taaaaatgtg caaaggacct
gaattagaca cttctcaaaa gaagaaattc 27180 agatggtcaa caaacatatg
aaaaggtgct caacaactct tagcatcagg gcgatgcaaa 27240 tcaaaaccac
actaagatat caccccccac ctgttgggat ggctattatc caaaagaaaa 27300
aagataagcg ttagtgagga tgtgggagaa aagaaatccc ttgtacactg ttggtgggaa
27360 tgtaaattat tacagctatt gtgggaaaca gtacaaaggt tcctcaaaat
taaaaataga 27420 actactggcc gggcatggtg gctcatgcct gtaatcccag
cactttggga ggttgagaca 27480 ggtggatcac atgaggccag gagttcgaga
ccagactggg caacatgcag aaaccccatc 27540 tcaactaaaa ttacaaaaat
tagctgggca tggtggtgca catctgtaat cccagctact 27600 ccgtaggctg
aggcaggata attgcttgaa ttcaggaggt ggaagttgta gtgagctgag 27660
agatcatgcc actgcacatc agcctgcgtg acacagcaag actctgtctc aaaagaaaaa
27720 aaaaaaaaat agaactgtaa gattcaacaa tgtcatttct tggtatatat
ccaaaggatt 27780 ttaaagcagg cttgtgaaga gatatttgca ctcccatgtt
cattgcagca ttattcacaa 27840 tagtcaagat ctggaaatag cctaaatgtc
ctttgacaga tgaatggata gggaaaaagt 27900 ggtatatata cacagtggaa
tagtattcaa ccttaagaaa aagaaggaaa ttctgccatt 27960 taggacaatg
tggatgaacc tggaggacat gttaagtaaa ataagcaagt cacagaaaga 28020
gaaatactgc gtgatctcac ttacatgtgg aatctataat agttaaattt aatagaagca
28080 gaccatagca tcatagttgc caggggctgc agggatgcgg gtgcgggggt
gtatgtagag 28140 atgtttatca aaaggtccaa aaattacggc agcaaaagaa
aaaaaaggta caaaaatgac 28200 aactatgaag aggtgacaga tatattaatt
agctgaaatg tgataatttc acaacgtata 28260 tcaaaacatc aagttgtata
actcaaatat ataacatttt tatatgtcaa attaaattgt 28320 cacaaaataa
aatatgtaaa agataaagca ataaatcagg agagggggct atgatagatg 28380
aaacaagata ggctctatgt aaatgatggt tgaaactagg tgaggcattc acagggttgg
28440 gagggggctt attgtactgt tgtctatagt tttgtgtctt tgaaaacatc
aattcagatt 28500 tgtgttttaa agggaaaaaa ataatagtgg acttggatta
atgccttacc tttgtagtaa 28560 ggattttcag cagagagaat tttaatcatt
tgcatatact agtcaatgta aggcatggag 28620 aatgtaggca ttaatttcgt
atggcttttc taaggatact ggaaggaaga gaatttaaaa 28680 tcatgttctt
ttcaaaagac tgaagtccaa ctctgtggct gtccgcgctg acaggtgggg 28740
acagagcccc acctgactca gcctcacact cacttggcat gtatcaggct acttttactg
28800 gggctttagg tgagagtgat aaaacggttc ttacttgatt tggggccctt
aagcaggttg 28860 ctgctctggg attaggttgt cttataaaaa ggtaactcac
cacgcataac aatgcttttt 28920 cctgtttaaa gagagaccta gttcttctat
cagatgattt cttaaggcag catgaagtag 28980 ccagacacaa aaagagttaa
gatccaagct ctaccagtta ttagctgtgt gatccctaac 29040 aagttattta
acttagaact ctggtttctt tctctgttta aaatggagat taaaagatct 29100
aattattggg gatgttatgt gaaagtggtt tatgcctgtg gttgtcttct gttaggcact
29160 tcatgaattg gaatgtttcc ttttcggtct atgataatgt ttttttcaaa
ataaataact 29220 taggccgggc gcggtggctc acgcctgtaa tcccagcact
ttgggaggcc gaggcgggcg 29280 gatcacgagg tcaggagatc gagaccatcc
cggctaaaac ggtgaaaccc cgtctctact 29340 aaaaatacaa aaaattagcc
gggcgtagtg gcgggcgcct gtagtcccag ctacttggga 29400 ggctgaggca
ggagaatggc gtgaacccgg gaggcggagc ttgcagtgag ccgagatccc 29460
gccactgcac tccagcctgg gcgacagagc gagactccgt ctcaaaaaaa aaaaaaaaaa
29520 ataaataaat aaataaataa ataaataaat aaataacttg ttcttttaaa
ttgggatttt 29580 tcttaatttt gaaaatttct taactttgaa attttattgt
tggaactaac atctattgta 29640 gaaaaggaaa ttgttcagtt ttcagaatgt
cattttataa ttataaaagg agatctcact 29700 ttggagaatg agtgagcctt
gtttaatgta gaaataatgc cttttgccca tattcacctt 29760 ttagtatctg
gcagcgaatc ttttatgtaa caaataatca ataactttac ctttttttct 29820
tatttataca gatagtacac atcctaaata tgacttctgc aaagataatt tcttttctgt
29880 taccacctga tgaaagtctt cattcactac agtctcgtat tgagcgtgaa
actggaataa 29940 atactggttc tcaagaactt ctttcagaga caggaatttc
tctggatcct cggaaaccag 30000 cctctcaatg tgttctagat ggagttgtaa
gaaaattaat tataatatcc ctagagtatg 30060 tgaaatctag agggattgcg
tgccctgcaa tttttatgca tacttggaat tccttgggag 30120 gccgcacatt
ttgttttggt tttgtccttc tcccaactct tatcattgct ttgcaaggtg 30180
aacaggatag aaatgaaaca tttgttgttt tgaactttgc aagacttttt ctttggtgcc
30240 agttttctgc tcctgttccc agaccaaact gagggtcagg ctgcttactc
tcgcggccca 30300 ataacgagat gcagatgaat tgggagagat gggagttttt
atttctgtaa ccagttacag 30360 gtagaaggcc tggaaattac cgccagacca
actcaaaatt acaaagtttt tccgtagttt 30420 atttatcttc taagctatat
gtctatgtgt aagtttgcat tcatctaaag acataagtga 30480 ttaacttctt
ttaatctgta gctgaggtct gagtcttgaa gacattcctc tggagcctca 30540
gtaaatttac ttactctaaa tgggtccagg tgctggggtg attaccctta tcttgtctcc
30600 tattaaatca cagaggttta aggagttcct tcagaccccc aataaacttg
tttgtggagg 30660 cctggggttt cttcagaccc ccaataaaac ttatttaatt
ctgaacgggt cctgttaaga 30720 attcctttgt tattttgctt ttaggcctgg
gaaaggcctg ggcagaactc ttggtaggct 30780 ttggctacat ttcagccttt
gtgtaagggc actggctctt ccagtttttt ttttttttag 30840 acagtcttgc
tctgttgccc aggctgaagt gcaatggcac aatctcagct cactgcaacc 30900
tccgcctcct gggttcaagt gattctccct gccacagcct cccaagtagc tgggattata
30960 ggtgcccacc accatgccta gctaattttt tgtatcttta gtagagacgg
ggttttgcca 31020 ctcctgaccg acctcaggcg atctgcctac ctcggcctcc
tgaagtgttg ggattacagg 31080 cgtgagccac tgcacccggc cactctttca
gcttttaata tttaacttca ccactcagtc 31140 gtgctgaaac agttgttatt
taggcctgca ttagtgagac ctggcctgcc acactcccag 31200 ttaatattcc
ttcttggctt ggtgcagtgg cccatgcctg taatcccggg actttgggaa 31260
cccaaggtgg gtggattgct gagttcaaga gttcgagacc tgcctgagca acatgttgaa
31320 gccctgtctc tacaaaaact acaaaaatta gcggggtttg gtggtgcacc
cctgtagtcc 31380 tagctacttg ggagattgag gtgggagaat cacttgagcc
cagggaggtt gaggctgcaa 31440 tgagccatga tcacgccgct gcactccagc
atgggtgaca gagcaagacc ctgtctcaaa 31500 aaaaaaaatt tctttttgta
tttgaagccc agtactctga aactactgtg agaacagcag 31560 tctcataggt
cctgccaaag gataaatggg tggtaaaata tgactgtggt catgactttt 31620
tcccccattt ttctttttat gactttattg aggcatatta tcatgttagt tcccatttaa
31680 ggtgtacaat atagtgattt tttatttctt ttttattaac ttgtcactga
cttgatgctc 31740 aattacagtg attttttttt aaagtaaatt taccaaattt
tgcagctgtc accataaatc 31800 agcttaaaaa catttccatt aacagtccat
taactgttaa tctccattct gtccacccca 31860 ggaaaccagg aatctatttt
ttatctccac aagtttgcct tttctgaaca ttccatataa 31920 atggaattat
gtaatatata gtcttgtgtc tgactttttt tacttgcctt gtatttttga 31980
agcccgtgca tgttgtagca tgtatcagta cttcattcgt tttgattacc gattagtttt
32040 ttcactgtat atatcttttt gcatttgttc tttgggtttt ggtgattgtt
cactttagcc 32100 catacaaaag tttgggtctt ttttcctcaa caaaagtttg
gactctgtta tctgtaagag 32160 atatggtaat acacattgat tttgttcatt
tgcatttcaa agtaggaaat ctggtgcata 32220 agtatttaga aaacctagga
ttttcaaaga tccttgaaca ggattttaac tgtttgtttc 32280 tttttcagag
aggctgtgat agctatatgg tttatttgtt tgataaaagt aaaactgtat 32340
atgaagggcc atttgcttcc agaagtttat ctgattgtgt aaattatatt ggtaagtcca
32400 gtccatttag tggctgagct ttaaacatga attaatataa ctttgactct
cagtatgatt 32460 agtagaaatg gacttttggt gttgtgttct gtatctcata
cttgaagttt ctatatggag 32520 ttatatgaga actctgaaaa atggatttat
atagtttgga aggtgaaaag gtagatatta 32580 aatctccatg agatcattca
agagaagatt tactttcttg aggatagggc tctcacagga 32640 gaatgaaact
tcactattag ttgaacctgc tttttaattg tctcttcata gtctaagcat 32700
gttcaagtat ttatttcatt tttcctgtga atactgcttg atgttcaaat gttttaattt
32760 atcctgtgaa aattattctt ttcttttgaa caagttaagt
gtttgatcac tgtatatctg 32820 tccaaataaa gtaaagcaca aaattgtaaa
atcaagaaca aaattgagca gaggggctaa 32880 aatgtatgta atcttacttt
tttaaaagtc tttttattag ttttcttttg aatttttctt 32940 ttaaattgag
gcagtttaca tagagtgaaa ttcacagatc atatagtttg ataacttttg 33000
acaaatttag atacttgtgt aatccacacc acagtcatca tgtagaacat ttctttcacc
33060 ttagaaagtt ccctcatgtc tcttttcagt caactcccct tgccccagac
tttattacta 33120 ttttactaat ggactttttt tttttaagaa tggttttaga
tttgccaaaa aattgggcag 33180 atagtacaga gttctcatat aactcccctc
actcacgcat gtagtttttc ctattgttaa 33240 cattttacct gggaggcgga
ggttgcagtg agccaatgtc acaccactgc acttcagcct 33300 gggcaacaga
gtgagacctt tctcaaaaaa ataaataaaa aacataaaat atttaaaaat 33360
attaaaaagt aaaaaaaaat ctattttaca ttagtatgat gtatttgtta catttaatga
33420 acccatttat tattaattga agcccatact ttattcacat ttccttcatt
tttacctaat 33480 atcctttttc tgttctaaga tctcatctta gaacacatta
catttggttg tcatgtctct 33540 ttagctcctc tttgctgtga cattttgtca
gacttcaaat agtccttgtt tttgatgacc 33600 ttgacaatga cctgaggaat
actagtcaag tattttgtaa gatgtccctc tgctgaaatt 33660 tgtctttttg
tttttgtttt tgttttgttt ttgagatgga gtttcactct tgtcgcccag 33720
gctggagtgc aatggcacaa tcttggctca ctgcaacccc cacctcccgg gtacaagcga
33780 ttctcctgtc tcagtctccc gagtagctgg gattataggt gcacaccacc
acgcccagct 33840 aatttttgca tttttagtag agacagggtt tcaccgtgtt
ggccaggctg ctcttgaact 33900 cctgacctca agggatctgc ccgccttggc
ttcccaaagt gctgggatta caggcgtgag 33960 ccatcacacc tggctcttgt
gtgtattctt aaacctggat tgttggtcca gaatatgaaa 34020 tttattattc
agaggtagga aagactcaga ggagatggga cttgatttag aacttgaaag 34080
aaataatttg gattagcaga gagaaaaaga gtattccagg ttgcagaaaa gcaagcaaaa
34140 aagacagata caggaataat ctcaagctct gtgagtgtag cgattgtgtc
atttttattt 34200 ccccccttgt atgtggagcg gggcttaagt attgcaggtg
ctctgtaaat gtatattgaa 34260 ctgaattaaa tggaaacaag tggttctttt
agcatcagtt tctagcatct atgggaatcc 34320 tgtttgtgtg gacaggaaat
cacaaagaat gataaatcaa ctctaaaatc ctcaaaagta 34380 actaacctcc
cttgcccttt ttctcatctt cttgtatata gtacaggaca gcaaaataca 34440
gcttccaatt atacagctgc gtaaagtgtg ggctgaagca gtgcactatg tgtctggact
34500 aaaagaagac tatagcaggc tctttcaggg acaaagggca gcaatgtaag
tggattctgt 34560 tgtttataag cacaatgcaa tgtgcatcat atactcaaga
attaatcttg ccggttttca 34620 ctaatcaata ttgcatgtaa tagtaacata
ctgggccaat ttgaagtgta atgttctgat 34680 aaacttgtag aattcttact
aatggactga gaccattcta ttgtaatggc ttgagggtaa 34740 taattcccat
acggtcctag tataaaaaga ttgtttatca caaaatatta taggcaggaa 34800
gtctttagag atcagattct agagctcagt tatttctcat agattgataa tagaaactta
34860 ggctttggcc ttgtggcagt ttatgttttt acattattgt tcaatgtttc
tgaatgctga 34920 taatatcttt tttctttttc ttttttttct tgctgtggtt
taggttaagt cttcttagat 34980 ataatgctaa cttaacaaaa atgaagaaca
ctttgatctc agcatcacaa caactgaaag 35040 ctaaattgga gttttttcac
aaaagcattc agcttgactt ggagagatac agcgagcaga 35100 tgacgtatgg
gatatgtaag tgtctgtgta atgtatttga aaggagcatc tggtttcttg 35160
aagccatctg ttattttgcc ttcttaactc agccagttgt tttacttacc ttcacattag
35220 aacaagagac aggacaactt aaattaacat aaaccatgtt tgttttgaat
gttacctctt 35280 tttttaaatt tgcttagttg agcaatttag cttcacaatt
gggctaaaat tatttagctc 35340 taaaaaaata caggtgcgat ttaataatct
tgaatttcaa acctgatttt aaaataaata 35400 caaaatctgg acttcttagc
cagtgctgtg gactgaatgt atccccccca aaatacatat 35460 gttgaaattc
tgaccctgaa ggtgaaggtg ttggtaggtg gagcctttgg gaagtgatta 35520
ggtcatgaga gtagagcctt cctgtttggg attaatgctc ttacaaaaga gacctgaatg
35580 agatccctca tcccttctgc atgtgaggac agagcaagaa gatggtcatc
taggaaccag 35640 aaagcaggcc ctcaaatctg ccttgttctt ggacttccca
gcctccagaa ctataagaaa 35700 taaatttctg ttgtttttat ggcacttagt
ctatggtatt ttgttatagt agtccaaatg 35760 gactaagaca accaggggag
tcttaaactg cttacttttc tgtaaataat gacacaaata 35820 ttctaaggct
atgttttttt caaaccatct actgtggagt cctactattt tttagtgtac 35880
acagtctcca aattttccat atttgactcc aagcttatct ccttcagatg tactaaggaa
35940 gtgtaaaaaa acagagttaa aatatgagta aattgaagga gtcatactac
cagcatttct 36000 gttttctttt ataaaacaaa tcctttttag gaaacaaata
cttgtttcct aaaaacatat 36060 acttgtcctg agcaaactct gtgtatctca
gagtaaacaa tcaatagtgt ccagttgggt 36120 gaatgaattt actctccaga
actggtttta ttaatattat tgatatttta tttctattta 36180 tatattattg
atattttatt tctatttgta tattattgac attctggtga gtgataaaac 36240
attataatta tttcattaaa atatcttaga cttgtcaaag gcttattata caattgttaa
36300 agaaaaatga ggtagatcta tatgtgcttt tattaaaaga tcacctaagt
ctttagttga 36360 cagttttaaa ttgtgggaaa aaataaagag tatacagatg
ttttcacttg tgagggagca 36420 ggctatgtgt ctatgtaaca gataatcata
gtaaggtccc ttgaggtgga tggtagaggg 36480 gtagaagaga gagtcttttc
tgtttatatc cttttatagt atttaatttt taaaaatcat 36540 attcaagtat
taaatttctt ttctggccag gtggtgactc acgcctgtaa tcccaacact 36600
ttgggaggcc aaagcaggtg gatcccttga gctcaggaat tcaagactaa cctgggcaac
36660 atggggtaga acgccatctc tacaaaaaat gtaaaaatta gccgggcatg
gtggtgcaca 36720 cctcagctat tcaggaggct aaggtgggag gatcgcttga
gccctggaga tagaggttgc 36780 ggtgagctga gatcatgcca ctgcactcca
gcctgggtga cagagtgaga ctctacctca 36840 aaaaaaaatt ttttttgttg
ttttttcctt tatccaccca tctgcacact ggtataaata 36900 tatatatttt
taagaaaaaa gtcatctgta ttcatttcct aagggttcca taacaaaatc 36960
ccataaactg agtggcttaa gcaatggaaa cttactgtct cacagccctg gaggctaaaa
37020 gtccaaaatt gaggtgttgg cagggccatg ttttctctga aatctgtaga
gaagaatctt 37080 tccttgtttc tctctagctt cttatggtct gctagccatc
attggcattc cttggcttac 37140 agctgcagca cctcaatctg tctccattgt
cacagagcat tctcccctgt gtatctgtat 37200 cctgtgtctc ttctaagtac
actggtcgta ctggactaag ggcccagacc attccagtat 37260 gagtattaat
ttatctgatt acttctgtaa tgaccttatt tccaaatagg tcacattctg 37320
aggtattagg ggttaggaat tccacatatc ttttggggga ccacaattca actcataaca
37380 catctttgat gttgatatgg caaggaggca ggcagccagt aattaaagat
atggctattt 37440 ctgagataac caatttagac ttttctttta gcttcagaaa
aaatgctaaa agcatggaaa 37500 gaaatggaag aaaaggccat ccactatgct
gaggtaaaat cattgacgtc attctgtata 37560 cttattaatt cttgatgtct
ccattagaaa ctatctttgg ttttgatata taaagagcat 37620 ttttcatgtc
attcctgaaa ttcagacaaa attctagtag atgctttgta tataaagagg 37680
gatgtatcat agtgattaaa aacaagggct ttgggaaaat tacctagtgc ctcactgagc
37740 ttcagtttcc tttatttatt tattttttga aacagggtct cactctgttg
cccaggttgg 37800 agtgtagtgg catgatcaaa gctcactgca gcctcgactt
cccaggctca agcagtcctc 37860 ccatcttagc ctcctgtgta gctgggacaa
gaggtgcacc accatgcctg gctaactttt 37920 acattttttg tagagacggg
gtctcactat gttgtccagg cttgtatcaa actcctgggc 37980 ttaagcaatc
ctcccacctc agcctcccat agagctggga ttataggcat gagccaccac 38040
atccagctag tttccatatt atagggacaa tttcttttct agtagggtta tgaagattaa
38100 atgggagtca gtaatggtag ttgtcactat tatttttgtg actggtttag
ataggatggc 38160 tgcacatcct ggtttacctg ggactgtccc agtttatact
taatgtcttg ccataattat 38220 cagtagtagt ccctttcact ctcaaaatta
tgccggtttg gacaataaat tatatggtgt 38280 ccctaggtat agctcattct
ctaggtggca gattttttcc ccttatcctg tacatttgtc 38340 cacggtccta
agttattaca ctccttaatg tcagtactta ccctgaggat gtgggtgggg 38400
atacccatgg tgtcctgggg tgggaaatcc aagttcttta tctttttgtg tccctattca
38460 aggtagtggg gaccaggggg cagtcaaaat catagagcca agatacaagt
aataccaatg 38520 gttttggtgg tcctttttct ttcttgttag ttgatacctg
ttgtagcttt ttgcttctcc 38580 ggaggtaaaa gcaaatacta gtagtttact
gttgtcagga gagaaaagag agaagactaa 38640 gactaagtat gttcttatgt
gaaagattaa gtgtcagaat ctgaagaata gaaaggcttt 38700 aacaaaatca
gtgtttcgta ttatattatt tagttctcta atgaagcagc aagtcattct 38760
gtatccttaa gatctgaatt ttgtttccta tgatgtagtg gtaagccttt catttccttt
38820 gtattttcat catattttag gttggtgtca ttggatacct ggaggatcag
attatgtctt 38880 tgcatgctga aatcatggag ctacagaaga gcccctatgg
aagacgtcag ggagacttga 38940 tggaatctct gtaaggatgc acttgtgttg
ttggtcttga catctgtaat attttgtatt 39000 ctttctgctt acatctggca
aattaaacct ctagtggttc ttttctatag ttttcctttc 39060 cccaaatata
ttggcattat atttaaaatg gaagcttttt ctcagtcaca ggaaacttcg 39120
gaaataattc tttgggttgg ccttttttgg tttttaataa aagcatttat ttcataggag
39180 tgttaagaat tttccaagta taactaagtt acaaactatt tccctactca
ttatacctct 39240 ggtccaaggt attgactctt aaatgcccta aaacttgaaa
ttttttctga attaaagttc 39300 tggattaaaa acttcagata ttttctcgaa
ttataggtca taatttttgt ctttttaagt 39360 gtctgtggtg cagagatgtc
cccatgatag atcagaatag atatgaacta tattaaactt 39420 tgcattttaa
gaacttgtat agttcttatt actcaataca atgctctgtg aaatacttgt 39480
tggcaatatg aacttgcttt aaaatgggtt cattattcaa ataaaatatt gtttcaagtt
39540 gtagattatt gaagtcatta aattaccttc acttatcaga gcgtgttttc
tcttttatag 39600 ggaacagcgt gccattgatc tatataagca gttaaaacac
agaccttcag gtaagacact 39660 tctaccacag tgcttgaagg cattttaggt
aactatcact agcctaacaa atgtggggga 39720 tctaatattt ttaagttaaa
aattttctta aagctgcaga ataaattcca gttttggtat 39780 tgccattgag
acatggtata ataagcttta agattgtgag aatttaggag atgataaaga 39840
gaaagtaaca gagaacgtga ggtagaaaaa ttttagaagg ggacgtgaaa agaaagaatg
39900 gaagacagaa aaaaggaact cagagaggaa aaagcagcaa gaaaatgttt
agaaagatgg 39960 aaaaaataag tgttgcttaa gattagagag gttgggccag
ttgtggtggc tcatgcctgt 40020 aatcccagca ctttgggagg ccaaggtggg
tggatcacct gaggtcagga gttcaaaacc 40080 agcttggcca acatggtgaa
accccatctc tactaaaaat agaaaagtta gctgggcatg 40140 ttggcatgtg
cctgtaatcc cagctacttg ggaggatgag gcaggagaat cgcttgaacc 40200
tgggaagagg aggttgcagt gagccaagat cacaccactg cactccagcc tgggtgacag
40260 agcgagattc tgtctcaaaa caaaacaaaa aaatattata gaggttggaa
agaatttgag 40320 gcagtgggtt caggtatgga atggttagag ggaaggcctg
tgctgttcta ggccaaagca 40380 gattgctttg aaaataatgc cttcatctgc
actggtcact gatgacttgt tttgttggca 40440 aaataattcc agttgaacat
ctgctaaata ctgagcaagg tctgctggaa agagtactgg 40500 atggggaatc
agaggcttga attctagatt gagtaattta ctgaccgtta cctcaggcaa 40560
atctgcgttt cccatttcct ccttctgcaa aatggggttg aaacctaact ttcctactca
40620 gtgagtatat tgtgacagtg tatttaagag gatgtttgtg aaaatgcttt
gaaaactata 40680 aactattatg tacttgcaag atattatcat ttagttaaaa
atttgttttg aagggtaaca 40740 gaagaaataa agtctgtgcc ctaatgtgca
ttatacttaa atgcatgtta aaaaaagata 40800 tacttttaca ctttaaaatt
tgtagacaca tagcaacaca tgcaaattaa tatacatgtg 40860 aaattgtatt
tcatatacta ttttcacatt taaaaaataa cattatctcc tcttatactg 40920
cttttatttt actgaaatgt attcagttta gtctgaccag cagttctcaa aatgtggcat
40980 atggacctgt gagaatcctt gaaactttgt cagagagtct gtgaggttag
aaccattttc 41040 ataataagat gttatttgca tttttcacca tgttgacatt
ttcattgatg gtgcaacata 41100 aaatggcttt caccttagta tgaatcaagg
cactagcgcc aaactatgct agtagccgtt 41160 atatttttta ccaccatgca
gtcacagttt ttaaaaaatg acagtttcac ttacaaatgt 41220 ccttgataaa
gcagtaaaaa tattatttta tttaaatctc aacgcttgag tatgggtttt 41280
taatattttg tgtgatgaaa tgggaagtac tcataaagtg tttcttctat acagtgaagt
41340 atgatggttg tcttaaggaa aaccacttca gtaattgttt acgttgctag
ttgtctggcc 41400 tctttcaggg aagataccat ttttacttgg aagagcaact
gtcaggcaaa ctatgattat 41460 tcagatttgc atacgtgaga taatttctaa
gaaattaaat gaaaataaaa agtatagcac 41520 acttattact ttaagggaaa
aaagcagtat ttgttgtcaa tgataaaatt ccatgtttca 41580 agcaaaaaga
attttagaaa acttgtattt accagctgga atgacagctt ctcagtactt 41640
aaaaactttt ctgatgagat tagtggtgat atcagaaagg tttgcaaaaa tgtaaaacag
41700 tgcaagtgtt ctcactcaaa tttttttgtt ttagaaaata tcgttttttg
gtttttgttt 41760 ttagaggtgg gctcttgcta tgttgaccag gctggccttg
gactccctgg gttcaagtga 41820 tccttccact tcaggcttcc aagtatctca
agtagcagta accagtgaaa ggaggtggct 41880 ctcgtagata tgagtatact
gtgtagacac atgtatgtat agcacttagc atgtctgagg 41940 aaatgcagta
tgactggagt gtagagtttc aggagagaga aaacctaaat ataggagaga 42000
gagaaaacgt aaatataggt ttttttgttt gtttttaata aaaacatgtt aatgggcaat
42060 aggtttgtta tttttaaatg aattaagaca cgagtatttt taaatttctg
tttaattcct 42120 gaaaaaccct ttggggttct caagaatttt ctagtgtaaa
ggggtcctga gaccaaaaag 42180 tttgagaact gcttatctag actattctca
gaagcttggc tgctcaggaa agaagagagg 42240 tgggtgagta gctaaaagtt
gatgtagagt caagggaggt cacttctcag aaggaagact 42300 taaacatgag
gtgccacatg atgatgtttt ggttaacaac aaattgtgta tactatggta 42360
gtcccataag attatactac tgtattttta ctatgtattt tctatgttta catgcacaaa
42420 tacttaccat tgtgttacag ttgcctacag tattcagtac agtcacttgc
tgaacaggta 42480 tgtagcctat tgtcaaaagg ctataccata cagcttaagt
gtatagcagg ctgtaccatc 42540 agggtttgta taagtattca cacagtgacg
aaagtgccca atgatgcact tctcagaaca 42600 tatgctcagg gccaaaaaag
cctgtggaga gggaggtccc tgaacaagtt gagagattgg 42660 ctccaaagaa
cagtaggagg gattagcctt gatactattg atattggaaa gagaatgagt 42720
gtggatacag atactttttt gggagtagag gttggaagat cagggtctta ctgccttatg
42780 gcctcttatc ctgcagtggt aagtgaggtt aactgaaatg gaagaggtga
cgacctcata 42840 gatttgaata gagtgctaaa gacttctaaa agccactgtg
tgcagtggaa gatgttgttg 42900 acccaaaaca ttcattaaga atccaggcca
ggtggctcat acctgtgata ctagcgcttt 42960 gggaggctga gacaggagga
tcgtttgagg ccaacagttt gagaccagtc tgggcaacat 43020 agtgagaccc
tgtctctaca aaaaaataaa tcaataaaat ttttaaaaag ctgggtgtgg 43080
tagtgtatgc ctgtagtccc agctactcag gaggctgaag tagcaggatc gcttgaggcc
43140 caggaggtcg aggctgcagt gagccaagat tacactactg cacttcagcc
tgggttgaca 43200 gagcaagact ttcttaaaaa aaaaaaaaaa aaaaacccag
agcagaaatg aaaaacctgt 43260 gaattagcag tgacaaatag tgaacagtaa
tattaatttt ttgttgataa tttagtagct 43320 tggatacagg aactgataga
gcagatggtt ggattaatcc aaggttagga ttttgctgat 43380 ggaataagag
gataaaaagg ttgaagatgt taacaaatag tgatttaagt gatagaagtt 43440
gtagctagag taaaatatta aaatctaaga tttcaaatag agcaagtcca attctttttc
43500 tttaatcaat atttctgggc caggcgtcgt ggcacatgcc tgtaatccca
gccactttgg 43560 aggctgaggc aggagaattg cttgaaccag ggaggcagag
gttgcagtga accaagactg 43620 tgccactgca ctctagcctg ggtgacagag
ctagactcca tttcaaaaaa aaaattctga 43680 ctcccagttt tctggctaat
gaatcagata ccacagatac acagtgatag catttgttta 43740 ggttatattt
tttaaaactt tgaggcagcc ctgtcaacat accatgaagc tcagaaataa 43800
actgctaccc aggtagcttc tttgagtgtc accttgagtg acaaactgta acaagattgt
43860 gtctgtcatt ggggcctatt atataatcac acttgatttg tggcctctat
tgtcttttct 43920 agcttagagt tgacttgaat ctcttgttca tctgaaatgg
ttaaaaccag ctttcttcat 43980 tatcctggag aaggctaaag ttctgagggc
tttcactgtc ttaaggagct agttatatga 44040 gaaagctctt ggcttggtcc
atggtattta ggaaggaatg ctgtttggag aagaaccttt 44100 ttggagcctt
gcatttgatt gttggtgagg aggagacaac agagatgtgt ggggaataaa 44160
gttgggctat gggggaagta tttggagtga tagataacaa aggacacaac gttttttaaa
44220 cctcctttga tgctcatcat ttcctttttg gtaaaatggg ggtaacaaaa
gtagtcttct 44280 cacatgactg ctgtgaggat taaatgggaa aattataagc
agagagtagc aaatattatt 44340 attattgcat ttaggcttaa ttattgtcca
aagaaaatac cagcatgatg cataagcaac 44400 tttcaactcc tttagccatt
ttctgaaata taacaattaa tgattttttt ttaaaatcga 44460 ggtcacattt
taaactgtag aacccattgt gaccaacagc catcccattt gagggtatct 44520
ggctgctgcc agttggcttt aagatgtcat actgccctca aatggaactt tcaggatgtc
44580 tacaggtccc tttactagtt ggtttcaatg agcttttttc aaagtttttc
tgggttgtaa 44640 acagacggat aaacatgaaa ttccaagcaa cctagttcaa
gggcctcctt cctttaggta 44700 atgaaagttt gatgaattgt cctactgttt
ctaaatatgt ggttaatcat ccaagaaaaa 44760 ttgttccctt ggccagaggc
tgcatttgtg tctagggctt gggttagtat gggttaaaga 44820 tagccaagac
tattgttctc aagcacaggc aactaaggtc tcttactgat aaatttatat 44880
atacattata cacctgcaaa ctttattttt tatatataac tagattttat ttataggtaa
44940 tctttatatc ttttatctta acccaatttt taatttaaat ggagatgttt
attaattaaa 45000 attagtttca gagtttcagg gatttttgag gaattttttg
tttggaggct gatatgttac 45060 agaagaaagg ttatatccac tgactgatgt
cttttataga tcactcctac agtgacagca 45120 cagagatggt gaaaatcatt
gtgcacactg tgcagagtca ggaccgtgtg ctcaaggagc 45180 tgtttggtca
tttgaggtag gaaaattgct tcatttccta ctgaaaaacc aggatacgtt 45240
tatcttgttc ctctcaaagt acaaaagccc tctttggtaa agtatactta cccaggaaga
45300 attctgaagg taccttgggt agcttggcca tagctatctg catttatgtg
tccttttgta 45360 tgcattactt tggagataag tcattaaatc tgatgctgat
tgtggttcat ttcctcagct 45420 gggttttggg aatatggtgt acagtcttgg
ggcatgtgaa agtgtctgac actgatttaa 45480 tctctccagc aagttgttgg
gctgtaagca gaagattatt gatctactcc ctaaggtgga 45540 agtggccctc
agtaatatca aagaagctga caatactgtc atgttcatgc agggaaaaag 45600
gcagaaagaa atatggcatc tccttaaaat tgcctgtgta agtaattatt aatgaattat
45660 ttaatgtgac attgagttgc tattattcct tgcaaagggg atttttatca
gcatgaggtg 45720 ggccctcttg aagacatgta tattttgcat tgggatgaca
tccatgttcc ttgcttggtg 45780 tccagactag caattgagat gcaggactta
ttatcatctc tcctttccaa tttctctgtt 45840 gttatactcc tcttagccct
tgactactgt ggtattccaa cctaacttgg ggatagatgt 45900 acctgcctag
ataatgaaat tgaccatcta ttcaactggg tttttttttt ctttcttcaa 45960
ctaaaatggt ataagatgag tggatatatt tggttttagt caatgaagaa agggtccaag
46020 gacaggttac taatggtgtg taaaggtggg aatagttttg tactgaaata
ctctggttgt 46080 gcatataagt agtaacggaa atgtaagtct taggaatatg
ataagaaatt gtgtgaatca 46140 acagagatgc ttgcctgtaa tcttttgatg
tttttttctt ttccttcaga cacagagttc 46200 tgcccggtcc cttgtaggat
ccagtctaga aggtgcagta acccctcaga catcagcatg 46260 gctgcccccg
acttcagcag aacatgatca ttctctgtca tgtgtggtaa ctcctcaaga 46320
tgggtgggtt tactttgtaa caatagatag ctgtgttttc atagcagtgg tataggaatg
46380 aggcagtagt tgttaaggta gtcttctgtg ccttcagacc agggttgagt
cctgactatc 46440 cctttgtaat aatcttaggc aacttacttg acttttttga
tgttcatatt tcctcacctg 46500 taaaacagga atgtctcata gaactgctgt
ggaaatttga ttagacaatg tatataaagt 46560 gcttgggaaa taaatgttca
gcaaatgttg cgtattatta gctcaaatgt tgcttattat 46620 tagctaagtc
caagttctaa gtcctttctt ttattggctt attactttct tttactcaaa 46680
ccttactcct tcattaacag tttatgagaa attatgctat cattttgcat ttatcatttt
46740 ggggatcctt gattaccctt ctaataaatt tttgagatct attttataac
tttgtatttc 46800 taagattaga cttttttcag tgttttaaat cagttttgca
agaactcaga ttccaagccc 46860 tatattttat taattcctac tcttttgcta
aaagctgaat gactatccat ttctgttatt 46920 ttctcagctt aagatgtgcc
gtatccaaag ccagtggctc tttgctagct tgtgtattgt 46980 cgcttttctc
acaactcatg gagacctcta cagcaccagc atttcctgga agctctgccc 47040
ataattagtt ttgcagaagt gcagccatgt tacatcactg ctcaggaggt catgcagttg
47100 cacattctgt actctacttt tatgagagaa aagcacatgg caggaaggca
agctgatatc 47160 aggtatttta tttttttaaa gaaagcaagt tccagggtat
atatcttggg gtcattatac 47220 atgtggccca atgcagtaca cctacaactg
tagcacaggc tataattaaa tatatagact 47280 ttacaaaata ctccagtaat
cagagttgaa ggaagtaaaa tttaatactt tacttacttt 47340 tttgccttta
ttaaaggcat ttaagttttt tagcggatgt taatgtttac aaatgtcacc 47400
acaatacagg taatttccaa tttgaagtgt agttgcttgc aaaagtatat ctgacagaca
47460 ttttcaatgt attttctaga aggaacacat tatattttgt tttgagacgg
agtctcattc 47520 tgtcgcccag gctggagtgc agtggcacag tcttggctca
ttgcaacctc cgcctcccag 47580 attcaagcag ttctcctgct tcagcttccc
gagtagctgg gatttcaggt gcgtgccacc 47640 atacctggct aatttttgta
ttttagtaaa gacggagttt caccatgttg gccaggctgg 47700 tctcgaactc
ctgacctcaa gtaatccacc tgcctcggcc tccaaaagtg ctggaattac 47760
aggcatgagc caccatgtcc agccagaata cattgtataa gaaaattgat ttcaattata
47820 atgtacctgt ggtacttaac taccaaggat tgggaaccgt
ttgaccataa tttttccatc 47880 tgttggataa gagctgaatc agaatgtcat
tctaagctaa tagtggctat catccatgac 47940 ctaaaacgga ctcttgttag
gggactgaag attggcaaag ggagaagcta gaggccatat 48000 agaagaggaa
agcatgaagg tagaaaccac aatcaggact gccttcaaat atgaacttcc 48060
agggatactt cttggttctg ggtagctccc ccaccacact gttttctttt gctgtcctca
48120 ggaatcagct tgcttgtttc gtttcattgc tatcatctgt agtttctaat
gagtttttca 48180 ctttctgtga tctctacaga acagatttta cttttctctc
ttctccccta catttgtgtc 48240 tgtgaagtat gcatgctggt gtttttacta
cttttaagga aagagaagta tttagaaaca 48300 aaaacgtatt tcatctgtct
ttgccacaag gattttttcc tccctagtta cattttcacg 48360 ggaaaaacaa
gttgtacata agtgcatctt acatttcaga ggaaagtcag actggagtta 48420
tagtggactt cttatcatta gttaggtaga ggtagaggca gcagcctggt cttaacagga
48480 ctctgcctct gtagggaggg atgaggaaac taaagactac aggagtcaac
tgcactgggg 48540 tactgggtta caggaaggct agtgatcttg ctaacctaga
agactagaaa gtagaggaaa 48600 ttcatatgcc cttttcctga gtttgaaaag
gctttgttct ttcatctcac agggagactt 48660 cagcacaaat gatagaagaa
aatttgaact gccttggcca tttaagcact attattcatg 48720 aggcaaatga
ggaacagggc aatagtatga tggtaagttt tgtgtggata tgggtgcctg 48780
ctttggctat gttgggtgca aaaggtttaa ctttccatgc tagccttatc tggcatttgg
48840 gatgcatatg ggaaatagaa gaactcaaga ggaaagagca tttggggaat
atcctcaacc 48900 ttaaatcctt atctgccgtt actcagggat atactaggat
tatgtcatca attatcttca 48960 ataatagcat ttttggtcaa attaaatgag
tggtaagctt cttcacaatg tgaccattga 49020 aattgaatgg tttgttctgt
acctttttgc ttcagcaatc aattttctcc attaagatgg 49080 gacttgtact
ttaattcaga tatggtacct cccgaataga aaataaatta tgttaatata 49140
gttgtaataa taagtgtgtg ttaagatttg gttactataa actactgatt tgttaaaact
49200 tgaggaaatt accataaaat gtctactgaa tcaatttttc ctgcatttag
tcttaatgtc 49260 aattctgtac atttcctctt tcattaagaa aaatagcagt
ggccaggcat ggtggctcac 49320 gcctgtaatc ctagcacttt gggaggccaa
ggcaggtgga ttgcttgagc ccaggagttt 49380 gagactagcc tggccaacat
gggaaaccct gtctttataa aaaatataaa aattggccag 49440 gtgtggtggc
acacacctgt ggtcccagct acttgggagg ctgaggcagg aagatcgctt 49500
gagttcaaga gtttcaggct gcagtgagcc gtaatcctgc cattgcactc cagcctgtga
49560 cagagtgaga ctttgtctcg gggaaaaaaa aaaaaaaaaa ggataatggt
ggccagccat 49620 atgacatgta cctgtagtcc gagttactag ggaggctgag
gcagaaggat tgcatgagcc 49680 caggagttca aggctgcagt gcattatgat
tggacttgtg aatagccact gtactccagc 49740 ttggcaacat agcaagatcc
tgctctctta agaaaaaaaa aaaaaaagaa aagaaaagga 49800 aagaataatt
gtttacttca aatatttatg aaaaaaactc tgaaattttt ttaaatcagg 49860
aaataaggta aatgaaatga tttttcaact tttgattatg aaatgtccaa acagaaaact
49920 tgcaaaaatg aacacccata tacttataac ttatagtcaa catctaattg
taactttttc 49980 atgcctggtt ttagaatctt gattggagtt ggttaacaga
atgagttgtc acttgttcac 50040 tgtccccaaa cctatggaag ttgttgctat
acatgttgga aatgtgtttt tcccccatga 50100 aaccattctt cagacatcag
tcaatggaag aaatggctat gaacagaaac tacatttcta 50160 ctatgatcag
aagaacatga ttttacaagt ataacagttt tgagtaattc aagcctctaa 50220
acagacagga atttagaaaa agtcaatgta cttgtttgaa tatttgtttt aataccacag
50280 ctatttagaa gcatcatcac gacacatttg ccttcagtct tggtaaaaca
ttacttattt 50340 aactgattaa aaataccttc tatgtattag tgtcaacttt
taacttttgg gcgtaagacc 50400 aaatgtagtt ttgtatacag agaagaaaac
ctcaagtaat aggcatttta agtaaaagtc 50460 tacctgtgtt tttttctaaa
aaggctgctc acaagttcta tttcttgaag aataaattct 50520 acctccttgt
gttgcactga acaggttctc ttcctggcat cataaggagt tggtgtaatc 50580
attttaaatt ccactgaaaa tttaacagta tccccttctc atcgaaggga ttgtgtatct
50640 gtgcttctaa tattagttgg ctttcataaa tcatgttgtt gtgtgtatat
gtatttaaga 50700 tgtacattta ataatatcaa agagaagatg cctgttaatt
tataatgtat ttgaaaatta 50760 catgtttttt catttgtaaa aatgagtcat
ttgtttaaac aatctttcat gtcttgtcat 50820 acaaatttat aaaggtctgc
actcctttat ctgtaattgt aattccaaaa tccaaaaagc 50880 tctgaaaaca
aggtttccat aagcttggtg acaaaattca tttgcttgca atctaatctg 50940
aactgacctt gaatcttttt atcccattta gtgtgaatat tcctttattt tgctgcttga
51000 tgatgagagg gagggctgct gccacagact gtggtgaggg ctggttaatg
tagtatggta 51060 tatgcacaaa actacttttc taaaatctaa aatttcataa
ttctgaaaca acttgcccca 51120 agggtttcag agaaaggact gtggacctct
atcatctgct aagtaattta gaagatatta 51180 tttgtcttaa aaaatgtgaa
atgcttttat attctaatag tttttcactt tgtgtattaa 51240 atggttttta
aattactttc ttgatctcta ttcattataa aaatcagatt ataataaaac 51300
agttgaatat ggcttaggaa aatatgaagg ttccatgaag tggaattaag agcatagaat
51360 aactgtactt tccttaggaa taataggact tatggtaaag gtagtattgg
gcaacttctt 51420 taagagtgtt ttcctctgaa atgtcctatc accactatct
acatctaaaa aacatgctcg 51480 attcttgccc tataaactga tgtcacagcc
ccaccatccc catttttgct agtggtatca 51540 ttttcctagt caaccaaatt
ttttagtcat cacatatcac tgttcgtcat ctattataat 51600 aggcagtctc
tctcttgttt ccctggtttt agaagatttt agtaataaac atttattggg 51660
tacctgttac ttggagggta ttaagctaga tggcaagatc tgaaacaaca taggcagtat
51720 agtaaaagtg cttatctggg agtctgaaca ttacaagcca tccaagcatt
gcaattattg 51780 ttaaggatta ttttcaatgg tcatgcattt tctaatattt
taataattgg ttaaagattt 51840 gttatagcgt gggggccgct gctggtgtgt
ggctggggtt atgtcagggc agcctgatct 51900 atataatttg ggcagatggt
gtgagtcaga acagactatt atagtgggat ccccaaactt 51960 gcttttgatg
catgagttgg accactattt tttggtgggt aacaactttt cagaggggaa 52020
tggcagttgt gaattgtata catttcattc tttaagcaat attatgaaaa acttcagaga
52080 atgtctacag aaaacagggt atggagcaag ttattttcca ttctttttgc
tttttggaag 52140 ttaaaatagc taagctctgc aaatatcatt tatttggaac
agataaggtc ccagacattc 52200 ctagataata aaaatcaaat gaatgataca
ggtaagtgta tttattgaag ggtgtgggta 52260 gagtggtctg ggaagtcttg
ctttaatgaa gacggattga ttctatttac ctcttcttac 52320 ttccactact
aagtaaccct gggcctgcag tttcctaacc acttgacttc cttgccaaca 52380
gtctgtctcc actttcagcc gttttataga ttcattggcc ctaaagcact gctttcccat
52440 acttccctac ccaagagggc aggggtcccc aagccctggg ccgcggaccg
gaaccagaca 52500 gtggcctgtt aggaaccaag ccggatagca ggaggtgagc
ggcgggcgag tgagcattat 52560 cgccttgagc tctgcctgct gtcagattag
cgagtggcat cagattctca taggcacgcg 52620 aaccctattg tgaactgcac
atgtgaggtt tgccggctcc ctgtgagaat ctaatgcctg 52680 atgatctgaa
gtggaacagt ttcatcccca acccacacac acccccaccc cgtccgctgg 52740
gttagaggac tttacaaccc tagtccacct tgtccagtta tagttccacc tctagccttt
52800 caaggcttaa accattaatg tccttaattt ctcttgtatt catctatctc
ccaactatac 52860 ctttttcctt ccctttttta tttttggcaa tatgtgccca
tggtttttta atttaaaacg 52920 aacagaatat gtagtgacgc ctaccatagc
actccttccc atcacacaag ccttcaagga 52980 aactgaagta cttactttgg
tctccctgga ggattccctc cgcctcccgc cccatgtgct 53040 tagcaattct
gttcctgtag tctggatggc cttcataaag ccctcactgg accagcattc 53100
tggcacataa taggaacact taaaaaacga gagaatgatg ccgttatatc taatatcttc
53160 ctcttctgaa ctttcacagc actttatttg caacaagttt agttgctcct
gaagggcagg 53220 atctctgccg gaagcaagtg cgtgccacac agtggggctc
cgcatacact gcaaaaggac 53280 aaataaaccg aacagctacc gtttgagagt
gagcgagtgg gttctctgca caagaacaaa 53340 ccaaccagtc ccttgtccga
aagggcgtct ccttttctct gcttcgctgc tcactccaga 53400 ctgcgggctg
tcctcttccg aagcagttaa ccagcagtgt acagaaagcg acttgcctcc 53460
aaaggagcct gcgcggcccg cggctaggag aattttgtcc catgcgctcc ccgtctcact
53520 agccgcgggc cggggctacg ccgtgtgcgt ccccgcgcag ccgcagtgct
gggcgagtgg 53580 gcggggccgg ctgttggcgg cggttggctc ggcgcgggag
tcggctgcac gtgcgggcgg 53640 gggcgatgcg tcactgatcg gtgaggcgcg
gccgaggggt cggctttcct cgcgagcctg 53700 cggctgggct tcttctcagt
tagtgccttc cacccgggag cgacccttgg gagagggagt 53760 ttcaggaagc
tcaccgagca ggggcggccc actggcctcc gggggcggag gagttggcaa 53820
ggggtcagcg ggctcagcca gaagggaaga atgaggggac aggggtactg gactccccgg
53880 ctcagcctgc gagagagcgc caagtttccg gagggagagg gtagaaactg
gagggggtgg 53940 acctgtcact cacgggactg agggtccttt tctcccgctc
ccaggaggaa cgagaatgaa 54000 tatgactcaa gcccgggttc tggtggctgc
agtggtgggg ttggtggctg tcctgctcta 54060 cgcctccatc cacaagattg
aggagggcca tctggctgtg tactacaggt gagcggcatg 54120 tgcagtcagt
tagggctcta gagcagatta aaagggtact ccagagtgga gtctgggaag 54180
ttgttccctc tgcaacgttc gggggcgcat gtcccatctc tagggagagc acgggggtcg
54240 aagcgggctt ttgcggagca ccctccagat gttggacagt tttagtgcta
tgttcgtttt 54300 gttgcagaga ttaagtctcc ctacggttat tcctatacac
agcaaactca attatttgta 54360 gaaaaatgaa gaaggaagaa aggaagtagt
cactctacct ctaactgaag agatggataa 54420 attatagttc gttcaactgt
acaggcttta gtttgccgct gtagatgctg tatgtttatg 54480 gttgtttctt
agactttttc atttaaaagc ttccagtgtc tggttacgtc ttgtaacacc 54540
atgatgactg tgatggtgct tatgtccctg gacctccagg gacataagca caacactgat
54600 gggccatgct cctccctcct tactgttgct gttcaagttg gggacaggag
tggtgtggga 54660 gctgttgctc tcttgttgag ctcgcttagc ttttccttca
gcctaattac cacaagttcc 54720 ttttgataca gactagtaag agaggccagt
ttgtggagaa attgggataa tggtgtattc 54780 aaaaagagcg tcaacttttt
gcttgtttcc actatccctc aaaactgata tttctacttt 54840 caaatccatc
ctgaatccta gaaaaagagc aattttaatg gtatatctta gcagatggag 54900
ctctacataa gaaattacag attagtccag atttccttcc agtttaaaag cattgtcttt
54960 ctgacagtaa aaccagagaa aggacatttc aaggaatgac tcaaagaatt
agtaatgaac 55020 ctcaaacagt tcatctttcc ttgttattat gaccttgcaa
atgtgtttat attttcaaga 55080 ccggtagcta agttctatga atttacaagg
tgcaaaatag gactttgttt tctttgctcc 55140 ccaaatgcct cctttttata
atttccaggg cttgctttca tagttttcac attccaggat 55200 ttggttaaca
agcccttatg ctatttattt gtatagtctt gtcacccttc atcttttcat 55260
gctaattttt tgtttttata tagaaatttt atgttcccag tattttaatt accttgtaga
55320 tagatcgaaa gaattataag cctcagaaat tttctcttta cttcaaagtt
tgcattgtct 55380 ttttttcaac ctggtgatta aatgtattat atattttgat
ttggctcttg ttatgtacta 55440 ttaatgtacc tctactgaaa gccattagaa
ttttttaaac aaatatttct cagaaggtta 55500 aagaccgagg agaatccttt
gaaccttaga gcagtgctgg agacccagag gtacggttgg 55560 ttgagcttat
gaaacaaggt gtaacattgg cagtttaatt tgatcttttt ccctttttag 55620
gtgtggaggt attttaagca gttgttcatt atgagcaatt catattagtg aggctttagt
55680 aggtcagcag catgagcaag agtgagtatt tattatagca ttttagattt
ggaaagataa 55740 tttttctttt tataattttg tcaaattgaa acatcttttt
gcaggggagg agctttacta 55800 actagcccca gtggaccagg ctatcatatc
atgttgcctt tcattactac gttcagatct 55860 gtgcaggtga gtgattccta
ggggaagcct ccataccaga tagacaggca tgacaggagt 55920 taccacactt
gtgggtggag atagatttcc ttattgctta agcagatggt ggtatttact 55980
taaatgcaga agttaacgtt cagagtacag ttgttacaac acaagttatg atgttaaacc
56040 acctcactta cttgaaatga attaaaggta tgaaccagga agcttggaag
gactgacttc 56100 taccatctat atcgtggtat atagcaaatt ttctgttgct
gctgtcacaa taaaagtaag 56160 aattagaaac acaactagac atacaaacca
gtgattatgt ggtttagaac agtgtttagt 56220 ttccatagag gcaggctcct
gggctactgt taacatggag gatcaaaatt tttatacaca 56280 gcctttctgc
ctcatgtttt tttgcctggt atattaagca caattttatg tggaacctta 56340
gtgaatgtta tattttgatg gcatcccggg atatttgaaa taagtgtcta atcctcgaaa
56400 agtgtcttgg ccatgggaga caataaaata agtatctttc aatgtaacaa
gatgacaaaa 56460 ccaggattag gctgtgacca ctttcgcaag cccagtgtga
tcataaatct ttggattcag 56520 aagtttgtct gccatctgct gctaattttt
caggactcag gaaaattctg tgttatttaa 56580 tcttgtaatc ttatgtctag
gcttcctgct aagtgagagt ttaattataa attcttaaat 56640 cccctaggtt
ttcttcctgt ccccttttag atggcttagg gaaagaattg tgtctactga 56700
agtgaaatgt attcttcccc taatcagact gatttccata tgttgttcca tatttatttt
56760 acccattctt tccttagttc atgccatcac ttctttatct gccatctcac
tctccattcc 56820 ttctctcctg tctaaatctt gccaggcttt taaaggctca
tcccagattc ttcctcttct 56880 gatctatagc tcttaaagtc tatattgtaa
attttggccc tttgtcttat tttctttaat 56940 attgttgtct tgcattgttt
gctaagtttt tttatgtgtt ttctttccag aaaggctcca 57000 gttttcttaa
aggcaggagt tgtaacacat ttatatttct ggctttctat ttatggtgga 57060
agttgttaaa ttgagctgat ttctcaggaa gcaatgtggt gtaatgaaca tggggaccca
57120 gcgttaccgc cagttggtgg catgactttg gaaaaattgc ttaactgtca
tagacttcag 57180 ttagtcttct gtgaaaggag gaattttaat tacataacct
catcaacagc cgaaacaatc 57240 tataataatg ttagcaatgg cagcattaga
cccttaaaaa tcaagactct aaggcccggc 57300 gcagtggctc atgcctgtaa
tccgagcact ttgggaggct gaagcaggtg gatcacttga 57360 gcctaggagt
ttgagaccag cctgggcaac atggggaaac tccgtctctt aaaaaaaaaa 57420
aaaaaaaaaa atcaaggctc taacaaatag atcttgttca aaaccaggta gatctgtctc
57480 tgccttcatg cttagtatgt taagtcatac tgatgcaaaa atataaataa
aggagatata 57540 gataatagta aacagatttt ggagtttaat tgtgtatata
tataacaaat atagtgtgtg 57600 tatatattta ataataaact atcataagag
atgtaaatga aattagtaca gagacttcag 57660 tgtaaactaa aatacttcgc
atctacaaaa aagttttaca tgggctagag ccagccagtg 57720 tgactaaata
ggaatgttct ttatgtgtaa aacggtaaca ttctaggaaa taattatatt 57780
aataaaacca tatttaaaaa gtgttcttgg ccgggcgtgg cggctcacgc ctgtaatccc
57840 agcactttgg gaggccgagg caggcagatc acttgaggtc aggagttcaa
gaccagcctg 57900 gccaacctgg tcaaacgctg tgtctacaaa aatacaaaaa
ttagctgggc gtggttgtgc 57960 gtgcctgtaa tcccagctat ttgggaggct
gaggcaggag aatcgcttga acctgggaga 58020 ccgaggttgc agtgagccga
gatcgtaccc ttgcactcca gcctgggcaa cagaggaaga 58080 ctccgtctca
aaaaaaaaaa aaaaaaaaga gtgttcttaa gagagtaaga cataaattta 58140
tttttaggaa tttttggaac atattagaag acatagtcca gataaacaaa tggtttaaca
58200 aacttggcaa ttgaaaggaa tgtatataaa tgtgaaattc catatatgat
gtaaaaagaa 58260 aaagcaatgg agaaattata tgaaaacctt cagccttcat
aaagtaacca tagattcact 58320 ttttaaataa attttctgtc atactgggaa
agtaattttt aagaggacat aaaagaaata 58380 taaaagtaca taaagatagg
gtgttaaatg gaagatttaa catttgaacc ctttcctagt 58440 tcttctgagt
ttgaaaattg gctagagaat atccttttgg tttaaatagg aacgtgtatt 58500
taaagttggt gtggaatatg gaaactgaat taatgttata aaggaaataa atataatttg
58560 tctcttcatc aatcccctta acctagaact gccaccaatg tttactgtca
ttctaatgtc 58620 tcaacctaag agttattttt tattctttcc cctcctggaa
ttttatatca accaagtatc 58680 agatacctcc tctattatgt ctggattgac
agctttcatg gcctttactg aattaatcta 58740 aataaattaa ttatttagcc
tcccagctgg ttttcttacc tccctataag atggagaatg 58800 agaactacat
cacatgtgag aaaaggaatt aagtaaacat atttttgaat ggttcgtttt 58860
gtgactgtat ataaggtgaa ctagagagat cccaaactcc ccagccagca ttcagggccc
58920 tctgtctagt attaccatgg tctcttgtca gaatttcatc tttcatccaa
attgtccctt 58980 ctgctccagg caagcctatc tactttcgtt cgcctataca
catgttgtct tcaactgtgt 59040 gtatttcagt aggatattta ttgagcttaa
gatgttcctc tgcctcatct gttcttattt 59100 ctcccactct agacctggct
tataaatcct tcacctcaga gagcttttca aactactatt 59160 tttttcactt
tctgtaattt accacttaac agttcagttt ataatttgtg tttttggttt 59220
tgttttgttt tgttttgtac ctgttgatgg attgattgca ttgtaacatt ttgtctcctg
59280 tgctggattg tgaactcttt ggaaacaggg actgtgattc ctttttcctt
tgctttgtgc 59340 ttgataagat gcagtatgtc tagaatgtac tcattaggtg
ctgacctatt tgaggcctta 59400 gagtacagca ggaagtcatt cattctggat
ctgaacaaag gtgtgacaag ggcaaagcaa 59460 ttaagtacca gattgttggt
tcaccaggag gaagcagtac tggtggtagc acttctatta 59520 aaaaggaaac
tgaataggac atgtgagaga tcactaggta ttaaactaaa atgttgacca 59580
cagctttaga gaagaaaggt tgctgttgga ttcaagggca gctttgagtc ttagagtact
59640 tgtatgcata aaatctcttt tatcttcaac ataaatagta atattagcag
tggaatcaaa 59700 gttcagagtg caagttgtag ccagagtatc tatgtgagca
agctgtgttg tttacaaccc 59760 ctggccagct ccaggcaggt aagggactct
ggaagagttt tctacttatt agcagactga 59820 acattgaacc tttcccagtg
aatcttaatt caaacattcg ctgtaggcca ggcacagtgg 59880 ctcacgcctg
taatcccagc actttgggag gccaaggtgg gtggatcact tgaggccagg 59940
agttcaagac cagcctggcc aacatggtga aacctcatct ttactaaaaa tacaaaaatt
60000 agctgagcgt agtggtgcat acctgtaatc ccagctactt gggaggctgg
ggcacaagaa 60060 ccgcctgaac ccaggaggca gaggttgcag tgagccaaga
ttgcaccact gcattccagc 60120 ctgggtgaca aagcaagact ctgtctccaa
aaaaaaaaaa aaattgctgt aaaatatttt 60180 ctgtctcaag tctttcattt
ctactagaat acaaaaagaa atgactgttc tgagagctaa 60240 tctttgggaa
ctgaaattgt gatggatctc tgctgtactt actgtcagca gctttccaga 60300
gatttgcctc tggtaccagt agttttttga caatgtgtgt cttgagtgtt gatgagatat
60360 cttttcctcc ctagacaaca ctacaaactg atgaagttaa aaatgtgcct
tgtggaacaa 60420 ggtaagcttt ctctttgcct acgagcctcc ctttagctca
gacagagttt ctgttcctac 60480 agctatgatc acttagacat atctcaaata
ggcatgatag ttaacccaag gggacttctg 60540 atctgagcat tgtttgagca
aaggcctatc tccaaatagg agcttttcca tcttaagaac 60600 cattattgtt
cttatggtat tattggaaaa aactggagtt ttagagcaag tagacctaac 60660
tgtgtattag cactgctgca tctgctttat tgactatttt ttaaagactt tattttttac
60720 agcagtttta ggttcatagt gaaattgacc aaaaagtaca gacagttttt
atatacctct 60780 ttccccaaca cattccttta ttgacacttt aaagcttagt
ttcctccttt ataaaggcag 60840 taaaatctac cttgcagagt tgttgtaagg
attagaatta ctgtatatga agtgcctaac 60900 cagaacttgg cccagaatag
gtgcttaata aaacatagtt taaaattaat catcccaccg 60960 tcttctcatc
tgtagtgtta aatgccatct aatttagagt ttctgattca ataggtctgg 61020
ggtgaggccc gagaatttgc atttctaaca aaagttcctg ggtgatgctg atactgctgg
61080 tctgaagacc tcactttgag aattgctgtc ctaaagaggt ttagggaaag
aaaggcaagc 61140 agggcccaaa ttatttctag ggtataatca ctgcattgaa
ttgtatcaaa agaagaatgt 61200 gaagttcaag gtatttttta aattacttat
tttaaatgac tattacaagt gtaaaaaaga 61260 cattgacatc tatagaggga
cagaaatgct atgaaaaagg aagcgaactc tttctgtgtt 61320 ttaagtgaa 61329
39 31394 DNA Homo sapien misc_feature 5165, 5166, 5167, 5168, 5169,
5170, 5171, 5172, 5173, 5174, 5175, 5176, 5177, 5178, 5179, 5180,
5181, 5182, 5183, 5184, 5185, 5186, 5187, 5188, 5189, 5190, 5191,
5192, 5193, 5194, 5195, 5196, 5197, 5198, 5199, 5200, 5201, 5202,
5203 n = A,T,C or G misc_feature 5204, 5205, 5206, 5207, 5208,
5209, 5210, 5211, 5212, 5213, 5214, 5215, 5216, 5217, 5218, 5219,
5220, 5221, 5222, 5223, 5224, 5225, 5226, 5227, 5228, 5229, 5230,
5231, 5232, 5233, 5234, 5235, 5236, 5237, 5238, 5239, 5240, 5241,
5242 n = A,T,C or G misc_feature 5243, 5244, 5245, 5246, 5247,
5248, 5249, 5250, 5251, 5252, 5253, 5254, 5255, 5256, 5257, 5258,
5259, 5260, 5261, 5262, 5263, 5264, 5265, 5266, 5267, 5268, 5269,
5270, 5271, 5272, 5273, 5274, 5275, 5276, 5277, 5278, 5279, 5280,
5281 n = A,T,C or G misc_feature 5282, 5283, 5284, 5285, 5286,
5287, 5288, 5289, 5290, 5291, 5292, 5293, 5294, 5295, 5296, 5297,
5298, 5299, 5300, 5301, 5302, 5303, 5304, 5305, 5306, 5307 n =
A,T,C or G 39 tgcacacaaa actctctaca ctgctagtct ttaattctca
aaaacagttc cctgagagag 60 ggaacataat tgcctctatt ttacagatga
caaaaccagg cttagaagtt acacaaattg 120 ccttgcacag tggctcacac
ctataatccc acacattggg aggctgaggc aggaggattg 180 agttagaaac
cagcctggtc aacatagcaa gacccccatc tctacaaaag aaaagataaa 240
tttgctgggc atagtggtgc acacctatac tcctgcttgg gaggctgagg caggaggatc
300 gcttgagccc aggaggtcga ggctacagtg atccatgatc gcaccattgc
actccagcct 360 gggcgacaga gcgaggccct atcgcttaga aaacaaaaag
ttacatgaag ttcccccaag 420 gtcccgtagc tagcatgtga tgtggcaggg
agagagcctg tccgcttcac tctctcacat 480 gggaccagaa aggggtgatg
ctgtgcgtgg acaggacatc tggcaagtgg tgtgtccagg 540 acacaggaag
cagcgtagtg gccaagctgc ctccagctgt cacatctgtg tgtctctgca 600
gtaaggatga gtctgccttc atggtccacc cgctgacggc acagggagtg
gccgtggtaa 660 tagtggctta cggcatcgcc cccaaaggta ataggagtgg
ttgctgcagg tccgagggcc 720 ggtgggcttt aggaggaagt gcatccctga
cccagctcat gctctctgtg caggcaccct 780 ggaccacatg gtagaccagg
tgacccgcag cgttgcgttt gtccagaagc ggtatccaag 840 caacaagtgg
gtgttgccag tagattttct tcctgttgga cctcagtggg tgggaggaca 900
gtgcaatcag tacctaggat acagccaagc tgcccctctg gcctgtggag cagaaagcaa
960 attgggactc ttcgaagggg gcctgatgtt gtggggaaac tgcgtgcaaa
tccagctctc 1020 tgctctgacc cctcccaggg gaatttacct gtgtggacac
tcagccgggg cccacctggc 1080 tgccatgatg ctcctggccg actggaccaa
gcatggggtc acgcccaacc tcagaggttt 1140 ccatgggagc tacagcctgg
ctgggcaacc ttcatctccc catgagcctt ggggtttggg 1200 ccaactgctt
gaaatcctcc cggccatgag tggcttgacc gcagggcttc gagccctctg 1260
agcaaggcct tctctgcctc ctctgtgccc cttcccctgg tcctgcccct ctggcggtgg
1320 gggtgggctg gccctgcctt gctccgcctg gagcctggcc ctgtgacaat
tctgtacctt 1380 cacaggcttt ttcctggtga gtggggtctt tgacctggag
cccatcgtgt atacttcaca 1440 gaacgttgct ctccagctga ccctgtgagt
tacttggcca ccacctcccc tggctcacca 1500 ggagcctggc tcctctctgt
cctgacccct gtccactccc caccccaggg aggacgctca 1560 gaggaatagc
ccccagctga aggtggccca ggcacagccg gtggacccca cctgccgtgt 1620
gctggtggtc gtgggccagt tcgactcccc cgaattccac cgacagtcct gggagtttta
1680 ccaggtactc ccagtgcagg tttgtggcca gaggtcgagg gtcatgtggg
ctcattattt 1740 tcctctttct tttacccaag tggacaagac cctccatcat
ttatttaaca cgtactgagt 1800 gaaccctgac ctgtgccagg catggcccca
tgtcctgccc cacctcccct cccactgctc 1860 aggcccctct tcccatgtct
cccctgccca gaccctgtgt caaggagagt ggaaagcctc 1920 atttgaagag
ctccacgatg tggaccactt tgaaattgtt gagaatctga cccagaagga 1980
caacgtgctc acccaggtgg ggcctcatcc ctggcagccc tttcatggta gacagcacag
2040 gtcctgtcgc agccccttag atgtgcaaac caggagttca agaccagcct
ggccaacatg 2100 gagaaactcc gtgtctacta aaaatacaaa acaaattagc
cagatgtgat ggtacgtgcc 2160 tgtaatccca gctactcagg aggctgaggc
atgagaattg ctttaaccca ggaggcagag 2220 attgcagtga gccaagattg
caccactgca ctccggcctg agcgacagag tataagactc 2280 tgtctcaaaa
aaaaaaagac ctgagaacct aggaaagaaa gaacctaaga aagaccatct 2340
tgtttctttc tttttttttt tctttagaga cagggtctcc tctcttgcac ccaggctgga
2400 gtatagtggc aaaatcttag ctctctgctg cctcctgggc tcaagtggtt
atcctgcccc 2460 agcctcttga gtagctggga ctacaaaggc gtgccactgt
gccctgctaa ttattcaatg 2520 tttttgtaga gatggggtct ctctatgttc
cccaggctgg tcgtcgtcgt cttccttctt 2580 cttcctcctt cctccttcat
cttctttctt ctcctcctcc ccctaccccc ccttcttctt 2640 cttcttcctt
catctcagca tattatacct ctaaccagac tgtcttcaac tcctgagttc 2700
aagtgatctt cccacctcag cctcccgaag ggttggggtt gcagacgtga gccactgcat
2760 ccagccctgt ctcatttcct taatcaaatt cctgcaaaga gctgctctga
ttctggggct 2820 tttgtgtctt ctcttcctgt tccagattat cttgaaaaca
atcttccagt agttctgacg 2880 atacttggag cctggtccac gtgcatccca
ccttgggaag cctctccaaa gagctttcgg 2940 agctgacact gacagcttca
gtttccccca gcacccagga gagccttgct gtgtctgtct 3000 gcccggcaag
agtccattct cactgctggg acactcatga aaatctccac gtcctccctc 3060
ttcccagcct ggatggagct ccagggctgg ggaacgtccg caagtcaatg ctcagagatg
3120 cccggagctg cctcttagac tcgtctggcc catctacctg ctgacagagc
atgacaaaga 3180 tgacgctcaa aagtaatgcc attacttctt tttttttttt
tttttttttt ttttttgaga 3240 aggagtctta ctctgtcacc caggctggag
tgcaggggcg tgatcttggc tcactgcaac 3300 ctccacctcc cggggtcaag
ctattctcct tcttcagcct cctgagcctt tgggactggg 3360 ggcgcctgcc
atcatgcctg gctaattttt gtttttttag gggagacggg ggttcaacct 3420
attggccagg ctggtctcaa actcctaacc ttgtgattcg cccgccttgg cctcccaaag
3480 tgctgggatt ccaggcgtga gccactgcgc ccggccagaa tggcattata
tttaaatagt 3540 tcataaagaa gcacaaaaga atattatttc ataacatgta
aaaattatat aaaacgtaaa 3600 tttccatgtt gataaataaa gttgtattgg
aacacggccc tgctcgttca tttacgtatt 3660 gtctagggca gctttcaagc
tgcagggtgg agctgagtgg tggtgacagt gacctcctgg 3720 cccgtagtgc
ctgaagcatt tcctatctgg cccttttcag gaaaagtttg cggacccctg 3780
ggtaagggct tccccaagcc agctaccctc tgccttctgc ttcagtctcc tgtctgctct
3840 gtcctccatc tccctgacat ttgggtctgt ccctgaatgc cccggggtct
cagcgttggg 3900 gccaacacct tgaccttgac aggacaatgc tacccacatg
taggacaagc ttcctctgca 3960 cccgagagtg gtgtgcatgt cccaagaagc
cagctatatc catccaccac ccaccccacc 4020 acaccgcctc tttctctcac
accgaatagc attcaagaat ttccactgtg ggctgggcgc 4080 aggggctagt
gcctgtaatc ccagcacttt aggaggccga ggtgggcgaa tcacctgagg 4140
tcaggagttc aagaccagcc tggccaacat ggcaaaaccc tgtctctact aaaaatacaa
4200 aaattagcca ggcgtggtgg ctggcgcctg taatcctaga tacgtgggag
gctgagttag 4260 gagaatctct tgaatccggg aggcagaggt tgcagtgagc
tgagattgtg cccctgcact 4320 ccagcctggg tgacaaagcc agactccatc
tcaaagaaaa aaaaaaaaat tccactgtat 4380 gtgtgcagca aaccatcatg
acacacattt acctgtgtaa caaacctgca catcctacac 4440 atataccctg
gaacttaaag taaaagttgg gggggggggt aaaaaagaat ttccaccgtg 4500
acattattga gtatagcaaa aaaaaaaaaa acaagaaaca gcctagtgtt cattagggaa
4560 taaacgcatt caagcagcat caaaccctgc agccattaca aagagatcta
tgttgaccat 4620 gtggaatatc tccaagagcc acagtagcct cccttatctg
taggattcac tccaagaccc 4680 tctgaaacca tggataatac tgaaccctat
atacactatg ttttttcttg tatatacata 4740 cctacgataa agtttaattt
ataaattggc aaagggtata taaatattcc ttctaagaga 4800 ttaacaataa
ctaataaagt agaacgatta aaacaatata ctgtgatcaa agttatgtga 4860
agccaggtgc tgtggctcat gcctgtaatc ccagcacttt gggaggctga gacaggtgga
4920 tcacctgagg tcaggagttg gagaccagcc tggccaacat gacaaaaccc
cgtctctact 4980 aaagataaaa aaaattagcc gggcatggtg acacatgcct
gtaatcccag ctacttggga 5040 ggctgaggca ggagaatcgc ttgaacctgg
gaggcggagg ttgcagtgag ctaagatcac 5100 accattgcac tccagcctgg
gcaacaagag tgaaactctg tctcaaaaca aaacaaaaca 5160 aaacnnnnnn
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 5220
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
5280 nnnnnnnnnn nnnnnnnnnn nnnnnnnact tcatctcaaa aaaaaaaaaa
gaaaaaagaa 5340 gtccagaaac ccgcggagac actatcacac tatccccccc
aggctggtag tgcagtggcc 5400 caatcatggc tcactgcagc ctcgacctcc
caggatcaag tgatccttcc acctcagcct 5460 cccgagtagc tggaagtata
ggtgcacgcc cgactgattt tttttttttt tttttagacg 5520 gagtctcact
cttgttgctc tggctggagt gcaatggcag gatctcggct cactgcaacc 5580
tctgcctctt agattcaagc gattctcgtg cctcagcctc ccgagtagct gggattacag
5640 gtgcccacca ccatgcccgg ataatttttt gtatttttaa tagagacagg
gtttcaccat 5700 attggtcagg ctggtctcaa actcctgacc tcaggtgatc
cacctgcctc agcctcccaa 5760 actgctggga ttacaggcgt gagccaccgg
gcatggcctt tcctggctaa ttttttaaat 5820 ttttgataga gatggggtct
cagtgttgcc caggctgatc ttgaactcct agattcaagt 5880 gatcctccct
ccttggtctc ccaaagtgct gagattacag gcgtgagcca ccgccccggg 5940
ctggaaaata cttttttaaa cgagggcaat gtgaatctga aatgccattt gaggaaagat
6000 ctgttcgcct gacatcctgt ttgagcctgg gtggacagga cagcacctgc
cagcatcggg 6060 aagcactgca gatgggaaga ggcttggtca ctctccaaag
gtggcaggag ttggaggggg 6120 tgagctgaag gtaaggagaa aggaggtggg
gacccaggag acaggggctg cgcagcgggc 6180 tcggggctga cacccccacg
gatacagttc actggggctc aaacataaaa ggaacccaac 6240 tattgtggga
ggaaaagact cttctgcctt tctgcctttt ctttttttct ttttctttct 6300
ttcttttttt tttttttttt ttgagacaga gtcttgctct atcgcccagg ctggagtgca
6360 gtggcgtgat ctcggctcac tgcaagctct gcctcccggg atcacgccat
tctcctgcct 6420 caacctcccg agcagctggg actacaggcg cctgccacca
cacccggcta tttttttgta 6480 ttttttagta gagatggggt ttcaccatgt
tagccaggac ggtctcgatc tcctgacctt 6540 gtgatccgcc cgcctcggcc
tcccaaagtg ctgggattac aggcgtgagc caccgcgcct 6600 ggctcttttt
tctttctttt tttttttccg agacagagtt tcactcttgt tgcccaggct 6660
ggagtgcagt ggcgcgatct tggctcactg caacctccac ctccagggtt caagcgattc
6720 tcctgcctca gcctcctgag tagctgggac tgcaggcgcg caccaccacg
cctggctaat 6780 ttttgtattt ttagtagaga cagggtttca ccatattggc
caggctggtc tcgaactcct 6840 gaccttgtga tctgcccacc tcagcctccc
aaagtcctgg gattacaggc gtgagccacc 6900 gtgcccagcc tgacccctct
gccctttcaa aaactatgtt cgttctctca cagccttctc 6960 ttgtcatatt
aagtccacac cgcaggccta atttgtccag tgaatgctat gcaaatattt 7020
catgcacctg ctgatcgcag gaatgatatg tacttggtac gcactgatcg tacctcgggg
7080 tgggagaaga gagggcaagg aagcaaagaa tagccccctc ctttcctggt
gcaccttcag 7140 atgtgccgat ggggcccagg ctcgctgcag atggccccct
tcccagagac aggggaggat 7200 cctccaccca ctccccagcc tccaggacca
tcctgactcc tgccttcagg cactcaagtt 7260 atgcgtctag acatgcggat
atattcaagc tgggcacagc acagcagccc caccccaggc 7320 agcttgaaat
cagagctggg gtccaaaggg accacacccc gagggactgt gtgggggtcg 7380
gggcacacag gccactgctt ccccccgtct ttctcagcca ttcctgaagt cagcctcact
7440 ctgcttctca gggatttcaa atgtgcagag actctggcac ttttgtagaa
gccccttctg 7500 gtcctaactt acacctggat gctgtggggc tgcagctgct
gctcgggctc gggaggatgc 7560 tgggggcccg gtgcccatga gcttttgaag
ctcctggaac tcggttttga gggtgttcag 7620 gtccaggtgg acacctgggc
tgtccttgtc catgcatttg atgacattgt gtgcagaagt 7680 gaaaaggagt
taggccgggc atgctggctt atgcctgtaa tcccagcact ttgggaggct 7740
gaggcgggtg gatcacgagg tcaggagttc aataccagcc tggccaagat ggtgaaaccc
7800 cgtctctact aaaaatacaa aaaaattagc cgggcatggt ggcgggcgca
tgtaatccca 7860 gctactgggg gggctgaggc agagaattgc tggaacccag
gagatggagg ttgcagtgag 7920 ccaagattgt gccactgcac tgcactccag
cctggcgaca gagcaagact ctgtctcaaa 7980 aaaaaaaaaa aaaagtgaaa
aggagttgtt cctttcctcc ctcctgaggg caggcaactg 8040 ctgcggttgc
cagtggaggt ggtgcgtcct tggtctgtgc ctgggggcca ccccagcaga 8100
ggccatggtg gtgccagggc ccggttagcg agccaatcag caggacccag gggcgacctg
8160 ccaaagtcaa ctggatttga taactgcagc gaagttaagt ttcctgattt
tgatgattgt 8220 gttgtggttg tgtaagagaa tgaagtattt cggggtagta
tggtaatgcc ttcaacttac 8280 aaacggttca ggtaaaccac ccatatacat
acatatacat gcatgtgata tatacacata 8340 cagggatgtg tgtgtgttca
catatatgag gggagagaga ctaggggaga gaaagtaggt 8400 tggggagagg
gagagagaaa ggaaaacagg agacagcgag agagcgggga gtagagagag 8460
ggaaggggta agagagggag aggaggagag aaagggagga agaagcagag agtgaatgtt
8520 aaaggaaata ggcaaaacat aaacagaaaa tctgggtgaa gggtatatga
gtattctttg 8580 tactattctt gcaattatct tttatttaaa ttgacatcgg
gccgggcgca gtggctcaca 8640 tctgtaatcc cagcactttg ggaggccgag
gcaggcagat cacttgaggt caggagtttg 8700 agaccagcct ggcaaacatg
gtgaaacccc atctctacta aaaatacaaa aattagcctg 8760 gtgtggtggt
gcatgccttt aatctcagct actcgggagg ctgaggcagg agaatcgctt 8820
gaacccgtgg cggggaggag gttgcagtga gctgagatca tgccactgca ctccagcctg
8880 ggcgatagag cgagactcag tttcaaataa ataaataaac atcaaaataa
aaagttactg 8940 tattaaagaa tgggggcggg gtgggagggg tggggagagg
ttgcaaaaat aaataaataa 9000 ataaataaac cccaaaatga aaaagacagt
ggaggcacca ggcctgcgtg gggctggagg 9060 gctaataagg ccaggcctct
tatctctggc catagaacca gagaagtgag tggatgtgat 9120 gcccagctcc
agaagtgact ccagaacacc ctgttccaaa gcagaggaca cactgatttt 9180
ttttttaata ggctgcagga cttactgttg gtgggacgcc ctgctttgcg aagggaaagg
9240 aggagtttgc cctgagcaca ggcccccacc ctccactggg ctttccccag
ctcccttgtc 9300 ttcttatcac ggtagtggcc cagtccctgg cccctgactc
cagaaggtgg ccctcctgga 9360 aacccaggtc gtgcagtcaa cgatgtactc
gccgggacag cgatgtctgc tgcactccat 9420 ccctcccctg ttcatttgtc
cttcatgccc gtctggagta gatgcttttt gcagaggtgg 9480 caccctgtaa
agctctcctg tctgactttt tttttttttt tagactgagt tttgctcttg 9540
ttgcctaggc tggagtgcaa tggcacaatc tcagctcact gcaccctctg cctcccgggt
9600 tcaagcgatt ctcctgcctc agcctcccga gtagttggga ttacaggcat
gcaccaccac 9660 gcccagctaa tttttgtatt tttagtagag acaaggtttc
accgtgatgg ccaggctggt 9720 cttgaactcc aggactcaag tgatgctcct
gcctaggcct ctcaaagtgt tgggattaca 9780 ggcgtgagcc actgcacccg
gcctgcacgc gttctttgaa agcagtcgag ggggcgctag 9840 gtgtgggcag
ggacgagctg gcgcggcgtc gctgggtgca ccgcgaccac gggcagagcc 9900
acgcggcggg aggactacaa ctcccggcac accccgcgcc gccccgcctc tactcccaga
9960 aggccgcggg gggtggaccg cctaagaggg cgtgcgctcc cgacatgccc
cgcggcgcgc 10020 cattaaccgc cagatttgaa tcgcgggacc cgttggcaga
ggtggcggcg gcggcatggg 10080 tgccccgacg ttgccccctg cctggcagcc
ctttctcaag gaccaccgca tctctacatt 10140 caagaactgg cccttcttgg
agggctgcgc ctgcaccccg gagcgggtga gactgcccgg 10200 cctcctgggg
tcccccacgc ccgccttgcc ctgtccctag cgaggccact gtgactgggc 10260
ctcgggggta caagccgccc tcccctcccc gtcctgtccc cagcgaggcc actgtggctg
10320 ggccccttgg gtccaggccg gcctcccctc cctgctttgt ccccatcgag
gcctttgtgg 10380 ctgggcctcg gggttccggg ctgccacgtc cactcacgag
ctgtgctgtc ccttgcagat 10440 ggccgaggct ggcttcatcc actgccccac
tgagaacgag ccagacttgg cccagtgttt 10500 cttctgcttc aaggagctgg
aaggctggga gccagatgac gaccccatgt aagtcttctc 10560 tggccagcct
cgatgggctt tgttttgaac tgagttgtca aaagatttga gttgcaaaga 10620
cacttagtat gggagggttg ctttccaccc tcattgcttc ttaaacagct gttgtgaacg
10680 gatacctctc tatatgctgg tgccttggtg atgcttacaa cctaattaaa
tctcatttga 10740 ccaaaatgcc ttggggtgga cgtaagatgc ctgatgcctt
tcatgttcaa cagaatacat 10800 cagcagaccc tgttgttgtg aactcccagg
aacgtccaag tgcttttttt gagatttttt 10860 aaaaaacagt ttaattgaaa
tataacctac acagcacaaa aattaccctt tgaaagtgtg 10920 cacttcacac
tttcggaggc tgaggcgggc ggatcacctg aggtcaggag ttcaagacct 10980
gcctggccaa cttggcgaaa ccccgtctct actaaaaata caaaaattag ccgggcatgg
11040 tagcgcacgc ccgtaatccc agctactcgg gaggctaagg caggagaatc
gcttgaacct 11100 gggaggcgga ggttgcagtg agccgagatt gtgccaatgc
actccagcct cggcgacaga 11160 gcgagactcc gtcataaaaa taaaaaattg
aaaaaaaaaa aagaaagaaa gcatatactt 11220 cagtgttgtt ctggattttt
ttcttcaaga tgcctagtta atgacaatga aattctgtac 11280 tcggatggta
tctgtctttc cacactgtaa tgccatattc ttttctcacc tttttttctg 11340
tcggattcag ttgcttccac agctttaatt tttttcccct ggagaatcac cccagttgtt
11400 tttctttttg gccagaagag agtagctgtt ttttttctta gtatgtttgc
tatggtggtt 11460 atactgcatc cccgtaatca ctgggaaaag atcagtggta
ttcttcttga aaatgaataa 11520 gtgttatgat attttcagat tagagttaca
actggctgtc tttttggact ttgtgtggcc 11580 atgttttcat tgtaatgcag
ttctggtaac ggtgatagtc agttatacag ggagactccc 11640 ctagcagaaa
atgagagtgt gagctagggg gtcccttggg gaacccgggg caataatgcc 11700
cttctctgcc cttaatcctt acagtgggcc gggcacggtg gcttacgcct gtaataccag
11760 cactttggga ggccgaggcg ggcggatcac gaggtcagga gatcgagacc
atcttggcta 11820 atacggtgaa accccgtctc cactaaaaat acaaaaaatt
agccgggcgt ggtggtgggc 11880 gcctgtagtc ccagctactc gggaggctga
ggcaggagaa tggcgtgaac ccaggaggcg 11940 gagcttgcag tgagccgaga
ttgcaccact gcactccagc ctgggcgaca gaatgagact 12000 ccgtctcaaa
aaaaaaaaaa aaagaaaaaa atctttacag tggattacat aacaattcca 12060
gtgaaatgaa attacttcaa acagttcctt gagaatgttg gagggatttg acatgtaatt
12120 cctttggaca tataccatgt aacacttttc caactaattg ctaaggaagt
ccagataaaa 12180 tagatacatt agccacacag atgtgggggg agatgtccac
agggagagag aaggtgctaa 12240 gaggtgccat atgggaatgt ggcttgggca
aagcactgat gccatcaact tcagacttga 12300 cgtcttactc ctgaggcaga
gcagggtgtg cctgtggagg gcgtggggag gtggcccgtg 12360 gggagtggac
tgccgcttta atcccttcag ctgcctttcc gctgttgttt tgatttttct 12420
agagaggaac ataaaaagca ttcgtccggt tgcgctttcc tttctgtcaa gaagcagttt
12480 gaagaattaa cccttggtga atttttgaaa ctggacagag aaagagccaa
gaacaaaatt 12540 gtatgtattg ggaataagaa ctgctcaaac cctgttcaat
gtctttagca ctaaactacc 12600 tagtccctca aagggactct gtgttttcct
caggaagcat tttttttttt tttctgagat 12660 agagtttcac tcttgttgcc
caggctggag tgcaatggtg caatcttggc tcactgcaac 12720 ctctgcctct
cgggttcaag tgattctcct gcctcagcct cccaagtaac tgggattaca 12780
gggaagtgcc accacaccca gctaattttt gtatttttag tagagatggg gtttcaccac
12840 attgcccagg ctggtcttga actcctgacc tcgtgattcg cccaccttgg
cctcccaaag 12900 tgctgggatt acaggcgtga accaccacgc ctggcttttt
tttttttgtt ctgagacaca 12960 gtttcactct gttacccagg ctggagtggg
gtggcctgat ctcggatcac tgcaacctcc 13020 gcctcctggg ctcaagtgat
ttgcctgctt cagcctccca agtagccgag attacaggca 13080 tgtgccacca
cacccaggta atttttgtat ttttggtaga gacgaggttt caccatgttg 13140
gccaggctgg tcttgaactc ctgacctcag gtgatccacc cgcctcagcc tcccaaagtg
13200 ctgagattat aggtgtgagc caccacacct ggcctcagga agtattttta
tttttaaatt 13260 tatttattta tttgagatgg agtcttgctc tgtcgcccag
gctagagtgc agcgacggga 13320 tctcggctca ctgcaagctc cgccccccag
gttcaagcca ttctcctgcc tcagcctccc 13380 gagtagctgg gactacaggc
gcccgccacc acacccggct aatttttttg tatttttagt 13440 agagacgggt
tttcaccgtg ttagccagga gggtctcgat ctcctgacct cgtgatctgc 13500
ctgcctcggc ctcccaaagt gctgggatta caggtgtgag ccaccacacc cggctatttt
13560 tatttttttg agacagggac tcactctgtc acctgggctg cagtgcagtg
gtacaccata 13620 gctcactgca gcctcgaact cctgagctca agtgatcctc
ccacctcatc ctcccaagta 13680 attgggacta caggcgcacc ccaccatgcc
caccttattt atttatttat ttatttattt 13740 atttattttc atagagatga
gggttccctg tgttgtccag gctggtcttg aactcctgag 13800 ctcaagggat
ccttttgcct gggcctccca aagtgctgag attacaggca tgagccaccg 13860
tgcccagcta ggaatcattt ttaaagcccc taggatgtct gtgtgatttt aaagctcctg
13920 gagtgtggcc ggtataagta tataccggta taagtaaatc ccacattttg
tgtcagtatt 13980 tactagaaac ttagtcattt atctgaagtt gaaatgtaac
tgggctttat ttatttattt 14040 atttatttat ttatttttaa tttttttttt
tgagacgagt ctcactttgt cacccaggct 14100 ggagtgcagt ggcacgatct
cggctcactg caacctctgc ctcccggggt caagcgattc 14160 tcctgcctta
gcctcccgag tagctgggac tacaggcacg caccaccatg cctggctaat 14220
ttttgtattt ttagtagacg gggtttcacc atgctggcca agctggtctc aaactcctga
14280 ccttgtgatc tgcccgcttt agcctcccag agtgctggga ttacaggcat
gagccaccat 14340 gcgtggtctt tttaaaattt tttgattttt tttttttttg
agacagagcc ttgctctgtc 14400 gcccaggctg gagtgcagtg gcacgatctc
agctcactac aagctccgcc tcccgggttc 14460 acgccattct tctgcctcag
cctcctgagt agctgggact acaggtgccc accaccacgc 14520 ctggctaatt
ttttttggta tttttattag agacaaggtt tcatcatgtt ggccaggctg 14580
gtctcaaact cctgacctca agtgatctgc ctgcctcggc ctcccaaagc gctgagatta
14640 caggtgtgat ctactgcacc aggcctgggc gtcatatatt cttatttgct
aagtctggca 14700 gccccacaca gaataagtac tgggggattc catatccttg
tagcaaagcc ctgggtggag 14760 agtcaggaga tgttgtagtt ctgtctctgc
cacttgcaga ctttgagttt aagccagtcg 14820 tgctcatgct ttccttgcta
aatagaggtt agacccccta tcccatggtt tctcaggttg 14880 cttttcagct
tgaaaattgt attcctttgt agagatcagc gtaaaataat tctgtcctta 14940
tatgtggctt tattttaatt tgagacagag tgtcactcag tcgcccaggc tggagtgtgg
15000 tggtgcgatc ttggctcact gcgacctcca cctcccaggt tcaagcgatt
ctcgtgcctc 15060 aggctcccaa gtagctgaga ttataggtgt gtgccaccag
gcccagctaa cttttgtatt 15120 tttagtagag acagggtttt gccatgttgg
ctaagctggt ctcgaactcc tggcctcaag 15180 tgatctgccc gccttggcat
cccaaagtgc tgggattaca ggtgtgaacc accacacctg 15240 gcctcaatat
agtggctttt aagtgctaag gactgagatt gtgttttgtc aggaagaggc 15300
cagttgtggg tgaagcatgc tgtgagagag cttgtcacct ggttgaggtt gtgggagctg
15360 cagcgtggga actggaaagt gggctgggga tcatcttttt ccaggtcagg
ggtcagccag 15420 cttttctgca gcgtgccata gaccatctct tagccctcgt
gggtcagagt ctctgttgca 15480 tattgtcttt tgttgttttt cacaaccttt
tagaaacata aaaagcattc ttagcccgtg 15540 ggctggacaa aaaaaggcca
tgacgggctg tatggatttg gcccagcagg cccttgcttg 15600 ccaagccctg
ttttagacaa ggagcagctt gtgtgcctgg aaccatcatg ggcacagggg 15660
aggagcagag tggatgtgga ggtgtgagct ggaaaccagg tcccagagcg
ctgagaaaga 15720 cagagggttt ttgcccttgc aaatagagca actgaaatct
gacaccatcc agttccagaa 15780 agccctgaag tgctggtgga cgctgcgggg
tgctccgctc tagggttaca gggatgaaga 15840 tgcagtctgg tagggggagt
ccactcacct gttggaagat gtgattaaga aaagtagact 15900 ttcagggccg
ggcatggtgg ctcacgcctg taatcccagc actttgggag gccgaggcgg 15960
gtggatcacg aggtcaggag atcgagacca tcctggctaa catggtgaaa ccccgtcttt
16020 actaaaaata caaaaaatta gctgggcgtg gtggcgggcg cctgtagtcc
cagctactcg 16080 ggaggctgag gcaggagaat ggcgtgaacc tgggaggtgg
agcttgcagt gagccgagat 16140 cgcgccactg cactccagcc tgggcgacag
agcgagactc cgtctcaaaa aaaaaaaaaa 16200 aagtaggctt tcatgatgtg
tgagctgaag gcgcagtagg cagaagtaga ggcctcagtc 16260 cctgcaggag
acctctcggt ctctatctcc tgatagtcag acccagccac actggaaaga 16320
ggggagacat tacagcctgc aagaaaagta gggagattta aaaactgctt ggcttttatt
16380 ttgaactgtt ttttttgttt gtttgttttc cccaattcag aatacagaat
acttttatgg 16440 atttgttttt attactttaa ttttgaaaca atataatctt
ttttttgttg tttttttgag 16500 acggggtctt actctgtcac ccaggctgag
tgcagtggtg tgatcttggc tcacctcagc 16560 ctcgaccccc tgggctcaaa
tgattctccc acctcagctt cccaagtagc tgggaccaca 16620 ggtgcgtgtg
ttgcgctata caaatcctga agacaaggat gctgttgctg gtgatgctgg 16680
ggattcccaa gatcccagat ttgatggcag gatgcccctg tctgctgcct tgccagggtg
16740 ccaggagggc gctgctgtgg aagctgaggc ccggccatcc agggcgatgc
attgggcgct 16800 gattcttgtt cctgctgctg cctcggtgct tagcttttga
aacaatgaaa taaattagaa 16860 ccagtgtgaa aatcgatcag gaaataaatt
taatgtggaa ataaactgaa caacttagtt 16920 cttcataaga gtttacttgg
taaatacttg tgatgaggac aaaacgaagc actagaagga 16980 gaggcgagtt
gtagacctgg gtggcaggag tgttttgttt gttttctttg gcagggtctt 17040
gctctgttgc tcaggctgga gtacagtggc gcaatcacag ctcactatag cctcgacctc
17100 ctggactcaa gcaatcctcc tgcctcagcc tcccagtagc tgggactaca
ggcgcatgcc 17160 accatgcctg gctaatttta aatttttttt tttctctttt
ttgagatgga atctcactct 17220 gtcgcccagg ctggagtgca gtggcgtgat
ctcggctgac ggcaagctcc gcctcccagg 17280 ttcactccat tcgcctgcct
cagcctccca agtagctggg actacaggcg ctgggattac 17340 aaacccaaac
ccaaagtgct gggattacag gcgtgagcca ccgcacccgg cctgttttgt 17400
ctttcaatag caagagttgt gtttgcttcg cccctacctt tagtggaaaa atgtataaaa
17460 tggagatatt gacctccaca ttggggtggt taaattatag catgtatgca
aaggagcttc 17520 gctaatttaa ggcttttttg aaagagaaga aactgaataa
tccatgtgtg tatatatatt 17580 ttaaaagcca tggtcatctt tccatatcag
taaagctgag gctccctggg actgcagagt 17640 tgtccatcac agtccattat
aagtgcgctg ctgggccagg tgcagtggct tgtgcctgaa 17700 tcccagcact
ttgggaggcc aaggcaggag gattcattga gcccaggagt tttgaggcga 17760
gcctgggcaa tgtggccaga cctcatctct tcaaaaaata cacaaaaaat tagccaggca
17820 tggtggcacg tgcctgtagt ctcagctact caggaggctg aggtgggagg
atcactttga 17880 gccttgcagg tcaaagctgc agtaagccat gatcttgcca
ctgcattcca gcctggatga 17940 cagagcgaga ccctgtctct aaaaaaaaaa
aaaccaaacg gtgcactgtt ttcttttttc 18000 ttatcaattt attattttta
aattaaattt tcttttaata atttataaat tataaattta 18060 tattaaaaaa
tgacaaattt ttattactta tacatgaggt aaaacttagg atatataaag 18120
tacatattga aaagtaattt tttggctggc acagtggctc acacctgtaa tcccagcact
18180 ttgggaggcc gtggcgggca gatcacatga gatcatgagt tcgagaccaa
cctgaccaac 18240 atggagagac cccatctcta ctaaaaatac aaaattagcc
ggggtggtgg cgcatgcctg 18300 taatcccagc tactcgggag gctgaggcag
gagaatctct tgaacccggg aggcagaggt 18360 tgcggtgagc caagatcgtg
cctttgcaca ccagccaagg caacaagagc gaaagtccgt 18420 ctcaaaaaaa
aagtaatttt ttttaagtta acctctgtca gcaaacaaat ttaacccaat 18480
aaaggtcttt gttttttaat gtagtagagg agttagggtt tataaaaaat atggtaggga
18540 agggggtccc tggatttgct aatgtgattg tcatttgccc cttaggagag
agctctgtta 18600 gcagaatgaa aaaattggaa gccagattca gggagggact
ggaagcaaaa gaatttctgt 18660 tcgaggaaga gcctgatgtt tgccagggtc
tgtttaactg gacatgaaga ggaaggctct 18720 ggactttcct ccaggagttt
caggagaaag gtagggcagt ggttaagagc agagctctgc 18780 ctagactagc
tggggtgcct agactagctg gggtgcccag actagctggg gtgcctagac 18840
tagctgggta ctttgagtgg ctccttcagc ctggacctcg gtttcctcac ctgtatagta
18900 gagatatggg agcacccagc gcaggatcac tgtgaacata aatcagttaa
tggaggaagc 18960 aggtagagtg gtgctgggtg cataccaagc actccgtcag
tgtttcctgt tattcgatga 19020 ttaggaggca gcttaaacta gagggagttg
agctgaatca ggatgtttgt cccaggtagc 19080 tgggaatctg cctagcccag
tgcccagttt atttaggtgc tctctcagtg ttccctgatt 19140 gttttttcct
ttgtcatctt atctacagga tgtgactggg aagctctggt ttcagtgtca 19200
tgtgtctatt ctttatttcc aggcaaagga aaccaacaat aagaagaaag aatttgagga
19260 aactgcgaag aaagtgcgcc gtgccatcga gcagctggct gccatggatt
gaggcctctg 19320 gccggagctg cctggtccca gagtggctgc accacttcca
gggtttattc cctggtgcca 19380 ccagccttcc tgtgggcccc ttagcaatgt
cttaggaaag gagatcaaca ttttcaaatt 19440 agatgtttca actgtgctct
tgttttgtct tgaaagtggc accagaggtg cttctgcctg 19500 tgcagcgggt
gctgctggta acagtggctg cttctctctc tctctctctt ttttgggggc 19560
tcatttttgc tgttttgatt cccgggctta ccaggtgaga agtgagggag gaagaaggca
19620 gtgtcccttt tgctagagct gacagctttg ttcgcgtggg cagagccttc
cacagtgaat 19680 gtgtctggac ctcatgttgt tgaggctgtc acagtcctga
gtgtggactt ggcaggtgcc 19740 tgttgaatct gagctgcagg ttccttatct
gtcacacctg tgcctcctca gaggacagtt 19800 tttttgttgt tgtgtttttt
tgtttttttt tttttggtag atgcatgact tgtgtgtgat 19860 gagagaatgg
agacagagtc cctggctcct ctactgttta acaacatggc tttcttattt 19920
tgtttgaatt gttaattcac agaatagcac aaactacaat taaaactaag cacaaagcca
19980 ttctaagtca ttggggaaac ggggtgaact tcaggtggat gaggagacag
aatagagtga 20040 taggaagcgt ctggcagata ctccttttgc cactgctgtg
tgattagaca ggcccagtga 20100 gccgcggggc acatgctggc cgctcctccc
tcagaaaaag gcagtggcct aaatcctttt 20160 taaatgactt ggctcgatgc
tgtgggggac tggctgggct gctgcaggcc gtgtgtctgt 20220 cagcccaacc
ttcacatctg tcacgttctc cacacggggg agagacgcag tccgcccagg 20280
tccccgcttt ctttggaggc agcagctccc gcagggctga agtctggcgt aagatgatgg
20340 atttgattcg ccctcctccc tgtcatagag ctgcagggtg gattgttaca
gcttcgctgg 20400 aaacctctgg aggtcatctc ggctgttcct gagaaataaa
aagcctgtca tttcaaacac 20460 tgctgtggac cctactgggt ttttaaaata
ttgtcagttt ttcatcgtcg tccctagcct 20520 gccaacagcc atctgcccag
acagccgcag tgaggatgag cgtcctggca gagacgcagt 20580 tgtctctggg
cgcttgccag agccacgaac cccagacctg tttgtatcat ccgggctcct 20640
tccgggcaga aacaactgaa aatgcacttc agacccactt atttctgcca catctgagtc
20700 ggcctgagat agacttttcc ctctaaactg ggagaatatc acagtggttt
ttgttagcag 20760 aaaatgcact ccagcctctg tactcatcta agctgcttat
ttttgatatt tgtgtcagtc 20820 tgtaaatgga tacttcactt taataactgt
tgcttagtaa ttggctttgt agagaagctg 20880 gaaaaaaatg gttttgtctt
caactccttt gcatgccagg cggtgatgtg gatctcggct 20940 tctgtgagcc
tgtgctgtgg gcagggctga gctggagccg cccctctcag cccgcctgcc 21000
acggcctttc cttaaaggcc atccttaaaa ccagaccctc atggctacca gcacctgaaa
21060 gcttcctcga catctgttaa taaagccgta ggcccttgtc taagtgcaac
cgcctagact 21120 ttctttcaga tacatgtcca catgtccatt tttcaggttc
tctaagttgg agtggagtct 21180 gggaagggtt gtgaatgagg cttctgggct
atgggtgagg ttccaatggc aggttagagc 21240 ccctcgggcc aactgccatc
ctggaaagta gagacagcag tgcccgctgc ccagaagaga 21300 ccagcaagcc
aaactggagc ccccattgca ggctgtcgcc atgtggaaag agtaactcac 21360
aattgccaat aaagtctcat gtggttttat ctactttttt tttctttttc tttttttttg
21420 agacaaggcc ttgccctccc aggctggagt gcagtggaat gaccacagct
caccgcaacc 21480 tcaaattctt gcgttcaagt gaacctccca ctttagcctc
ccaagtagct gggactacag 21540 gcgcacgcca tcacacccgg ctaattgaaa
aatttttttt tttgtttaga tggaatctca 21600 ctttgttgcc caggctggtc
tcaaactcct gggctcaagt gatcatcctg cttcagcgtc 21660 cgacttgttg
gtattatagg cgtgagccac tgggcctgac ctagctacca ttttttaatg 21720
cagaaatgaa gacttgtaga aatgaaataa cttgtccagg atagtcgaat aagtaacttt
21780 tagagctggg atttgaaccc aggcaatctg gctccagagc tgggccctca
ctgctgaagg 21840 acactgtcag cttgggaggg tggctatggt cggctgtctg
attctaggga gtgagggctg 21900 tctttaaagc accccattcc attttcagac
agctttgtca gaaaggctgt catatggagc 21960 tgacacctgc ctccccaagg
ctttcataga tcctctctgt acattgtaac cttttatttt 22020 gaaatgaaaa
ttcacaggaa gttgtaaggc tagtacaggg gatcccgcat accgttctcc 22080
tccttttccc cgttgtgagg acagtgcact gtattcactt tcaacatcca aggtctgtgt
22140 tgagagggtg agttgtgcgg cccagcttgc cccaagcccg tcttgccatc
acgaaggatt 22200 tcctgactca gctgaatcta gctcaacaaa gcaaaggact
aactaagctg cagattttct 22260 ctccgctggg cttagcctac attatttaca
cctgttttca gctggatggg accaaactgg 22320 ggccttgact tctggggttt
ccacgtgatc cccaaagagg agacatccat tcattcactc 22380 aggtatttgc
ttgtgcacct actgtgcgcc aggtgccaga tccaggaatt gaagcccagg 22440
aacaactctt gcagtgagct ccccgggttt ttgttttctg aacggtggga agacaggtca
22500 gcaaatagcc atgcacgtct gcacatgtgt ggctggtgtc tgcaggagta
ccgagaagac 22560 ggtgagctct gccctggatg gcaacaggaa agggcatccc
tgtgcagctg tttgtcctgg 22620 cacaggtggc aggctgccag cgggaccagg
aagggaaggg gaggatgcac aggtcggagg 22680 gaacggaacg ggcactgcag
gtgcgctctg cgatgtgagg aaggaagcaa aggctggttc 22740 cgggatgatg
tcggagaagc cttgcagcct agggcactgc ccattccacg ttacccagtg 22800
agtcaaagcc tcccagaaag acatttagtc actcaaagcc agtgcaggtc tcagctttgt
22860 ccggggagga actttcactc tagaaaacac gaactcattt ccctttgagg
aagttgtttc 22920 tccaggatat catttccagc caccatccag gatgctgggc
aggtgctgca gtgcccaagg 22980 cacggctgcc gaagaagctt gtcccttaga
aaccccttgt ggcctcggag gcgcctgcct 23040 gcctccctga gaatggaacc
ctgttgggcc gccaggctcg ctgggcaggt gctggggaag 23100 gctgggtcac
cgtgctgagc ccctcacagg cctgggagtt gtgcttgggg aggagagctc 23160
ccagctcttt ggctgcctcc caatcatggc cacatctcac aaacaatacc actgtccacg
23220 aaaaatgaaa aggtgcctgg ttctaaccca tggccccaac catgaaagag
cccttagaat 23280 gtgggggttt gactcgaaca gtggctttga gtctcccagg
ggggattagt gtcctaatgc 23340 cctgagttaa tgaattgact tgttggacag
ggaaactcag tgaggttcat tttctgtggg 23400 gggacgtttg tcaaatgctg
tgaaatcagt tgtagcatac gcgacgctca gctgctccga 23460 cagtgactca
gccacggctc cagctgcttc cccagccctg tagtggaggt ggcagatggt 23520
gtcatcagcc tggcgcagag ggctgtgccc actggcatca caagggccag gcctgctggt
23580 cctccagcca ctggatccag gtagttacat atgggtttaa aagaagagag
gcagggccag 23640 ttgcggtggc tcatgcctgt aatcccagca ctttgggaag
ccgaggtggg tgatcatttg 23700 aggttaggag ttcaagatca gcctgaccaa
catggcgaaa cccagcctcc actaaaaata 23760 caaaaaaatt agctgggcat
agtgctgggt gcctgtagtc ccagctactc aggaggctga 23820 ggcaggagaa
ttgcttgaac ccggaaggca gaggttgcaa tgagtcagga tcatgccact 23880
gcactccagc ctgggcaaca gaacgagact ctaaaaaaaa aagatcaata aataaaataa
23940 aaataagaga ggcagggtgg gagtattact tgaggccagg agtttgagac
cagcctgggt 24000 aacaaagcaa gaccctgtct ctaccaaaat aataatttaa
aaaattaccc gggaatggtg 24060 tgacatgcct gtggtcccag ctactctgga
ggctgaggtg ggaggaacac tcgcacatag 24120 gaagttgagg ctgcagtgag
ccgtgatcac accactgcac cactctagcc tgggtgacag 24180 agcaagacct
catctctggg gaaaaaaaaa aaaagaagag gcttggaagc cacattgttt 24240
tttgtcactt ataactagag tttcgtccat caaaagccca ttctgcactg atttcagtga
24300 taggggtctg atgctggctt ggaagagggc ctgacagctt agagacaggc
ttggagttcc 24360 aagttgatga aagaacgatg aataatgaaa aataatgtgt
tgttctaaag tagtgccttt 24420 tcatactgag ctagggtata atttgggtcc
atttcaaggg aaatatacta gcagaattca 24480 tcaaagtagc ccctcgaaga
ctggtgcctg ctgcctacgg gacaatttaa tcagtggttc 24540 cagctgcaca
tctagagttg ggaacactga acccaggctt tctcatgcag atggggtgtg 24600
gattgctaag gtggctgtct cctagcgctg cagcatgagt gaaacacgtg tccattctac
24660 agagacgcgg ctccgtgatg ctgaaacctc cttctttttt tttttttttg
agacggagtc 24720 ttgctctgtc gcccaggctg gagtgcagtg gcgatatctt
ggctcactgc aacctccgcc 24780 tcctgggttc atgcaattct cctccctcag
cctcctgagt agctgggact acaggcgcgt 24840 gccaccatgc ctggctaatt
tttgtatttt tagtagagac agggtttcac cgtgttagcc 24900 aggatggtct
cgatctcctg acctcgtgat ctgcctgcct cggcctccca aagtgccggg 24960
attacaggcg tgagccactg caccccgctg aaacctcctt ctagataaca gcggcgttaa
25020 gcattttttt ttttttttga gatagagtgt tgctctgttg cccaggctgg
agtgcagtga 25080 tgcgatctcg gctcactgca acctccacct cccaggttca
agcaattctt ctgcctcagc 25140 ctcccgagta gctgggacta caggcacacg
ccaccaggcc cagctaattt ttgtattttt 25200 agtagagacg gggtttcagt
atattggcca ggctggtctc gaactcctga cctcatgatc 25260 cacccgtctc
agcctcccaa agtgctggga ttacaggcgt gagccactgc acctggccaa 25320
aacctcctcc tagataacag tggcgttaag catttttttt ttttttgaga tggagtcttg
25380 ctctgttgcc caggctggag tgcagtggtg tgatctcagt tcactgcaac
ctctgcctcc 25440 cgggttcaag tgattcttct gcctcaggct cctgagtagc
tgggataaca ggcatgagcc 25500 accactccca gctaattttt gtatttttag
tagagacagg gttttgccat gttggccagg 25560 ctggtctgga actcctgacc
tcaagtgatc ctcccacctc ggcctcccaa agtgctgaga 25620 ttacaggcgt
gagccactgc acctggcctt tttttgtttt ttaagacagg gtctcactct 25680
gttgatcaca gccacagctc actgcagcct caacctcctg cctgggttca agcaatcctc
25740 ccaccttggc ctcccaggta gctgagacca caggcacaag ccaccatatc
tggccctacg 25800 taagctttat gtgggtgcaa taacagatct caaaccagct
ctcagctcaa gagaagggga 25860 agctgggcac agtggctcac gcctataatc
ccagcacttt gggaagccga ggcaggagga 25920 tcacttgaac ccaggagttc
gagaccagcc tgggcaacat agggagaccc ctgccctccc 25980 tgtctctgaa
aaaaaaaaaa aaaaaaaaag aaaggagagg ctgggcttgg tggctcacac 26040
ctgtaatccc agcactttgg gaagcccagg caggaggatc acttgaaccc tggagtttga
26100 gtccagcctg ggcaacatag ggagaccttg tctctaaaaa aataaataga
gtgagagaga 26160 gaaggggaag tcaccgggag gacacacatg gagcacgcag
cccaaatggg gctcctctgt 26220 tctccctgca gagtcagttc gtacctttca
catgcacaaa ggtgttgatc caaccgaagc 26280 aatattgatt cttgttatat
tgctttattg tggggtcagt gtcagagtac cacgagtttt 26340 catcacgcac
cgttctttcc ccagctgcag tgtgtttagg gggggaacga ggccccaccc 26400
cctatcccta tcgcctctgc ctttataacg gctgctactc tctgagcacc tgttgagttc
26460 tttcaccttc cagatctaat gtgctatccc cacccctaga gaacagggct
gctggatgcc 26520 acatggctcc cagagagggg ctgtggtcag ggaagcccca
gctccaaggg ggctggaaaa 26580 ccccagagct gcccacgtgc ggcaacacag
tctggtgctg caggtgtgag gacggctaca 26640 aaatacctga gccattctgt
cactctgtct gaactctgtt tgaaatattg tttcaataac 26700 tgctaggctg
gtttttcctt cctgactata ttcctcaaac caacaagagc ctagcagagg 26760
aaagcatggc caaaacgccc caagaagaga gccttggctt cacctccaag ggccaccatc
26820 cgggaggatg tcctcagacc tctgatcccc tctctgcagg ctctgcccag
ccctgtgtgg 26880 caacccagag gaagcctccc cttcgtttga gatttaaccc
cagaccttag gcgatggctg 26940 cccagcctgt cccttccgcc tgtgtggctg
cccacgcggg cgttgctcat ggggctagtc 27000 ctgggtggat gggtgggggc
ctctcgccgg ctcctctgcc tcccaccccc actggcaccc 27060 cacgcctgtc
ctagaaggtt ctttctgcct gttttctccg tgagtgactg gacaggcaga 27120
ggccggcctt gctggagggg gcatttgtaa ttatgagtga atccaaaaca aggttttttc
27180 cttccgcagc cccccgcccg gctgtggggc ccagccactg cacttcaccg
gatgccgtct 27240 ggttggtcct caggactgat acagaccagg accccagggc
cagcccgtgc caggctccta 27300 tgcttccagg agcacgggtg ggtggtcctg
ctgcctggcc ggccatcctc ctggggtcgg 27360 tctctggccg atcctccctc
ctcctctcaa gccctgcaca gcccggccag gcaggtgcat 27420 cttgtttggc
tgctgaggag ccgggggttc agggaaatta aggaacgtgc ccagggaccc 27480
ggggccagcc cgtggggacg ctgggattgg agcccaagcc ccaggttcgc cgcgcggctc
27540 tcgacttcct ctcctttccc ccaggggcga gctcagcgac cgcagagagg
tggggtcgat 27600 ctccctgcga ccccaggggg cccgcgaggc cagtgcgcgg
gcaggagcgg ggacgtgctc 27660 agaagagccg ggcgccgccg cgcccgcccg
ccccccgtcc cccggctccc ggctccgcgc 27720 gccccccgcc gcccccgggg
ccctgctacc cccgacccgt ccccacccgc cggccgcccc 27780 catggcccgg
ctgggcgcgc tgctcctggc cgccgccctg ggtgcactgc tcagcttcgc 27840
gctcctggcc gccgcggtcg ccagcgacta ctggtacatc ctggaggtgg cggacgccgg
27900 caatggcagc gcctggcccg ggcgcgcaga gctgctctcc tcgcactcgg
ggctctggcg 27960 catctgcgaa ggtaaccggc caccgcgccg gccctcctcc
ctccgcgacc tcgtccctcc 28020 gacaccccct taaccccgcc ctgctcgtgt
tgccccgcca gacccccttc caggacccct 28080 ctcttgcatc ccgctctgcc
ccagggtgtc tctccgctcc ctccccatca ccctccgtct 28140 tcccccaaaa
ctgacagccc aagggctaca ggagggaggg agcccagttc ggggcccctc 28200
acagccggag gagggggctg tggggcgaca gtgggggagg gaagcctaga ggtgtgtagt
28260 tggggggcct catccaagtc accaggggtt gtttcttgat cacgctcccc
ggggtttggg 28320 cctggcagcc ccttgtcccc gtccctgtca ggcactgtca
gagctgttca ccccacacct 28380 cctgatgccg cgggggcagg ggttccaaat
gtgtacagag gcttcagagt ccggggagag 28440 agaaggggat cccagcaggc
tggagggtcc acagggcccc ctccgttccc cccagctccc 28500 tcctccggag
ctggggccag cctggggagc ttccccttca cagcgcgagg gctggcaggg 28560
caggggtgtg tgtgtgagtg agcatgtgtg tgcatctgaa tatgtgtgca tgtgctgaac
28620 atgaacatgt gtgagcacct ggctgtgtat gtaaatatga atatgtgagt
gtgagtgtat 28680 gtagttgtgt gtgcatgtga atatgagtgt gtgagcatgt
cagtgtgcat gtgtgtgcat 28740 gtgagagtgt gtgggcatgt atatgagtga
gtgcatgtgt gggtggtgca catgtgctgg 28800 tggcttcctc agtgcaactg
gggaggagtt aggagccttg tgttcatcca acagactgcc 28860 taactgtgct
ccaaagacct cctgacatgc acctgtgtgc acgtgtgcac acgcctgtgc 28920
ctatgcaggt ggctctgcac gcgtgtgaga accagagagt atgtatgccc acatgacacg
28980 agtcaggaag attccagagc gaggcttccc aggagcccgt tttgtacacc
ctttttcctt 29040 tcaggcaaag cctgagttcc acacacaaac gcattacaag
gacccctgcc tgagggactc 29100 tgagggggcc tccatggagc gtttgaaagt
ttaaacatgc acctgtgcag gcataacttg 29160 cacgtgaaaa taaacaaggt
gaaggctggg cccggtggcc cacgcctgta atcccagcac 29220 tttgggaggc
cgaggcgggt ggatcacgag gtcaggagat cgagaccaca gtgaaacccc 29280
gtctctacta aaaatacaaa aaattagccg ggcacggtgg cgggtgcctg tagtcccagc
29340 tacttgggag gctgaggcag gagaatggcg cgaaaccggg aggcggagct
tgcagtgaac 29400 cgagatcgcg ccactgcact ccagcctggg gtgacagagc
gagactccat ctcaaaaaaa 29460 aaaaaaagaa aaagaaaata aacaaggcta
aaggtcttgg gccttttcat cccacttgga 29520 gtcccagccc tgagtttcag
cagaagagaa gctggcaggg ccctgtggtg ccagggggtc 29580 ccctgggctc
gggtaggctg ctgggtgcac cacagcgtcc aggccccagg cttcgaggcc 29640
tatgtttccc agggcagaac ggctgcatcc cgctggtcga cccttttgcc agtgagagcc
29700 tggacgtctc cacctcggtg cagcacctca tctgtgagtc ctgggtgggg
ccacctcccc 29760 atcctttcca aatattcagg caaggcagat cccagccatc
cccatcccca tcccgcagca 29820 ctgcttccac tgcccctgcg tttccaagag
gacgtttcca cgcagacctg tcccagtgct 29880 gtgctgctgt ccctaaccac
aggtgcccag ctcccagcct atcctgtttt cccctgtctc 29940 agtttccctc
tcagtggtga tggctcatcc ctttgttatt ggggaagcca ggcagctccc 30000
agcttggctg gatcctgcgg gccttgcaga ctcctggctc cccatttagg actcgattct
30060 tccagatggc ctgctgtcta cctggctggc accttccacc tggcgggttg
ggggacatgt 30120 gaggcctggg gatgggggct gggagtgttc tggggacgcc
tcccctcctg gccttaggaa 30180 gcccctgcca gagtcagagg gggccactgg
gagggtccag tggtgtccac agagatgggc 30240 gtcagccgct tttcctgaga
agacgaagca ccacgtcact gtcctccggc agacaagtgc 30300 tgaagggccc
acctggacca gaattctcac ctgagccccc tttcctgcag tgctgcaccg 30360
tgcagtcatt gtggtcctgc ccctgagcct ggtccttctc gtgtgtggct ggatctgcgg
30420 cctgctcagc tccctggccc agagcgtgtc tctgctgctt ttcaccggct
gctacttcct 30480 gctggggagt gagtctgggg ccctggggga atggctccaa
agatgggagc tgggccacag 30540 gtcccgggag tggggtgctg cctctcctgt
tgccctcgtc ccctgctccc ttctgggcgg 30600 gtgctgagtc tggcgatgga
gccgtggcag gcagctccct gtggttccag aaggtaccca 30660 tgtatatttg
ttctcacgtt gctataaaga aatgcctgag actgcgtaat ttataaagaa 30720
aagaggttta ggccgggagc agtggctcat gcctgtgatc ccagcacttt
gggaggctga 30780 ggtgggtgga tcatctgagg tcaggagttc aagaccagcc
tgaccaacat ggtaaaaccc 30840 catctttact aaaaacacaa aaattagctg
ggcttataaa gaaaagaggt ttaattggct 30900 catggttctg catgctgtag
aggaagcatg atgcttggct tctggggagg cctcaggaaa 30960 atgacaatca
caggcggaag gccaaggcag ggagccagca tttcacatgg ctgggacagg 31020
aggaagagag tagggaggtg ttacacactt tttttttttt ttgagatgga gtttcgctct
31080 tgttgcccag gctggagtgc aatcgcatga tctcgactca ctgcaacctc
cgcatcctgg 31140 gttcgagcaa ttctcctgcc tcagcctcct gagtagctgg
gattacaggc atgcgccacc 31200 acacccagct aattttgtat ttttggtaga
gacggggtct ctccatgctg gtgaggctgg 31260 tcttgaactc ctgacctcag
gtaatccacc tgcctcagcc tccctaagtg ctgggattat 31320 aggtgtgagg
caccgttccc agctgggtgc tacatagttt tttttttttt ttttttgaga 31380
cggagtctcg ctct 31394 40 24915 DNA Homo sapien 40 ttggttcttt
gaaaagatga accaaattgg caaaccttta gctacaggaa gaaaaaggtg 60
agaagacaca aataactaaa atcataaagg aaagtgtaca tattaccacc aaacttacag
120 aaataaaaag gattataata ctgtgaacaa ttgtatgtca acaaattaga
taacttagat 180 gaatagaaaa gctgaacaga cctataacaa gtaaagagat
tcaatcagta atcaaaacat 240 tccaacaaag aaaagtccag gagtagatca
cttctctggt gaataatatc caacatttaa 300 agaattaaca ctgttccttt
tcaaactctc aataaataga cctgagagaa cactctctaa 360 ctcattctat
taagctagta ctctgatacc aaggcagata atgacatcac aagaaaagca 420
aattacagat attgcacacc catgtttata gcagcacttt tcacaatagc caagaggtgg
480 aagcaaccca aatatctatc aacaggtgaa tggataaaaa aaatgtgtta
tctacataca 540 atggaatatc attcagcctt aagaggaagg aaatcctgtc
aaatgctaca atgtggatga 600 accttgaggt cattatccta aatgaaataa
gccagtgaca gacaaatact gtatgattcc 660 acttacatga ggtatattaa
ggagtcaaat tcctagaaac agaaagtaga gtggtgttta 720 ccaggggctg
gggacagagg gaaaaggcca attgtttaat gggcatagag ttatagtttt 780
gcaaggtgaa aacgttttgg agatctgttt cacaactatt tgaatatgtt tcacactact
840 aaactgtact tttgaaaatg gttaatatgg taagttttat gttatatgct
tttaccacta 900 taaaaaaatt gcagaccagt atcccttacg aatatagata
caaaagttct caaaaagggc 960 cgggcaccgt gactcatgcc tgtaatccca
gcactttggg aggccgaggt gggtggatca 1020 tgaggtcagg agttcaagac
catcctggcc cagatggtga aaccccgtct ctactaaaac 1080 tacaaaaatt
agccaggtgc agtggcaggc atctgtaatc ccagctactt gggaggctga 1140
ggcaagataa ttgcctgagc tggggtagca gagtttgcag tgaattgaga tcatgcaact
1200 gcactccagc ctgggtgaca aagtgacact ccatctcaaa aaaaaagaat
ccgtctcaaa 1260 aaaaaaaatt ctcaaaaact actagtaaac ccaaaccaac
agcatattaa aaggattata 1320 taccattacc acatgaggtt tatcccagga
atgcaagggt agctcaacat aagaaagtca 1380 attagtataa tatgccacat
tagtaaaaca agggaaaaag acaaacattt catttcaatt 1440 gagcagaata
ggcattttat tccctaataa aaaccagcag aaaactggga atagaaggga 1500
acttcctcaa cttgataaag ggtataatga aaaacccaca gctaacatca agcaacaaca
1560 acaaaataga taaattggat ttcatcaaaa ttaaaaactt ttgggcatca
aagaacattg 1620 taaagaaagt aaaaagacaa tttacagaat ggaagaaaat
atttgtaaat catatatctg 1680 ataaagattt aacatacaga atatataaag
aacttctaca accccacaac aaaaagaacc 1740 cagttaaaaa aagtactgat
caaaggactt gaacagacat ttctcaaaag aagatataca 1800 aatagccaac
atgcagttga aaatatgctc tacaccatta gttattaggg aaatgcaaat 1860
caaaatcaca atgagatacc atttcacatc tactaggatg gcaataataa tataataata
1920 atacagaaaa taacaagtgt tggcaaggat atggaaaaac ttaaatcctt
taacattgct 1980 agtgggaatg taaaataatg gagccattat ggaaggtagt
ttagagattc ctcaaaaagt 2040 taaagaagaa ccatatgacc tagcaatcct
gcttctaggt atatatccca aataatttaa 2100 agactcagat acttgtacac
caagtataca ccaagtgtgc tgtttcatcg tagcgctatt 2160 aaccatagcc
aaaaagtaga aacaactcaa ttgtccataa atggataaac aaaatgtagt 2220
tatacatatc tttaattatt actcagctat gaaaaggaat gaagttctga tatgtgctac
2280 aacatgatga ttcttgaaat aatgctaagt gttttaaaat aatgtttgaa
aaaacactaa 2340 atgaaataag ccagacacaa gtaaaaagac aatctataga
atgggagaaa atatttgatt 2400 tttcaggggc ttgggagatg ggggaattag
gtgttactgc ttaatggaaa gtttctgatt 2460 gtggttatga aaaagttttg
gaaatagatg gtagtgatga ttacacaata ttgtgaatat 2520 aattaatagt
cactgaattg tatatataac atagttaaaa tggcaaattg tatattatat 2580
gtatcttaac caccttaaaa gaacccccct caaaaagaat attaatactt gacagtcttc
2640 accttcccca caacaaaaag tcacattggt cttcattagg ggactaaatc
tgtagaataa 2700 attacagata attcctgtag tttattaagt tggactctct
ggaagcaatc agggtagcag 2760 ttcttaagta aataattctc tttatgtatt
tttatgtata gagagaggga gagacagagt 2820 ctcactgttg cccaggctgg
tctcaaactc ttggcctcat gtgatccttt cacctggcct 2880 cccaaagcac
tgggatttca ggtgtgagcc accacacctg gcccttgcct ttatgtctta 2940
agtggggctc tttaaggtct taaaaggtga atttttcaag tgctgagaaa attaaagtag
3000 aaaagcttag aagacactgg gtctgtgagg tagaggacac ctaagccctt
ttggccccca 3060 tgttccagct gatagccaaa aggtagcgat agatgtgaaa
ggaggaggtt ctgctgccag 3120 ctaaacccca atattccagg acctatggta
agactttgag gacaccatct agtccagtac 3180 tttgttctca gatctgattg
aacaagtcca ggtgcttcag ataatcgctt cttgtgacaa 3240 cttcaggttg
tgccattctt tactccctga aaggaatatt aaagtggaaa acctcaagta 3300
gtattgcatt aagaatgagc tagtgtaccc tcttctaggt gcagaaagta gagttgacag
3360 tgtaggcatt gttaggcttc catgaggatt tcttttgaaa agatgttccc
tgcttaaaaa 3420 ataaatgaca aatttagagt attctcagag ttcaaaagaa
aaagtttatt ttccccctat 3480 tttctatgtt ggcaaatggg ccacataaag
gtgcagttct gttcatttaa gcttttggag 3540 tattgcaaca ggtacctcaa
agttggggaa atgttacttg attttgaaaa gcatgaggca 3600 taggaggtaa
aagatgacac caaaaagcta acctaaatca acttaagagg gatacaggct 3660
catggtttgt tagtggtcag agttatagaa tgtaggcata gtgcagctgt ggtgtttctc
3720 tctgtttctt catcattccc aggcacacta aatttcataa acaactttat
tttcatagag 3780 tgactcttgc ctggggatta agggtactcc ctaggaacct
aagggaatga gtaaaatgac 3840 ccccacagga agacagggct actctccctg
gtgtgagaat gacttcatca atttgtaacc 3900 caggccgact aaatatatca
atcatgtgtt gtttttataa cagctagaaa ctgggaagaa 3960 tagagttcat
aaggttttca gtattgttac ctcacaaaat aatccttcag aaattccaat 4020
tcaaaacata gaaggagtct attgtttgtt tcatattgct tattttgtta cattactctg
4080 ttcttcctta gctagcaact ttgagccccc ataggctatg ctaattaggc
tctttattta 4140 tttttttgag acacagtctc ttccagcctg gagtgcagtg
gcgtgatcat ggctcactgc 4200 agccttgacc tcctgggctc aagggatcct
cctgcctcaa cctcctgagt agctgggatt 4260 gcaggtgtga gctactgtat
ccagccctag gctcactata ttacagagcc cagaaacact 4320 cagtgtaatt
ccaaggctct ttaagactgt cactggctca aggatccaaa tccaggcctc 4380
tgactaaaac atacttagaa tggctcattc tctcactccc ttcaagtgtt gctcaaatgt
4440 tacctattca gtgctatctt actccctatt taaaactgca cccatgatct
cacctctgac 4500 actccacatc cctctttctg gttttatttt tctcaatagc
gcttatatca tctaatgtac 4560 tatatatttt acttcattct cttgtttatt
atctgtatcc cttccatgat gacagggatt 4620 tttgtctgtc ttgttctttg
ttgtagctgt aacacctaga tcagcacctg gcacatagtg 4680 cctgcatata
agacatgctt aatatatatt ttgatggacg atatttcgaa tgaaaattta 4740
cctacataac cacatcaagt aaaattgaat ttggactcta ggacaactgc ttgggtccaa
4800 atcctggttc agcgacttat ggctgttgac cttgggcaag ttatataacc
tctctctgcc 4860 tcagttttct catgtgtaaa atgagactaa taataattgt
tcttattata tagactttgt 4920 tgtgatgatg aacatgaatt ggtattataa
agcactttga aaaatacctg gcacattgga 4980 aacaccatgt attaataaat
gttaacttgt agtggtagaa ggagttgcat ttaactgtat 5040 ctgagttagt
gctatgtttt ataaataaag catttattta gattttatga tatccccctt 5100
gtgtgtagga gccaatttgt atgaaaataa ggatgacagg agtctgatgt gtgagttagg
5160 ccctttaata attcaagaaa gtctgtacaa ttaagcatct tatgaaaaat
agttcaatgg 5220 aaagtataat tcaagatcgc atatatttct atgaaagcct
accgttactc ctttataata 5280 cttatcacaa taatttttac ttattcaata
tgtcttctat gtctagattc taagctaagc 5340 agattcaaag ttccatggga
acaaaataga tttgtcttgt tcaccactaa aactcatgtc 5400 atatttagca
catggtaggt gcttggttca tatttggtta aaggaatatg tattcttggc 5460
tactcagaat acgaggagcc tatactgaac aagttatctt tttatccacc agaagcaaga
5520 attcattact tccacatcta aaaatgttat ctcacccctc ttttcttcta
tccggttttt 5580 gtttactaga actcatcatg actgctttta cctcatggca
tctgttgttt gcttttccat 5640 ctgtatctta acttttgccc atcattacta
gctaatgact ttctctgtgt attgaagtca 5700 aattctcaac agagacccta
cctggtttta attaatgaga attattggtc ttggacagag 5760 ttctcctatc
attggctgct ctcaagtata gaaattgact gcccttgggt caggtggcca 5820
tttttgacct aatcagatat ggctagggtg atctggttca gaataagttt atctgtttgt
5880 aaagaacaac aaggacttat tccctcagta gatcttctgg gtttaagaaa
gaatggggaa 5940 agctggcttt ctgaggctgg ggctgcccag catgccccaa
attacaatca ttaatatttg 6000 ctatgtgtta gagggataca gtgtggggta
tagatataag accctatcct tgccctcata 6060 tagctcacaa actagaggag
gcaggcaagc agattgaata tagagtaatg gtattgctgg 6120 agttgaacac
tttgtgccat gggaacatgc aggacaaaca ctcagtctgg actgggattg 6180
tgaaataacc tccatcttta ctgaggtttc tctgttccaa gtatatgtta aagagaggaa
6240 gttctgtgtc acaatgatgc ccttcagtat ccttttttta aaaaaaatct
tgtttaaaag 6300 tttctaaatt aaaacccagt gaaggcattg agcatatact
cctttaccta aaagacattt 6360 tgttttcaag tttaggaaga gtactgttct
ttgactttca ggtgttttct ccagtgaata 6420 tagtgagtgt gccagaatca
tttgataaga acagttacac tacctgtaat ctgagagatg 6480 tggctttgaa
actgggctga catttaggga gcacttcctt gcctgtcctc cttcagagtt 6540
ctactactcc ttcatttcac agcagtccta actactaatt gtttaagctc cctctttcat
6600 tcctggaaat cttgattaca tggcagggag gacttctcgc aatccatcat
tctaatggaa 6660 tcttcaatga ggatatatat ggatgagaga agccctctgg
aacactatgc tttatccatg 6720 actattgatt tccaccattt gcttcaaact
ttcatcttct gccactgtct tgcatactct 6780 tcttttccag tcaattctcc
agtaaaatct gttccagaca atagcttgaa ggttggtcat 6840 cgggcctgaa
ggcaatatac accaccctgc atattagcac ccactgagtg agagtataac 6900
cctactctag atgaccatcc aatgctgaag atgatgatgc tattcttggt atttcgattc
6960 agacccaaga tgagcctgag agtttgtatc catttgagga aattcttaat
cacctaaata 7020 tctctaccaa tagtgttcat cattaaatac ttgctgtctg
gcacatagtg cttttttaaa 7080 attatttttt gttcaataaa tgaaagcagt
tttagtggat cggttcaaga ggagtctttg 7140 tattagcttt gtttagtgtg
gagtttggaa ctgaagaagc atttaataat taacatttta 7200 aagtgtagac
tacggtatat cagacctttc tggaacactc cctagacact gaaatgcggc 7260
aaagacactg cattggaaac ctccgaaaga gtctgtatcc ttggccagca gatggcattc
7320 ttatactata gtttagagaa agcaagtagc aaaaggtaaa gaagcacttc
ttaatgatga 7380 agtgaccatc tacctccttt ttcccttccc catcaccaca
taaccttatc tcacttccat 7440 tttatcccag gcaactatct tttccatcca
tccattcacc cttctttagg tgcctagaga 7500 tgtcttgcta ccattctctc
tagttgctca taggaaaata tcttttgtac ttattaatag 7560 tcaatccata
gaaataggac aagccacata attttcaggg ccctgtgtaa aatgaaaatg 7620
cagaagttgt tcaaaaagta ttgagaattt taggacagta agaaaagagc attaaaccaa
7680 gcacagggcc ctctgagcct gcacaagcca tacacccatg aaaccagcca
tactttataa 7740 tgtaaatatt tccctcttcc catcctacct tccattgttc
taatgtcctg tcccctgtga 7800 cccacaccaa aaaggctgct taagaaagga
aggaatcact aaatgtcatt cagacatcat 7860 agaatcttag cactgaaaga
aaatttggag atctagttca atcctctcat tttaaagatg 7920 attaaactac
attccaaaga gatgaagaga ctctcccaaa gtccccagct agttatgtca 7980
cagactggac tggaactaag tattctcatc taccaggcca aggctcactc tctcacgcag
8040 actccatgag aggtatttta agaaaatagt tttcccatct actcaggata
tactttgccc 8100 cattttctct agaggagaag ggtacatgtt tatgttacat
ttgagtacca ggtatatgcc 8160 aggcattaca ctcgatgttt tatacataat
acctcaattg caacaagctc agaaaatcat 8220 tacattttcc ccatttttca
aatgagaaaa atagagaatc agaacttgaa ttttagtcag 8280 taaatgggca
gacaagaatt tgggcaagtc tgtgattcta aaacccattc tttttcacca 8340
catcattcct aaattttagg aaagctgagt cagcctgtgt ttcctgaaca ttggtcttgt
8400 ttttggtcag ctctgggtga tttggggact ctttccaggc tatgttcttc
aaatttacca 8460 ggtttgctac tgccttggtg tctttgctcc agggtgtttc
ctctgcttga aacactcttc 8520 ccccagatat ttcatgaaaa acaggaccac
gtgttggaga ggtaacaggg catctgatca 8580 tatagggtct tacaggtcat
tgtaagaaca ttggcttttt cctctgagtg aaaatggaag 8640 ccattgcagc
agtttgagca gataagcgat gtgatcagat ttagattttt aaaggatcct 8700
tctggctgct gtgtagaaaa agagactaaa ggggtggagt agggaaggta gaagttcttg
8760 ggctttttag agagttgtaa cagtaatcca ggaaaaagat aatggtggct
catactaagg 8820 tgatagcagt ggaagtgatg ggaagtggtt ggattctgga
gttatatgtg tatgtaagta 8880 ctatattttt aacttcttgt tatggaaaat
ctcaaataca tacagaagta cagagatgaa 8940 tataatgaat tcctgtgttc
ctattcagtg aacctatcat atgttattat gtgtcaggca 9000 ctttgcaaca
gctgcaagaa ttgccaactt gtatcaatct tcctccccaa ccctagattg 9060
gggagaaagc aatttgaagc aaatcccaga taccatattg catgtgtaaa tatttcagct
9120 tctactactc caactgcttc tgactgcccc gacattctgc tatctaccct
atggttactg 9180 cttgttcata gtttcaggta ctcatgactt ctacttattg
tctgctctat ttgtcctctt 9240 taagcctttt tccttgctat caactactaa
ctgtgtactt ctcaaattca aactccctag 9300 aaagaggttc ttgcaggttc
agttagttgt catttttcct tctacacaga gctcatgatc 9360 tggccaaatt
gtggtagatt ggctgctctt gggtcaggga ccctcttttg gccaagcagg 9420
gtggtgtaat gacaaggttg gttttcccga aaaacggttg ttgaaatgga agtcacttaa
9480 caaataatag gctgttctcc aatactaata aatagtggaa atttaattct
agaaaggata 9540 taaatggtga aaaaagtcag cctgcaatga gactagaaat
agataagtgt ctaatcactc 9600 aacaccatta ttttacagtc tgacctgact
catttagagt gctattcttg ggtgccactg 9660 aggaattaag gacatttttt
ttcttaatat atttttgttt ataacaggct gagtaatagc 9720 atagatcata
atgatctagg ctgaaagtca agacctacat ttaattcttc tgttcatcat 9780
ggactttgta caatattaga caagctaata aacttctttg tgcctttgtc tctctcagtt
9840 tttgtattga gattaatacc tgccttttcc tgattcatat atggtagaca
cataagatta 9900 gagggataaa aatacattga gcttttatga agagaagcat
gtgaaaatct aaataattac 9960 tatttcctat gtgaagattt ttatgaaatt
ataaaatgaa catagattta aggtattttt 10020 atataaattt aaggagtaca
agtatagttt tgttacattg atatattgca tagtggtaaa 10080 gtctgggctg
taaccatcac cccctgaata atgtacattg tacccattaa gtaaattatc 10140
atcgccccct gccaaccctc caactcttcc aagtctccag tgtctattat tctacactct
10200 ggtgtgtcca tgtgtataca ttatttagct cccacttata agtgagaaca
cgcagtattc 10260 agctttcaaa gattacatac tctctattca ttttcatcag
gtactcaaac atttttcttc 10320 ctacatttta atatatatgt catatgtata
taccacttag aacagtgtgt ggcaagtagt 10380 aagaactatg taaatgttaa
gctgttatct acatcagtgt gaaaaagaat ttgtcaaaag 10440 tctatagagt
tgcagctgca aatgcttaaa aattgtaccc cctctccatc ccctaatatg 10500
taccctgcta aaatacaatt atgtgaagca cataaaattt tacgtcatgt gaaaatatga
10560 ataaaaggaa actttttgag ccctggaaaa ttggcaatgt atgtgtttat
gtgaaaaaac 10620 ccacaacaac aaaaaagaag ggggtggtta aaaataacat
tgataactga tatctccatt 10680 gttctcaaag acaagttagt ggacaggctg
catcagaatc accaggaaat tttctagaat 10740 tcttggttct attagcgatt
ctgatataat agaataacag tggagattaa gaatgcagaa 10800 attgtttaag
tttctcagat gatcctgatt ttcagccaag cgtgggaatg actgatgtgt 10860
acaaaaatgt aatacaataa tttaatttaa tgtagtacat catttatttc ctatcatcct
10920 attcaaagtg gtatgtattg ggaaatatct gcccccaggg aagagagctg
agaggatatc 10980 tatcaaaatc agaatggcaa ttatctctgg attatgagtt
atggaaatac ttattttaca 11040 tttgtgtttc ttatttctaa gttttctggt
tgagcatata ttatttttat aataatagat 11100 attacttaca accagtgcca
gaaaattatt cacaaccagt tcaagatgtt aaagactcat 11160 atgcagccta
tgcttttagt ttatcaaatg taaagaaaaa aaacaacttg agaggctgag 11220
gcaggaggat tgcttgaggc caggggttca agaccagcct gggcaatata gcaagacccc
11280 atctctaaaa aaaataaaaa gaaaacaaaa caaaaattag ctgggcactg
tggaatgtac 11340 ccatagtcct agctactcag aagcctgagg caggaagatt
gcatgagtcc aggagtttga 11400 ggttacagtg agctatgatc tctccactac
attccagcct gagtgaaaga gtaagaccct 11460 ctctctacga aaacagtctg
tcaattcata ttcacatact ttttctttta aaaggcaagg 11520 ctacatataa
agcaacatgg acgcaatatt tatttttgtt ttttaaaatg tcacaaattc 11580
ttttagtaaa tatatatctt agagaatata aaacaaatta aatcctaatt agcttttact
11640 tcttttgctt tggcatgaat ggaaatatac caatgtttat aaaaggacca
atcaatcagt 11700 aaccttatac ttgatggggt gaaataaata gttctggtac
aatatagatc ttctgtccaa 11760 tttgtatttt ggactgtgcg ccaagaataa
tgctcaatcc tttttaaaga tccttttggc 11820 ttctactcct tcatacacat
aagtctgaaa ggtgttaaca aacagaagaa tttatcttta 11880 aggtttacat
ctaagttaag aaaaagtagt attatagttt tatactatat gagtttatac 11940
tatttttata ttatatattc atatatatta tatgagtatg agtttataca attttttctt
12000 gtacttgata gcaagattta tacagatatg ggatacagcc attttaaaac
acatgttggt 12060 ttgtagtcat ttatattttc agttaaaata ggcattgatt
atttgacata tataagttgt 12120 ctttagtaaa taggcatcag aaaatagcta
tacaatattt ttaaaatcag taagatttat 12180 gagaaaagaa gtagtcagtt
tactgaagat ctggctcagc agtgacaaaa attattgttt 12240 tgtagtaatt
tttgcctttt gaaaatagct ataaatttga caactcacct ggactagttc 12300
atcacctata atttctcgga cagtattcag ttcgtttcca ttgtctgtcc gtttgaattt
12360 tccaataagt ttatttccct caaggctcca ggtcccctac ataataataa
taataataat 12420 aataataata ataatggcat ttattagggt cttacacatg
tcaggcactg ttctgagttc 12480 tgtagacaca ctttttcatt taatcttcat
ggtgacccta tgaaactgat actattatct 12540 ctactttgca gaagaggaga
ctgtaaaatg gcacagtgat tcacctaaag tcacagagtg 12600 aggattgaac
ctaggccatt gggctacaga gtatgtgttg ataaccatta tgcttgttct 12660
cttcaataat ttccatacgc tgaattttaa ggcagatgag cagcacttct ttctgaatct
12720 ttcatgttac gtgtccagaa atatagtcat ggttaaaatt agtttcatgg
agatgttaac 12780 aactctgtgc ttgttttaat ttgtgtgatt gtggtactaa
ttaaagaaag acataactta 12840 tgataaaaca ttcagtagtt aaattgtcat
tacaaacatg taatttaaat agaaaaatgt 12900 tgaggctaaa aaaagttttt
aaaaaatttg ttgaaaatga tgacttccta aaatgtgtta 12960 tcttaaacat
gaaaaggtac actcccaatg acctacaata catcaaatct ttccatttgg 13020
taactcactc atttgttcat ctattcattt cttttattat ttcaacatgt taaacattac
13080 agttaagtta catattaact ctaaaggctt cttttgctgt actgtttaaa
aacacagttt 13140 taaatttctt tatgaatagg tctatattca gtcattcaaa
agatagtttt ctaattctta 13200 ctgttcaatt cagattcttc acagatgcgt
ataaaaataa aaacatattt aaatatcctg 13260 ccaatttgtg caatatataa
tgtagacaaa cttcattggc ttcttcagtt agtgaaggag 13320 acctgcatta
attaaaccat ccaatgaaat agagcagaaa tcattttaat attgggtaga 13380
aaaatcaaga atgcattgct cataaaaaaa aaaattctta ccctgagttc agttccgtct
13440 gctagattgt aattaaaggt gacaccaagt tcaaaaacaa cttcaatgtt
tcgaaaagcg 13500 cttgattctt tgactgtgaa tttatttcct tcttgtgtaa
ttgtcagctt caaattgtca 13560 tgagctgcaa gcttcctttt cactatatta
acacctgtaa aaggtaagac aatggagaaa 13620 ataaagtcaa atcccatagg
aagtgtttat ttttccaagt tatgttttgt ttgtttattt 13680 tgagggtggg
aagaaaactc ggtagcattg cctttgcaca agaacattca gattgcttct 13740
acagaagttc aggttcaatc cgatcaagag aactgattaa agttgttttt ccataacttg
13800 agaaaaatta agtagagtac agaattatag aatcttagag ctgagcaacc
ttaataagat 13860 taacaaactg ttcatttcag tgttaagtaa atcaaattct
agaaatatca aatgaaggat 13920 taaaatgtat attcatatat ttataaggaa
gcacatcttt aattccacat atacagtatt 13980 cttttttctt aaatgcccct
ttgtttttct tagtggcttt aacattttga actttatagt 14040 tctatttgct
tcttgtggca tttctttctg acacaaaaat gagaaaatta gcaaatatta 14100
agccaaaatc actataagaa tctacaactt ttataaattt aaacttagag ttcctaaact
14160 gtaagctata attgatgcag atttttcttt taggggaatg acacaaattt
gtttgcctat 14220 cttgtatagt atatgggctg ccagagtatt ggtatcaaat
tataaatcag ttaaattaaa 14280 cgccctggag aaaactacac agcttctcaa
gattattcca aaactttttt aagttagaag 14340 aaatcagaat taaatctaac
ttattcagcg ctataaattt taagagaacc ttttccatta 14400 ctttcttata
tggtctttgt ttttccatct cctagaattt aacattttca gatctggaag 14460
ttatcattac acgctgccat tttcttactg taaacaacaa aaaaagtcat ttttcagtta
14520 aagagtgaag gaaacatttc tggatctttt tggtgtgttt cagagaacag
cactcaaaga 14580 atcttttctt ccattagtga gatagtggat tcttcctaat
gtaagaaggg agaatatggt 14640 ttccctcctc tgaacatctt ctggatcctc
tctaccacat caggagctat ctgtaagcta 14700 tctgtaatct cctagagatt
taggagttag aaagaaaaat gtttgtaaga aagcaaagaa 14760 tgagccacaa
agaaataaag tctttaccca ttttttccat gaacttgtca tagttttcac 14820
tccggtctac cttccaagtg ctgtcaaacg ccatgatttc agttgagtca gcctctaggc
14880 agctagagat tcaggtctgt ccttgggcga gaatttatta tatttcttat
cttgaaccaa 14940 cttcatactg tgatgtggaa gcttaaagtt caggaaatca
cctaactact ctatgtcaag 15000 caattaatta attcttattc ttattaattc
ttaattctcg ttctgtacat tcctgagatc 15060 ttctgaaatt caactgaatt
aaaacatgag aagcatacct attctgtctt aaattacagt 15120 ttgtgggcat
acttatttca agaattagaa tgcatgccgt ctgaagattg tttttctaag 15180
ttcaaagtgc agactatgtt ggaggtaata taatttatag catatatcaa ataacatctg
15240 ggaaatgaga ccatgacctt tccctttttc acaacagcaa ttatcttgta
aagtaagact 15300 ttatgatgaa atacagttga aaagttaaag ttaataagat
tcgttttcta atatgtcttt 15360 ttgcattgtg tttagcattg ccaggagttt
ggataaatat cctggcactt tgtagggtgc 15420 atgacacaca ataagtgctc
attgattttt gtcttgatcc tgaatggtga taggccactg 15480 agcaaaaatt
caagaggtac aaatgaggct ggatggtgta aaaacttgca taaaagaaag 15540
aatgagcttt aaacatctgc catgtgcaga gagaacaaaa tccagtttta gctgtgccat
15600 taattaaaac cactccttca ttttactctc aatgcctctc atccccaaac
ccagtagagt 15660 cagaagccac agctcaatta tgtatgtggg gtcagggtat
agagggtagc tagaaaggta 15720 caaagaaaag gtcaccatat ctctcctttc
cctacttcag gctgccagcc aggaacaggc 15780 caagcttggg gaggaaactt
ggtgtgaagt aaagagtttt aattactagt cttaagatgt 15840 ttattactaa
aatgagtcta ttattgtgac tgaatgtgac cagataacat cttattaact 15900
aagggtcagg aaagaagctc tagggcttgc ccaagatatt acttagggtc aagggaagat
15960 aaatctaaca gatttgaaca gaactttggg gaacagaatc ccagttttta
aattttgata 16020 ttattttata ttttgatttt taaaaatctt ttaaacatgt
tagatattgt ttgatttttt 16080 atttcgagtt gcccactttt tattttatta
agtaagcact ttaaaaagca aactattttc 16140 tactagctct attatttcat
taaaaaagaa aaagaggccg ggcgcggtgg ctcgcgcctg 16200 taatcccagc
actttgggag gccgaggcgg gcggatcacg aggtcaggag atcgagacca 16260
tcctggctaa cacggtgaaa ccccgtctct actaaaaatg caaaaaaaat tagccgggcg
16320 tggtggtggg cgcctgtagt cccagctgct cgggaggctg aggcaggaga
atggcgtgaa 16380 cccgggaggc ggagcttgca gtgagccgag atcgtgccac
tgcactccag cctgggcgac 16440 agagcgagac tccgtctcaa aaaacaaaaa
aacaaaaaaa caaaaaaaca aaaaacaaaa 16500 aaaaacaaaa aagaaaaaga
gttgcctggt gcagtggttc ttgcctgtaa tcccagctat 16560 tagagaggct
gaggcaggag aatcacttca ggccaggagt tcgaaaccag cagtttgagg 16620
ccagcctaag caacataggg agagcctggc tgtaacaatt ataataaaat aaattagcca
16680 ggtgtaatgg cacatgcctg tagtcccagc tacttgggag gctaaggcgg
gaagatctct 16740 tgatcccagg agttggagat tgcaatgagt tgtgatcatg
ccagtacact ccagcctggg 16800 caacagagag atcccatctc taaaaaataa
ataatttttt aaatgtcaaa aaaggaaaga 16860 aaacacatat acacactcat
gagttgggga caaaggtcat aatgggaagt cctttacatt 16920 cataatttag
ctttactttc aaaattaggt acattgtgat tttttaatct tttttcaaag 16980
aattaaaact gtatcacttg ggagctatca acctaataca ctttctttct ttttttatta
17040 cactttaagt tttagggcac atgtgcacaa cgtgcaggtt tgttacatat
gtacacatgt 17100 gccatgttgg tgtgctgcac ccattaactc gtcatttaac
attaggtata cctcctaatg 17160 ctatccctac cccctccccc caccccataa
caggccccgg tgtgtgatgt tccccttcct 17220 gtgtctaagt gttctcattg
ttcaattccc acctataagt gagaacatgc ggtctttggt 17280 tttttgtcct
tgcgatagtt tgctgagaat gatggcttcc agcttcatcc atgtccctac 17340
aaaggacatg aactcatcat tttttatggc tgcatagtat tccatggtgt atatgtgcca
17400 cattttctta atccagtcta tcattgttgg acatttgggt tggttccaag
tctttgctat 17460 tgtgaatagt gcccctataa acatatgtgt gcatgtgtct
ttatagcagc atgttttata 17520 atcctttggt tatataccca gtaatgggat
ggctgggtca aatggtattt ctagttctag 17580 atccctgagg aatcgccaca
ctgacttcca caagggttga actagtttac agtcccctca 17640 acagtgtaaa
agtgttccta tttctccaca tcctctccag cacctgttgt ttcctgactt 17700
tttaatgatt gccattctaa ctggtgtgag atggtatctc attgtggttt tgatttgcat
17760 ttctctgatg gccagtgatg atgagcattt tttcatgtgt cttttggctg
cataaatgtc 17820 ttcttttgag aagtgtctgt tcatatcctt cgcccacttg
ttgatgtggt tgtttatttt 17880 tttcctgtaa atttgtttga gttcattgta
gattctggat attagccctt tgtcagatga 17940 gtagattgca aaaattttcg
cccattctgt aggttgcctg ttcaccctga tggtagtttc 18000 ttttgctgtg
cagaagctct ttagtttaat tagatctcat ttgtcaattt tggcttttgt 18060
tgccattgct tttggtgttt ttagtcatga agtacttgcc catgcctatg tcctgaatgg
18120 tattgcctag gttttcttct agggtttttg tggttttagg tctaacattt
aagtctttaa 18180 tctatcttga attaattttt gtataaggtg taaggaaggg
atccagtttc agctttctac 18240 atatggctag ccagttttcc cagcaccatt
tattaaacag ggaatccttt ctccatttct 18300 tgtttttgtc aggtttgtca
aagatcagat cgttgtagat aagcagcatt atttctgagg 18360 gctctgttct
gttccattgg tctatatctc tattttggta ccagtactat gctgttttgg 18420
ttactgtagc cttgtagtat agtttgaagt caggtagcgt gatgcctcca gctttgttct
18480 tttggcttag gattgacttg gcaatgtggg cttttttggt tccatatgaa
ctttcaagta 18540 gttttttcca attctgtgaa gaaagtcatt ggtagcttga
tggggatggc attgaatcta 18600 taaattactt tgggaggatg gccattttca
cgatattgat tcttcctacc catgagcatg 18660 gaatgttctt ccatttgttt
gtatcctctt ttatttcatt gagcagtggt ttgtagttct 18720 ccttgaagac
gtccttcata tcccatgtaa gttggatttc tgggtatttt attctctttg 18780
aagcaattgt gaatgggact tcactcatga tttggctctc tgtttgtctg ttattggtgt
18840 ataagaatgt ttgtgatttt tgcacattga ttttgtatcc tgagactttg
ctgaagttgc 18900 ctatcagctt aaggagattt tgggctgaga caatggggtt
ttctagatat acaatcatgt 18960 catctgcaaa cagggacaat ttggcttcct
cttttcctaa ttgaatgtcc tttatttcct 19020 tctcctgcct gattgccctg
gccagaactt ccaacactat gttgaatagg agtggtgaga 19080 gagggcatcc
ctgtcttgtg ccagttttca aagggaatgc ttccagtttt tgcccattca 19140
gtatgatatt ggctgtgggt ttatcataga tagctcttat tattttgaga tacgtcccat
19200 caatacctaa tatattgaga gtttttagca tgaaggttgt tgaattttgt
caaaggcctt 19260 ttctgcatct attgagataa tcatgtggtt tttgttgttg
gttctgttta tatgctggat 19320 tacatttatt gatttgtgta tgttgaacca
gccttgcatc ccagggatga agcccacttg 19380 atcatggtgg ataaactttt
tgatgtgctg ctggagttgg tttgccagta ttttattgag 19440 gatttttgca
tcgatgttca tcagggatat tggtctaaaa tctctttttt tgttgtgtct 19500
ctgacaggct ttggtatcag gatgatgctg gcctcatgaa atgagttagg gaggattccc
19560 tctttttcta ttgtttggaa tagtttcaga aggaatggta ccagctcctc
cttgtacctc 19620 tggtagaatt tggctgtgaa tccatctggt cctggacttt
ttttggttgg taagctatta 19680 attattgcct caatttagga gcctgttact
ggtctattca gagattcaac ttcttcctgg 19740 tttagtcttg ggaggatgca
tgtgccgagg aatttatcca tttcttctag attttctagt 19800 ttatttgcgt
agaggtgttt atagtattct ctgatggtag tttgtatttc tgtgggattg 19860
gtggtggtat cccctttatc attttttatt gcatccattt gattcttctc tcttttcttc
19920 tttattagtc ttgctagtgg tccatcaatt ttgttgatct tttcaaaaaa
ccagctccag 19980 aattcattga ttttttgaag ggttttttat gtctctatta
cactttcaac ttggagggaa 20040 gtagaaaact ttgtttaaag ctgaggactc
aacagtctct caggtagttg actggctgtg 20100 gtgatttgtg aactcagaag
cctatggata atgaatccaa tctttatttc taggtcagaa 20160 aactacatgt
atctggtcac tgaaataaac gtatggtaga gtgaaaagaa catgtgtttt 20220
agaaacaaga cccattgact tgggtttcag tgctgactaa acatacatta ctctgcagaa
20280 tcttcgtcac attacttact caatctctct gagcctcagt tttctcatca
ataaaatgaa 20340 gacaataata atacctgata tgtatatttt atgaacaaat
tacataaagc acccacctga 20400 aacaacttat agataacagg tcctcaacaa
acctttgttt ctctcctaat tctctgagaa 20460 aggaaatctg ggagcaataa
caatgtttta gaagcatcct aggtctcaaa ccagtgatat 20520 tttgttaaga
aaacccatgt cattctggtg tttatgagaa agtcacctaa aagttactta 20580
ggtattttat gatttgcact agtgatcaac ttgggactgg ctatgccttg gatttgcctg
20640 ttaaggatag tattgcactg tatcaccctc tgggaataca tgtttccact
atgtgatctc 20700 atagcaattg aaattaaagt gtctgattca agaaggaagg
ttggcaggaa gaaactaaag 20760 tggcttctta ccattttcac catgacctaa
cccgtgtctc atggccattt taagcttcgc 20820 agaaggatcc aaataggatc
aatataaagc aaatagccct tgggctcaca gaaggctggt 20880 aacaaaataa
gcatgttaaa ctttcagtct taaatttatc agtggcagtg acctaattta 20940
accttaggta ggacatcctg ttatctctgt taagtagcat cattagtttt tttttttatt
21000 aatgctcaga ttacattttt aacattacac ttttccttct ccagctaccc
aattctgtag 21060 cccctcggtt tttatcactt atctgtagtc catctagaaa
ataatatacg gaacttaggg 21120 accaatttct ctacagacct tttctactga
tgatatgatt gagccaacca gctaagggca 21180 gacccagccc tttcttttcc
attttgataa tattaaaaga cttgagtatt gcagtttggg 21240 ggtcgtgagt
ggactaagta tatgtgtgca tgtggatgca gatgctatgg aggtagagct 21300
atagctgtac cctctctctc tgggctctac agtcaacatc acccacggag tggtctggaa
21360 gaagtggaaa tcagcagaaa ctcctgtgga tccctattcc attcctccaa
gggaaagtac 21420 aaaccacaat ccttagcatg gtttacaaag ccactactta
aatctccagt tgatttttcc 21480 attccctact gtgtactgca agatctagat
ctattaaact tcttgcagtc cccaatcccc 21540 tgaatgtcca tgcctctgtt
gcttcaggtc tcagcttgga tgctgattgc tcaaagattc 21600 ttgcctcgaa
ctatcaagag tgagtcaggc aacccttctt ctcccaggtg taatgacctt 21660
ctaaaggtct cagagtagct cataaaagac aagaacagct gattcatgat aaccaaagga
21720 atattgttat catgaagccc agcgacatca ttagcaaggg caataggatg
ttaaaatctc 21780 atctcaaagg aataatgttt tagagatcac aaaaatcctt
ttaccccatt gtataataga 21840 agggagtgtt tttgtagagt attattcttt
atatatcctt attgtgaaat agcattttaa 21900 ggcatattct aatcatctca
gaatttatgt atcctatttt ttgatcttaa gttgtttagc 21960 tttttatatg
tcaaaaaata gaggtctcta tcagaaatat atgagagaag gatttgaggg 22020
acagatagaa ggagtaacag gaaagtctta aggttactgt caaatcccta gcagtgaatg
22080 gggaatgaga gagaattctg aagtttgccg acaaagctta tttcattctg
ttttaatgat 22140 tagcctctaa attttagaaa tattatgacc tatttaatta
ggttttagat tttgtatctg 22200 ggatgactgg ttggttttct ttcaaagaga
ttataaatgt acagcagtgt gcaggagaat 22260 gtctaccata gagtggtgag
tgtggcttgc taattctcat taataacaaa acagcctaag 22320 aaaacagcca
ttttagaaga ttaatgtgtc cattgaatga tcttgcatga agaatttatg 22380
actccttctt agtctgggtt tatcccagaa atgatgagcc actttcaagc aatgtagata
22440 agaggttgtt catctcctat gcccctcata ccacaggtag aaaaagatgg
atttgcatta 22500 tcacagctcc ctactgccac aaccaaacca ttatgtttct
gcctttttca aaacaaaata 22560 ttgttttctg agaccactcc catttggctg
tggtccagcc tacttcatca ttgttatcta 22620 ccatccactc cctctagcat
tatcatctac catctatcac tgaaaatctt ggtacttggc 22680 tcaccatcat
ctgtgtcctg ccatcatcct gggtgacctc aaagtccatg tggataagcc 22740
acttgtcagt ctggcttcac agctccttaa ccccctcata tctaatgacc tttacttctt
22800 ttatgcttta tctacttaca tccagggctg tgtcctgtga tctgtcatca
ttaaagctgc 22860 tccttctgtt gtatttaaca tgccactctg accacaactt
tccatgctct cagctccttt 22920 attttattct gcacattagt tattggctgg
gagagaaatt ttcttagttg ttgcctctat 22980 ttcctctcaa gttaaagccc
cctcctgttt gtttaaattc tttattcaat tgatatcctg 23040 tttccaattt
taaaactctc tcactttttc ctgcactgac ctaggtgcta ttaggcatgc 23100
atggtttcta gtcttagagt aattatcagc actccctgga atgacttctg tgtatttcaa
23160 tcagcttcct ccacatttgc gacaatggct gtgcctttag tcaccctcct
gtagcctttt 23220 acctttgccc tcagcagaca acctgcacat tacttcacag
aggaaagaaa ccatcaaaca 23280 attccctcaa tttatgcctt ctcaattaca
aagttaccta ctattacttt ctttactcct 23340 attaattgtt acactcacgt
atgaggttgt ttttgcattg ctataaagaa atacctgaga 23400 ctgggtaatt
tataaagaaa agaggtttaa tttgctcatg attatgcagg ctatacagga 23460
agcatggtgg catgtgcttc tggggaggcc tcaggaagct tccgatcatg gtggaaggca
23520 aagggggagc tggcacatca catggcaaaa gcaagcgaga gggggaaggt
gccacatact 23580 tttaaatgac cagatctcgt gaaaactcac cattgcgagt
acagtaccaa cggaatggta 23640 aaccattcat gagaaatctg cccccttgag
ccaatcacct cccaccaggc ccaacctcca 23700 acattgggga ttacatttca
acatgagatt tgggtgggga caacatcaaa ccatgtcaac 23760 ccacttactc
tttcaatgtg tattttaatt cctcctatgg tctaggcact gtgccacatt 23820
ataaacagaa aaatagacac agtttcagtc aagggaaagc aaggcttgaa attgcaatag
23880 tgtgtcacat gtaaagtttt ttttttttct ccctgtttct tcctatcacc
ctctctttct 23940 tttttattta tttgcttatt tggatgaggt gggatgtgaa
gacgatcaag tctctcccat 24000 tataagacat aaagatacct aaattctgtt
gacatttcta gaacctactc aatctttctc 24060 cttctctttg tagccaggct
tcttgaaaaa ataatttgta ctttttgtct tcacatgctt 24120 atccttcatt
aatacctcaa tctactgcaa catggctttc atattcacca gtccactgaa 24180
actactcact aaggtcatag atgaattttt agttgccaaa cccagtgcac atcttggccc
24240 ttattttatg tcattgtagc attgcatacc attgactctt cttcttcttc
ttcttctttt 24300 taatttctcc ttttcttttg ctgacttttt tctttcttga
cattcctctt ctctggatac 24360 ttcttttcag gctcttccat aagcaacttt
tccttaactg acagatacct taaatgttgg 24420 aattgcatct ttaaatcact
ttttattcca tacaatctac ctcgatcatc tttttcattc 24480 tcatgtctta
atgttcaaac catatgtctc taaaatttag agatccagcc cagacatctt 24540
tcctactggg taggaaaagt gctgtaatcc cagcactttg ggaggccgaa gtgggcagat
24600 cacctgaggt tgcaagttca agaccagcct ggccaacatg gcgaaatccc
gtctctacta 24660 caaatacaaa aattagctag gcatggtgtg cctgtaatcc
tagctacttg ggaggctgtg 24720 gcttctcatt cagaagcaga acagaaaatg
atgcataatg tctaaaacta tagatgtgag 24780 atatatatac atatatatta
ggaataacta gttaagtgag aggtaaaggt ttggggtttg 24840 cttaatttta
gaggttatat tcatgttgag tacctgtttc atggcttcat gtaccatcca 24900
tgtaaccagt gattc 24915 41 201 DNA Homo sapien 41 catataggtc
gatggatact cactgtcaca tgtgatccag gtctcctccg aggggctggc 60
cagtctcttc tcccatggag atgatgggca ctgccacagc rtgtcaggtc cttttggaca
120 ctgaacattg tccacccaca tgggaatgga catggccttg tctaaagatg
cagggttgat 180 tttccctttg tctgcacagc c 201 42 201 DNA Homo sapien
42 aaacatcttg gctgcttcaa agtaaggtca acaggtcctc ctctctgctg
agggtcccaa 60 ttcaacatca actgtagcca gttttccatg ggttctacta
ytaaactaga aaacatacaa 120 aatagggtga aaatcaaatc attatgttcc
aatttccctt tatactgtta gaaaggtaat 180 tttgcaggtt gtccattttc t 201 43
201 DNA Homo sapien 43 ggatgtgact gggaagctct ggtttcagtg tcatgtgtct
attctttatt tccaggcaaa 60 ggaaaccaac aataagaaga aagaatttga
ggaaactgcg ragaaagtgc gccgtgccat 120 cgagcagctg gctgccatgg
attgaggcct ctggccggag ctgcctggtc ccagagtggc 180 tgcaccactt
ccagggttta t 201 44 201 DNA Homo sapien 44 ctcataaaaa aaaaaattct
taccctgagt tcagttccgt ctgctagatt gtaattaaag 60 gtgacaccaa
gttcaaaaac aacttcaatg tttcgaaaag ygcttgattc tttgactgtg 120
aatttatttc cttcttgtgt aattgtcagc ttcaaattgt catgagctgc aagcttcctt
180 ttcactatat taacacctgt a 201 45 23 DNA Homo sapien 45 cagttttcca
tgggttctac tac 23 46 23 DNA Homo sapien 46 cagttttcca tgggttctac
tat 23 47 25 DNA Homo sapien 47 ttatgaaatg gtacagacaa gtgat 25 48
16 DNA Homo sapien 48 cacggcgcac tttctt 16 49 16 DNA Homo sapien 49
cacggcgcac tttctc 16 50 26 DNA Homo sapien 50 tgttttttcc tttgtcatct
tatcta 26 51 18 DNA Homo sapien 51 tccaaaagga cctgacat 18 52 18 DNA
Homo sapien 52 tccaaaagga cctgacac 18 53 18 DNA Homo sapien 53
ggctgcagaa tggaattt 18 54 20 DNA Homo sapien 54 cacagtcaaa
gaatcaagcg 20 55 21 DNA Homo sapien 55 tcacagtcaa agaatcaagc a 21
56 24 DNA Homo sapien 56 aaattcttac cctgagttca gttc 24
* * * * *