U.S. patent application number 13/644229 was filed with the patent office on 2013-05-30 for tumor therapy with replication competent sindbis viral vectors.
This patent application is currently assigned to NEW YORK UNIVERSITY. The applicant listed for this patent is New York University. Invention is credited to Daniel Meruelo, Jen-Chieh Tseng.
Application Number | 20130137754 13/644229 |
Document ID | / |
Family ID | 40986218 |
Filed Date | 2013-05-30 |
United States Patent
Application |
20130137754 |
Kind Code |
A1 |
Meruelo; Daniel ; et
al. |
May 30, 2013 |
TUMOR THERAPY WITH REPLICATION COMPETENT SINDBIS VIRAL VECTORS
Abstract
Disclosed herein are methods for treating a mammal harboring a
solid tumor which expresses higher levels of High Affinity Laminin
Receptors (LAMR) than normal cells of the same lineage comprising
systematically administering to a mammal in need of such treatment
a therapeutically effective amount of a Replication Competent (RC)
Sindbis virus vector, wherein said vector encodes a suicide
gene.
Inventors: |
Meruelo; Daniel;
(Scarborough, NY) ; Tseng; Jen-Chieh; (Woodside,
NY) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
New York University; |
New York |
NY |
US |
|
|
Assignee: |
NEW YORK UNIVERSITY
New York
NY
|
Family ID: |
40986218 |
Appl. No.: |
13/644229 |
Filed: |
October 3, 2012 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
12390096 |
Feb 20, 2009 |
8282916 |
|
|
13644229 |
|
|
|
|
61030367 |
Feb 21, 2008 |
|
|
|
Current U.S.
Class: |
514/44R ;
435/320.1 |
Current CPC
Class: |
A61K 38/00 20130101;
C12N 7/00 20130101; A61K 48/0058 20130101; C12Y 207/01021 20130101;
A61K 35/768 20130101; A61K 35/768 20130101; C12N 9/1211 20130101;
C12N 2830/007 20130101; A61K 38/45 20130101; C12N 2770/36132
20130101; G01N 2800/52 20130101; A61P 35/00 20180101; C12N 15/86
20130101; C12N 2770/36145 20130101; C12N 2770/36143 20130101; A61K
51/0491 20130101; C12N 9/78 20130101; G01N 33/574 20130101; C12N
2770/36171 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
514/44.R ;
435/320.1 |
International
Class: |
A61K 48/00 20060101
A61K048/00; C12N 15/86 20060101 C12N015/86; A61K 38/17 20060101
A61K038/17; C12N 15/85 20060101 C12N015/85 |
Goverment Interests
[0002] The United States government has certain rights to this
invention, by virtue of the funding received from grant CA 100687
from the National Institutes of Health.
Claims
1-32. (canceled)
33. A Replication competent mut-4 Sindbis virus vector comprising
SEQ. ID NOS.: 2-8.
34. The RC mut-4 Sindbis virus vector of claim 33 further
comprising a suicide gene.
35. The RC mut-4 Sindbis virus vector of claim 34 wherein said
suicide gene is HSVtk.
36. The RC mut-4 Sindbis virus vector of claim 34 wherein said
suicide gene is cytosine deaminase.
37. The RC mut-4 Sindbis virus vector of claim 34 wherein said
suicide gene is Vericella Zoster virus thymidine kinase.
38. The RC mut-4 Sindbis virus vector of claim 34 wherein the
genome of said vector is in a single component.
39. A pharmaceutical formulation or dosage form for administration
to a mammal suffering from a solid tumor which expresses higher
levels of LAMR than normal cells of the same lineage comprising a
mut-4 RC Sindbis virus vector comprising SEQ. ID NOS.: 2-8 and a
pharmaceutically acceptable carrier or diluent, wherein said vector
further comprises a suicide gene.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application claims the benefit of U.S.
Provisional Patent Application Ser. No. 61/030,367, filed Feb. 21,
2008, the contents of which are hereby incorporated by reference in
their entirety.
FIELD OF THE INVENTION
[0003] The present invention is directed to methods to treat
mammals suffering from tumors and to monitor anticancer therapy
using Sindbis viral vectors and pharmaceutical formulations for use
in the methods. In particular, the vectors are replication
competent Sindbis Viral Vectors and the tumors are solid tumors
expressing increased levels of High Affinity Laminin Receptors
(LAMR) compared to normal cells of the same lineage.
BACKGROUND OF THE INVENTION
[0004] One type of gene therapy of tumors, gene-directed
enzyme-prodrug therapy (GDEPT), holds considerable promise,
although practical considerations limit its clinical applicability.
These include the lack of acceptable noninvasive methods that are
adaptable to humans for selective tumor targeting of the
therapeutic genetic material. Sindbis virus is an oncolytic,
alphavirus that selectively targets tumors through the 67-kDa
laminin receptor (LAMR).
[0005] Gene therapy targets the genome of tumor cells as a basis
for a highly selective and nontoxic anticancer therapy. To enhance
selectively and specificity to the killing of cancer cells, several
enzyme/prodrug systems--such as carboxylesterase/CPT-11 (1),
cytosine deaminase/5-fluoro-cytosine (2), and herpes simplex virus
thymidine kinase type 1 (HSVtk)/ganciclovir (GCV) (3,4)--have been
developed for gene-directed enzyme-prodrug therapy (GDEPT). In this
strategy, tumor cells are transduced with therapeutic genes that
encode enzymes for specific conversion/activation of prodrugs,
which are toxicologically inert at relatively high doses, into
highly toxic metabolites for tumor killing.
[0006] In addition to a proper vector system, cancer GDEPT therapy
would greatly benefit from a means to noninvasively monitor the
GDEPT enzyme activity after vector treatments in vivo. Such
capability could improve the Sindbis-based HSVtk/GCV GDEPT in
clinical settings by providing important information to address 2
critical questions: (i) Do the vectors systemically target tumor
cells and spare normal tissues? (i) Do the tumors have sufficient
expression levels of the enzyme for tumor eradication by subsequent
prodrug activation? In addition, monitoring during the therapy
could facilitate optimizing the dose and dosing schedule of the
prodrug to reduce unwanted side affects.
[0007] U.S. Pat. No. 7,306,712 discloses that vectors based on
Sindbis virus, a blood-borne alphavirus transmitted through
mosquito bites, infect tumor cells specifically and systemically
throughout the body. The tumor specificity of Sindbis vectors may
be mediated by the 67-kDa high-affinity laminin receptor (LAMR),
which is over expressed in several types of human tumors. Another
advantageous property of Sindbis vectors for cancer therapy is
that, without carrying cytotoxic genes, they have been shown to
induce apoptosis in mammalian cells. Furthermore, as Sindbis
vectors are capable of expressing very high levels of their
transduced suicide genes in infected tumor cells, the efficient
production of the enzymes for sufficient prodrug conversion is
ensured.
[0008] U.S. patent application Ser. No. 10/920,030 discloses
methods and compositions for detecting cancer cells and monitoring
cancer therapy using replication defective Sindbis virus
vectors.
[0009] U.S. Pat. No. 7,303,798 discloses novel defective Sindbis
virus vectors and their use in treating tumors in mammals.
SUMMARY OF THE INVENTION
[0010] The present inventors have unexpectedly discovered that
replication competent (RC) Sindbis viral vectors have enhanced
anti-tumor and cancer therapy monitoring activities when used with
tumors which express higher levels of LAMR than normal cells of the
same lineage. The RC Sindbis virus vectors are based on the mut-4
replication defective Sindbis virus vector disclosed in the '798
patent.
[0011] In one aspect, the present invention provides a method for
treating a mammal harboring a solid tumor which expresses higher
levels of High Affinity Laminin Receptors (LAMR) than normal cells
of the same lineage comprising systematically administering to a
mammal in need of such treatment a therapeutically effective amount
of a Replication Competent (RC) Sindbis virus vector, wherein said
vector encodes a suicide gene.
[0012] In another aspect, the present invention provides a method
for monitoring anti-cancer therapy in a mammal harboring a solid
tumor which expresses higher levels of LAMR than normal cells of
the same lineage comprising administering to a mammal in need of
such treatment a diagnostically effective amount of a Replication
Competent (RC) Sindbis virus vector comprising a gene encoding a
detectable label, and determining the amount of cancer cells in the
body of said mammal, wherein the amount of cancer cells is
proportional to the amount of label produced by said cancer cells
and said vector encodes a suicide gene.
[0013] In a further aspect, the present invention provides a method
for identifying cancer cells which expresses higher levels of LAMR
than normal cells of the same lineage in the body of a mammal
comprising administering to a mammal in need of such treatment a
diagnostically effective amount of a mut-4 RC Sindbis virus vector
comprising a gene encoding a detectable label, and assaying for
said label, wherein said cell is a cancer cell if it expresses said
label and said vector encodes a suicide gene.
[0014] In a still further aspect, the present invention provides a
method for determining the amount of cancer cells which expresses
higher levels of LAMR than normal cells of the same lineage in the
body of a mammal comprising the steps of (a) administering to a
mammal in need of such treatment a diagnostically effective amount
of a mut-4 RC Sindbis virus vector comprising a gene encoding a
detectable label, and (b) determining the amount of said detectable
label, wherein the amount of cancer cells in the body of said
mammal is proportional to the amount of said label and said vector
encodes a suicide gene.
[0015] In yet another aspect, the present invention provides a
Replication Competent mut-4 Sindbis virus vector, wherein said
vector encodes a suicide gene.
[0016] In a still further aspect, the present invention provides a
pharmaceutical formulation or dosage form for administration to a
mammal suffering from a solid tumor which expresses higher levels
of LAMR than normal cells of the same lineage comprising a mut-4 RC
Sindbis virus vector and a pharmaceutically acceptable carrier or
diluent, wherein said vector further comprises a suicide gene.
[0017] These and other aspects of the present invention will be
apparent to those of ordinary skill in the art in light of the
present description and drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0018] FIG. 1 (A-D). Various Sindbis vector systems. (A) The
wild-type Sindbis virus RNA genome has two major groups of genes:
non-structural genes on the 5' side and structural genes on the 3'
side. (B) The conventional two-component replication defective (RD)
system contains a replicon RNA for therapeutic gene expression, and
a helper RNA to provide structural genes for vector production. (C)
A replication-capable (RC) vector system with integrated structural
genes. (D) a RC vector system with a suicide gene fused in-frame
with the ns3 gene to achieve "controlled" vector propagation and
replication in tumor cells. The ns3 gene encodes a protein which is
critical for viral replication and survival.
[0019] FIG. 2 (A-E). Replication-capable (RC) Sindbis virus vectors
show superior tumor targeting and killing over conventional
replication-defective (RD) systems. (A and B) Design of
conventional RD-Sindbis/Fluc and a prototype RC-Sindbis/Fluc both
carrying the firefly luciferase gene as reporter. (C)
Bioluminescent imaging of tumor-bearing mice that received two
consecutive daily treatments of RD- or RC-Sindbis/Fluc. The first
dose was on day 0 and the last dose was on day 1. Subcutaneous BIM
tumors were implanted on the right hind limb. (D) Quantitative
representation of tumor signals after two consecutive treatments on
day 2. (E) Tumor volume measured on day 10.
[0020] FIG. 3. (A-D). A suicide gene further enhances the
therapeutic efficacy of Sindbis virus vectors. (A) Design of a
conventional RD vector capable of expressing HSVtk for prodrug
activation. (B) Bioluminescent imaging of ES2/Fluc ovarian cancer
cells that express firefly luciferase for monitoring disease
progression. SCID mice were inoculated with ES2/Fluc cells on day
0. Daily treatments with RD-Sindbis/HSVtk and GCV started on day 3.
The combination of Sindbis virus vectors and the prodrug
ganciclovir (GCV) significantly improved anti-tumor efficacy. (C
and D) Quantitative representations of the imaging data.
[0021] FIG. 4 is a map of the pSP6-R/NS3-HSVtk/Fluc-Mut4 plasmid.
pSP6-R/NS3-HSVtk/Fluc-Mut4 is a Sindbis RC vector construction
based on pSP6-R and Mut-4, which provides the replicase genes and
structural genes respectively.
DETAILED DESCRIPTION OF THE INVENTION
[0022] The instant invention takes advantage of the natural
affinity of an alphavirus, particularly Sindbis virus, for tumor
cells, in particular, for solid tumors that express higher levels
of high affinity laminin receptors (alternatively referred to
herein as LAMR or HALR), as compared to normal cells of the same
lineage. The term "high affinity laminin receptor" or "LAMR" has
its ordinary meaning in the art, i.e., the Mr 67,000 laminin
receptor that can function as the receptor for Sindhis virus entry
into cells (Wang et al., J. Virol. 1992, 66:4992-5001; Strauss et
al., Arch. Virol. Suppl. 1994, 9:473-84).
[0023] Accordingly, the present invention provides a method for
treating a mammal (e.g., human) suffering from a tumor that
expresses greater levels of high affinity laminin receptor (LAMR)
compared to normal cells of the same lineage. The method comprises
administering to a mammal harboring such a tumor an amount of a
vector effective to treat the tumor, wherein the vector has a
preferential affinity for LAMR and the vector genome comprises a
single component.
[0024] While not bound by any particular theory, three sets of
observations may account for the remarkable anti-tumor efficiency
of Sindbis vector-based therapy of the present invention. First,
the LAMR can function as the receptor for Sindbis virus entry into
cells of most species (Wang et al., J. Virol., 1992, 66:4992-5001;
and Strauss et al., Arch. Virol. Suppl., 1994, 9:473-484). Second,
it is widely recognized that expression of the LAMR is markedly
elevated in many types of cancers (Menard et al., Breast Cancer
Res. Treat, 1998, 52:137-145). In fact, a significant correlation
has been established between the increased expression of Mr 67,000
LAMR and cancers of the breast (Menard et al., 1998, supra; Paolo
Viacava et al., J. Pathol., 1997, 182:36-44; Martignone et al., J.
Natl. Cancer Inst., 1993, 85:398-402), thyroid (Basolo et al.,
Clin. Cancer Res., 1996, 2:1777-1780), colon (San Juan et al., J.
Pathol., 1996, 179:376-380), prostate (Menard S et al., Breast
Cancer Res. Treat, 1998, 52:137-145), stomach (de Manzoni et al.,
Jpn J. Clin. Oncol., 1998, 28:534-537), pancreas (Pelosi et al., J.
Pathol., 1997, 183:62-69), ovary (Menard et al., Breast Cancer Res.
Treat, 1998, 52:137-145; and van den Brule et al., Eur J Cancer,
1996, 32A:1598-1602.), melanocytes (Taraboletti et al., J. Natl.
Cancer Inst., 1993, 85:235-240), lung (Menard et al., Breast Cancer
Res. Treat, 1998, 52:137-145), liver (Ozaki et al., Gut, 1998,
43:837-842), endometrium, and uterus (van den Brute et al., Hum
Pathol, 1996, 27:1185-1191). Indeed, data on more than 4000 cases
of different tumors from diverse organs studied by
immunohistochemistry are all concordant with a role for HALR in
invasiveness, metastasis, and tumor growth (Menard et al., Breast
Cancer Res. Treat., 1998, 52:137-145). Sindbis vectors, which are
naturally blood-borne, can easily travel through the circulation
and specifically home to and target growing and metastatic tumors
expressing increased levels of LAMR. Finally, Sindbis virus is well
known to be highly apoptotic for mammalian cells (Levine et al.,
Nature 1993, 739-742; Jan et al. J. Virol., 1999: 10296-10302; Jan
et al. J Virol 2000 6425-6432). Cell death begins within a few
hours of infection and by 48-96 hours virtually all infected cells
are dead (Sawai et al., Mol Genet Metab. 1999, 67:36-42; Griffin et
al., Ann. Rev., 1997, Microbiol. 51:565-592).
[0025] The Sindbis vectors of the present invention, do not infect
normal cells to the same extent in vivo compared to tumor cells.
This allows for a differential effect in vector therapy, e.g.,
infection by Sindbis vectors results in the death of tumor cells
leading to tumor elimination without apparent deleterious effects
to other tissues and organs of the treated subjects. This
phenomenon may be explained by the observation that an increased
number of LAMR in tumors versus normal cells leads to a high number
of exposed or unoccupied receptors on tumor cells (Liotta, L.A.
Cancer Research, 1986, 46:1-7; Aznavoorian et al., 1992, Molecular
Aspects of Tumor Cell Invasion and Metastasis, pp. 1368-1383). For
example, it has been demonstrated that breast carcinoma and colon
carcinoma tissues contain a higher number of exposed (unoccupied)
LAMR compared to benign lesions (Liotta et al., 1985, Exp. Cell
Res., 156:117-26; Barsky et al., Breast Cancer Res. Treat., 1984,
4:181-188; Terranova et al., Proc. Natl. Acad. Sci. USA, 1983, 80:
444-448). These excess unoccupied LAMR receptors on tumor cells,
which are not found in normal cells, may be available for Sindbis
virus binding, infection, and induction of cell death.
[0026] The invention advantageously provides a method for treating
a mammal suffering from a tumor, in which the cells of the tumor
express greater levels of LAMR compared to normal cells of the same
lineage. The different levels of LAMRs result in target-mediated
delivery, i.e., preferential binding of vectors of the invention to
tumor cells. "Greater levels" of expression generally refer herein
to levels that are expressed by tumor cells (as compared to
non-tumor cells) and result in such preferential binding, e.g., at
least a 3-fold greater binding, preferably at least a 30-fold
greater binding and, most preferably at least a 300-fold greater
binding. The increased level of expression in tumor cells can be
evaluated on an absolute scale, i.e., relative to any other LAMR
expressing non-tumor cells described, or on a relative scale, i.e.,
relative to the level expressed by untransformed cells in the same
lineage as the transformed cancer cells (e.g., melanocytes in the
case of melanoma; hepatocytes in the case of hepatic carcinoma;
ovarian endothelial cells in the case of ovarian adenocarcinoma,
renal endothelial or epithelial cells in the case of renal
carcinoma).
[0027] As used herein, the term "infectious", "replication
competent" or "replication capable", when used to describe a
Sindbis virus vector RNA molecule, means an RNA molecule which is
self-replicating and provides for transcription in a host cell. The
term "replication", when used in conjunction with a Sindbis virus
genomic RNA vector RNA molecule means production of full-length
equivalents of (+)-strand RNA using (-)-strand RNA as a
template.
[0028] As used herein, the term "tumor" refers to a malignant
tissue comprising transformed cells that grow uncontrollably.
Tumors include leukemias, lymphomas, myelomas, plasmacytomas, and
the like; and solid tumors. Examples of solid tumors that can be
treated according to the invention include sarcomas and carcinomas
such as, but not limited to: fibrosarcoma, myxosarcoma,
liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma,
angiosarcoma, endotheliosarcoma, lymphangiosarcoma,
lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's
tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma,
pancreatic cancer, breast cancer, ovarian cancer, prostate cancer,
squamous cell carcinoma, basal cell carcinoma, epidermoid
carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland
carcinoma, papillary carcinoma, papillary adenocarcinomas,
cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma,
renal cell carcinoma, hepatoma, bile duct carcinoma,
choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor,
cervical cancer, testicular tumor, lung carcinoma, small cell lung
carcinoma, bladder carcinoma, epithelial carcinoma, .cndot.glioma,
astrocytoma, medulloblastoma, craniopharyngioma, ependymoma,
pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma,
meningioma, melanoma, neuroblastoma, neuroglioma, and
retinoblastoma. As noted above, the method of the invention depends
on expression of LAMRs by cells of the tumor targeted for
treatment.
[0029] The term "about" or "approximately" usually means within an
acceptable error range for the type of value and method of
measurement. For example, it can mean within 20%, more preferably
within 10%, and most preferably still within 5% of a given value or
range. Alternatively, especially in biological systems, the term
"about" means within about a log (i.e., an order of magnitude)
preferably within a factor of two of a given value.
[0030] The phrase "pharmaceutically acceptable", as used in
connection with compositions of the invention, refers to molecular
entities and other ingredients of such compositions that are
physiologically tolerable and do not typically produce untoward
reactions when administered to a human. Preferably, as used herein,
the term "pharmaceutically acceptable" means approved by a
regulatory agency of the Federal or a state government or listed in
the U.S. Pharmacopeia or other generally recognized pharmacopeia
for use in mammals, and more particularly in humans.
[0031] The term "therapeutically effective" when applied to a dose
or an amount refers to that quantity of a compound or
pharmaceutical composition that is sufficient to result in a
desired activity upon administration to a mammal in need thereof.
As used herein with respect to viral vectors of the invention, the
term "therapeutically effective amount/dose" refers to the
amount/dose of a vector or pharmaceutical composition containing
the vector that is sufficient to produce an effective anti-tumor
response upon administration to a mammal.
[0032] As disclosed in Ser. No. 10/920,030 the present inventors
previously discovered and devised methods for detecting tumor cells
and monitoring cancer therapy. The methods are based on the natural
preference of Sindbis virus to infect human cancer cells that
express higher level of 37/67-KDa laminin receptor (LAMR) than
normal, non-cancerous cells.
[0033] Various Sindbis viral vectors for cancer gene therapy have
been designed based on the RNA genome of wild-type virus (FIG. 1A).
The wild-type genome contains two major parts. The first part,
located on the 5' side of the genome, carries all of the
non-structural genes (ns1, ns2, ns3, and ns4) and a packaging
signal in the ns1 region. The expression of the ns1-4 genes leads
to synthesis of the non-structural proteins nsp 1-4 that form the
viral replicase. The replicase is necessary for expression of the
structural genes in the second part of the genome at 3' side for
virus formation. In order to do so, the replicase specifically
recognizes a short stretch of sequences between the non-structural
and structural genes, called the sub-genomic promoter (P.sub.SG).
One structural protein, the capsid protein, specifically recognizes
the packaging signal at the ns1 region and picks it up to form
viral particles with the rest of the structural proteins.
[0034] FIG. 1B depicts the conventional design of a replication
defective Sindbis viral vector that is capable of efficient
delivery of therapeutic genes while being unable to propagate and
generate more vector particles. This particular system has two
components. Replicon RNA contains all of the non-structural genes,
while the structural genes are replaced with the therapeutic gene
to be delivered. In order to generate vector particles, the
structural genes are provided in trans using a helper RNA, the
second component of this system. Unlike replicon RNA, the helper
RNA contains the non-structural genes and lacks the packaging
signal. Therefore, the produced vector particles only carry
replicon RNA with the gene of interest.
[0035] One major advantage of such conventional dual-component
vector systems is that the produced vector particle is
replication-defective (RD) and is safer for clinical use. However,
this advantage may become a significant drawback for cancer genes
therapy. The goal of cancer gene therapy is to infect the majority
of tumor cells and deliver the therapeutic genes for tumor
detection or eradication. To achieve this goal using a
replication-defective system may require a repetitive treatment
regime and high doses of vectors. In some cases, such high doses
may not be easily obtained using a replication defective
system.
[0036] On the other hand, a Sindbis vector system that is capable
of "controlled" replication and propagation is of great interest
for cancer gene therapy. Such a replication-capable (RC) system
should comprise a single-component to ensure efficient propagation
of the vector in the tumor. That is, the system does not require a
helper component for replication. One major benefit of such a
system is that fewer treatments would be required and a lower dose
should be sufficient to achieve successful therapeutic outcomes
while retaining the same tumor targeting capability as RD vectors.
Preferably, a safety mechanism is incorporated in the vector to
ensure that the vector is eliminated in order to prevent unwanted
toxicity, if any, associated with the propagation of the
vector.
[0037] The present invention provides a replication-capable (RC)
Sindbis viral vector (FIG. 1C) which was tested to see if a
single-component system performed better than the conventional
dual-component system. Instead of using a separate helper RNA
vector to carry the structural genes, the structural genes are
directly inserted into a conventional replicon vector, along with a
dedicated sub-genomic promoter following the therapeutic gene. The
dedicated second promoter guarantees that the efficient expression
of the structural genes for high-level vector production occurs. A
simple dual-promoter RC system was first developed in order to
study the function of the sub-genomic promoter in mammalian cells
(5,6). This type of RC system was later used to deliver antigen
encoding genes for vaccination purposes (7-9). Since Sindbis virus
infects mosquito cells, a simple RC system was also used to study
Sindbis virus spreading in mosquitoes (10). In light of the present
inventors' discovery that RD Sindbis vectors can target tumors in
living animals, a single-component RC Sindbis vector has been used
to detect/treat tumors in mouse tumor models (11). However, instead
of using a dedicated sub-genomic promoter, the prior art RC system
used a cleavable component to release the reporter protein from the
structural proteins and, therefore, significantly reduced the titer
of vector production (12).
[0038] FIG. 2 shows the results of an experiment comparing the
anti-tumor activity of replication defective (RD, FIG. 2A) and
replication competent (RC, FIG. 2B) Sindbis virus vectors. In this
set of experiments, a subcutaneously induced BHK tumor model (on
the right hind limb of SCID mice) was used to test and compare
these two vector systems. In order to evaluate specific tumor
targeting and elimination in this model, both vector systems carry
a firefly luciferase gene as a reporter. The bioluminescent signals
generated in the tumors can be easily detected and analyzed using
the IVIS.TM. imaging system. Mice received only two consecutive
treatments of RD or RC vectors via systemic (intravenous)
injections on a day 0 and day 1. No further treatment was
administrated. FIG. 2C depicts the imaging result on day 2 and FIG.
2D quantitatively shows the level of bioluminescent signals in
tumors that directly reflects vector infection level. The data
indicate that the RC vector system has about a 30 fold increase in
infectivity compared to the conventional RD vector system. Higher
infectivity is also reflected in enhanced tumor killing as
evidenced by tumor size reduction on day 10 (FIG. 2E).
[0039] These data provide proof-of-concept results in support of
the use of a RC Sindbis vector system for cancer gene therapy. The
capability of the RC vector to propagate and spread to the tumor
dramatically enhances the ability of Sindbis vectors to target and
kill cancer cells. The data also show that the same level of tumor
detection can be achieved using a lower range of effective doses of
the RC vector compared to conventional RD vectors. In the example
depicted in FIG. 2, 10.sup.6 RD or RC vector particles were
intravenously administered into mice. The signals from tumors
treated with the RC vectors were about 30 fold higher than the RD
signals. Therefore, the same imaging intensity in tumors was
achieved using a lower dose (10.sup.4 to 10.sup.5) of RC vectors.
Interestingly, no toxicity was associated with this prototype RC
vector administered at 10.sup.6 level and no bioluminescent
signals, except in residual tumors, were observed in animals up to
20 days after vector injections, suggesting that using the same
level of RC vector, as high as 10.sup.6 per dose, should not cause
side effects. The higher dose is useful not only for detecting
small lesions but also for better tumor killing and therapeutic
effects. Since humans are about 1000 times the body weight of mice,
the expected range of effective amounts for humans will range
between about 10.sup.7 and about 10.sup.10 particles per dose. The
lower range (10.sup.7-10.sup.8) is sufficient to detect tumor
masses and higher range (10.sup.8-10.sup.10) are better for tumor
eradication.
[0040] For use in the present invention, the RC Sindbis virus
vectors can be produced as described in U.S. Pat. Nos. 7,306,712
and 7,303,798 and in the example below. This involves the described
in vitro transcription/electroporation method.
[0041] However, as mentioned above, a safety mechanism
significantly reduces the risk, if any, of toxicity by controlling
the propagation of the RC vector system. In a preferred embodiment,
a "suicide gene" is incorporated into one of the Sindbis virus
non-structural genes that are essential for viral propagation and
survival. FIG. 1C depicts the concept of such a design. A suicide
gene, which encodes an enzyme capable of activating inert prodrugs
into cytotoxic metabolites, is fused in frame with the ns3 gene to
ensure co-expression with non-structural protein 3, an essential
component of viral replicase. A particular region of the ns3 gene
has been shown to be suitable for fusion without compromising the
function of nsp3. Therefore, by such design the RC vector is
genetically tagged with this safety mechanism which can shut-off
vector propagation by killing the vector producing cell during or
after the treatment regime.
[0042] In addition to serving as a safety feature, the fused
suicide gene provides another advantage. The tumor cells that are
selectively infected by the vector are more susceptible and
sensitive to the prodrug treatment, since they would not only face
the killing imposed by Sindbis infection, but also are exposed to
toxic metabolites as a result of prodrug activation. In this
regard, it has been discovered that activated toxic metabolites can
passively diffuse to neighboring uninfected tumor cells to further
enhance tumor killing. This is called a "bystander effect". The
bystander effect plays an important role in the eradication of
surrounding untransduced (uninfected) tumor cells. This is caused
by transmission of the activated prodrug from the transduced tumor
cells (which may be only a small fraction of total tumor mass) to
uninfected tumor cells. In the HSVtk/GCV system, the activated GCV
is not membrane permeable because of its highly charged phosphate
groups. However, it, can be transferred to uninfected cells via the
gap junctions or through the exchange of apoptotic vesicles that
kill the surrounding untransduced tumor cells (14).
[0043] Several suicide genes and their appropriate prodrugs are
available and suitable for use with the Sindbis virus vector in
this embodiment. For example, as disclosed herein, a conventional
Sindbis virus vector carrying a thymidine kinase gene isolated from
herpes simplex virus (HSVtk) significantly enhanced tumor killing
(FIG. 3A) (Horsburgh B C et al, Recurrent acyclovir-resistant
herpes simplex in an immunocompromised patient; can strain
differences compensate for loss of thymidine kinase in
pathogenesis? J. Infect. Dis., 178 (3), 618-625, 1998). A specific
prodrug, ganciclovir (GCV), has been developed to target HSVtk, and
has been clinically approved for treatment of cytomegalovirus and
herpes simplex virus infection in humans. (Ganciclovir, GCV, is
marketed under the trade name CYTOVENE.TM. by Roche Laboratories
Inc.)
[0044] Additional examples of suicide genes are thymidine kinase of
Varicella Zoster virus (VZV-tk) (disclosed in Lacey S F et al,
Analysis of mutations in the thymidine kinase genes of
drug-resistant varicella-zoster virus populations using the
polymerase chain reaction, J. Gen. Virol. 72 (PT 3), 623-630, 1991)
and the bacterial gene cytosine deaminase (Perna N T et al, Genome
sequence of enterohaemorrhagic Escherichia coli O157:H7, Nature 409
(6819), 529-533, 2001).
[0045] The prodrugs useful in the methods of the present invention
are any that can be converted to a toxic product, i.e., toxic to
tumor cells. A preferred prodrug is ganciclovir, which is converted
in vivo to a toxic compound by HSV-tk (Chen et al., Cancer Res.
1996, 56: 3758-3762). Other representative examples of prodrugs
include acyclovir, FIAU
[1-(2-deoxy-2-fluoro-.beta.-D-arabinofuranosyl)-5-iodouracil]
(FIALURIDINE.TM., Moravek Biochemicals and Radiochemicals),
6-methoxypurine arabinoside (converted by VZV-tk), and
5-fluorocytosine (converted by cytosine deaminise)
(5-fluorocytosine, Roche).
[0046] Prodrugs, may be readily administered to patients by
physicians having ordinary skill in the art. Using methods known in
the field, such physicians would also be able to determine the most
appropriate dose and route for the administration of the prodrug.
For example, ganciclovir is preferably administered systemically
(e.g. orally or parenterally) in a dose of about 1-20 mg/day/kg
body weight; acyclovir is administered in a dose of about 1-100
mg/day/kg body weight, and FIAU is administered in a dose of about
1-50 mg/day/kg body weight.
[0047] In the example below, SCID mice were intraperitoneally
implanted with ES2 human ovarian cancer cells. In order to track
and monitor disease progression, the ES2 cells were genetically
engineered to express a firefly luciferase gene for bioluminescent
imaging. Therefore, the bioluminescent signal intensity is
proportional to the tumor load in these animals (FIG. 3B).
Tumor-bearing mice were either mock treated or received daily
treatments with a conventional RD-Sindbis/HSVtk vector. Some mice
also received daily GCV treatments to determine if the prodrug
enhanced tumor killing in conjunction with Sindbis vector
treatment. Quantitative analysis indicated that, without HSVtk
expression, unactivated GCV confers no therapeutic effect on tumor
loading (FIG. 3C). On the contrary, GCV dramatically enhanced the
Sindbis/HSVtk vector treatments and suppressed tumor growth (FIG.
3D).
[0048] In an alternate embodiment, the vectors of the present
invention can be used to detect cancer cells and monitor
anti-cancer therapy. Previously, the present inventors used an
optical bioluminescence imaging system and RD Sindbis virus vectors
to detect tumor-specific targeting of Sindbis virus vectors in
small animals (15, 16 and Ser. No. 10/920,030). The advantages of
bioluminescent imaging include short imaging time, low costs, and
ease of use. However, optical imaging methods suffer from very
substantial attenuation of the light signal and, thus, are not
amenable to applications in large animals and in patients. Recent
advances in optical and radionuclide imaging technology provide
several methods for non-invasive monitoring of marker gene
expression in living animals. On the other hand, radionuclide
imaging methods such as g-camera, SPECT, and PET have excellent
depth sensitivity and can detect accumulation of gene expression
within the transfected tumors anywhere in the body and on the basis
of gene expression imaging (17,18,19). A major advantage of PET is
the ability to generate quantitative high spatial resolution,
3-dimensional images. When combined with other forms of tomographic
imaging, such as CT or MRI, fusion images of functional and
anatomic data provides more detailed in situ information of marker
genes' expression and localization.
[0049] As disclosed in copending Ser. No. 10/920,030, the present
inventors have discovered that imaging can be translated into
photon counts produced by the detectable label delivered to cancer
cells and that these are proportional to the amount of tumor cells
that remain alive. Therefore, the present invention can be used to
monitor anti-cancer therapy as follows. Patients can be
administered a diagnostically-effective amount of the RC Sindbis
vector of the present invention comprising a detectable label
before the onset of treatment, and this value can be compared to
one obtained upon administration of a diagnostically effective
amount of a Sindbis virus comprising a detectable label after
therapy has been completed. In this way, it is possible to
determine the extent of tumor kill. Since only living tumor cells
would contain the label, therapy would continue only until a
minimal amount of label is detected.
[0050] Since Sindbis virus vectors are gene transfer vectors, the
cancer cells are labeled using genetic markers incorporated into
the RC Sindbis virus vectors. In this embodiment, the genes useful
for live tumor monitoring or labeling include but are not limited
to the Green Fluorescence Protein (GFP) gene, [Cormack, B. P. et
al. (1966) FACS-optimized mutants of the green fluorescent protein
(GFP). Gene 173:33-38] the Firefly luciferase (Fluc) gene, [de Wet,
J. R., et al. (1987) Firefly luciferase gene: structure and
expression in mammalian cells Mol. Cell. Biol. 7 (2), 725-737], the
Renilla luciferase (Rluc) gene [Lorenz, W. W. et al. (1991)
Isolation and expression of a cDNA encoding Renilla reinformis
luciferase, Proc. Natl. Acad. Sci. U.S.A. 88 (10), 4438-4442] and
the dopamine-2 receptor (D.sub.2R) gene. The use of the D.sub.2R
gene as a reporter gene in living animals is disclosed in MacLaren
et al. (Gene Therapy 6:785-791 (1999)) and Yaghoubi et al. (Gene
Therapy 8:1072-1080 (2001)) These genes can be incorporated into
the Sindbis virus vectors of the present invention using techniques
well known to those of ordinary skill in the art, as described in
Bredenbeek P. J. et al. (1993) (Sindbis virus expression vectors:
packaging of RNA replicons by using defective helper RNAs, J.
Virol.; 67(11):6439-46.)
[0051] Cells expressing the genetic markers of the present
invention can be identified as follows: for the HSV-tk gene, the
subject can be administered radiolabeled
9-[(4[.sup.18F]fluoro-3-hydroxymethylbutyl)guanine (FHBG),
administered intravenously, about 6000 .mu.Ci/Kg body weight of the
recipient, (commercially available from PET Imaging Science Center,
U. of South California). Expression of HSV-tk activity in tumor
cells results in the accumulation of radiolabeled FHBG and can be
monitored by Positron Emission Tomography (PET). In vivo GFP
expressing tumor cells can be monitored by fluoresence microscopic
examination of tissue sections. Tissue sections of Fluc or Rluc
expressing tumor cells can be monitored by Cooled Charge-Coupled
Device (CCD) cameras in vivo (commercially available from Xenogen
Corp., Alamenda, Calif.). D.sub.2R activity can be identified by
administering 3-(2-[.sup.18F]fluoroethyl)spiperone ([.sup.18F]FESP)
and monitored by PET.
[0052] A subject to whom the diagnostic compound of the present
invention has been administered as an effective diagnostic monitor
for a disease or disorder is preferably a human, but can be any
animal, including a laboratory animal in the context of a clinical
trial or screening or activity experiment. Thus, as can be readily
appreciated by those of ordinary skill in the art, the methods and
compositions of the present invention are particularly suited to
administration to any animal, particularly a mammal, and including,
but by no means limited to, domestic animals, such as feline or
canine subjects, farm animals, such as but not limited to bovine,
equine, caprine, ovine, and porcine subjects, wild animals (whether
in the wild or in a zoological garden), research animals, such as
mice, rats, rabbits, goats, sheep, pigs, dogs, cats, etc., avian
species, such as chickens, turkeys, songbirds, etc., i.e., for
veterinary medical use.
[0053] In summary, the single-component RC Sindbis vector system of
the present invention dramatically enhances the tumor targeting,
monitoring and killing capability of replication-capable Sindbis
vectors, and incorporation of suicide genes provides an additional
layer of protection to achieve "controlled" propagation in tumors
and enhances tumor cell killing by RC Sindbis virus vectors.
[0054] In a preferred embodiment, the RC vectors are derived from
the RD mut-4 vector disclosed in U.S. Pat. No. 7,303,798. The mut-4
vector is similar to the SinRep5 system (Invitrogen Corp.), except
for changes in the amino acid sequences in the E2 protein. Since
this protein is directly involved in vector binding and targeting
to tumor cells, it is expected that RC vectors derived from the
mut-4 vector will have the same improved binding capability as the
RD vectors.
[0055] The construction of the RC mut-4 vector containing the
HSV-tk gene is shown in Paper Example 1 below.
[0056] The present invention also provides pharmaceutical
formulations or dosage forms for administration to mammals.
[0057] When formulated in a pharmaceutical composition, the vectors
of the present invention can be admixed with a pharmaceutically
acceptable carrier or excipient. The phrase "pharmaceutically
acceptable" refers to molecular entities and compositions that are
"generally regarded as safe", e.g., that are physiologically
tolerable and do not typically produce an allergic or similar
untoward reaction, such as gastric upset, dizziness and the like,
when administered to a human. Preferably, as used herein, the term
"pharmaceutically acceptable" means approved by a regulatory agency
of the Federal or a state government or listed in the U.S.
Pharmacopeia or other generally recognized pharmacopeia for use in
animals, and more particularly in humans. The term "carrier" refers
to a diluent, adjuvant, excipient, or vehicles with which the
compound is administered. Such pharmaceutical carriers can be
sterile liquids, such as water and oils, including those of
petroleum, animal, vegetable or synthetic origin, such as peanut
oil, soybean oil, mineral oil, sesame oil and the like. Water or
aqueous saline solutions and aqueous dextrose and glycerol
solutions are preferably employed as carriers, particularly for
injectable solutions. Alternatively, the carrier can be a solid
dosage form carrier, including but not limited to one or more of a
binder (for compressed pills), an encapsulating agent, a flavorant,
and a colorant. Suitable pharmaceutical carriers are described in
"Remington's Pharmaceutical Sciences" by E. W. Martin.
[0058] The preferred route of administration of the vectors of the
present invention, for treatment and monitoring, is parenteral and
most preferably systemic. This includes, but is not limited to
intravenous, intraperitoneal, intra-arteriole, intra-muscular,
intradermal, subcutaneous, intranasal and oral. These routes of
administration will permit homing of the vector to tumor cells
wherein, only Sindbis virus is a blood-borne virus. Therefore, gene
therapy vectors based on this virus have an advantage over other
viral vectors that are not adapted to travel in the bloodstream.
This property is largely responsible for the observation that
systemic administration of Sindbis viral vectors by i.p. or i.v.
injections, target and infect only tumors expressing greater
amounts of LAMR than normal cells of the same lineage growing s.c.,
i.p., intrapancreatically, or in the lungs. Thus, the blood-borne
nature of Sindbis viral vectors provides them with the capacity to
treat malignancies and monitor cancer therapy.
[0059] The present invention is described below in examples which
are intended to further describe the invention without limiting the
scope therapy.
Materials and Methods
Vector Construction
[0060] The prototype RC-Sindbis/Fluc vector was constructed using
the pSinRep5/Fluc plasmid as a backbone. In order to make the
vector replication competent, a DNA segment containing a
sub-genomic promoter and the Sindbis viral structural genes was
excised from the ptRNA-DHBB plasmid (Invitrogen Corp., Carlsbad,
Calif.) using NsiI and BamHI enzymes and then inserted into
pSinRep5/Fluc at the StuI site. Therefore the constructed
pSinRep5/Fluc-tBB plasmid has two independent sub-genomic promoters
to drive expression of firefly luciferase and Sindbis viral
structural proteins.
[0061] For construction of a RC vector with a suicide gene in the
HSVtk gene was fused in-frame with the Ns3 gene at the SpeI site in
pSinRep5/Fluc-tBB. The HSVtk gene (from the pORF-HSVtk plasmid,
Invivogen, San Diego, Calif.) was inserted at this site to generate
the pSinRep5-nsp3-HSVtk/Fluc-tBB plasmid.
Vector Preparation
[0062] The RC-Sindbis/Fluc vector was prepared using an in vitro
transcription/electroporation method as described U.S. Pat. Nos.
7,306,712 and 7,303,798. The plasmid pSinRep5/Fluc-tBB was
linearized using the NotI restriction enzyme. The linearized
plasmid DNA was then used as template for in vitro transcription.
The in vitro transcription was done in a total volume of 20 .mu.L
using a commercially available SP6 in vitro transcription kit
(Ambion Inc., Austin, Tex.). Transcribed RNA (20 .mu.L) was then
electroporated into 6.times.10.sup.6 BHK cells and cultured at
37.degree. C. in a 10 cm dish containing 10 mL of aMEM (Invitrogen
Corp.) with 10% FCS. The next day, the culture media was replaced
with 9 mL of OptiMEM (Invitrogen). The OptiMEM was then harvested
and stored at -80.degree. C.
In Vivo Imaging
[0063] BHK tumors were induced in female SCID mice (Taconic,
Germantown, N.Y.) by subcutaneous injection of 2 million BHK cells.
Ten days later, on day 0, mice were split into two groups. One
group of five mice received the first intravenous injection of
10.sup.6 particles of RD-Sindbis/Fluc, and the other group received
the first i.v. injection of 10.sup.6 particles of RC-Sindbis/Fluc.
The next day (day 1), both groups received a second dose (10.sup.6)
of i.v. treatments. Twenty-four hours later (day 2), tumor
luminescence signals were measured using the IVIS.RTM. spectrum
imaging system (Caliper LifeSciences, Hopkinton, Mass.) and tumor
specific signals were analyzed using Living Image 3.0 software.
Five minutes before imaging, 03 mL of 15 mg/mL D-luciferin
(Promega, Madison, Wis.) was i.p. injected in order to generate
bioluminescent signals. Tumor sizes were measured using calipers
and volumes were calculated using the formula:
4.pi./3.times.length.times.width.times.height.
[0064] An ES-2/Fluc ovarian cancer model was used to test if the
HSVtk suicide gene enhanced the therapeutic effects of Sindbis
vectors. The prodrug GCV (CYTOVENE-IV.degree., The Roche
Laboratories Inc.) enhanced the killing of Sindbis/tk-infected
ES-2/Fluc cells in vivo, as determined by the IVIS.RTM. system,
which is capable of non-invasive detection of bioluminescent signal
generated by ES-2/Fluc tumors. SCID mice were inoculated with
ES-2/Fluc on day 0.
[0065] Daily GDEPT treatments involving i.p. injections of
RD-Sindbis/tk and GCV (25 mg/Kg of body weight) were started on day
3. The Sindbis/tk -GCV group (n=5) received Sindbis/tk treatments
but no GCV. The Sindbis/tk +GCV group (n=5) received both
Sindbis/tk and GCV treatments. The Control group (n=5) was neither
treated with Sindbis/tk nor GCV. The Control +GCV group (n=5)
received no Sindbis/tk but was treated with GCV. Disease
progression was monitored and the whole body photon counts were
determined using the IVIS.RTM. system on days 1, 3, 6, 8, 12 and
14. Representative images of each treatment group are shown in FIG.
2.
[0066] In FIG. 2, tumor luciferase signals were higher and tumor
volumes were lower using the RC vector compared to the RD vector.
The average tumor luciferase signal in RC-treated mice was
43,970,000 photons, which was much higher than the average of
1,776,000 photons in RD-treated animals. The average tumor size of
RC-treated tumors was 410 mm3 and for RD-treated tumor the average
size was 1609 mm3.
Paper Example 1
[0067] The plasmid pSP6-R/NS3-HSVtk/Fluc-Mut4 enclodes a Sindbis
virus RC vector construction based on pSP6-R and Mut-4, which
provide replicase genes and structural genes respectively. Its
sequence is set forth in Appendix A and a map of the plasmid is
shown in FIG. 4. In an alternate embodiment, the promoter is T7
(Ambun, Austin, Tex. ??) Its construction is described as
follows.
[0068] A DNA segment containing a sub-genomic promoter and the
Sindbis viral structural genes is excised from the pSP6-Mut4
plasmid (disclosed in U.S. Pat. No. 7,303,798) using NsiI and BamHI
enzymes and then inserted into pSP6-R/Fluc at the PmII site. In
addition, the HSVtk gene fragment (from the pORF-HSVtk plasmid
(Invivogen, San Diego, Calif.) is inserted at the SpeI site in the
ns3 region on pSP6-R/Fluc-tBB to generate the
pSP6-R/nsp3-HSVtk/Fluc-tBB plasmid. A map showing the
pSP6-R/nsp3-HSVtk/Fluc-tBB plasmid is shown in FIG. 4 and its
sequence is set forth in Appendix A below.
[0069] In FIG. 4, the location of the genes is set forth below:
[0070] Sindbis non-structural genes locations (bp):
TABLE-US-00001 NS1 60-1979 (SEQ. ID NO.: 2) NS2 1980-4100 (SEQ. ID
NO.: 3) NS3 4101-5261 and 6399-6878 (SEQ. ID NO.: 4) NS4 6879-8729
(SEQ. ID NO.: 5) First Psg 8722-8734 (SEQ. ID NO.: 6) Second Psg
10766-10789 (SEQ. ID NO. 7) Structural Genes: 10833-14567 (SEQ. ID
NO.: 8)
[0071] In addition, the vector may also comprise a suicide gene,
such as the thymidine kinase (TK) gene located within the NS3 gene
(Nucleotide 5262-6398).
REFERENCES
[0072] 1. Danks M K, Morton C L, Krull E J, et al., Comparison of
activation of CPT-11 by rabbit and human carboxylesterases for use
in enzyme/prodrug therapy. Clin Cancer Res. 1999; 5:917-924. [0073]
2. Austin E A, Huber B E. A first step in the development of gene
therapy for colorectal carcinoma: cloning, sequencing, and
expression of Escherichia coli cytosine deaminase. Mol. Pharmacol.
1993:43:380-387. [0074] 3. Caruso M. Panis Y. Gagandeep S, Houssin
D, Salzmann J L, Klatzmann D. Regression of established macroscopic
liver metastases after in situ transduction of a suicide gene. Proc
Natl Acad Sci U.S.A. 1993; 90:7024-7028. [0075] 4. Sterman D H,
Treat J, Litzky L A, et al. Adenovirus-mediated herpes simplex
virus thymidine kinase/ganciclovir gene therapy in patients with
localized malignancy: results of a phase I clinical trial in
malignant mesothelioma. Hum Gene Ther. 1998; 9:1083-1092. [0076] 5.
Levis, R., Schlesinger, S. & Huang, H. V. Promoter for Sindbis
virus RNA-dependent subgenomic RNA transcription. J Virol 64,
1726-33 (1990). Raju, R. & Huang, H. V. Analysis of Sindbis
virus promoter recognition in vivo, using novel vectors with two
subgenomic mRNA promoters. J Virol 65, 2501-10 (1991). [0077] 7.
Hahn, C. S., Hahn, Y. S., Braciale, T. J. & Rice, C. M.
Infectious Sindbis virus transient expression vectors for studying
antigen processing and presentation. Proc Natl Acad Sci U.S.A. 89,
2679-83 (1992). [0078] 8. Pugachev, K. V., Mason, P. W., Shope, R.
E. & Frey, T. K. Double-subgenomic Sindbis virus recombinants
expressing immunogenic proteins of Japanese encephalitis virus
induce significant protection in mice against lethal JEV infection.
Virology 212, 587-94 (1995). [0079] 9. Tsuji, M. et al. Recombinant
Sindbis viruses expressing a cytotoxic T-lymphocyte epitope of a
malaria parasite or of influenza virus elicit protection against
the corresponding pathogen in mice. J Virol 72, 6907-10 (1998).
[0080] 10. Pierro, D. J., Myles, K. M., Foy, B. D., Beaty, B. J.
& Olson, K. E. Development of an orally infections Sindbis
virus transducing system that efficiently disseminates and
expresses green fluorescent protein in Aedes aegypti. Insect Mol
Biol 12, 107-16 (2003). [0081] 11. Unno, Y. et al. Oncolytic viral
therapy for cervical and ovarian cancer cells by Sindbis virus
AR339 strain. Clin Cancer Res 11, 4553-60 (2005). [0082] 12.
Thomas, J. M., Klimstra, W. B., Ryman, K. D. & Heidner, H. W.
Sindbis virus vectors designed to express a foreign protein as a
cleavable component of the viral structural polyprotein. J Virol
77, 5598-606 (2003). [0083] 13. Frolova, E. et al. Formation of
nsP3-specific protein complexes during Sindbis virus replication. J
Virol 80, 4122-34 (2006). [0084] 14. Dilber M S, Abedi M R,
Christensson B, et al. Gap junctions promote the bystander effect
of herpes simplex virus thymidine kinase in vivo. Cancer Res. 1997;
57:1523-1528. [0085] 15. Tseng J C, Levin B, Hirano T, Yee H,
Pampeno C, Meruelo D. In vivo antitumor activity of sindbis viral
vectors. J Natl Cancer Inst. 2002; 94:1790-1802. [0086] 16. Tseng J
C, Hurtado A, Yee H, et al. Using sindbis viral vectors for
specific detection and suppression of advanced ovarian cancer in
animal models. Cancer Res. 2004; 64:6684-6692. [0087] 17. Serganova
I, Doubrovin M, Vider J, et al. Molecular imaging of temporal
dynamics and spatial heterogeneity of hypoxia-inducible factor-1
signal transduction activity in tumors n living mice. Cancer Res.
2004; 64:6101-6108. [0088] 18. Blsasberg R G, Gelovani J.
Molecular-genetic imaging: a nuclear medicine-based perspective.
Mol. Imaging. 2002; 1:280-300. [0089] 19. Wen B, Burgman P,
Zanzonico P, et al. A preclinical model for noninvasive imaging of
hypoxia-induced gene expression: comparison with an exogenous
marker of tumor hypoxia. Eur J Nucl Med Mol. Imaging. 2004;
31:1530-1538.
TABLE-US-00002 [0089] APPENDIX A pSP6-R/nsp3-HSVtk/Fluc-Mut4 1
ATTGACGGCG TAGTACACAC TATTGAATCA AACAGCCGAC CAATTGCACT TAACTGCCGC
ATCATGTGTG ATAACTTAGT TTGTCGGCTG GTTAACGTGA 51 ACCATCACAA
TGGAGAAGCC AGTAGTAAAC GTAGACGTAG ACCCCCAGAG TGGTAGTGTT ACCTCTTCGG
TCATCATTTG CATCTGCATC TGGGGGTCTC 101 TCCGTTTGTC GTGCAACTGC
AAAAAAGCTT CCCGCAATTT GAGGTAGTAG AGGCAAACAG CACGTTGACG TTTTTTCGAA
GGGCGTTAAA CTCCATCATC 151 CACAGCAGGT CACTCCAAAT GACCATGCTA
ATGCCAGAGC ATTTTCGCAT GTGTCGTCCA GTGAGGTTTA CTGGTACGAT TACGGTCTCG
TAAAAGCGTA 201 CTGGCCAGTA AACTAATCGA GCTGGAGGTT CCTACCACAG
CGACGATCTT GACCGGTCAT TTGATTAGCT CGACCTCCAA GGATGGTGTC GCTGCTAGAA
251 GGACATAGGC AGCGCACCGG CTCGTAGAAT GTTTTCCGAG CACCAGTATC
CCTGTATCCG TCGCGTGGCC GAGCATCTTA CAAAAGGCTC GTGGTCATAG 301
ATTGTGTCTG CCCCATGCGT AGTCCAGAAG ACCCGGACCG CATGATGAAA TAACACAGAC
GGGGTACGCA TCAGGTCTTC TGGGCCTGGC GTACTACTTT 351 TATGCCAGTA
AACTGGCGGA AAAAGCGTGC AAGATTACAA ACAAGAACTT ATACGGTCAT TTGACCGCCT
TTTTCGCACG TTCTAATGTT TGTTCTTGAA 401 GCATGAGAAG ATTAAGGATC
TCCGGACCGT ACTTGATACG CCGGATGCTG CGTACTCTTC TAATTCCTAG AGGCCTGGCA
TGAACTATGC GGCCTACGAC 451 AAACACCATC GCTCTGCTTT CACAACGATG
TTACCTGCAA CATGCGTGCC TTTGTGGTAG CGAGACGAAA GTGTTGCTAC AATGGACGTT
GTACGCACGG 501 GAATATTCCG TCATGCAGGA CGTGTATATC AACGCTCCCG
GAACTATCTA CTTATAAGGC AGTACGTCCT GCACATATAG TTGCGAGGGC CTTGATAGAT
551 TCATCAGGCT ATGAAAGGCG TGCGGACCCT GTACTGGATT GGCTTCGACA
AGTAGTCCGA TACTTTCCGC ACGCCTGGGA CATGACCTAA CCGAAGCTGT 601
CCACCCAGTT CATGTTCTCG GCTATGGCAG GTTCGTACCC TGCGTACAAC GGTGGGTCAA
GTACAAGAGC CGATACCGTC CAAGCATGGG ACGCATGTTG 651 ACCAACTGGG
CCGACGAGAA AGTCCTTGAA GCGCGTAACA TCGGACTTTG TGGTTGACCC GGCTGCTCTT
TCAGGAACTT CGCGCATTGT AGCCTGAAAC 701 CAGCACAAAG CTGAGTGAAG
GTAGGACAGG AAAATTGTCG ATAATGAGGA GTCGTGTTTC GACTCACTTC CATCCTGTCC
TTTTAACAGC TATTACTCCT 751 AGAAGGAGTT GAAGCCCGGG TCGCGGGTTT
ATTTCTCCGT AGGATCGACA TCTTCCTCAA CTTCGGGCCC AGCGCCCAAA TAAAGAGGCA
TCCTAGCTGT 801 CTTTATCCAG AACACAGAGC CAGCTTGCAG AGCTGGCATC
TTCCATCGGT GAAATAGGTC TTGTGTCTCG GTCGAACGTC TCGACCGTAG AAGGTAGCCA
851 GTTCCACTTG AATGGAAAGC AGTCGTACAC TTGCCGCTGT GATACAGTGG
CAAGGTGAAC TTACCTTTCG TCAGCATGTG AACGGCGACA CTATGTCACC 901
TGAGTTGCGA AGGCTACGTA GTGAAGAAAA TCACCATCAG TCCCGGGATC ACTCAACGCT
TCCGATGCAT CACTTCTTTT AGTGGTAGTC AGGGCCCTAG 951 ACGGGAGAAA
CCGTGGGATA CGCGGTTACA CACAATAGCG AGGGCTTCTT TGCCCTCTTT GGCACCCTAT
GCGCCAATGT GTGTTATCGC TCCCGAAGAA 1001 GCTATGCAAA GTTACTGACA
CAGTAAAAGG AGAACGGGTA TCGTTCCCTG CGATACGTTT CAATGACTGT GTCATTTTCC
TCTTGCCCAT AGCAAGGGAC 1051 TGTGCACGTA CATCCCGGCC ACCATATGCG
ATCAGATGAC TGGTATAATG ACACGTGCAT GTAGGGCCGG TGGTATACGC TAGTCTACTG
ACCATATTAC 1101 GCCACGGATA TATCACCTGA CGATGCACAA AAACTTCTGG
TTGGGCTCAA CGGTGCCTAT ATAGTGGACT GCTACGTGTT TTTGAAGACC AACCCGAGTT
1151 CCAGCGAATT GTCATTAACG GTAGGACTAA CAGGAACACC AACACCATGC
GGTCGCTTAA CAGTAATTGC CATCCTGATT GTCCTTGTGG TTGTGGTACG 1201
AAAATTACCT TCTGCCGATC ATAGCACAAG GGTTCAGCAA ATGGGCTAAG TTTTAATGGA
AGACGGCTAG TATCGTGTTC CCAAGTCGTT TACCCGATTC 1251 GAGCGCAAGG
ATGATCTTGA TAACGAGAAA ATGCTGGGTA CTAGAGAACG CTCGCGTTCC TACTAGAACT
ATTGCTCTTT TACGACCCAT GATCTCTTGC 1301 CAAGCTTACG TATGGCTGCT
TGTGGGCGTT TCGCACTAAG AAAGTACATT GTTCGAATGC ATACCGACGA ACACCCGCAA
AGCGTGATTC TTTCATGTAA 1351 CGTTTTATCG CCCACCTGGA ACGCAGACCA
TCGTAAAAGT CCCAGCCTCT GCAAAATAGC GGGTGGACCT TGCGTCTGGT AGCATTTTCA
GGGTCGGAGA 1401 TTTAGCGCTT TTCCCATGTC GTCCGTATGG ACGACCTCTT
TGCCCATGTC AAATCGCGAA AAGGGTACAG CAGGCATACC TGCTGGAGAA ACGGGTACAG
1451 GCTGAGGCAG AAATTGAAAC TGGCATTGCA ACCAAAGAAG GAGGAAAAAC
CGACTCCGTC TTTAACTTTG ACCGTAACGT TGGTTTCTTC CTCCTTTTTG 1501
TGCTGCAGGT CTCGGAGGAA TTAGTCATGG AGGCCAAGGC TGCTTTTGAG ACGACGTCCA
GAGCCTCCTT AATCAGTACC TCCGGTTCCG ACGAAAACTC 1551 GATGCTCAGG
AGGAAGCCAG AGCGGAGAAG CTCCGAGAAG CACTTCCACC CTACGAGTCC TCCTTCGGTC
TCGCCTCTTC GAGGCTCTTC GTGAAGGTGG 1601 ATTAGTGGCA GACAAAGGCA
TCGAGGCAGC CGCAGAAGTT GTCTGCGAAG TAATCACCGT CTGTTTCCGT AGCTCCGTCG
GCGTCTTCAA CAGACGCTTC 1651 TGGAGGGGCT CCAGGCGGAC ATCGGAGCAG
CATTAGTTGA AACCCCGCGC ACCTCCCCGA GGTCCGCCTG TAGCCTCGTC GTAATCAACT
TTGGGGCGCG 1701 GGTCACGTAA GGATAATACC TCAAGCAAAT GACCGTATGA
TCGGACAGTA CCAGTGCATT CCTATTATGG AGTTCGTTTA CTGGCATACT AGCCTGTCAT
1751 TATCGTTGTC TCGCCAAACT CTGTGCTGAA GAATGCCAAA CTCGCACCAG
ATAGCAACAG AGCGGTTTGA GACACGACTT CTTACGGTTT GAGCGTGGTC 1801
CGCACCCGCT AGCAGATCAG GTTAAGATCA TAACACACTC CGGAAGATCA GCGTGGGCGA
TCGTCTAGTC CAATTCTAGT ATTGTGTGAG GCCTTCTAGT 1851 GGAAGGTACG
CGGTCGAACC ATACGACGCT AAAGTACTGA TGCCAGCAGG CCTTCCATGC GCCAGCTTGG
TATGCTGCGA TTTCATGACT ACGGTCGTCC 1901 AGGTGCCGTA CCATGGCCAG
AATTCCTAGC ACTGAGTGAG AGCGCCACGT TCCACGGCAT GGTACCGGTC TTAAGGATCG
TGACTCACTC TCGCGGTGCA 1951 TAGTGTACAA CGAAAGAGAG TTTGTGAACC
GCAAACTATA CCACATTGCC ATCACATGTT GCTTTCTCTC AAACACTTGG CGTTTGATAT
GGTGTAACGG 2001 ATGCATGGCC CCGCCAAGAA TACAGAAGAG GAGCAGTACA
AGGTTACAAA TACGTACCGG GGCGGTTCTT ATGTCTTCTC CTCGTCATGT TCCAATGTTT
2051 GGCAGAGCTT GCAGAAACAG AGTACGTGTT TGACGTGGAC AAGAAGCGTT
CCGTCTCGAA CGTCTTTGTC TCATGCACAA ACTGCACCTG TTCTTCGCAA 2101
GCGTTAAGAA GGAAGAAGCC TCAGGTCTGG TCCTCTCGGG AGAACTGACC CGCAATTCTT
CCTTCTTCGG AGTCCAGACC AGGAGAGCCC TCTTGACTGG 2151 AACCCTCCCT
ATCATGAGCT AGCTCTGGAG GGACTGAAGA CCCGACCTGC TTGGGAGGGA TAGTACTCGA
TCGAGACCTC CCTGACTTCT GGGCTGGACG 2201 GGTCCCGTAC AAGGTCGAAA
CAATAGGAGT GATAGGCACA CCGGGGTCGG CCAGGGCATG TTCCAGCTTT GTTATCCTCA
CTATCCGTGT GGCCCCAGCC 2251 GCAAGTCAGC TATTATCAAG TCAACTGTCA
CGGCACGAGA TCTTGTTACC CGTTCAGTCG ATAATAGTTC AGTTGACAGT GCCGTGCTCT
AGAACAATGG 2301 AGCGGAAAGA AAGAAAATTG TCGCGAAATT GAGGCCGACG
TGCTAAGACT TCGCCTTTCT TTCTTTTAAC AGCGCTTTAA CTCCGGCTGC ACGATTCTGA
2351 GAGGGGTATG CAGATTACGT CGAAGACAGT AGATTCGGTT ATGCTCAACG
CTCCCCATAC GTCTAATGCA GCTTCTGTCA TCTAAGCCAA TACGAGTTGC 2401
GATGCCACAA AGCCGTAGAA GTGCTGTACG TTGACGAAGC GTTCGCGTGC CTACGGTGTT
TCGGCATCTT CACGACATGC AACTGCTTCG CAAGCGCACG 2451 CACGCAGGAG
CACTACTTGC CTTGATTGCT ATCGTCAGGC CCCGCAAGAA GTGCGTCCTC GTGATGAACG
GAACTAACGA TAGCAGTCCG GGGCGTTCTT 2501 GGTAGTACTA TGCGGAGACC
CCATGCAATG CGGATTCTTC AACATGATGC CCATCATGAT ACGCCTCTGG GGTACGTTAC
GCCTAAGAAG TTGTACTACG 2551 AACTAAAGGT ACATTTCAAT CACCCTGAAA
AAGACATATG CACCAAGACA TTGATTTCCA TGTAAAGTTA GTGGGACTTT TTCTGTATAC
GTGGTTCTGT 2601 TTCTACAAGT ATATCTCCCG GCGTTGCACA CAGCCAGTTA
CAGCTATTGT AAGATGTTCA TATAGAGGGC CGCAACGTGT GTCGGTCAAT GTCGATAACA
2651 ATCGACACTG CATTACGATG GAAAGATGAA AACCACGAAC CCGTGCAAGA
TAGCTGTGAC GTAATGCTAC CTTTCTACTT TTGGTGCTTG GGCACGTTCT 2701
AGAACATTGA AATCGATATT ACAGGGGCCA CAAAGCCGAA GCCAGGGGAT TCTTGTAACT
TTAGCTATAA TGTCCCCGGT GTTTCGGCTT CGGTCCCCTA 2751 ATCATCCTGA
CATGTTTCCG CGGGTGGGTT AAGCAATTGC AAATCGACTA TAGTAGGACT GTACAAAGGC
GCCCACCCAA TTCGTTAACG TTTAGCTGAT 2801 TCCCGGACAT GAAGTAATGA
CAGCCGCGGC CTCACAAGGG CTAACCAGAA AGGGCCTGTA CTTCATTACT GTCGGCGCCG
GAGTGTTCCC GATTGGTCTT 2851 AAGGAGTGTA TGCCGTCCGG CAAAAAGTCA
ATGAAAACCC ACTGTACGCG TTCCTCACAT ACGGCAGGCC GTTTTTCAGT TACTTTTGGG
TGACATGCGC 2901 ATCACATCAG AGCATGTGAA CGTGTTGCTC ACCCGCACTG
AGGACAGGCT TAGTGTAGTC TCGTACACTT GCACAACGAG TGGGCGTGAC TCCTGTCCGA
2951 AGTGTGGAAA ACCTTGCAGG GCGACCCATG GATTAAGCAG CTCACTAACA
TCACACCTTT TGGAACGTCC CGCTGGGTAC CTAATTCGTC GAGTGATTGT 3001
TACCTAAAGG AAACTTTCAG GCTACTATAG AGGACTGGGA AGCTGAACAC ATGGATTTCC
TTTGAAAGTC CGATGATATC TCCTGACCCT TCGACTTGTG 3051 AAGGGAATAA
TTGCTGCAAT AAACAGCCCC ACTCCCCGTG CCAATCCGTT TTCCCTTATT AACGACGTTA
TTTGTCGGGG TGAGGGGCAC GGTTAGGCAA 3101 CAGCTGCAAG ACCAACGTTT
GCTGGGCGAA AGCATTGGAA CCGATACTAG GTCGACGTTC TGGTTGCAAA CGACCCGCTT
TCGTAACCTT GGCTATGATC 3151 CCACGGCCGG TATCGTACTT ACCGGTTGCC
AGTGGAGCGA ACTGTTCCCA GGTGCCGGCC ATAGCATGAA TGGCCAACGG TCACCTCGCT
TGACAAGGGT 3201 CAGTTTGCGG ATGACAAACC ACATTCGGCC ATTTACGCCT
TAGACGTAAT GTCAAACGCC TACTGTTTGG TGTAAGCCGG TAAATGCGGA ATCTGCATTA
3251 TTGCATTAAG TTTTTCGGCA TGGACTTGAC AAGCGGACTG TTTTCTAAAC
AACGTAATTC AAAAAGCCGT ACCTGAACTG TTCGCCTGAC AAAAGATTTG 3301
AGAGCATCCC ACTAACGTAC CATCCCGCCG ATTCAGCGAG GCCGGTAGCT TCTCGTAGGG
TGATTGCATG GTAGGGCGGC TAAGTCGCTC CGGCCATCGA 3351 CATTGGGACA
ACAGCCCAGG AACCCGCAAG TATGGGTACG ATCACGCCAT GTAACCCTGT TGTCGGGTCC
TTGGGCGTTC ATACCCATGC TAGTGCGGTA 3401 TGCCGCCGAA CTCTCCCGTA
GATTTCCGGT GTTCCAGCTA GCTGGGAAGG ACGGCGGCTT GAGAGGGCAT CTAAAGGCCA
CAAGGTCGAT CGACCCTTCC 3451 GCACACAACT TGATTTGCAG ACGGGGAGAA
CCAGAGTTAT CTCTGCACAG CGTGTGTTGA ACTAAACGTC TGCCCCTCTT GGTCTCAATA
GAGACGTGTC 3501 CATAACCTGG TCCCGGTGAA CCGCAATCTT CCTCACGCCT
TAGTCCCCGA GTATTGGACC AGGGCCACTT GGCGTTAGAA GGAGTGCGGA ATCAGGGGCT
3551 GTACAAGGAG AAGCAACCCG GCCCGGTCGA AAAATTCTTG AACCAGTTCA
CATGTTCCTC TTCGTTGGGC CGGGCCAGCT TTTTAAGAAC TTGGTCAAGT 3601
AACACCACTC AGTACTTGTG GTATCAGAGG AAAAAATTGA AGCTCCCCGT TTGTGGTGAG
TCATGAACAC CATAGTCTCC TTTTTTAACT TCGAGGGGCA 3651 AAGAGAATCG
AATGGATCGC CCCGATTGGC ATAGCCGGTG CAGATAAGAA TTCTCTTAGC TTACCTAGCG
GGGCTAACCG TATCGGCCAC GTCTATTCTT 3701 CTACAACCTG GCTTTCGGGT
TTCCGCCGCA GGCACGGTAC GACCTGGTGT GATGTTGGAC CGAAAGCCCA AAGGCGGCGT
CCGTGCCATG CTGGACCACA 3751 TCATCAACAT TGGAACTAAA TACAGAAACC
ACCACTTTCA GCAGTGCGAA AGTAGTTGTA ACCTTGATTT ATGTCTTTGG TGGTGAAAGT
CGTCACGCTT 3801 GACCATGCGG CGACCTTAAA AACCCTTTCG CGTTCGGCCC
TGAATTGCCT CTGGTACGCC GCTGGAATTT TTGGGAAAGC GCAAGCCGGG ACTTAACGGA
3851 TAACCCAGGA GGCACCCTCG TGGTGAAGTC CTATGGCTAC GCCGACCGCA
ATTGGGTCCT CCGTGGGAGC ACCACTTCAG GATACCGATG CGGCTGGCGT 3901
ACAGTGAGGA CGTAGTCACC GCTCTTGCCA GAAAGTTTGT CAGGGTGTCT TGTCACTCCT
GCATCAGTGG CGAGAACGGT CTTTCAAACA GTCCCACAGA 3951 GCAGCGAGAC
CAGATTGTGT CTCAAGCAAT ACAGAAATGT ACCTGATTTT CGTCGCTCTG GTCTAACACA
GAGTTCGTTA TGTCTTTACA TGGACTAAAA 4001 CCGACAACTA GACAACAGCC
GTACACGGCA ATTCACCCCG CACCATCTGA GGCTGTTGAT CTGTTGTCGG CATGTGCCGT
TAAGTGGGGC GTGGTAGACT 4051 ATTGCGTGAT TTCGTCCGTG TATGAGGGTA
CAAGAGATGG AGTTGGAGCC TAACGCACTA AAGCAGGCAC ATACTCCCAT GTTCTCTACC
TCAACCTCGG
4101 GCGCCGTCAT ACCGCACCAA AAGGGAGAAT ATTGCTGACT GTCAAGAGGA
CGCGGCAGTA TGGCGTGGTT TTCCCTCTTA TAACGACTGA CAGTTCTCCT 4151
AGCAGTTGTC AACGCAGCCA ATCCGCTGGG TAGACCAGGC GAAGGAGTCT TCGTCAACAG
TTGCGTCGGT TAGGCGACCC ATCTGGTCCG CTTCCTCAGA 4201 GCCGTGCCAT
CTATAAACGT TGGCCGACCA GTTTTACCGA TTCAGCCACG CGGCACGGTA GATATTTGCA
ACCGGCTGGT CAAAATGGCT AAGTCGGTGC 4251 GAGACAGGCA CCGCAAGAAT
GACTGTGTGC CTAGGAAAGA AAGTGATCCA CTCTGTCCGT GGCGTTCTTA CTGACACACG
GATCCTTTCT TTCACTAGGT 4301 CGCGGTCGGC CCTGATTTCC GGAAGCACCC
AGAAGCAGAA GCCTTGAAAT GCGCCAGCCG GGACTAAAGG CCTTCGTGGG TCTTCGTCTT
CGGAACTTTA 4351 TGCTACAAAA CGCCTACCAT GCAGTGGCAG ACTTAGTAAA
TGAACATAAC ACGATGTTTT GCGGATGGTA CGTCACCGTC TGAATCATTT ACTTGTATTG
4401 ATCAAGTCTG TCGCCATTCC ACTGCTATCT ACAGGCATTT ACGCAGCCGG
TAGTTCAGAC AGCGGTAAGG TGACGATAGA TGTCCGTAAA TGCGTCGGCC 4451
AAAAGACCGC CTTGAAGTAT CACTTAACTG CTTGACAACC GCGCTAGACA TTTTCTGGCG
GAACTTCATA GTGAATTGAC GAACTGTTGG CGCGATCTGT 4501 GAACTGACGC
GGACGTAACC ATCTATTGCC TGGATAAGAA GTGGAAGGAA CTTGACTGCG CCTGCATTGG
TAGATAACGG ACCTATTCTT CACCTTCCTT 4551 AGAATCGACG CGGCACTCCA
ACTTAAGGAG TCTGTAACAG AGCTGAAGGA TCTTAGCTGC GCCGTGAGGT TGAATTCCTC
AGACATTGTC TCGACTTCCT 4601 TGAAGATATG GAGATCGACG ATGAGTTAGT
ATGGATCCAT CCAGACAGTT ACTTCTATAC CTCTAGCTGC TACTCAATCA TACCTAGGTA
GGTCTGTCAA 4651 GCTTGAAGGG AAGAAAGGGA TTCAGTACTA CAAAAGGAAA
ATTGTATTCG CGAACTTCCC TTCTTTCCCT AAGTCATGAT GTTTTCCTTT TAACATAAGC
4701 TACTTCGAAG GCACCAAATT CCATCAAGCA GCAAAAGACA TGGCGGAGAT
ATGAAGCTTC CGTGGTTTAA GGTAGTTCGT CGTTTTCTGT ACCGCCTCTA 4751
AAAGGTCCTG TTCCCTAATG ACCAGGAAAG TAATGAACAA CTGTGTGCCT TTTCCAGGAC
AAGGGATTAC TGGTCCTTTC ATTACTTGTT GACACACGGA 4801 ACATATTGGG
TGAGACCATG GAAGCAATCC GCGAAAAGTG CCCGGTCGAC TGTATAACCC ACTCTGGTAC
CTTCGTTAGG CGCTTTTCAC GGGCCAGCTG 4851 CATAACCCGT CGTCTAGCCC
GCCCAAAACG TTGCCGTGCC TTTGCATGTA GTATTGGGCA GCAGATCGGG CGGGTTTTGC
AACGGCACGG AAACGTACAT 4901 TGCCATGACG CCAGAAAGGG TCCACAGACT
TAGAAGCAAT AACGTCAAAG ACGGTACTGC GGTCTTTCCC AGGTGTCTGA ATCTTCGTTA
TTGCAGTTTC 4951 AAGTTACAGT ATGCTCCTCC ACCCCCCTTC CTAAGCACAA
AATTAAGAAT TTCAATGTCA TACGAGGAGG TGGGGGGAAG GATTCGTGTT TTAATTCTTA
5001 GTTCAGAAGG TTCAGTGCAC GAAAGTAGTC CTGTTTAATC CGCACACTCC
CAAGTCTTCC AAGTCACGTG CTTTCATCAG GACAAATTAG GCGTGTGAGG 5051
CGCATTCGTT CCCGCCCGTA AGTACATAGA AGTGCCAGAA CAGCCTACCG GCGTAAGCAA
GGGCGGGCAT TCATGTATCT TCACGGTCTT GTCGGATGGC 5101 CTCCTCCTGC
ACAGGCCGAG GAGGCCCCCG AAGTTGTAGC GACACCGTCA GAGGAGGACG TGTCCGGCTC
CTCCGGGGGC TTCAACATCG CTGTGGCAGT 5151 CCATCTACAG CTGATAACAC
CTCGCTTGAT GTCACAGACA TCTCACTGGA GGTAGATGTC GACTATTGTG GAGCGAACTA
CAGTGTCTGT AGAGTGACCT 5201 TATGGATGAC AGTAGCGAAG GCTCACTTTT
TTCGAGCTTT AGCGGATCGG ATACCTACTG TCATCGCTTC CGAGTGAAAA AAGCTCGAAA
TCGCCTAGCC 5251 ACAACTCTAT TACTAGTGCC TCGTACCCCG GCCATCAACA
CGCGTCTGCG TGTTGAGATA ATGATCACGG AGCATGGGGC CGGTAGTTGT GCGCAGACGC
5301 TTCGACCAGG CTGCGCGTTC TCGCGGCCAT AGCAACCGAC GTACGGCGTT
AAGCTGGTCC GACGCGCAAG AGCGCCGGTA TCGTTGGCTG CATGCCGCAA 5351
GCGCCCTCGC CGGCAGCAAG AAGCCACGGA AGTCCGCCCG GAGCAGAAAA CGCGGGAGCG
GCCGTCGTTC TTCGGTGCCT TCAGGCGGGC CTCGTCTTTT 5401 TGCCCACGCT
ACTGCGGGTT TATATAGACG GTCCCCACGG GATGGGGAAA ACGGGTGCGA TGACGCCCAA
ATATATCTGC CAGGGGTGCC CTACCCCTTT 5451 ACCACCACCA CGCAACTGCT
GGTGGCCCTG GGTTCGCGCG ACGATATCGT TGGTGGTGGT GCGTTGACGA CCACCGGGAC
CCAAGCGCGC TGCTATAGCA 5501 CTACGTACCC GAGCCGATGA CTTACTGGCG
GGTGCTGGGG GCTTCCGAGA GATGCATGGG CTCGGCTACT GAATGACCGC CCACGACCCC
CGAAGGCTCT 5551 CAATCGCGAA CATCTACACC ACACAACACC GCCTCGACCA
GGGTGAGATA GTTAGCGCTT GTAGATGTGG TGTGTTGTGG CGGAGCTGGT CCCACTCTAT
5601 TCGGCCGGGG ACGCGGCGGT GGTAATGACA AGCGCCCAGA TAACAATGGG
AGCCGGCCCC TGCGCCGCCA CCATTACTGT TCGCGGGTCT ATTGTTACCC 5651
CATGCCTTAT GCCGTGACCG ACGCCGTTCT GGCTCCTCAT ATCGGGGGGG GTACGGAATA
CGGCACTGGC TGCGGCAAGA CCGAGGAGTA TAGCCCCCCC 5701 AGGCTGGGAG
CTCACATGCC CCGCCCCCGG CCCTCACCCT CATCTTCGAC TCCGACCCTC GAGTGTACGG
GGCGGGGGCC GGGAGTGGGA GTAGAAGCTG 5751 CGCCATCCCA TCGCCGCCCT
CCTGTGCTAC CCGGCCGCGC GGTACCTTAT GCGGTAGGGT AGCGGCGGGA GGACACGATG
GGCCGGCGCG CCATGGAATA 5801 GGGCAGCATG ACCCCCCAGG CCGTGCTGGC
GTTCGTGGCC CTCATCCCGC CCCGTCGTAC TGGGGGGTCC GGCACGACCG CAAGCACCGG
GAGTAGGGCG 5851 CGACCTTGCC CGGCACCAAC ATCGTGCTTG GGGCCCTTCC
GGAGGACAGA GCTGGAACGG GCCGTGGTTG TAGCACGAAC CCCGGGAAGG CCTCCTGTCT
5901 CACATCGACC GCCTGGCCAA ACGCCAGCGC CCCGGCGAGC GGCTGGACCT
GTGTAGCTGG CGGACCGGTT TGCGGTCGCG GGGCCGCTCG CCGACCTGGA 5951
GGCTATGCTG GCTGCGATTC GCCGCGTTTA CGGGCTACTT GCCAATACGG CCGATACGAC
CGACGCTAAG CGGCGCAAAT GCCCGATGAA CGGTTATGCC 6001 TGCGGTATCT
GCAGTGCGGC GGGTCGTGGC GGGAGGACTG GGGACAGCTT ACGCCATAGA CGTCACGCCG
CCCAGCACCG CCCTCCTGAC CCCTGTCGAA 6051 TCGGGGACGG CCGTGCCGCC
CCAGGGTGCC GAGCCCCAGA GCAACGCGGG AGCCCCTGCC GGCACGGCGG GGTCCCACGG
CTCGGGGTCT CGTTGCGCCC 6101 CCCACGACCC CATATCGGGG ACACGTTATT
TACCCTGTTT CGGGCCCCCG GGGTGCTGGG GTATAGCCCC TGTGCAATAA ATGGGACAAA
GCCCGGGGGC 6151 AGTTGCTGGC CCCCAACGGC GACCTGTATA ACGTGTTTGC
CTGGGCCTTG TCAACGACCG GGGGTTGCCG CTGGACATAT TGCACAAACG GACCCGGAAC
6201 GACGTCTTGG CCAAACGCCT CCGTTCCATG CACGTCTTTA TCCTGGATTA
CTGCAGAACC GGTTTGCGGA GGCAAGGTAC GTGCAGAAAT AGGACCTAAT 6251
CGACCAATCG CCCGCCGGCT GCCGGGACGC CCTGCTGCAA CTTACCTCCG GCTGGTTAGC
GGGCGGCCGA CGGCCCTGCG GGACGACGTT GAATGGAGGC 6301 GGATGGTCCA
GACCCACGTC ACCACCCCCG GCTCCATACC GACGATATGC CCTACCAGGT CTGGGTGCAG
TGGTGGGGGC CGAGGTATGG CTGCTATACG 6351 GACCTGGCGC GCACGTTTGC
CCGGGAGATG GGGGAGGCTA ACACTAGTAT CTGGACCGCG CGTGCAAACG GGCCCTCTAC
CCCCTCCGAT TGTGATCATA 6401 GGACAGTTGG TCGTCAGGAC CTAGTTCACT
AGAGATAGTA GACCGAAGGC CCTGTCAACC AGCAGTCCTG GATCAAGTGA TCTCTATCAT
CTGGCTTCCG 6451 AGGTGGTGGT GGCTGACGTT CATGCCGTCC AAGAGCCTGC
CCCTATTCCA TCCACCACCA CCGACTGCAA GTACGGCAGG TTCTCGGACG GGGATAAGGT
6501 CCGCCAAGGC TAAAGAAGAT GGCCCGCCTG GCAGCGGCAA GAAAAGAGCC
GGCGGTTCCG ATTTCTTCTA CCGGGCGGAC CGTCGCCGTT CTTTTCTCGG 6551
CACTCCACCG GCAAGCAATA GCTCTGAGTC CCTCCACCTC TCTTTTGGTG GTGAGGTGGC
CGTTCGTTAT CGAGACTCAG GGAGGTGGAG AGAAAACCAC 6601 GGGTATCCAT
GTCCCTCGGA TCAATTTTCG ACGGAGAGAC GGCCCGCCAG CCCATAGGTA CAGGGAGCCT
AGTTAAAAGC TGCCTCTCTG CCGGGCGGTC 6651 GCAGCGGTAC AACCCCTGGC
AACAGGCCCC ACGGATGTGC CTATGTCTTT CGTCGCCATG TTGGGGACCG TTGTCCGGGG
TGCCTACACG GATACAGAAA 6701 CGGATCGTTT TCCGACGGAG AGATTGATGA
GCTGAGCCGC AGAGTAACTG GCCTAGCAAA AGGCTGCCTC TCTAACTACT CGACTCGGCG
TCTCATTGAC 6751 AGTCCGAACC CGTCCTGTTT GGATCATTTG AACCGGGCGA
AGTGAACTCA TCAGGCTTGG GCAGGACAAA CCTAGTAAAC TTGGCCCGCT TCACTTGAGT
6801 ATTATATCGT CCCGATCAGC CGTATCTTTT CCACTACGCA AGCAGAGACG
TAATATAGCA GGGCTAGTCG GCATAGAAAA GGTGATGCGT TCGTCTCTGC 6851
TAGACGCAGG AGCAGGAGGA CTGAATACTG ACTAACCGGG GTAGGTGGGT ATCTGCGTCC
TCGTCCTCCT GACTTATGAC TGATTGGCCC CATCCACCCA 6901 ACATATTTTC
GACGGACACA GGCCCTGGGC ACTTGCAAAA GAAGTCCGTT TGTATAAAAG CTGCCTGTGT
CCGGGACCCG TGAACGTTTT CTTCAGGCAA 6951 CTGCAGAACC AGCTTACAGA
ACCGACCTTG GAGCGCAATG TCCTGGAAAG GACGTCTTGG TCGAATGTCT TGGCTGGAAC
CTCGCGTTAC AGGACCTTTC 7001 AATTCATGCC CCGGTGCTCG ACACGTCGAA
AGAGGAACAA CTCAAACTCA TTAAGTACGG GGCCACGAGC TGTGCAGCTT TCTCCTTGTT
GAGTTTGAGT 7051 GGTACCAGAT GATGCCCACC GAAGCCAACA AAAGTAGGTA
CCAGTCTCGT CCATGGTCTA CTACGGGTGG CTTCGGTTGT TTTCATCCAT GGTCAGAGCA
7101 AAAGTAGAAA ATCAGAAAGC CATAACCACT GAGCGACTAC TGTCAGGACT
TTTCATCTTT TAGTCTTTCG GTATTGGTGA CTCGCTGATG ACAGTCCTGA 7151
ACGACTGTAT AACTCTGCCA CAGATCAGCC AGAATGCTAT AAGATCACCT TGCTGACATA
TTGAGACGGT GTCTAGTCGG TCTTACGATA TTCTAGTGGA 7201 ATCCGAAACC
ATTGTACTCC AGTAGCGTAC CGGCGAACTA CTCCGATCCA TAGGCTTTGG TAACATGAGG
TCATCGCATG GCCGCTTGAT GAGGCTAGGT 7251 CAGTTCGCTG TAGCTGTCTG
TAACAACTAT CTGCATGAGA ACTATCCGAC GTCAAGCGAC ATCGACAGAC ATTGTTGATA
GACGTACTCT TGATAGGCTG 7301 AGTAGCATCT TATCAGATTA CTGACGAGTA
CGATGCTTAC TTGGATATGG TCATCGTAGA ATAGTCTAAT GACTGCTCAT GCTACGAATG
AACCTATACC 7351 TAGACGGGAC AGTCGCCTGC CTGGATACTG CAACCTTCTG
CCCCGCTAAG ATCTGCCCTG TCAGCGGACG GACCTATGAC GTTGGAAGAC GGGGCGATTC
7401 CTTAGAAGTT ACCCGAAAAA ACATGAGTAT AGAGCCCCGA ATATCCGCAG
GAATCTTCAA TGGGCTTTTT TGTACTCATA TCTCGGGGCT TATAGGCGTC 7451
TGCGGTTCCA TCAGCGATGC AGAACACGCT ACAAAATGTG CTCATTGCCG ACGCCAAGGT
AGTCGCTACG TCTTGTGCGA TGTTTTACAC GAGTAACGGC 7501 CAACTAAAAG
AAATTGCAAC GTCACGCAGA TGCGTGAACT GCCAACACTG GTTGATTTTC TTTAACGTTG
CAGTGCGTCT ACGCACTTGA CGGTTGTGAC 7551 GACTCAGCGA CATTCAATGT
CGAATGCTTT CGAAAATATG CATGTAATGA CTGAGTCGCT GTAAGTTACA GCTTACGAAA
GCTTTTATAC GTACATTACT 7601 CGAGTATTGG GAGGAGTTCG CTCGGAAGCC
AATTAGGATT ACCACTGAGT GCTCATAACC CTCCTCAAGC GAGCCTTCGG TTAATCCTAA
TGGTGACTCA 7651 TTGTCACCGC ATATGTAGCT AGACTGAAAG GCCCTAAGGC
CGCCGCACTA AACAGTGGCG TATACATCGA TCTGACTTTC CGGGATTCCG GCGGCGTGAT
7701 TTTGCAAAGA CGTATAATTT GGTCCCATTG CAAGAAGTGC CTATGGATAG
AAACGTTTCT GCATATTAAA CCAGGGTAAC GTTCTTCACG GATACCTATC 7751
ATTCGTCATG GACATGAAAA GAGACGTGAA AGTTACACCA GGCACGAAAC TAAGCAGTAC
CTGTACTTTT CTCTGCACTT TCAATGTGGT CCGTGCTTTG 7801 ACACAGAAGA
AAGACCGAAA GTACAAGTGA TACAAGCCGC AGAACCCCTG TGTGTCTTCT TTCTGGCTTT
CATGTTCACT ATGTTCGGCG TCTTGGGGAC 7851 GCGACTGCTT ACTTATGCGG
GATTCACCGG GAATTAGTGC GTAGGCTTAC CGCTGACGAA TGAATACGCC CTAAGTGGCC
CTTAATCACG CATCCGAATG 7901 GGCCGTCTTG CTTCCAAACA TTCACACGCT
TTTTGACATG TCGGCGGAGG CCGGCAGAAC GAAGGTTTGT AAGTGTGCGA AAAACTGTAC
AGCCGCCTCC 7951 ATTTTGATGC AATCATAGCA GAACACTTCA AGCAAGGCGA
CCCGGTACTG TAAAACTACG TTAGTATCGT CTTGTGAAGT TCGTTCCGCT GGGCCATGAC
8001 GAGACGGATA TCGCATCATT CGACAAAAGC CAAGACGACG CTATGGCGTT
CTCTGCCTAT AGCGTAGTAA GCTGTTTTCG GTTCTGCTGC GATACCGCAA 8051
AACCGGTCTG ATGATCTTGG AGGACCTGGG TGTGGATCAA CCACTACTCG TTGGCCAGAC
TACTAGAACC TCCTGGACCC ACACCTAGTT GGTGATGAGC 8101 ACTTGATCGA
GTGCGCCTTT GGAGAAATAT CATCCACCCA TCTACCTACG TGAACTAGCT CACGCGGAAA
CCTCTTTATA GTAGGTGGGT AGATGGATGC 8151 GGTACTCGTT TTAAATTCGG
GGCGATGATG AAATCCGGAA TGTTCCTCAC CCATGAGCAA AATTTAAGCC CCGCTACTAC
TTTAGGCCTT ACAAGGAGTG 8201 ACTTTTTGTC AACACAGTTT TGAATGTCGT
TATCGCCAGC AGAGTACTAG TGAAAAACAG TTGTGTCAAA ACTTACAGCA ATAGCGGTCG
TCTCATGATC 8251 AAGAGCGGCT TAAAACGTCC AGATGTGCAG CGTTCATTGG
CGACGACAAC
TTCTCGCCGA ATTTTGCAGG TCTACACGTC GCAAGTAACC GCTGCTGTTG 8301
ATCATACATG GAGTAGTATC TGACAAAGAA ATGGCTGAGA GGTGCGCCAC TAGTATGTAC
CTCATCATAG ACTGTTTCTT TACCGACTCT CCACGCGGTG 8351 CTGGCTCAAC
ATGGAGGTTA AGATCATCGA CGCAGTCATC GGTGAGAGAC GACCGAGTTG TACCTCCAAT
TCTAGTAGCT GCGTCAGTAG CCACTCTCTG 8401 CACCTTACTT CTGCGGCGGA
TTTATCTTGC AAGATTCGGT TACTTCCACA GTGGAATGAA GACGCCGCCT AAATAGAACG
TTCTAAGCCA ATGAAGGTGT 8451 GCGTGCCGCG TGGCGGACCC CCTGAAAAGG
CTGTTTAAGT TGGGTAAACC CGCACGGCGC ACCGCCTGGG GGACTTTTCC GACAAATTCA
ACCCATTTGG 8501 GCTCCCAGCC GACGACGAGC AAGACGAAGA CAGAAGACGC
GCTCTGCTAG CGAGGGTCGG CTGCTGCTCG TTCTGCTTCT GTCTTCTGCG CGAGACGATC
8551 ATGAAACAAA GGCGTGGTTT AGAGTAGGTA TAACAGGCAC TTTAGCAGTG
TACTTTGTTT CCGCACCAAA TCTCATCCAT ATTGTCCGTG AAATCGTCAC 8601
GCCGTGACGA CCCGGTATGA GGTAGACAAT ATTACACCTG TCCTACTGGC CGGCACTGCT
GGGCCATACT CCATCTGTTA TAATGTGGAC AGGATGACCG 8651 ATTGAGAACT
TTTGCCCAGA GCAAAAGAGC ATTCCAAGCC ATCAGAGGGG TAACTCTTGA AAACGGGTCT
CGTTTTCTCG TAAGGTTCGG TAGTCTCCCC 8701 AAATAAAGCA TCTCTACGGT
GGTCCTAAAT AGTCAGCATA GTACATTTCA TTTATTTCGT AGAGATGCCA CCAGGATTTA
TCAGTCGTAT CATGTAAAGT 8751 TCTGACTAAT ACTACAACAC CACCACCTCT
AGCCCGGGCT CGAGATCTGC AGACTGATTA TGATGTTGTG GTGGTGGAGA TCGGGCCCGA
GCTCTAGACG 8801 GATCTAAGTA AGCTTGGCAT TCCGGTACTG TTGGTAAAGC
CACCATGGAA CTAGATTCAT TCGAACCGTA AGGCCATGAC AACCATTTCG GTGGTACCTT
8851 GACGCCAAAA ACATAAAGAA AGGCCCGGCG CCATTCTATC CGCTGGAAGA
CTGCGGTTTT TGTATTTCTT TCCGGGCCGC GGTAAGATAG GCGACCTTCT 8901
TGGAACCGCT GGAGAGCAAC TGCATAAGGC TATGAAGAGA TACGCCCTGG ACCTTGGCGA
CCTCTCGTTG ACGTATTCCG ATACTTCTCT ATGCGGGACC 8951 TTCCTGGAAC
AATTGCTTTT ACAGATGCAC ATATCGAGGT GGACATCACT AAGGACCTTG TTAACGAAAA
TGTCTACGTG TATAGCTCCA CCTGTAGTGA 9001 TACGCTGAGT ACTTCGAAAT
GTCCGTTCGG TTGGCAGAAG CTATGAAACG ATGCGACTCA TGAAGCTTTA CAGGCAAGCC
AACCGTCTTC GATACTTTGC 9051 ATATGGGCTG AATACAAATC ACAGAATCGT
CGTATGCAGT GAAAACTCTC TATACCCGAC TTATGTTTAG TGTCTTAGCA GCATACGTCA
CTTTTGAGAG 9101 TTCAATTCTT TATGCCGGTG TTGGGCGCGT TATTTATCGG
AGTTGCAGTT AAGTTAAGAA ATACGGCCAC AACCCGCGCA ATAAATAGCC TCAACGTCAA
9151 GCGCCCGCGA ACGACATTTA TAATGAACGT GAATTGCTCA ACAGTATGGG
CGCGGGCGCT TGCTGTAAAT ATTACTTGCA CTTAACGAGT TGTCATACCC 9201
CATTTCGCAG CCTACCGTGG TGTTCGTTTC CAAAAAGGGG TTGCAAAAAA GTAAAGCGTC
GGATGGCACC ACAAGCAAAG GTTTTTCCCC AACGTTTTTT 9251 TTTTGAACGT
GCAAAAAAAG CTCCCAATCA TCCAAAAAAT TATTATCATG AAAACTTGCA CGTTTTTTTC
GAGGGTTAGT AGGTTTTTTA ATAATAGTAC 9301 GATTCTAAAA CGGATTACCA
GGGATTTCAG TCGATGTACA CGTTCGTCAC CTAAGATTTT GCCTAATGGT CCCTAAAGTC
AGCTACATGT GCAAGCAGTG 9351 ATCTCATCTA CCTCCCGGTT TTAATGAATA
CGATTTTGTG CCAGAGTCCT TAGAGTAGAT GGAGGGCCAA AATTACTTAT GCTAAAACAC
GGTCTCAGGA 9401 TCGATAGGGA CAAGACAATT GCACTGATCA TGAACTCCTC
TGGATCTACT AGCTATCCCT GTTCTGTTAA CGTGACTAGT ACTTGAGGAG ACCTAGATGA
9451 GGTCTGCCTA AAGGTGTCGC TCTGCCTCAT AGAACTGCCT GCGTGAGATT
CCAGACGGAT TTCCACAGCG AGACGGAGTA TCTTGACGGA CGCACTCTAA 9501
CTCGCATGCC AGAGATCCTA TTTTTGGCAA TCAAATCATT CCGGATACTG GAGCGTACGG
TCTCTAGGAT AAAAACCGTT AGTTTAGTAA GGCCTATGAC 9551 CGATTTTAAG
TGTTGTTCCA TTCCATCACG GTTTTGGAAT GTTTACTACA GCTAAAATTC ACAACAAGGT
AAGGTAGTGC CAAAACCTTA CAAATGATGT 9601 CTCGGATATT TGATATGTGG
ATTTCGAGTC GTCTTAATGT ATAGATTTGA GAGCCTATAA ACTATACACC TAAAGCTCAG
CAGAATTACA TATCTAAACT 9651 AGAAGAGCTG TTTCTGAGGA GCCTTCAGGA
TTACAAGATT CAAAGTGCGC TCTTCTCGAC AAAGACTCCT CGGAAGTCCT AATGTTCTAA
GTTTCACGCG 9701 TGCTGGTGCC AACCCTATTC TCCTTCTTCG CCAAAAGCAC
TCTGATTGAC ACGACCACGG TTGGGATAAG AGGAAGAAGC GGTTTTCGTG AGACTAACTG
9751 AAATACGATT TATCTAATTT ACACGAAATT GCTTCTGGTG GCGCTCCCCT
TTTATGCTAA ATAGATTAAA TGTGCTTTAA CGAAGACCAC CGCGAGGGGA 9801
CTCTAAGGAA GTCGGGGAAG CGGTTGCCAA GAGGTTCCAT CTGCCAGGTA GAGATTCCTT
CAGCCCCTTC GCCAACGGTT CTCCAAGGTA GACGGTCCAT 9851 TCAGGCAAGG
ATATGGGCTC ACTGAGACTA CATCAGCTAT TCTGATTACA AGTCCGTTCC TATACCCGAG
TGACTCTGAT GTAGTCGATA AGACTAATGT 9901 CCCGAGGGGG ATGATAAACC
GGGCGCGGTC GGTAAAGTTG TTCCATTTTT GGGCTCCCCC TACTATTTGG CCCGCGCCAG
CCATTTCAAC AAGGTAAAAA 9951 TGAAGCGAAG GTTGTGGATC TGGATACCGG
GAAAACGCTG GGCGTTAATC ACTTCGCTTC CAACACCTAG ACCTATGGCC CTTTTGCGAC
CCGCAATTAG 10001 AAAGAGGCGA ACTGTGTGTG AGAGGTCCTA TGATTATGTC
CGGTTATGTA TTTCTCCGCT TGACACACAC TCTCCAGGAT ACTAATACAG GCCAATACAT
10051 AACAATCCGG AAGCGACCAA CGCCTTGATT GACAAGGATG GATGGCTACA
TTGTTAGGCC TTCGCTGGTT GCGGAACTAA CTGTTCCTAC CTACCGATGT 10101
TTCTGGAGAC ATAGCTTACT GGGACGAAGA CGAACACTTC TTCATCGTTG AAGACCTCTG
TATCGAATGA CCCTGCTTCT GCTTGTGAAG AAGTAGCAAC 10151 ACCGCCTGAA
GTCTCTGATT AAGTACAAAG GCTATCAGGT GGCTCCCGCT TGGCGGACTT CAGAGACTAA
TTCATGTTTC CGATAGTCCA CCGAGGGCGA 10201 GAATTGGAAT CCATCTTGCT
CCAACACCCC AACATCTTCG ACGCAGGTGT CTTAACCTTA GGTAGAACGA GGTTGTGGGG
TTGTAGAAGC TGCGTCCACA 10251 CGCAGGTCTT CCCGACGATG ACGCCGGTGA
ACTTCCCGCC GCCGTTGTTG GCGTCCAGAA GGGCTGCTAC TGCGGCCACT TGAAGGGCGG
CGGCAACAAC 10301 TTTTGGAGCA CGGAAAGACG ATGACGGAAA AAGAGATCGT
GGATTACGTC AAAACCTCGT GCCTTTCTGC TACTGCCTTT TTCTCTAGCA CCTAATGCAG
10351 GCCAGTCAAG TAACAACCGC GAAAAAGTTG CGCGGAGGAG TTGTGTTTGT
CGGTCAGTTC ATTGTTGGCG CTTTTTCAAC GCGCCTCCTC AACACAAACA 10401
GGACGAAGTA CCGAAAGGTC TTACCGGAAA ACTCGACGCA AGAAAAATCA CCTGCTTCAT
GGCTTTCCAG AATGGCCTTT TGAGCTGCGT TCTTTTTAGT 10451 GAGAGATCCT
CATAAAGGCC AAGAAGGGCG GAAAGATCGC CGTGTAATTC CTCTCTAGGA GTATTTCCGG
TTCTTCCCGC CTTTCTAGCG GCACATTAAG 10501 TAGAGGCGCG CCGATCTCAC
GATCCCCTGA AAAGGCTGTT TAAGTTGGGT ATCTCCGCGC GGCTAGAGTG CTAGGGGACT
TTTCCGACAA ATTCAACCCA 10551 AAACCGCTCC CAGCCGACGA CGAGCAAGAC
GAAGACAGAA GACGCGCTCT TTTGGCGAGG GTCGGCTGCT GCTCGTTCTG CTTCTGTCTT
CTGCGCGAGA 10601 GCTAGATGAA ACAAAGGCGT GGTTTAGAGT AGGTATAACA
GGCACTTTAG CGATCTACTT TGTTTCCGCA CCAAATCTCA TCCATATTGT CCGTGAAATC
10651 CAGTGGCCGT GACGACCCGG TATGAGGTAG ACAATATTAC ACCTGTCCTA
GTCACCGGCA CTGCTGGGCC ATACTCCATC TGTTATAATG TGGACAGGAT 10701
CTGGCATTGA GAACTTTTGC CCAGAGCAAA AGAGCATTCC AAGCCATCAG GACCGTAACT
CTTGAAAACG GGTCTCGTTT TCTCGTAAGG TTCGGTAGTC 10751 AGGGGAAATA
AAGCATCTCT ACGGTGGTCC TAAATAGTCA GCATAGTACA TCCCCTTTAT TTCGTAGAGA
TGCCACCAGG ATTTATCAGT CGTATCATGT 10801 TTTCATCTGA CTAATACTAC
AACACCACCA CCATGAATAG AGGATTCTTT AAAGTAGACT GATTATGATG TTGTGGTGGT
GGTACTTATC TCCTAAGAAA 10851 AACATGCTCG GCCGCCGCCC CTTCCCGGCC
CCCACTGCCA TGTGGAGGCC TTGTACGAGC CGGCGGCGGG GAAGGGCCGG GGGTGACGGT
ACACCTCCGG 10901 GCGGAGAAGG AGGCAGGCGG CCCCGATGCC TGCCCGCAAC
GGGCTGGCTT CGCCTCTTCC TCCGTCCGCC GGGGCTACGG ACGGGCGTTG CCCGACCGAA
10951 CTCAAATCCA GCAACTGACC ACAGCCGTCA GTGCCCTAGT CATTGGACAG
GAGTTTAGGT CGTTGACTGG TGTCGGCAGT CACGGGATCA GTAACCTGTC 11001
GCAACTAGAC CTCAACCCCC ACGTCCACGC CAGCCACCGC GCCAGAAGAA CGTTGATCTG
GAGTTGGGGG TGCAGGTGCG GTCGGTGGCG CGGTCTTCTT 11051 GCAGGCGCCC
AAGCAACCAC CGAAGCCGAA GAAACCAAAA ACGCAGGAGA CGTCCGCGGG TTCGTTGGTG
GCTTCGGCTT CTTTGGTTTT TGCGTCCTCT 11101 AGAAGAAGAA GCAACCTGCA
AAACCCAAAC CCGGAAAGAG ACAGCGCATG TCTTCTTCTT CGTTGGACGT TTTGGGTTTG
GGCCTTTCTC TGTCGCGTAC 11151 GCACTTAAGT TGGAGGCCGA CAGATTGTTC
GACGTCAAGA ACGAGGACGG CGTGAATTCA ACCTCCGGCT GTCTAACAAG CTGCAGTTCT
TGCTCCTGCC 11201 AGATGTCATC GGGCACGCAC TGGCCATGGA AGGAAAGGTA
ATGAAACCTC TCTACAGTAG CCCGTGCGTG ACCGGTACCT TCCTTTCCAT TACTTTGGAG
11251 TGCACGTGAA AGGAACCATC GACCACCCTG TGCTATCAAA GCTCAAATTT
ACGTGCACTT TCCTTGGTAG CTGGTGGGAC ACGATAGTTT CGAGTTTAAA 11301
ACCAAGTCGT CAGCATACGA CATGGAGTTC GCACAGTTGC CAGTCAACAT TGGTTCAGCA
GTCGTATGCT GTACCTCAAG CGTGTCAACG GTCAGTTGTA 11351 GAGAAGTGAG
GCATTCACCT ACACCAGTGA ACACCCCGAA GGATTCTATA CTCTTCACTC CGTAAGTGGA
TGTGGTCACT TGTGGGGCTT CCTAAGATAT 11401 ACTGGCACCA CGGAGCGGTG
CAGTATAGTG GAGGTAGATT TACCATCCCT TGACCGTGGT GCCTCGCCAC GTCATATCAC
CTCCATCTAA ATGGTAGGGA 11451 CGCGGAGTAG GAGGCAGAGG AGACAGCGGT
CGTCCGATCA TGGATAACTC GCGCCTCATC CTCCGTCTCC TCTGTCGCCA GCAGGCTAGT
ACCTATTGAG 11501 CGGTCGGGTT GTCGCGATAG TCCTCGGTGG AGCTGATGAA
GGAACACGAA GCCAGCCCAA CAGCGCTATC AGGAGCCACC TCGACTACTT CCTTGTGCTT
11551 CTGCCCTTTC GGTCGTCACC TGGAATAGTA AAGGGAAGAC AATTAAGACG
GACGGGAAAG CCAGCAGTGG ACCTTATCAT TTCCCTTCTG TTAATTCTGC 11601
ACCCCGGAAG GGACAGAAGA GTGGTCCGCA GCACCACTGG TCACGGCAAT TGGGGCCTTC
CCTGTCTTCT CACCAGGCGT CGTGGTGACC AGTGCCGTTA 11651 GTGTTTGCTC
GGAAATGTGA GCTTCCCATG CGACCGCCCG CCCACATGCT CACAAACGAG CCTTTACACT
CGAAGGGTAC GCTGGCGGGC GGGTGTACGA 11701 ATACCCGCGA ACCTTCCAGA
GCCCTCGACA TCCTTGAAGA GAACGTGAAC TATGGGCGCT TGGAAGGTCT CGGGAGCTGT
AGGAACTTCT CTTGCACTTG 11751 CATGAGGCCT ACGATACCCT GCTCAATGCC
ATATTGCGGT GCGGATCGTC GTACTCCGGA TGCTATGGGA CGAGTTACGG TATAACGCCA
CGCCTAGCAG 11801 TGGCAGAAGC AAAAGAAGCG TCATCGATGA CTTTACCCTG
ACCAGCCCCT ACCGTCTTCG TTTTCTTCGC AGTAGCTACT GAAATGGGAC TGGTCGGGGA
11851 ACTTGGGCAC ATGCTCGTAC TGCCACCATA CTGAACCGTG CTTCAGCCCT
TGAACCCGTG TACGAGCATG ACGGTGGTAT GACTTGGCAC GAAGTCGGGA 11901
GTTAAGATCG AGCAGGTCTG GGACGAAGCG GACGATAACA CCATACGCAT CAATTCTAGC
TCGTCCAGAC CCTGCTTCGC CTGCTATTGT GGTATGCGTA 11951 ACAGACTTCC
GCCCAGTTTG GATACGACCA AAGCGGAGCA GCAAGCGCAA TGTCTGAAGG CGGGTCAAAC
CTATGCTGGT TTCGCCTCGT CGTTCGCGTT 12001 ACAAGTACCG CTACATGTCG
CTTAAGCAGG ATCACACCGT TAAAGAAGGC TGTTCATGGC GATGTACAGC GAATTCGTCC
TAGTGTGGCA ATTTCTTCCG 12051 ACCATGGATG ACATCAAGAT TAGCACCTCA
GGACCGTGTA GAAGGCTTAG TGGTACCTAC TGTAGTTCTA ATCGTGGAGT CCTGGCACAT
CTTCCGAATC 12101 CTACAAAGGA TACTTTCTCC TCGCAAAATG CCCTCCAGGG
GACAGCGTAA GATGTTTCCT ATGAAAGAGG AGCGTTTTAC GGGAGGTCCC CTGTCGCATT
12151 CGGTTAGCAT AGTGAGTAGC AACTCAGCAA CGTCATGTAC ACTGGCCCGC
GCCAATCGTA TCACTCATCG TTGAGTCGTT GCAGTACATG TGACCGGGCG 12201
AAGATAAAAC CAAAATTCGT GGGACGGGAA AAATATGATC TACCTCCCGT TTCTATTTTG
GTTTTAAGCA CCCTGCCCTT TTTATACTAG ATGGAGGGCA 12251 TCACGGTAAA
AAAATTCCTT GCACAGTGTA CGACCGTCTG AAAGAAACAA AGTGCCATTT TTTTAAGGAA
CGTGTCACAT GCTGGCAGAC TTTCTTTGTT 12301 CTGCAGGCTA CATCACTATG
CACAGGCCGG GCCCGCACGC TTATACATCC GACGTCCGAT GTAGTGATAC GTGTCCGGCC
CGGGCGTGCG AATATGTAGG 12351 TACCTGGAAG AATCATCAGG GAAAGTTTAC
GCAAAGCCGC CATCTGGGAA ATGGACCTTC TTAGTAGTCC CTTTCAAATG CGTTTCGGCG
GTAGACCCTT 12401 GAACATTACG TATGAGTGCA AGTGCGGCGA CTACAAGACC
GGAACCGTTT CTTGTAATGC ATACTCACGT TCACGCCGCT GATGTTCTGG
CCTTGGCAAA
12451 CGACCCGCAC CGAAATCACT GGTTGCACCG CCATCAAGCA GTGCGTCGCC
GCTGGGCGTG GCTTTAGTGA CCAACGTGGC GGTAGTTCGT CACGCAGCGG 12501
TATAAGAGCG ACCAAACGAA GTGGGTCTTC AACTCACCGG ACTTGATCCG ATATTCTCGC
TGGTTTGCTT CACCCAGAAG TTGAGTGGCC TGAACTAGGC 12551 ACATGACGAC
CACACGGTCC AAGGGAAATT GCATTTGCCT TTCAAGTTGA TGTACTGCTG GTGTGCCAGG
TTCCCTTTAA CGTAAACGGA AAGTTCAACT 12601 TCCCGAGTAC CTGCATGGTC
CCTGTTGCCC ACGCGCCGAA TGTAATACAT AGGGCTCATG GACGTACCAG GGACAACGGG
TGCGCGGCTT ACATTATGTA 12651 GGCTTTAAAC ACATCAGCCT CCAATTAGAT
ACAGACCACT TGACATTGCT CCGAAATTTG TGTAGTCGGA GGTTAATCTA TGTCTGGTGA
ACTGTAACGA 12701 CACCACCAGG AGACTAGGGG CAAACCCGGA ACCAACCACT
GAATGGATCG GTGGTGGTCC TCTGATCCCC GTTTGGGCCT TGGTTGGTGA CTTACCTAGC
12751 TCGGAAAGAC GGTCAGAAAC TTCACCGTCG ACCGAGATGG CCTGGAATAC
AGCCTTTCTG CCAGTCTTTG AAGTGGCAGC TGGCTCTACC GGACCTTATG 12801
ATATGGGGAA ATCATGAGCC AGTGAGGGTC TATGCCCAAG AGTCAGCACC TATACCCCTT
TAGTACTCGG TCACTCCCAG ATACGGGTTC TCAGTCGTGG 12851 AGGAGACCCT
CACGGATGGC CACACGAAAT AGTACAGCAT TACTACCATC TCCTCTGGGA GTGCCTACCG
GTGTGCTTTA TCATGTCGTA ATGATGGTAG 12901 GCCATCCTGT GTACACCATC
TTAGCCGTCG CATCAGCTAC CGTGGCGATG CGGTAGGACA CATGTGGTAG AATCGGCAGC
GTAGTCGATG GCACCGCTAC 12951 ATGATTGGCG TAACTGTTGC AGTGTTATGT
GCCTGTAAAG CGCGCCGTGA TACTAACCGC ATTGACAACG TCACAATACA CGGACATTTC
GCGCGGCACT 13001 GTGCCTGACG CCATACGCCC TGGCCCCAAA CGCCGTAATC
CCAACTTCGC CACGGACTGC GGTATGCGGG ACCGGGGTTT GCGGCATTAG GGTTGAAGCG
13051 TGGCACTCTT GTGCTGCGTT AGGTCGGCCA ATGCTGAAAC GTTCACCGAG
ACCGTGAGAA CACGACGCAA TCCAGCCGGT TACGACTTTG CAAGTGGCTC 13101
ACCATGAGTT ACTTGTGGTC GAACAGTCAG CCGTTCTTCT GGGTCCAGTT TGGTACTCAA
TGAACACCAG CTTGTCAGTC GGCAAGAAGA CCCAGGTCAA 13151 GTGCATACCT
TTGGCCGCTT TCATCGTTCT AATGCGCTGC TGCTCCTGCT CACGTATGGA AACCGGCGAA
AGTAGCAAGA TTACGCGACG ACGAGGACGA 13201 GCCTGCCTTT TTTAGTGGTT
GCCGGCGCCT ACCTGGCGAA GGTAGACGCC CGGACGGAAA AAATCACCAA CGGCCGCGGA
TGGACCGCTT CCATCTGCGG 13251 TACGAACATG CGACCACTGT TCCAAATGTG
CCACAGATAC CGTATAAGGC ATGCTTGTAC GCTGGTGACA AGGTTTACAC GGTGTCTATG
GCATATTCCG 13301 ACTTGTTGAA AGGGCAGGGT ATGCCCCGCT CAATTTGGAG
ATCACTGTCA TGAACAACTT TCCCGTCCCA TACGGGGCGA GTTAAACCTC TAGTGACAGT
13351 TGTCCTCGGA GGTTTTGCCT TCCACCAACC AAGAGTACAT TACCTGCAAA
ACAGGAGCCT CCAAAACGGA AGGTGGTTGG TTCTCATGTA ATGGACGTTT 13401
TTCACCACTG TGGTCCCCTC CCCAAAAATC AAATGCTGCG GCTCCTTGGA AAGTGGTGAC
ACCAGGGGAG GGGTTTTTAG TTTACGACGC CGAGGAACCT 13451 ATGTCAGCCG
GCCGTTCATG CAGACTATAC CTGCAAGGTC TTCGGAGGGG TACAGTCGGC CGGCAAGTAC
GTCTGATATG GACGTTCCAG AAGCCTCCCC 13501 TCTACCCCTT TATGTGGGGA
GGAGCGCAAT GTTTTTGCGA CAGTGAGAAC AGATGGGGAA ATACACCCCT CCTCGCGTTA
CAAAAACGCT GTCACTCTTG 13551 AGCCAGATGA GTGAGGCGTA CGTCGAACTG
TCAGCAGATT GCGCGTCTGA TCGGTCTACT CACTCCGCAT GCAGCTTGAC AGTCGTCTAA
CGCGCAGACT 13601 CCACGCGCAG GCGATTAAGG TGCACACTGC CGCGATGAAA
GTAGGACTGC GGTGCGCGTC CGCTAATTCC ACGTGTGACG GCGCTACTTT CATCCTGACG
13651 GTATAGTGTA CGGGAACACT ACCAGTTTCC TAGATGTGTA CGTGAACGGA
CATATCACAT GCCCTTGTGA TGGTCAAAGG ATCTACACAT GCACTTGCCT 13701
GTCACACCAG GAACGTCTAA AGACTTGAAA GTCATAGCTG GACCAATTTC CAGTGTGGTC
CTTGCAGATT TCTGAACTTT CAGTATCGAC CTGGTTAAAG 13751 AGCATCGTTT
ACGCCATTCG ATCATAAGGT CGTTATCCAT CGCGGCCTGG TCGTAGCAAA TGCGGTAAGC
TAGTATTCCA GCAATAGGTA GCGCCGGACC 13801 TGTACAACTA TGACTTCCCG
GAATATGGAG CGATGAAACC AGGAGCGTTT ACATGTTGAT ACTGAAGGGC CTTATACCTC
GCTACTTTGG TCCTCGCAAA 13851 GGAGACATTC AAGCTACCTC CTTGACTAGC
AAGGATCTCA TCGCCAGCAC CCTCTGTAAG TTCGATGGAG GAACTGATCG TTCCTAGAGT
AGCGGTCGTG 13901 AGACATTAGG CTACTCAAGC CTTCCGCCAA GAACGTGCAT
GTCCCGTACA TCTGTAATCC GATGAGTTCG GAAGGCGGTT CTTGCACGTA CAGGGCATGT
13951 CGCAGGCCGC ATCAGGATTT GAGATGTGGA AAAACAACTC AGGCCGCCCA
GCGTCCGGCG TAGTCCTAAA CTCTACACCT TTTTGTTGAG TCCGGCGGGT 14001
CTGCAGGAAA CCGCACCTTT CGGGTGTAAG ATTGCAGTAA ATCCGCTCCG GACGTCCTTT
GGCGTGGAAA GCCCACATTC TAACGTCATT TAGGCGAGGC 14051 AGCGGTGGAC
TGTTCATACG GGAACATTCC CATTTCTATT GACATCCCGA TCGCCACCTG ACAAGTATGC
CCTTGTAAGG GTAAAGATAA CTGTAGGGCT 14101 ACGCTGCCTT TATCAGGACA
TCAGATGCAC CACTGGTCTC AACAGTCAAA TGCGACGGAA ATAGTCCTGT AGTCTACGTG
GTGACCAGAG TTGTCAGTTT 14151 TGTGAAGTCA GTGAGTGCAC TTATTCAGCA
GACTTCGGCG GGATGGCCAC ACACTTCAGT CACTCACGTG AATAAGTCGT CTGAAGCCGC
CCTACCGGTG 14201 CCTGCAGTAT GTATCCGACC GCGAAGGTCA ATGCCCCGTA
CATTCGCATT GGACGTCATA CATAGGCTGG CGCTTCCAGT TACGGGGCAT GTAAGCGTAA
14251 CGAGCACAGC AACTCTCCAA GAGTCGACAG TACATGTCCT GGAGAAAGGA
GCTCGTGTCG TTGAGAGGTT CTCAGCTGTC ATGTACAGGA CCTCTTTCCT 14301
GCGGTGACAG TACACTTTAG CACCGCGAGT CCACAGGCGA ACTTTATCGT CGCCACTGTC
ATGTGAAATC GTGGCGCTCA GGTGTCCGCT TGAAATAGCA 14351 ATCGCTGTGT
GGGAAGAAGA CAACATGCAA TGCAGAATGT AAACCACCAG TAGCGACACA CCCTTCTTCT
GTTGTACGTT ACGTCTTACA TTTGGTGGTC 14401 CTGACCATAT CGTGAGCACC
CCGCACAAAA ATGACCAAGA ATTTCAAGCC GACTGGTATA GCACTCGTGG GGCGTGTTTT
TACTGGTTCT TAAAGTTCGG 14451 GCCATCTCAA AAACATCATG GAGTTGGCTG
TTTGCCCTTT TCGGCGGCGC CGGTAGAGTT TTTGTAGTAC CTCAACCGAC AAACGGGAAA
AGCCGCCGCG 14501 CTCGTCGCTA TTAATTATAG GACTTATGAT TTTTGCTTGC
AGCATGATGC GAGCAGCGAT AATTAATATC CTGAATACTA AAAACGAACG TCGTACTACG
14551 TGACTAGCAC ACGAAGATGA CCGCTACGCC CCAATGATCC GACCAGCAAA
ACTGATCGTG TGCTTCTACT GGCGATGCGG GGTTACTAGG CTGGTCGTTT 14601
ACTCGATGTA CTTCCGAGGA ACTGATGTGC ATAAGTGAGC ATGCGTTTAA TGAGCTACAT
GAAGGCTCCT TGACTACACG TATTCACTCG TACGCAAATT 14651 ACTGGGCCCA
ATGTTCCCCA ATGATCCGAC CAGCAAAACT CGATGTACTT TGACCCGGGT TACAAGGGGT
TACTAGGCTG GTCGTTTTGA GCTACATGAA 14701 CCGAGGAACT GATGTGCATA
ATGCATCAGG CTGGTACATT AGATCCCCGC GGCTCCTTGA CTACACGTAT TACGTAGTCC
GACCATGTAA TCTAGGGGCG 14751 TTACCGCGGG CAATATAGCA ACACTAAAAA
CTCGATGTAC TTCCGAGGAA AATGGCGCCC GTTATATCGT TGTGATTTTT GAGCTACATG
AAGGCTCCTT 14801 GCGCAGTGCA TAATGCTGCG CAGTGTTGCC ACATAACCAC
TATATTAACC CGCGTCACGT ATTACGACGC GTCACAACGG TGTATTGGTG ATATAATTGG
14851 ATTTATCTAG CGGACGCCAA AAACTCAATG TATTTCTGAG GAAGCGTGGT
TAAATAGATC GCCTGCGGTT TTTGAGTTAC ATAAAGACTC CTTCGCACCA 14901
GCATAATGCC ACGCAGCGTC TGCATAACTT TTATTATTTC TTTTATTAAT CGTATTACGG
TGCGTCGCAG ACGTATTGAA AATAATAAAG AAAATAATTA 14951 CAACAAAATT
TTGTTTTTAA CATTTCAAAA AAAAAAAAAA AAAAAAAAAA GTTGTTTTAA AACAAAAATT
GTAAAGTTTT TTTTTTTTTT TTTTTTTTTT 15001 AAAAAAAAAA AAAATTTAAA
TTAATTAAGC GGCCGCCTCG AGGACGTCAG TTTTTTTTTT TTTTAAATTT AATTAATTCG
CCGGCGGAGC TCCTGCAGTC 15051 GTGGCACTTT TCGGGGAAAT GTGCGCGGAA
CCCCTATTTG TTTATTTTTC CACCGTGAAA AGCCCCTTTA CACGCGCCTT GGGGATAAAC
AAATAAAAAG 15101 TAAATACATT CAAATATGTA TCCGCTCATG AGACAATAAC
CCTGATAAAT ATTTATGTAA GTTTATACAT AGGCGAGTAC TCTGTTATTG GGACTATTTA
15151 GCTTCAATAA TATTGAAAAA GGAAGAGTAT GAGTATTCAA CATTTCCGTG
CGAAGTTATT ATAACTTTTT CCTTCTCATA CTCATAAGTT GTAAAGGCAC 15201
TCGCCCTTAT TCCCTTTTTT GCGGCATTTT GCCTTCCTGT TTTTGCTCAC AGCGGGAATA
AGGGAAAAAA CGCCGTAAAA CGGAAGGACA AAAACGAGTG 15251 CCAGAAACGC
TGGTGAAAGT AAAAGATGCT GAAGATCAGT TGGGTGCACG GGTCTTTGCG ACCACTTTCA
TTTTCTACGA CTTCTAGTCA ACCCACGTGC 15301 AGTGGGTTAC ATCGAACTGG
ATCTCAACAG CGGTAAGATC CTTGAGAGTT TCACCCAATG TAGCTTGACC TAGAGTTGTC
GCCATTCTAG GAACTCTCAA 15351 TTCGCCCCGA AGAACGTTTT CCAATGATGA
GCACTTTTAA AGTTCTGCTA AAGCGGGGCT TCTTGCAAAA GGTTACTACT CGTGAAAATT
TCAAGACGAT 15401 TGTGGCGCGG TATTATCCCG TATTGACGCC GGGCAAGAGC
AACTCGGTCG ACACCGCGCC ATAATAGGGC ATAACTGCGG CCCGTTCTCG TTGAGCCAGC
15451 CCGCATACAC TATTCTCAGA ATGACTTGGT TGAGTACTCA CCAGTCACAG
GGCGTATGTG ATAAGAGTCT TACTGAACCA ACTCATGAGT GGTCAGTGTC 15501
AAAAGCATCT TACGGATGGC ATGACAGTAA GAGAATTATG CAGTGCTGCC TTTTCGTAGA
ATGCCTACCG TACTGTCATT CTCTTAATAC GTCACGACGG 15551 ATAACCATGA
GTGATAACAC TGCGGCCAAC TTACTTCTGA CAACGATCGG TATTGGTACT CACTATTGTG
ACGCCGGTTG AATGAAGACT GTTGCTAGCC 15601 AGGACCGAAG GAGCTAACCG
CTTTTTTGCA CAACATGGGG GATCATGTAA TCCTGGCTTC CTCGATTGGC GAAAAAACGT
GTTGTACCCC CTAGTACATT 15651 CTCGCCTTGA TCGTTGGGAA CCGGAGCTGA
ATGAAGCCAT ACCAAACGAC GAGCGGAACT AGCAACCCTT GGCCTCGACT TACTTCGGTA
TGGTTTGCTG 15701 GAGCGTGACA CCACGATGCC TGTAGCAATG GCAACAACGT
TGCGCAAACT CTCGCACTGT GGTGCTACGG ACATCGTTAC CGTTGTTGCA ACGCGTTTGA
15751 ATTAACTGGC GAACTACTTA CTCTAGCTTC CCGGCAACAA TTAATAGACT
TAATTGACCG CTTGATGAAT GAGATCGAAG GGCCGTTGTT AATTATCTGA 15801
GGATGGAGGC GGATAAAGTT GCAGGACCAC TTCTGCGCTC GGCCCTTCCG CCTACCTCCG
CCTATTTCAA CGTCCTGGTG AAGACGCGAG CCGGGAAGGC 15851 GCTGGCTGGT
TTATTGCTGA TAAATCTGGA GCCGGTGAGC GTGGGTCTCG CGACCGACCA AATAACGACT
ATTTAGACCT CGGCCACTCG CACCCAGAGC 15901 CGGTATCATT GCAGCACTGG
GGCCAGATGG TAAGCCCTCC CGTATCGTAG GCCATAGTAA CGTCGTGACC CCGGTCTACC
ATTCGGGAGG GCATAGCATC 15951 TTATCTACAC GACGGGGAGT CAGGCAACTA
TGGATGAACG AAATAGACAG AATAGATGTG CTGCCCCTCA GTCCGTTGAT ACCTACTTGC
TTTATCTGTC 16001 ATCGCTGAGA TAGGTGCCTC ACTGATTAAG CATTGGTAAC
TGTCAGACCA TAGCGACTCT ATCCACGGAG TGACTAATTC GTAACCATTG ACAGTCTGGT
16051 AGTTTACTCA TATATACTTT AGATTGATTT AAAACTTCAT TTTTAATTTA
TCAAATGAGT ATATATGAAA TCTAACTAAA TTTTGAAGTA AAAATTAAAT 16101
AAAGGATCTA GGTGAAGATC CTTTTTGATA ATCTCATGAC CAAAATCCCT TTTCCTAGAT
CCACTTCTAG GAAAAACTAT TAGAGTACTG GTTTTAGGGA 16151 TAACGTGAGT
TTTCGTTCCA CTGAGCGTCA GACCCCGTAG AAAAGATCAA ATTGCACTCA AAAGCAAGGT
GACTCGCAGT CTGGGGCATC TTTTCTAGTT 16201 AGGATCTTCT TGAGATCCTT
TTTTTCTGCG CGTAATCTGC TGCTTGCAAA TCCTAGAAGA ACTCTAGGAA AAAAAGACGC
GCATTAGACG ACGAACGTTT 16251 CAAAAAAACC ACCGCTACCA GCGGTGGTTT
GTTTGCCGGA TCAAGAGCTA GTTTTTTTGG TGGCGATGGT CGCCACCAAA CAAACGGCCT
AGTTCTCGAT 16301 CCAACTCTTT TTCCGAAGGT AACTGGCTTC AGCAGAGCGC
AGATACCAAA GGTTGAGAAA AAGGCTTCCA TTGACCGAAG TCGTCTCGCG TCTATGGTTT
16351 TACTGGTCTT CTAGTGTAGC CGTAGTTAGG CCACCACTTC AAGAACTCTG
ATGACCAGAA GATCACATCG GCATCAATCC GGTGGTGAAG TTCTTGAGAC 16401
TAGCACCGCC TACATACCTC GCTCTGCTAA TCCTGTTACC AGTGGCTGCT ATCGTGGCGG
ATGTATGGAG CGAGACGATT AGGACAATGG TCACCGACGA 16451 GCCAGTGGCG
ATAAGTCGTG TCTTACCGGG TTGGACTCAA GACGATAGTT CGGTCACCGC TATTCAGCAC
AGAATGGCCC AACCTGAGTT CTGCTATCAA 16501 ACCGGATAAG GCGCAGCGGT
CGGGCTGAAC GGGGGGTTCG TGCACACAGC TGGCCTATTC CGCGTCGCCA GCCCGACTTG
CCCCCCAAGC ACGTGTGTCG 16551 CCAGCTTGGA GCGAACGACC TACACCGAAC
TGAGATACCT ACAGCGTGAG GGTCGAACCT CGCTTGCTGG ATGTGGCTTG ACTCTATGGA
TGTCGCACTC 16601 CTATGAGAAA GCGCCACGCT TCCCGAAGGG AGAAAGGCGG
ACAGGTATCC GATACTCTTT CGCGGTGCGA AGGGCTTCCC TCTTTCCGCC
TGTCCATAGG
16651 GGTAAGCGGC AGGGTCGGAA CAGGAGAGCG CACGAGGGAG CTTCCAGGGG
CCATTCGCCG TCCCAGCCTT GTCCTCTCGC GTGCTCCCTC GAAGGTCCCC 16701
GAAACGCCTG GTATCTTTAT AGTCCTGTCG GGTTTCGCCA CCTCTGACTT CTTTGCGGAC
CATAGAAATA TCAGGACAGC CCAAAGCGGT GGAGACTGAA 16751 GAGCGTCGAT
TTTTGTGATG CTCGTCAGGG GGGCGGAGCC TATGGAAAAA CTCGCAGCTA AAAACACTAC
GAGCAGTCCC CCCGCCTCGG ATACCTTTTT 16801 CGCCAGCAAC GCGGCCTTTT
TACGGTTCCT GGCCTTTTGC TGGCCTTTTG GCGGTCGTTG CGCCGGAAAA ATGCCAAGGA
CCGGAAAACG ACCGGAAAAC 16851 CTCACATGTG GGAGGCTAGA GTACATTTAG
GTGACACTAT AGAA GAGTGTACAC CCTCCGATCT CATGTAAATC CACTGTGATA
TCTT
[0090] The present invention is not to be limited in scope by the
specific embodiments described herein. Indeed, various
modifications of the invention in addition to those described
herein will become apparent to those skilled in the art from the
foregoing description and the accompanying figures. Such
modifications are intended to fall within the scope of the appended
claims.
[0091] It is further to be understood that all values are
approximate, and are provided for description. Patents, patent
applications, publications, product descriptions, and protocols are
cited throughout this application, the disclosures of which are
incorporated herein by reference in their entireties for all
purposes.
Sequence CWU 1
1
8116894DNAArtificial SequenceSynthetic polynucleotide 1attgacggcg
tagtacacac tattgaatca aacagccgac caattgcact accatcacaa 60tggagaagcc
agtagtaaac gtagacgtag acccccagag tccgtttgtc gtgcaactgc
120aaaaaagctt cccgcaattt gaggtagtag cacagcaggt cactccaaat
gaccatgcta 180atgccagagc attttcgcat ctggccagta aactaatcga
gctggaggtt cctaccacag 240cgacgatctt ggacataggc agcgcaccgg
ctcgtagaat gttttccgag caccagtatc 300attgtgtctg ccccatgcgt
agtccagaag acccggaccg catgatgaaa tatgccagta 360aactggcgga
aaaagcgtgc aagattacaa acaagaactt gcatgagaag attaaggatc
420tccggaccgt acttgatacg ccggatgctg aaacaccatc gctctgcttt
cacaacgatg 480ttacctgcaa catgcgtgcc gaatattccg tcatgcagga
cgtgtatatc aacgctcccg 540gaactatcta tcatcaggct atgaaaggcg
tgcggaccct gtactggatt ggcttcgaca 600ccacccagtt catgttctcg
gctatggcag gttcgtaccc tgcgtacaac accaactggg 660ccgacgagaa
agtccttgaa gcgcgtaaca tcggactttg cagcacaaag ctgagtgaag
720gtaggacagg aaaattgtcg ataatgagga agaaggagtt gaagcccggg
tcgcgggttt 780atttctccgt aggatcgaca ctttatccag aacacagagc
cagcttgcag agctggcatc 840ttccatcggt gttccacttg aatggaaagc
agtcgtacac ttgccgctgt gatacagtgg 900tgagttgcga aggctacgta
gtgaagaaaa tcaccatcag tcccgggatc acgggagaaa 960ccgtgggata
cgcggttaca cacaatagcg agggcttctt gctatgcaaa gttactgaca
1020cagtaaaagg agaacgggta tcgttccctg tgtgcacgta catcccggcc
accatatgcg 1080atcagatgac tggtataatg gccacggata tatcacctga
cgatgcacaa aaacttctgg 1140ttgggctcaa ccagcgaatt gtcattaacg
gtaggactaa caggaacacc aacaccatgc 1200aaaattacct tctgccgatc
atagcacaag ggttcagcaa atgggctaag gagcgcaagg 1260atgatcttga
taacgagaaa atgctgggta ctagagaacg caagcttacg tatggctgct
1320tgtgggcgtt tcgcactaag aaagtacatt cgttttatcg cccacctgga
acgcagacca 1380tcgtaaaagt cccagcctct tttagcgctt ttcccatgtc
gtccgtatgg acgacctctt 1440tgcccatgtc gctgaggcag aaattgaaac
tggcattgca accaaagaag gaggaaaaac 1500tgctgcaggt ctcggaggaa
ttagtcatgg aggccaaggc tgcttttgag gatgctcagg 1560aggaagccag
agcggagaag ctccgagaag cacttccacc attagtggca gacaaaggca
1620tcgaggcagc cgcagaagtt gtctgcgaag tggaggggct ccaggcggac
atcggagcag 1680cattagttga aaccccgcgc ggtcacgtaa ggataatacc
tcaagcaaat gaccgtatga 1740tcggacagta tatcgttgtc tcgccaaact
ctgtgctgaa gaatgccaaa ctcgcaccag 1800cgcacccgct agcagatcag
gttaagatca taacacactc cggaagatca ggaaggtacg 1860cggtcgaacc
atacgacgct aaagtactga tgccagcagg aggtgccgta ccatggccag
1920aattcctagc actgagtgag agcgccacgt tagtgtacaa cgaaagagag
tttgtgaacc 1980gcaaactata ccacattgcc atgcatggcc ccgccaagaa
tacagaagag gagcagtaca 2040aggttacaaa ggcagagctt gcagaaacag
agtacgtgtt tgacgtggac aagaagcgtt 2100gcgttaagaa ggaagaagcc
tcaggtctgg tcctctcggg agaactgacc aaccctccct 2160atcatgagct
agctctggag ggactgaaga cccgacctgc ggtcccgtac aaggtcgaaa
2220caataggagt gataggcaca ccggggtcgg gcaagtcagc tattatcaag
tcaactgtca 2280cggcacgaga tcttgttacc agcggaaaga aagaaaattg
tcgcgaaatt gaggccgacg 2340tgctaagact gaggggtatg cagattacgt
cgaagacagt agattcggtt atgctcaacg 2400gatgccacaa agccgtagaa
gtgctgtacg ttgacgaagc gttcgcgtgc cacgcaggag 2460cactacttgc
cttgattgct atcgtcaggc cccgcaagaa ggtagtacta tgcggagacc
2520ccatgcaatg cggattcttc aacatgatgc aactaaaggt acatttcaat
caccctgaaa 2580aagacatatg caccaagaca ttctacaagt atatctcccg
gcgttgcaca cagccagtta 2640cagctattgt atcgacactg cattacgatg
gaaagatgaa aaccacgaac ccgtgcaaga 2700agaacattga aatcgatatt
acaggggcca caaagccgaa gccaggggat atcatcctga 2760catgtttccg
cgggtgggtt aagcaattgc aaatcgacta tcccggacat gaagtaatga
2820cagccgcggc ctcacaaggg ctaaccagaa aaggagtgta tgccgtccgg
caaaaagtca 2880atgaaaaccc actgtacgcg atcacatcag agcatgtgaa
cgtgttgctc acccgcactg 2940aggacaggct agtgtggaaa accttgcagg
gcgacccatg gattaagcag ctcactaaca 3000tacctaaagg aaactttcag
gctactatag aggactggga agctgaacac aagggaataa 3060ttgctgcaat
aaacagcccc actccccgtg ccaatccgtt cagctgcaag accaacgttt
3120gctgggcgaa agcattggaa ccgatactag ccacggccgg tatcgtactt
accggttgcc 3180agtggagcga actgttccca cagtttgcgg atgacaaacc
acattcggcc atttacgcct 3240tagacgtaat ttgcattaag tttttcggca
tggacttgac aagcggactg ttttctaaac 3300agagcatccc actaacgtac
catcccgccg attcagcgag gccggtagct cattgggaca 3360acagcccagg
aacccgcaag tatgggtacg atcacgccat tgccgccgaa ctctcccgta
3420gatttccggt gttccagcta gctgggaagg gcacacaact tgatttgcag
acggggagaa 3480ccagagttat ctctgcacag cataacctgg tcccggtgaa
ccgcaatctt cctcacgcct 3540tagtccccga gtacaaggag aagcaacccg
gcccggtcga aaaattcttg aaccagttca 3600aacaccactc agtacttgtg
gtatcagagg aaaaaattga agctccccgt aagagaatcg 3660aatggatcgc
cccgattggc atagccggtg cagataagaa ctacaacctg gctttcgggt
3720ttccgccgca ggcacggtac gacctggtgt tcatcaacat tggaactaaa
tacagaaacc 3780accactttca gcagtgcgaa gaccatgcgg cgaccttaaa
aaccctttcg cgttcggccc 3840tgaattgcct taacccagga ggcaccctcg
tggtgaagtc ctatggctac gccgaccgca 3900acagtgagga cgtagtcacc
gctcttgcca gaaagtttgt cagggtgtct gcagcgagac 3960cagattgtgt
ctcaagcaat acagaaatgt acctgatttt ccgacaacta gacaacagcc
4020gtacacggca attcaccccg caccatctga attgcgtgat ttcgtccgtg
tatgagggta 4080caagagatgg agttggagcc gcgccgtcat accgcaccaa
aagggagaat attgctgact 4140gtcaagagga agcagttgtc aacgcagcca
atccgctggg tagaccaggc gaaggagtct 4200gccgtgccat ctataaacgt
tggccgacca gttttaccga ttcagccacg gagacaggca 4260ccgcaagaat
gactgtgtgc ctaggaaaga aagtgatcca cgcggtcggc cctgatttcc
4320ggaagcaccc agaagcagaa gccttgaaat tgctacaaaa cgcctaccat
gcagtggcag 4380acttagtaaa tgaacataac atcaagtctg tcgccattcc
actgctatct acaggcattt 4440acgcagccgg aaaagaccgc cttgaagtat
cacttaactg cttgacaacc gcgctagaca 4500gaactgacgc ggacgtaacc
atctattgcc tggataagaa gtggaaggaa agaatcgacg 4560cggcactcca
acttaaggag tctgtaacag agctgaagga tgaagatatg gagatcgacg
4620atgagttagt atggatccat ccagacagtt gcttgaaggg aagaaaggga
ttcagtacta 4680caaaaggaaa attgtattcg tacttcgaag gcaccaaatt
ccatcaagca gcaaaagaca 4740tggcggagat aaaggtcctg ttccctaatg
accaggaaag taatgaacaa ctgtgtgcct 4800acatattggg tgagaccatg
gaagcaatcc gcgaaaagtg cccggtcgac cataacccgt 4860cgtctagccc
gcccaaaacg ttgccgtgcc tttgcatgta tgccatgacg ccagaaaggg
4920tccacagact tagaagcaat aacgtcaaag aagttacagt atgctcctcc
accccccttc 4980ctaagcacaa aattaagaat gttcagaagg ttcagtgcac
gaaagtagtc ctgtttaatc 5040cgcacactcc cgcattcgtt cccgcccgta
agtacataga agtgccagaa cagcctaccg 5100ctcctcctgc acaggccgag
gaggcccccg aagttgtagc gacaccgtca ccatctacag 5160ctgataacac
ctcgcttgat gtcacagaca tctcactgga tatggatgac agtagcgaag
5220gctcactttt ttcgagcttt agcggatcgg acaactctat tactagtgcc
tcgtaccccg 5280gccatcaaca cgcgtctgcg ttcgaccagg ctgcgcgttc
tcgcggccat agcaaccgac 5340gtacggcgtt gcgccctcgc cggcagcaag
aagccacgga agtccgcccg gagcagaaaa 5400tgcccacgct actgcgggtt
tatatagacg gtccccacgg gatggggaaa accaccacca 5460cgcaactgct
ggtggccctg ggttcgcgcg acgatatcgt ctacgtaccc gagccgatga
5520cttactggcg ggtgctgggg gcttccgaga caatcgcgaa catctacacc
acacaacacc 5580gcctcgacca gggtgagata tcggccgggg acgcggcggt
ggtaatgaca agcgcccaga 5640taacaatggg catgccttat gccgtgaccg
acgccgttct ggctcctcat atcggggggg 5700aggctgggag ctcacatgcc
ccgcccccgg ccctcaccct catcttcgac cgccatccca 5760tcgccgccct
cctgtgctac ccggccgcgc ggtaccttat gggcagcatg accccccagg
5820ccgtgctggc gttcgtggcc ctcatcccgc cgaccttgcc cggcaccaac
atcgtgcttg 5880gggcccttcc ggaggacaga cacatcgacc gcctggccaa
acgccagcgc cccggcgagc 5940ggctggacct ggctatgctg gctgcgattc
gccgcgttta cgggctactt gccaatacgg 6000tgcggtatct gcagtgcggc
gggtcgtggc gggaggactg gggacagctt tcggggacgg 6060ccgtgccgcc
ccagggtgcc gagccccaga gcaacgcggg cccacgaccc catatcgggg
6120acacgttatt taccctgttt cgggcccccg agttgctggc ccccaacggc
gacctgtata 6180acgtgtttgc ctgggccttg gacgtcttgg ccaaacgcct
ccgttccatg cacgtcttta 6240tcctggatta cgaccaatcg cccgccggct
gccgggacgc cctgctgcaa cttacctccg 6300ggatggtcca gacccacgtc
accacccccg gctccatacc gacgatatgc gacctggcgc 6360gcacgtttgc
ccgggagatg ggggaggcta acactagtat ggacagttgg tcgtcaggac
6420ctagttcact agagatagta gaccgaaggc aggtggtggt ggctgacgtt
catgccgtcc 6480aagagcctgc ccctattcca ccgccaaggc taaagaagat
ggcccgcctg gcagcggcaa 6540gaaaagagcc cactccaccg gcaagcaata
gctctgagtc cctccacctc tcttttggtg 6600gggtatccat gtccctcgga
tcaattttcg acggagagac ggcccgccag gcagcggtac 6660aacccctggc
aacaggcccc acggatgtgc ctatgtcttt cggatcgttt tccgacggag
6720agattgatga gctgagccgc agagtaactg agtccgaacc cgtcctgttt
ggatcatttg 6780aaccgggcga agtgaactca attatatcgt cccgatcagc
cgtatctttt ccactacgca 6840agcagagacg tagacgcagg agcaggagga
ctgaatactg actaaccggg gtaggtgggt 6900acatattttc gacggacaca
ggccctgggc acttgcaaaa gaagtccgtt ctgcagaacc 6960agcttacaga
accgaccttg gagcgcaatg tcctggaaag aattcatgcc ccggtgctcg
7020acacgtcgaa agaggaacaa ctcaaactca ggtaccagat gatgcccacc
gaagccaaca 7080aaagtaggta ccagtctcgt aaagtagaaa atcagaaagc
cataaccact gagcgactac 7140tgtcaggact acgactgtat aactctgcca
cagatcagcc agaatgctat aagatcacct 7200atccgaaacc attgtactcc
agtagcgtac cggcgaacta ctccgatcca cagttcgctg 7260tagctgtctg
taacaactat ctgcatgaga actatccgac agtagcatct tatcagatta
7320ctgacgagta cgatgcttac ttggatatgg tagacgggac agtcgcctgc
ctggatactg 7380caaccttctg ccccgctaag cttagaagtt acccgaaaaa
acatgagtat agagccccga 7440atatccgcag tgcggttcca tcagcgatgc
agaacacgct acaaaatgtg ctcattgccg 7500caactaaaag aaattgcaac
gtcacgcaga tgcgtgaact gccaacactg gactcagcga 7560cattcaatgt
cgaatgcttt cgaaaatatg catgtaatga cgagtattgg gaggagttcg
7620ctcggaagcc aattaggatt accactgagt ttgtcaccgc atatgtagct
agactgaaag 7680gccctaaggc cgccgcacta tttgcaaaga cgtataattt
ggtcccattg caagaagtgc 7740ctatggatag attcgtcatg gacatgaaaa
gagacgtgaa agttacacca ggcacgaaac 7800acacagaaga aagaccgaaa
gtacaagtga tacaagccgc agaacccctg gcgactgctt 7860acttatgcgg
gattcaccgg gaattagtgc gtaggcttac ggccgtcttg cttccaaaca
7920ttcacacgct ttttgacatg tcggcggagg attttgatgc aatcatagca
gaacacttca 7980agcaaggcga cccggtactg gagacggata tcgcatcatt
cgacaaaagc caagacgacg 8040ctatggcgtt aaccggtctg atgatcttgg
aggacctggg tgtggatcaa ccactactcg 8100acttgatcga gtgcgccttt
ggagaaatat catccaccca tctacctacg ggtactcgtt 8160ttaaattcgg
ggcgatgatg aaatccggaa tgttcctcac actttttgtc aacacagttt
8220tgaatgtcgt tatcgccagc agagtactag aagagcggct taaaacgtcc
agatgtgcag 8280cgttcattgg cgacgacaac atcatacatg gagtagtatc
tgacaaagaa atggctgaga 8340ggtgcgccac ctggctcaac atggaggtta
agatcatcga cgcagtcatc ggtgagagac 8400caccttactt ctgcggcgga
tttatcttgc aagattcggt tacttccaca gcgtgccgcg 8460tggcggaccc
cctgaaaagg ctgtttaagt tgggtaaacc gctcccagcc gacgacgagc
8520aagacgaaga cagaagacgc gctctgctag atgaaacaaa ggcgtggttt
agagtaggta 8580taacaggcac tttagcagtg gccgtgacga cccggtatga
ggtagacaat attacacctg 8640tcctactggc attgagaact tttgcccaga
gcaaaagagc attccaagcc atcagagggg 8700aaataaagca tctctacggt
ggtcctaaat agtcagcata gtacatttca tctgactaat 8760actacaacac
caccacctct agcccgggct cgagatctgc gatctaagta agcttggcat
8820tccggtactg ttggtaaagc caccatggaa gacgccaaaa acataaagaa
aggcccggcg 8880ccattctatc cgctggaaga tggaaccgct ggagagcaac
tgcataaggc tatgaagaga 8940tacgccctgg ttcctggaac aattgctttt
acagatgcac atatcgaggt ggacatcact 9000tacgctgagt acttcgaaat
gtccgttcgg ttggcagaag ctatgaaacg atatgggctg 9060aatacaaatc
acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg
9120ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta
taatgaacgt 9180gaattgctca acagtatggg catttcgcag cctaccgtgg
tgttcgtttc caaaaagggg 9240ttgcaaaaaa ttttgaacgt gcaaaaaaag
ctcccaatca tccaaaaaat tattatcatg 9300gattctaaaa cggattacca
gggatttcag tcgatgtaca cgttcgtcac atctcatcta 9360cctcccggtt
ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt
9420gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc
tctgcctcat 9480agaactgcct gcgtgagatt ctcgcatgcc agagatccta
tttttggcaa tcaaatcatt 9540ccggatactg cgattttaag tgttgttcca
ttccatcacg gttttggaat gtttactaca 9600ctcggatatt tgatatgtgg
atttcgagtc gtcttaatgt atagatttga agaagagctg 9660tttctgagga
gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc
9720tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt
acacgaaatt 9780gcttctggtg gcgctcccct ctctaaggaa gtcggggaag
cggttgccaa gaggttccat 9840ctgccaggta tcaggcaagg atatgggctc
actgagacta catcagctat tctgattaca 9900cccgaggggg atgataaacc
gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag 9960gttgtggatc
tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg
10020agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa
cgccttgatt 10080gacaaggatg gatggctaca ttctggagac atagcttact
gggacgaaga cgaacacttc 10140ttcatcgttg accgcctgaa gtctctgatt
aagtacaaag gctatcaggt ggctcccgct 10200gaattggaat ccatcttgct
ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt 10260cccgacgatg
acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg
10320atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc
gaaaaagttg 10380cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc
ttaccggaaa actcgacgca 10440agaaaaatca gagagatcct cataaaggcc
aagaagggcg gaaagatcgc cgtgtaattc 10500tagaggcgcg ccgatctcac
gatcccctga aaaggctgtt taagttgggt aaaccgctcc 10560cagccgacga
cgagcaagac gaagacagaa gacgcgctct gctagatgaa acaaaggcgt
10620ggtttagagt aggtataaca ggcactttag cagtggccgt gacgacccgg
tatgaggtag 10680acaatattac acctgtccta ctggcattga gaacttttgc
ccagagcaaa agagcattcc 10740aagccatcag aggggaaata aagcatctct
acggtggtcc taaatagtca gcatagtaca 10800tttcatctga ctaatactac
aacaccacca ccatgaatag aggattcttt aacatgctcg 10860gccgccgccc
cttcccggcc cccactgcca tgtggaggcc gcggagaagg aggcaggcgg
10920ccccgatgcc tgcccgcaac gggctggctt ctcaaatcca gcaactgacc
acagccgtca 10980gtgccctagt cattggacag gcaactagac ctcaaccccc
acgtccacgc cagccaccgc 11040gccagaagaa gcaggcgccc aagcaaccac
cgaagccgaa gaaaccaaaa acgcaggaga 11100agaagaagaa gcaacctgca
aaacccaaac ccggaaagag acagcgcatg gcacttaagt 11160tggaggccga
cagattgttc gacgtcaaga acgaggacgg agatgtcatc gggcacgcac
11220tggccatgga aggaaaggta atgaaacctc tgcacgtgaa aggaaccatc
gaccaccctg 11280tgctatcaaa gctcaaattt accaagtcgt cagcatacga
catggagttc gcacagttgc 11340cagtcaacat gagaagtgag gcattcacct
acaccagtga acaccccgaa ggattctata 11400actggcacca cggagcggtg
cagtatagtg gaggtagatt taccatccct cgcggagtag 11460gaggcagagg
agacagcggt cgtccgatca tggataactc cggtcgggtt gtcgcgatag
11520tcctcggtgg agctgatgaa ggaacacgaa ctgccctttc ggtcgtcacc
tggaatagta 11580aagggaagac aattaagacg accccggaag ggacagaaga
gtggtccgca gcaccactgg 11640tcacggcaat gtgtttgctc ggaaatgtga
gcttcccatg cgaccgcccg cccacatgct 11700atacccgcga accttccaga
gccctcgaca tccttgaaga gaacgtgaac catgaggcct 11760acgataccct
gctcaatgcc atattgcggt gcggatcgtc tggcagaagc aaaagaagcg
11820tcatcgatga ctttaccctg accagcccct acttgggcac atgctcgtac
tgccaccata 11880ctgaaccgtg cttcagccct gttaagatcg agcaggtctg
ggacgaagcg gacgataaca 11940ccatacgcat acagacttcc gcccagtttg
gatacgacca aagcggagca gcaagcgcaa 12000acaagtaccg ctacatgtcg
cttaagcagg atcacaccgt taaagaaggc accatggatg 12060acatcaagat
tagcacctca ggaccgtgta gaaggcttag ctacaaagga tactttctcc
12120tcgcaaaatg ccctccaggg gacagcgtaa cggttagcat agtgagtagc
aactcagcaa 12180cgtcatgtac actggcccgc aagataaaac caaaattcgt
gggacgggaa aaatatgatc 12240tacctcccgt tcacggtaaa aaaattcctt
gcacagtgta cgaccgtctg aaagaaacaa 12300ctgcaggcta catcactatg
cacaggccgg gcccgcacgc ttatacatcc tacctggaag 12360aatcatcagg
gaaagtttac gcaaagccgc catctgggaa gaacattacg tatgagtgca
12420agtgcggcga ctacaagacc ggaaccgttt cgacccgcac cgaaatcact
ggttgcaccg 12480ccatcaagca gtgcgtcgcc tataagagcg accaaacgaa
gtgggtcttc aactcaccgg 12540acttgatccg acatgacgac cacacggtcc
aagggaaatt gcatttgcct ttcaagttga 12600tcccgagtac ctgcatggtc
cctgttgccc acgcgccgaa tgtaatacat ggctttaaac 12660acatcagcct
ccaattagat acagaccact tgacattgct caccaccagg agactagggg
12720caaacccgga accaaccact gaatggatcg tcggaaagac ggtcagaaac
ttcaccgtcg 12780accgagatgg cctggaatac atatggggaa atcatgagcc
agtgagggtc tatgcccaag 12840agtcagcacc aggagaccct cacggatggc
cacacgaaat agtacagcat tactaccatc 12900gccatcctgt gtacaccatc
ttagccgtcg catcagctac cgtggcgatg atgattggcg 12960taactgttgc
agtgttatgt gcctgtaaag cgcgccgtga gtgcctgacg ccatacgccc
13020tggccccaaa cgccgtaatc ccaacttcgc tggcactctt gtgctgcgtt
aggtcggcca 13080atgctgaaac gttcaccgag accatgagtt acttgtggtc
gaacagtcag ccgttcttct 13140gggtccagtt gtgcatacct ttggccgctt
tcatcgttct aatgcgctgc tgctcctgct 13200gcctgccttt tttagtggtt
gccggcgcct acctggcgaa ggtagacgcc tacgaacatg 13260cgaccactgt
tccaaatgtg ccacagatac cgtataaggc acttgttgaa agggcagggt
13320atgccccgct caatttggag atcactgtca tgtcctcgga ggttttgcct
tccaccaacc 13380aagagtacat tacctgcaaa ttcaccactg tggtcccctc
cccaaaaatc aaatgctgcg 13440gctccttgga atgtcagccg gccgttcatg
cagactatac ctgcaaggtc ttcggagggg 13500tctacccctt tatgtgggga
ggagcgcaat gtttttgcga cagtgagaac agccagatga 13560gtgaggcgta
cgtcgaactg tcagcagatt gcgcgtctga ccacgcgcag gcgattaagg
13620tgcacactgc cgcgatgaaa gtaggactgc gtatagtgta cgggaacact
accagtttcc 13680tagatgtgta cgtgaacgga gtcacaccag gaacgtctaa
agacttgaaa gtcatagctg 13740gaccaatttc agcatcgttt acgccattcg
atcataaggt cgttatccat cgcggcctgg 13800tgtacaacta tgacttcccg
gaatatggag cgatgaaacc aggagcgttt ggagacattc 13860aagctacctc
cttgactagc aaggatctca tcgccagcac agacattagg ctactcaagc
13920cttccgccaa gaacgtgcat gtcccgtaca cgcaggccgc atcaggattt
gagatgtgga 13980aaaacaactc aggccgccca ctgcaggaaa ccgcaccttt
cgggtgtaag attgcagtaa 14040atccgctccg agcggtggac tgttcatacg
ggaacattcc catttctatt gacatcccga 14100acgctgcctt tatcaggaca
tcagatgcac cactggtctc aacagtcaaa tgtgaagtca 14160gtgagtgcac
ttattcagca gacttcggcg ggatggccac cctgcagtat gtatccgacc
14220gcgaaggtca atgccccgta cattcgcatt cgagcacagc aactctccaa
gagtcgacag 14280tacatgtcct ggagaaagga gcggtgacag tacactttag
caccgcgagt ccacaggcga 14340actttatcgt atcgctgtgt gggaagaaga
caacatgcaa tgcagaatgt aaaccaccag 14400ctgaccatat cgtgagcacc
ccgcacaaaa atgaccaaga atttcaagcc gccatctcaa 14460aaacatcatg
gagttggctg tttgcccttt tcggcggcgc ctcgtcgcta ttaattatag
14520gacttatgat ttttgcttgc agcatgatgc tgactagcac acgaagatga
ccgctacgcc 14580ccaatgatcc gaccagcaaa actcgatgta cttccgagga
actgatgtgc ataagtgagc 14640atgcgtttaa actgggccca atgttcccca
atgatccgac cagcaaaact cgatgtactt 14700ccgaggaact gatgtgcata
atgcatcagg ctggtacatt agatccccgc ttaccgcggg 14760caatatagca
acactaaaaa ctcgatgtac ttccgaggaa gcgcagtgca taatgctgcg
14820cagtgttgcc acataaccac tatattaacc atttatctag cggacgccaa
aaactcaatg 14880tatttctgag gaagcgtggt gcataatgcc acgcagcgtc
tgcataactt ttattatttc 14940ttttattaat caacaaaatt ttgtttttaa
catttcaaaa aaaaaaaaaa aaaaaaaaaa 15000aaaaaaaaaa
aaaatttaaa ttaattaagc ggccgcctcg aggacgtcag gtggcacttt
15060tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
caaatatgta 15120tccgctcatg agacaataac cctgataaat gcttcaataa
tattgaaaaa ggaagagtat 15180gagtattcaa catttccgtg tcgcccttat
tccctttttt gcggcatttt gccttcctgt 15240ttttgctcac ccagaaacgc
tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 15300agtgggttac
atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
15360agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
tattatcccg 15420tattgacgcc gggcaagagc aactcggtcg ccgcatacac
tattctcaga atgacttggt 15480tgagtactca ccagtcacag aaaagcatct
tacggatggc atgacagtaa gagaattatg 15540cagtgctgcc ataaccatga
gtgataacac tgcggccaac ttacttctga caacgatcgg 15600aggaccgaag
gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
15660tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
ccacgatgcc 15720tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
gaactactta ctctagcttc 15780ccggcaacaa ttaatagact ggatggaggc
ggataaagtt gcaggaccac ttctgcgctc 15840ggcccttccg gctggctggt
ttattgctga taaatctgga gccggtgagc gtgggtctcg 15900cggtatcatt
gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
15960gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
taggtgcctc 16020actgattaag cattggtaac tgtcagacca agtttactca
tatatacttt agattgattt 16080aaaacttcat ttttaattta aaaggatcta
ggtgaagatc ctttttgata atctcatgac 16140caaaatccct taacgtgagt
tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 16200aggatcttct
tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
16260accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
ttccgaaggt 16320aactggcttc agcagagcgc agataccaaa tactggtctt
ctagtgtagc cgtagttagg 16380ccaccacttc aagaactctg tagcaccgcc
tacatacctc gctctgctaa tcctgttacc 16440agtggctgct gccagtggcg
ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 16500accggataag
gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
16560gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
gcgccacgct 16620tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
agggtcggaa caggagagcg 16680cacgagggag cttccagggg gaaacgcctg
gtatctttat agtcctgtcg ggtttcgcca 16740cctctgactt gagcgtcgat
ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 16800cgccagcaac
gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtg
16860ggaggctaga gtacatttag gtgacactat agaa 1689421920DNAArtificial
SequenceSynthetic polynucleotide 2atggagaagc cagtagtaaa cgtagacgta
gacccccaga gtccgtttgt cgtgcaactg 60caaaaaagct tcccgcaatt tgaggtagta
gcacagcagg tcactccaaa tgaccatgct 120aatgccagag cattttcgca
tctggccagt aaactaatcg agctggaggt tcctaccaca 180gcgacgatct
tggacatagg cagcgcaccg gctcgtagaa tgttttccga gcaccagtat
240cattgtgtct gccccatgcg tagtccagaa gacccggacc gcatgatgaa
atatgccagt 300aaactggcgg aaaaagcgtg caagattaca aacaagaact
tgcatgagaa gattaaggat 360ctccggaccg tacttgatac gccggatgct
gaaacaccat cgctctgctt tcacaacgat 420gttacctgca acatgcgtgc
cgaatattcc gtcatgcagg acgtgtatat caacgctccc 480ggaactatct
atcatcaggc tatgaaaggc gtgcggaccc tgtactggat tggcttcgac
540accacccagt tcatgttctc ggctatggca ggttcgtacc ctgcgtacaa
caccaactgg 600gccgacgaga aagtccttga agcgcgtaac atcggacttt
gcagcacaaa gctgagtgaa 660ggtaggacag gaaaattgtc gataatgagg
aagaaggagt tgaagcccgg gtcgcgggtt 720tatttctccg taggatcgac
actttatcca gaacacagag ccagcttgca gagctggcat 780cttccatcgg
tgttccactt gaatggaaag cagtcgtaca cttgccgctg tgatacagtg
840gtgagttgcg aaggctacgt agtgaagaaa atcaccatca gtcccgggat
cacgggagaa 900accgtgggat acgcggttac acacaatagc gagggcttct
tgctatgcaa agttactgac 960acagtaaaag gagaacgggt atcgttccct
gtgtgcacgt acatcccggc caccatatgc 1020gatcagatga ctggtataat
ggccacggat atatcacctg acgatgcaca aaaacttctg 1080gttgggctca
accagcgaat tgtcattaac ggtaggacta acaggaacac caacaccatg
1140caaaattacc ttctgccgat catagcacaa gggttcagca aatgggctaa
ggagcgcaag 1200gatgatcttg ataacgagaa aatgctgggt actagagaac
gcaagcttac gtatggctgc 1260ttgtgggcgt ttcgcactaa gaaagtacat
tcgttttatc gcccacctgg aacgcagacc 1320atcgtaaaag tcccagcctc
ttttagcgct tttcccatgt cgtccgtatg gacgacctct 1380ttgcccatgt
cgctgaggca gaaattgaaa ctggcattgc aaccaaagaa ggaggaaaaa
1440ctgctgcagg tctcggagga attagtcatg gaggccaagg ctgcttttga
ggatgctcag 1500gaggaagcca gagcggagaa gctccgagaa gcacttccac
cattagtggc agacaaaggc 1560atcgaggcag ccgcagaagt tgtctgcgaa
gtggaggggc tccaggcgga catcggagca 1620gcattagttg aaaccccgcg
cggtcacgta aggataatac ctcaagcaaa tgaccgtatg 1680atcggacagt
atatcgttgt ctcgccaaac tctgtgctga agaatgccaa actcgcacca
1740gcgcacccgc tagcagatca ggttaagatc ataacacact ccggaagatc
aggaaggtac 1800gcggtcgaac catacgacgc taaagtactg atgccagcag
gaggtgccgt accatggcca 1860gaattcctag cactgagtga gagcgccacg
ttagtgtaca acgaaagaga gtttgtgaac 192032121DNAArtificial
SequenceSynthetic polynucleotide 3cgcaaactat accacattgc catgcatggc
cccgccaaga atacagaaga ggagcagtac 60aaggttacaa aggcagagct tgcagaaaca
gagtacgtgt ttgacgtgga caagaagcgt 120tgcgttaaga aggaagaagc
ctcaggtctg gtcctctcgg gagaactgac caaccctccc 180tatcatgagc
tagctctgga gggactgaag acccgacctg cggtcccgta caaggtcgaa
240acaataggag tgataggcac accggggtcg ggcaagtcag ctattatcaa
gtcaactgtc 300acggcacgag atcttgttac cagcggaaag aaagaaaatt
gtcgcgaaat tgaggccgac 360gtgctaagac tgaggggtat gcagattacg
tcgaagacag tagattcggt tatgctcaac 420ggatgccaca aagccgtaga
agtgctgtac gttgacgaag cgttcgcgtg ccacgcagga 480gcactacttg
ccttgattgc tatcgtcagg ccccgcaaga aggtagtact atgcggagac
540cccatgcaat gcggattctt caacatgatg caactaaagg tacatttcaa
tcaccctgaa 600aaagacatat gcaccaagac attctacaag tatatctccc
ggcgttgcac acagccagtt 660acagctattg tatcgacact gcattacgat
ggaaagatga aaaccacgaa cccgtgcaag 720aagaacattg aaatcgatat
tacaggggcc acaaagccga agccagggga tatcatcctg 780acatgtttcc
gcgggtgggt taagcaattg caaatcgact atcccggaca tgaagtaatg
840acagccgcgg cctcacaagg gctaaccaga aaaggagtgt atgccgtccg
gcaaaaagtc 900aatgaaaacc cactgtacgc gatcacatca gagcatgtga
acgtgttgct cacccgcact 960gaggacaggc tagtgtggaa aaccttgcag
ggcgacccat ggattaagca gctcactaac 1020atacctaaag gaaactttca
ggctactata gaggactggg aagctgaaca caagggaata 1080attgctgcaa
taaacagccc cactccccgt gccaatccgt tcagctgcaa gaccaacgtt
1140tgctgggcga aagcattgga accgatacta gccacggccg gtatcgtact
taccggttgc 1200cagtggagcg aactgttccc acagtttgcg gatgacaaac
cacattcggc catttacgcc 1260ttagacgtaa tttgcattaa gtttttcggc
atggacttga caagcggact gttttctaaa 1320cagagcatcc cactaacgta
ccatcccgcc gattcagcga ggccggtagc tcattgggac 1380aacagcccag
gaacccgcaa gtatgggtac gatcacgcca ttgccgccga actctcccgt
1440agatttccgg tgttccagct agctgggaag ggcacacaac ttgatttgca
gacggggaga 1500accagagtta tctctgcaca gcataacctg gtcccggtga
accgcaatct tcctcacgcc 1560ttagtccccg agtacaagga gaagcaaccc
ggcccggtcg aaaaattctt gaaccagttc 1620aaacaccact cagtacttgt
ggtatcagag gaaaaaattg aagctccccg taagagaatc 1680gaatggatcg
ccccgattgg catagccggt gcagataaga actacaacct ggctttcggg
1740tttccgccgc aggcacggta cgacctggtg ttcatcaaca ttggaactaa
atacagaaac 1800caccactttc agcagtgcga agaccatgcg gcgaccttaa
aaaccctttc gcgttcggcc 1860ctgaattgcc ttaacccagg aggcaccctc
gtggtgaagt cctatggcta cgccgaccgc 1920aacagtgagg acgtagtcac
cgctcttgcc agaaagtttg tcagggtgtc tgcagcgaga 1980ccagattgtg
tctcaagcaa tacagaaatg tacctgattt tccgacaact agacaacagc
2040cgtacacggc aattcacccc gcaccatctg aattgcgtga tttcgtccgt
gtatgagggt 2100acaagagatg gagttggagc c 212141641DNAArtificial
SequenceSynthetic polynucleotide 4gcgccgtcat accgcaccaa aagggagaat
attgctgact gtcaagagga agcagttgtc 60aacgcagcca atccgctggg tagaccaggc
gaaggagtct gccgtgccat ctataaacgt 120tggccgacca gttttaccga
ttcagccacg gagacaggca ccgcaagaat gactgtgtgc 180ctaggaaaga
aagtgatcca cgcggtcggc cctgatttcc ggaagcaccc agaagcagaa
240gccttgaaat tgctacaaaa cgcctaccat gcagtggcag acttagtaaa
tgaacataac 300atcaagtctg tcgccattcc actgctatct acaggcattt
acgcagccgg aaaagaccgc 360cttgaagtat cacttaactg cttgacaacc
gcgctagaca gaactgacgc ggacgtaacc 420atctattgcc tggataagaa
gtggaaggaa agaatcgacg cggcactcca acttaaggag 480tctgtaacag
agctgaagga tgaagatatg gagatcgacg atgagttagt atggatccat
540ccagacagtt gcttgaaggg aagaaaggga ttcagtacta caaaaggaaa
attgtattcg 600tacttcgaag gcaccaaatt ccatcaagca gcaaaagaca
tggcggagat aaaggtcctg 660ttccctaatg accaggaaag taatgaacaa
ctgtgtgcct acatattggg tgagaccatg 720gaagcaatcc gcgaaaagtg
cccggtcgac cataacccgt cgtctagccc gcccaaaacg 780ttgccgtgcc
tttgcatgta tgccatgacg ccagaaaggg tccacagact tagaagcaat
840aacgtcaaag aagttacagt atgctcctcc accccccttc ctaagcacaa
aattaagaat 900gttcagaagg ttcagtgcac gaaagtagtc ctgtttaatc
cgcacactcc cgcattcgtt 960cccgcccgta agtacataga agtgccagaa
cagcctaccg ctcctcctgc acaggccgag 1020gaggcccccg aagttgtagc
gacaccgtca ccatctacag ctgataacac ctcgcttgat 1080gtcacagaca
tctcactgga tatggatgac agtagcgaag gctcactttt ttcgagcttt
1140agcggatcgg acaactctat tatggacagt tggtcgtcag gacctagttc
actagagata 1200gtagaccgaa ggcaggtggt ggtggctgac gttcatgccg
tccaagagcc tgcccctatt 1260ccaccgccaa ggctaaagaa gatggcccgc
ctggcagcgg caagaaaaga gcccactcca 1320ccggcaagca atagctctga
gtccctccac ctctcttttg gtggggtatc catgtccctc 1380ggatcaattt
tcgacggaga gacggcccgc caggcagcgg tacaacccct ggcaacaggc
1440cccacggatg tgcctatgtc tttcggatcg ttttccgacg gagagattga
tgagctgagc 1500cgcagagtaa ctgagtccga acccgtcctg tttggatcat
ttgaaccggg cgaagtgaac 1560tcaattatat cgtcccgatc agccgtatct
tttccactac gcaagcagag acgtagacgc 1620aggagcagga ggactgaata c
164151851DNAArtificial SequenceSynthetic polynucleotide 5tgactaaccg
gggtaggtgg gtacatattt tcgacggaca caggccctgg gcacttgcaa 60aagaagtccg
ttctgcagaa ccagcttaca gaaccgacct tggagcgcaa tgtcctggaa
120agaattcatg ccccggtgct cgacacgtcg aaagaggaac aactcaaact
caggtaccag 180atgatgccca ccgaagccaa caaaagtagg taccagtctc
gtaaagtaga aaatcagaaa 240gccataacca ctgagcgact actgtcagga
ctacgactgt ataactctgc cacagatcag 300ccagaatgct ataagatcac
ctatccgaaa ccattgtact ccagtagcgt accggcgaac 360tactccgatc
cacagttcgc tgtagctgtc tgtaacaact atctgcatga gaactatccg
420acagtagcat cttatcagat tactgacgag tacgatgctt acttggatat
ggtagacggg 480acagtcgcct gcctggatac tgcaaccttc tgccccgcta
agcttagaag ttacccgaaa 540aaacatgagt atagagcccc gaatatccgc
agtgcggttc catcagcgat gcagaacacg 600ctacaaaatg tgctcattgc
cgcaactaaa agaaattgca acgtcacgca gatgcgtgaa 660ctgccaacac
tggactcagc gacattcaat gtcgaatgct ttcgaaaata tgcatgtaat
720gacgagtatt gggaggagtt cgctcggaag ccaattagga ttaccactga
gtttgtcacc 780gcatatgtag ctagactgaa aggccctaag gccgccgcac
tatttgcaaa gacgtataat 840ttggtcccat tgcaagaagt gcctatggat
agattcgtca tggacatgaa aagagacgtg 900aaagttacac caggcacgaa
acacacagaa gaaagaccga aagtacaagt gatacaagcc 960gcagaacccc
tggcgactgc ttacttatgc gggattcacc gggaattagt gcgtaggctt
1020acggccgtct tgcttccaaa cattcacacg ctttttgaca tgtcggcgga
ggattttgat 1080gcaatcatag cagaacactt caagcaaggc gacccggtac
tggagacgga tatcgcatca 1140ttcgacaaaa gccaagacga cgctatggcg
ttaaccggtc tgatgatctt ggaggacctg 1200ggtgtggatc aaccactact
cgacttgatc gagtgcgcct ttggagaaat atcatccacc 1260catctaccta
cgggtactcg ttttaaattc ggggcgatga tgaaatccgg aatgttcctc
1320acactttttg tcaacacagt tttgaatgtc gttatcgcca gcagagtact
agaagagcgg 1380cttaaaacgt ccagatgtgc agcgttcatt ggcgacgaca
acatcataca tggagtagta 1440tctgacaaag aaatggctga gaggtgcgcc
acctggctca acatggaggt taagatcatc 1500gacgcagtca tcggtgagag
accaccttac ttctgcggcg gatttatctt gcaagattcg 1560gttacttcca
cagcgtgccg cgtggcggac cccctgaaaa ggctgtttaa gttgggtaaa
1620ccgctcccag ccgacgacga gcaagacgaa gacagaagac gcgctctgct
agatgaaaca 1680aaggcgtggt ttagagtagg tataacaggc actttagcag
tggccgtgac gacccggtat 1740gaggtagaca atattacacc tgtcctactg
gcattgagaa cttttgccca gagcaaaaga 1800gcattccaag ccatcagagg
ggaaataaag catctctacg gtggtcctaa a 1851613DNAArtificial
SequenceSynthetic oligonucleotide 6gtcctaaata gtc
13724DNAArtificial SequenceSynthetic oligonucleotide 7tctctacggt
ggtcctaaat agtc 2483735DNAArtificial SequenceSynthetic
polynucleotide 8atgaatagag gattctttaa catgctcggc cgccgcccct
tcccggcccc cactgccatg 60tggaggccgc ggagaaggag gcaggcggcc ccgatgcctg
cccgcaacgg gctggcttct 120caaatccagc aactgaccac agccgtcagt
gccctagtca ttggacaggc aactagacct 180caacccccac gtccacgcca
gccaccgcgc cagaagaagc aggcgcccaa gcaaccaccg 240aagccgaaga
aaccaaaaac gcaggagaag aagaagaagc aacctgcaaa acccaaaccc
300ggaaagagac agcgcatggc acttaagttg gaggccgaca gattgttcga
cgtcaagaac 360gaggacggag atgtcatcgg gcacgcactg gccatggaag
gaaaggtaat gaaacctctg 420cacgtgaaag gaaccatcga ccaccctgtg
ctatcaaagc tcaaatttac caagtcgtca 480gcatacgaca tggagttcgc
acagttgcca gtcaacatga gaagtgaggc attcacctac 540accagtgaac
accccgaagg attctataac tggcaccacg gagcggtgca gtatagtgga
600ggtagattta ccatccctcg cggagtagga ggcagaggag acagcggtcg
tccgatcatg 660gataactccg gtcgggttgt cgcgatagtc ctcggtggag
ctgatgaagg aacacgaact 720gccctttcgg tcgtcacctg gaatagtaaa
gggaagacaa ttaagacgac cccggaaggg 780acagaagagt ggtccgcagc
accactggtc acggcaatgt gtttgctcgg aaatgtgagc 840ttcccatgcg
accgcccgcc cacatgctat acccgcgaac cttccagagc cctcgacatc
900cttgaagaga acgtgaacca tgaggcctac gataccctgc tcaatgccat
attgcggtgc 960ggatcgtctg gcagaagcaa aagaagcgtc atcgatgact
ttaccctgac cagcccctac 1020ttgggcacat gctcgtactg ccaccatact
gaaccgtgct tcagccctgt taagatcgag 1080caggtctggg acgaagcgga
cgataacacc atacgcatac agacttccgc ccagtttgga 1140tacgaccaaa
gcggagcagc aagcgcaaac aagtaccgct acatgtcgct taagcaggat
1200cacaccgtta aagaaggcac catggatgac atcaagatta gcacctcagg
accgtgtaga 1260aggcttagct acaaaggata ctttctcctc gcaaaatgcc
ctccagggga cagcgtaacg 1320gttagcatag tgagtagcaa ctcagcaacg
tcatgtacac tggcccgcaa gataaaacca 1380aaattcgtgg gacgggaaaa
atatgatcta cctcccgttc acggtaaaaa aattccttgc 1440acagtgtacg
accgtctgaa agaaacaact gcaggctaca tcactatgca caggccgggc
1500ccgcacgctt atacatccta cctggaagaa tcatcaggga aagtttacgc
aaagccgcca 1560tctgggaaga acattacgta tgagtgcaag tgcggcgact
acaagaccgg aaccgtttcg 1620acccgcaccg aaatcactgg ttgcaccgcc
atcaagcagt gcgtcgccta taagagcgac 1680caaacgaagt gggtcttcaa
ctcaccggac ttgatccgac atgacgacca cacggtccaa 1740gggaaattgc
atttgccttt caagttgatc ccgagtacct gcatggtccc tgttgcccac
1800gcgccgaatg taatacatgg ctttaaacac atcagcctcc aattagatac
agaccacttg 1860acattgctca ccaccaggag actaggggca aacccggaac
caaccactga atggatcgtc 1920ggaaagacgg tcagaaactt caccgtcgac
cgagatggcc tggaatacat atggggaaat 1980catgagccag tgagggtcta
tgcccaagag tcagcaccag gagaccctca cggatggcca 2040cacgaaatag
tacagcatta ctaccatcgc catcctgtgt acaccatctt agccgtcgca
2100tcagctaccg tggcgatgat gattggcgta actgttgcag tgttatgtgc
ctgtaaagcg 2160cgccgtgagt gcctgacgcc atacgccctg gccccaaacg
ccgtaatccc aacttcgctg 2220gcactcttgt gctgcgttag gtcggccaat
gctgaaacgt tcaccgagac catgagttac 2280ttgtggtcga acagtcagcc
gttcttctgg gtccagttgt gcataccttt ggccgctttc 2340atcgttctaa
tgcgctgctg ctcctgctgc ctgccttttt tagtggttgc cggcgcctac
2400ctggcgaagg tagacgccta cgaacatgcg accactgttc caaatgtgcc
acagataccg 2460tataaggcac ttgttgaaag ggcagggtat gccccgctca
atttggagat cactgtcatg 2520tcctcggagg ttttgccttc caccaaccaa
gagtacatta cctgcaaatt caccactgtg 2580gtcccctccc caaaaatcaa
atgctgcggc tccttggaat gtcagccggc cgttcatgca 2640gactatacct
gcaaggtctt cggaggggtc taccccttta tgtggggagg agcgcaatgt
2700ttttgcgaca gtgagaacag ccagatgagt gaggcgtacg tcgaactgtc
agcagattgc 2760gcgtctgacc acgcgcaggc gattaaggtg cacactgccg
cgatgaaagt aggactgcgt 2820atagtgtacg ggaacactac cagtttccta
gatgtgtacg tgaacggagt cacaccagga 2880acgtctaaag acttgaaagt
catagctgga ccaatttcag catcgtttac gccattcgat 2940cataaggtcg
ttatccatcg cggcctggtg tacaactatg acttcccgga atatggagcg
3000atgaaaccag gagcgtttgg agacattcaa gctacctcct tgactagcaa
ggatctcatc 3060gccagcacag acattaggct actcaagcct tccgccaaga
acgtgcatgt cccgtacacg 3120caggccgcat caggatttga gatgtggaaa
aacaactcag gccgcccact gcaggaaacc 3180gcacctttcg ggtgtaagat
tgcagtaaat ccgctccgag cggtggactg ttcatacggg 3240aacattccca
tttctattga catcccgaac gctgccttta tcaggacatc agatgcacca
3300ctggtctcaa cagtcaaatg tgaagtcagt gagtgcactt attcagcaga
cttcggcggg 3360atggccaccc tgcagtatgt atccgaccgc gaaggtcaat
gccccgtaca ttcgcattcg 3420agcacagcaa ctctccaaga gtcgacagta
catgtcctgg agaaaggagc ggtgacagta 3480cactttagca ccgcgagtcc
acaggcgaac tttatcgtat cgctgtgtgg gaagaagaca 3540acatgcaatg
cagaatgtaa accaccagct gaccatatcg tgagcacccc gcacaaaaat
3600gaccaagaat ttcaagccgc catctcaaaa acatcatgga gttggctgtt
tgcccttttc 3660ggcggcgcct cgtcgctatt aattatagga cttatgattt
ttgcttgcag catgatgctg 3720actagcacac gaaga 3735
* * * * *