Selective Reduction Of Allelic Variants

Abstract

Disclosed herein are antisense compounds and methods for selectively reducing expression of an allelic variant of a gene containing a single nucleotide polymorphism (SNP). Such methods, compounds, and composition are useful to treat, prevent, or ameliorate diseases, including neurodegenerative diseases, such as Huntington's Disease (HD).

Description

CROSS REFERENCED TO RELATED APPLICATIONS

[0001] This application is a continuation of U.S. Ser. No. 13/577,616, filed Oct. 4, 2012, which is a U.S. National Phase filing under 35 U.S.C. .sctn.371 claiming priority to International Serial No. PCT/US2011/024103 filed Feb. 8, 2011, which claims priority to U.S. Provisional Application No. 61/371,635, filed Aug. 6, 2010, and U.S. Provisional Application No. 61/302,469, filed Feb. 8, 2010, each of which is incorporated herein by reference in its entirety.

SEQUENCE LISTING

[0002] The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled BIOL0124USC1SEQ_ST25.txt created Dec. 19, 2014, which is 344 Kb in size. The information in the electronic format of the sequence listing is incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

[0003] Embodiments of the present invention provide methods, compounds, and compositions for selectively reducing expression of an allelic variant of a gene containing a single nucleotide polymorphism (SNP). Such methods, compounds, and compositions are useful to treat, prevent, or ameliorate diseases.

BACKGROUND OF THE INVENTION

[0004] Genetic diseases are caused by abnormalities in genes or chromosomes. Such abnormalities may include insertions, deletions, and expansions. Huntington's Disease (HD) is one example of a genetic disease caused by an expansion. HD is a progressive neurodegenerative disorder that is inherited in a dominant fashion and results from a mutation that expands the polymorphic trinucleotide (CAG) tract in the huntingtin gene (HTT). The average CAG tract size in the general population is 17-26 repeats (wild type allele), however, in HD patients the CAG tract has expanded to 36 repeats or more (mutant allele) (Huntington's Disease Collaborative Research Group 1993. Cell 72(6):971-83). The HTT gene encodes the HTT protein and the expanded CAG tract results in a pathological increase in the polyglutamine repeats near the N-terminal of the protein. Individuals carry two copies of the HTT gene and one mutant allele is sufficient to result in HD.

[0005] HTT protein appears to have a role during development of the nervous system and a protective role in cells. In mouse models, constitutive knockout of the HTT gene is lethal during embryonic development (Nasir et al 1995. Cell 81(5):811-23), while adult inactivation of the HTT gene leads to progressive cell death in the brain and the testes (Dragatsis et al 2000. Nat. Genet 26:300-306). Reduction of huntingtin expression from the wild type allele may, therefore, have negative consequences.

[0006] Like HD, there are disorders for which a strategy of selective reduction of a mutant allele would be beneficial. Thus, there remains an unmet need to selectively reduce expression of mutant allelic variants like that of HTT, which are causative of disease, over the wild type variant, which appears to be necessary for normal cellular processes.

BRIEF DESCRIPTION OF THE DRAWINGS

[0007] FIG. 1A-EEEEEEE provides the mRNA and genomic HTT sequence showing SNP positions.

SUMMARY OF THE INVENTION

[0008] Provided herein are methods, compounds, and compositions for selectively reducing expression of an allelic variant of a gene containing a single nucleotide polymorphism (SNP). Such methods, compounds, and compositions are useful to treat, prevent, or ameliorate diseases. SNPs may be associated with a mutant allele, the expression of which causes disease. In certain embodiments, the expressed gene product of a mutant allele results in aggregation of the mutant proteins causing disease. In certain embodiments, the expressed gene product of a mutant allele results in gain of function causing disease.

[0009] In certain embodiments, selective reduction of mRNA and protein expression of a mutant allele is achieved by targeting a SNP located on the mutant allele with an antisense compound. In certain embodiments, the antisense compound is an antisense oligonucleotide

[0010] In certain embodiments, antisense compounds designed to selectively reduce an allelic variant of a gene containing a SNP are created based on potency and selectivity of the antisense compound as well as population genetics.

DETAILED DESCRIPTION OF THE INVENTION

[0011] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed. Herein, the use of the singular includes the plural unless specifically stated otherwise. As used herein, the use of "or" means "and/or" unless stated otherwise. Furthermore, the use of the term "including" as well as other forms, such as "includes" and "included", is not limiting. Also, terms such as "element" or "component" encompass both elements and components comprising one unit and elements and components that comprise more than one subunit, unless specifically stated otherwise.

[0012] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this application, including, but not limited to, patents, patent applications, articles, books, and treatises, are hereby expressly incorporated by reference for the portions of the document discussed herein, as well as in their entirety.

DEFINITIONS

[0013] Unless specific definitions are provided, the nomenclature utilized in connection with, and the procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques may be used for chemical synthesis, and chemical analysis. Where permitted, all patents, applications, published applications and other publications, GENBANK Accession Numbers and associated sequence information obtainable through databases such as National Center for Biotechnology Information (NCBI) and other data referred to throughout in the disclosure herein are incorporated by reference for the portions of the document discussed herein, as well as in their entirety.

[0014] Unless otherwise indicated, the following terms have the following meanings: "2'-O-methoxyethyl" (also 2'-MOE and 2'-O(CH.sub.2).sub.2--OCH.sub.3) refers to an O-methoxy-ethyl modification of the 2' position of a furosyl ring. A 2'-O-methoxyethyl modified sugar is a modified sugar.

[0015] "2'-O-methoxyethyl nucleotide" means a nucleotide comprising a 2'-O-methoxyethyl modified sugar moiety.

[0016] "5-methylcytosine" means a cytosine modified with a methyl group attached to the 5' position. A 5-methylcytosine is a modified nucleobase.

[0017] "Active pharmaceutical agent" means the substance or substances in a pharmaceutical composition that provide a therapeutic benefit when administered to an individual. For example, in certain embodiments an antisense oligonucleotide targeted to an allelic variant is an active pharmaceutical agent.

[0018] "Active target region" or "target region" means a region to which one or more active antisense compounds is targeted. "Active antisense compounds" means antisense compounds that reduce target nucleic acid levels or protein levels.

[0019] "Administered concomitantly" refers to the co-administration of two agents in any manner in which the pharmacological effects of both are manifest in the patient at the same time. Concomitant administration does not require that both agents be administered in a single pharmaceutical composition, in the same dosage form, or by the same route of administration. The effects of both agents need not manifest themselves at the same time. The effects need only be overlapping for a period of time and need not be coextensive.

[0020] "Administering" means providing a pharmaceutical agent to an individual, and includes, but is not limited to administering by a medical professional and self-administering.

[0021] "Allele" is one member of a pair of genes or one member of a series of different forms of a DNA sequences that can exist at a single locus or marker on a specific chromosome. For a diploid organism or cell or for autosomal chromosomes, each allelic pair will normally occupy corresponding positions (loci) on a pair of homologous chromosomes, one inherited from the mother and one inherited from the father. If these alleles are identical, the organism or cell is said to be `homozygous` for that allele; if they differ, the organism or cell is said to be `heterozygous` for that allele. "Major allele" refers to an allele containing the nucleotide present in a statistically significant proportion of individuals in the human population. "Minor allele" refers to an allele containing the nucleotide present in a relatively small proportion of individuals in the human population. "Wild type allele" refers to the genotype typically not associated with disease or dysfunction of the gene product. "Mutant allele" refers to the genotype associated with disease or dysfunction of the gene product.

[0022] "Allelic variant" refers to one of the pair of genes or DNA sequence existing at a single locus. For example, an allelic variant may refer to either the major allele or the minor allele.

[0023] "Amelioration" refers to a lessening of at least one indicator, sign, or symptom of an associated disease, disorder, or condition. The severity of indicators may be determined by subjective or objective measures, which are known to those skilled in the art.

[0024] "Animal" refers to a human or non-human animal, including, but not limited to, mice, rats, rabbits, dogs, cats, pigs, and non-human primates, including, but not limited to, monkeys and chimpanzees.

[0025] "Antibody" refers to a molecule characterized by reacting specifically with an antigen in some way, where the antibody and the antigen are each defined in terms of the other. Antibody may refer to a complete antibody molecule or any fragment or region thereof, such as the heavy chain, the light chain, Fab region, and Fc region.

[0026] "Antisense activity" means any detectable or measurable activity attributable to the hybridization of an antisense compound to its target nucleic acid. In certain embodiments, antisense activity is a decrease in the amount or expression of a target nucleic acid or protein encoded by such target nucleic acid.

[0027] "Antisense compound" means an oligomeric compound that is is capable of undergoing hybridization to a target nucleic acid through hydrogen bonding.

[0028] "Antisense inhibition" means reduction of target nucleic acid levels or target protein levels in the presence of an antisense compound complementary to a target nucleic acid compared to target nucleic acid levels or target protein levels in the absence of the antisense compound.

[0029] "Antisense oligonucleotide" means a single-stranded oligonucleotide having a nucleobase sequence that permits hybridization to a corresponding region or segment of a target nucleic acid.

[0030] "Bicyclic sugar" means a furosyl ring modified by the bridging of two ring atoms. A bicyclic sugar is a modified sugar.

[0031] "Bicyclic nucleoside" means a nucleoside having a sugar moiety comprising a bridge connecting two carbon atoms of the sugar ring, thereby forming a bicyclic ring system. In certain embodiments, the bridge connects the 4'-carbon and the 2'-carbon of the sugar ring.

[0032] "Cap structure" or "terminal cap moiety" means chemical modifications, which have been incorporated at either terminus of an antisense compound.

[0033] "cEt" or "constrained ethyl" means a bicyclic nucleoside having a sugar moiety comprising a bridge connecting the 4'-carbon and the 2'-carbon, wherein the bridge has the formula: 4'-CH(CH.sub.3)--O-2'.

[0034] "Chemically distinct region" refers to a region of an antisense compound that is in some way chemically different than another region of the same antisense compound. For example, a region having 2'-O-methoxyethyl nucleotides is chemically distinct from a region having nucleotides without 2'-O-methoxyethyl modifications.

[0035] "Chimeric antisense compound" means an antisense compound that has at least two chemically distinct regions.

[0036] "Co-administration" means administration of two or more pharmaceutical agents to an individual. The two or more pharmaceutical agents may be in a single pharmaceutical composition, or may be in separate pharmaceutical compositions. Each of the two or more pharmaceutical agents may be administered through the same or different routes of administration. Co-administration encompasses parallel or sequential administration.

[0037] "Complementarity" means the capacity for pairing between nucleobases of a first nucleic acid and a second nucleic acid.

[0038] "Contiguous nucleobases" means nucleobases immediately adjacent to each other.

[0039] "Differentiating polymorphism" means a variation in a nucleotide sequence that permits differentiation between a wild type and a mutant allele of a nucleic acid sequence. Differentiating polymorphisms may include insertions or deletions of one or a few nucleotides in a sequence, or changes in one or a few nucleotides in a sequence. A differentiating polymorphism or polymorphic allele can be in linkage disequilibrium with one or more other polymorphisms or polymorphic alleles.

[0040] "Diluent" means an ingredient in a composition that lacks pharmacological activity, but is pharmaceutically necessary or desirable. For example, the diluent in an injected composition may be a liquid, e.g. saline solution.

[0041] "Dose" means a specified quantity of a pharmaceutical agent provided in a single administration, or in a specified time period. In certain embodiments, a dose may be administered in one, two, or more boluses, tablets, or injections. For example, in certain embodiments where subcutaneous administration is desired, the desired dose requires a volume not easily accommodated by a single injection, therefore, two or more injections may be used to achieve the desired dose. In certain embodiments, the pharmaceutical agent is administered by infusion over an extended period of time or continuously. Doses may be stated as the amount of pharmaceutical agent per hour, day, week, or month.

[0042] "Effective amount" means the amount of active pharmaceutical agent sufficient to effectuate a desired physiological outcome in an individual in need of the agent. The effective amount may vary among individuals depending on the health and physical condition of the individual to be treated, the taxonomic group of the individuals to be treated, the formulation of the composition, assessment of the individual's medical condition, and other relevant factors.

[0043] "Fully complementary" or "100% complementary" means each nucleobase of a first nucleic acid has a complementary nucleobase in a second nucleic acid. In certain embodiments, a first nucleic acid is an antisense compound and a target nucleic acid is a second nucleic acid.

[0044] "Gapmer" means a chimeric antisense compound in which an internal region having a plurality of nucleosides that support RNase H cleavage is positioned between external regions having one or more nucleosides, wherein the nucleosides comprising the internal region are chemically distinct from the nucleoside or nucleosides comprising the external regions. The internal region may be referred to as the "gap" and the external regions may be referred to as the "wings."

[0045] "Gap-widened" means a chimeric antisense compound having a gap segment of 12 or more contiguous 2'-deoxyribonucleosides positioned between and immediately adjacent to 5' and 3' wing segments having from one to six nucleosides.

[0046] "Gene product" refers to a biochemical material, such as RNA or protein, resulting from expression of a gene.

[0047] "Haplotype" means a set of alleles of closely linked loci on a chromosome that are generally inherited together. For example, a polymorphic allele at a first site in a nucleic acid sequence on the chromosome may be found to be associated with another polymorphic allele at a second site on the same chromosome, at a frequency other than would be expected for a random associate (e.g. "linkage equilibrium"). These two polymorphic alleles may be described as being in "linkage disequilibrium." A haplotype may comprise two, three, four, or more alleles. The set of alleles in a haplotype along a given segment of a chromosome are generally transmitted to progeny together unless there has been a recombination event.

[0048] "High-affinity sugar modification" is a modified sugar moiety which when it is included in a nucleoside and said nucleoside is incorporated into an antisense oligonucleotide, the stability (as measured by Tm) of said antisense oligonucleotide: RNA duplex is increased as compared to the stability of a DNA:RNA duplex.

[0049] "High-affinity sugar-modified nucleoside" is a nucleoside comprising a modified sugar moiety that when said nucleoside is incorporated into an antisense compound, the binding affinity (as measured by Tm) of said antisense compound toward a complementary RNA molecule is increased. In certain embodiments of the invention at least one of said sugar-modified high-affinity nucleosides confers a .DELTA.Tm of at least 1 to 4 degrees per nucleoside against a complementary RNA as determined in accordance with the methodology described in Freier et al., Nucleic Acids Res., 1997, 25, 4429-4443, which is incorporated by reference in its entirety. In another aspect, at least one of the high-affinity sugar modifications confers about 2 or more, 3 or more, or 4 or more degrees per modification. In the context of the present invention, examples of sugar-modified high affinity nucleosides include, but are not limited to, (i) certain 2'-modified nucleosides, including 2'-substituted and 4' to 2' bicyclic nucleosides, and (ii) certain other non-ribofuranosyl nucleosides which provide a per modification increase in binding affinity such as modified tetrahydropyran and tricycloDNA nucleosides. For other modifications that are sugar-modified high-affinity nucleosides see Freier et al., Nucleic Acids Res., 1997, 25, 4429-4443.

[0050] "Hybridization" means the annealing of complementary nucleic acid molecules. In certain embodiments, complementary nucleic acid molecules include an antisense compound and a target nucleic acid.

[0051] "Immediately adjacent" means there are no intervening elements between the immediately adjacent elements.

[0052] "Individual" means a human or non-human animal selected for treatment or therapy.

[0053] "Internucleoside linkage" refers to the chemical bond between nucleosides.

[0054] "Linked nucleosides" means adjacent nucleosides which are bonded together.

[0055] "Mismatch" or "non-complementary nucleobase" refers to the case when a nucleobase of a first nucleic acid is not capable of pairing with the corresponding nucleobase of a second or target nucleic acid.

[0056] "Modified internucleoside linkage" refers to a substitution or any change from a naturally occurring internucleoside bond (i.e. a phosphodiester internucleoside bond).

[0057] "Modified nucleobase" refers to any nucleobase other than adenine, cytosine, guanine, thymidine, or uracil. An "unmodified nucleobase" means the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U).

[0058] "Modified nucleotide" means a nucleotide having, independently, a modified sugar moiety, modified internucleoside linkage, or modified nucleobase. A "modified nucleoside" means a nucleoside having, independently, a modified sugar moiety or modified nucleobase.

[0059] "Modified oligonucleotide" means an oligonucleotide comprising a modified internucleoside linkage, a modified sugar, or a modified nucleobase.

[0060] "Modified sugar" refers to a substitution or change from a natural sugar.

[0061] "Motif" means the pattern of chemically distinct regions in an antisense compound.

[0062] "Naturally occurring internucleoside linkage" means a 3' to 5' phosphodiester linkage.

[0063] "Natural sugar moiety" means a sugar found in DNA (2'-H) or RNA (2'-OH).

[0064] "Nuclease resistant modification" means a sugar modification or modified internucleoside linkage which, when incorporated into an oligonucleotide, makes said oligonucleotide more stable to degradation under cellular nucleases (e.g. exo- or endo-nucleases). Examples of nuclease resistant modifications include, but are not limited to, phosphorothioate internucleoside linkages, bicyclic sugar modifications, 2'-modified nucleotides, or neutral internucleoside linkages.

[0065] "Nucleic acid" refers to molecules composed of monomeric nucleotides. A nucleic acid includes ribonucleic acids (RNA), deoxyribonucleic acids (DNA), single-stranded nucleic acids, double-stranded nucleic acids, small interfering ribonucleic acids (siRNA), and microRNAs (miRNA).

[0066] "Nucleobase" means a heterocyclic moiety capable of pairing with a base of another nucleic acid.

[0067] "Nucleobase sequence" means the order of contiguous nucleobases independent of any sugar, linkage, or nucleobase modification.

[0068] "Nucleoside" means a nucleobase linked to a sugar.

[0069] "Nucleoside mimetic" includes those structures used to replace the sugar or the sugar and the base and not necessarily the linkage at one or more positions of an oligomeric compound such as for example nucleoside mimetics having morpholino, cyclohexenyl, cyclohexyl, tetrahydropyranyl, bicyclo or tricyclo sugar mimetics e.g. non furanose sugar units. Nucleotide mimetic includes those structures used to replace the nucleoside and the linkage at one or more positions of an oligomeric compound such as for example peptide nucleic acids or morpholinos (morpholinos linked by --N(H)--C(.dbd.O)--O-- or other non-phosphodiester linkage). Sugar surrogate overlaps with the slightly broader term nucleoside mimetic but is intended to indicate replacement of the sugar unit (furanose ring) only. The tetrahydropyranyl rings provided herein are illustrative of an example of a sugar surrogate wherein the furanose sugar group has been replaced with a tetrahydropyranyl ring system.

[0070] "Nucleotide" means a nucleoside having a phosphate group covalently linked to the sugar portion of the nucleoside.

[0071] "Oligomeric compound" or "oligomer" means a polymer of linked monomeric subunits which is capable of hybridizing to at least a region of a nucleic acid molecule.

[0072] "Oligonucleotide" means a polymer of linked nucleosides each of which can be modified or unmodified, independent one from another.

[0073] "Parenteral administration" means administration through injection or infusion. Parenteral administration includes subcutaneous administration, intravenous administration, intramuscular administration, intraarterial administration, intraperitoneal administration, or intracranial administration, e.g. intrathecal or intracerebroventricular administration.

[0074] "Peptide" means a molecule formed by linking at least two amino acids by amide bonds. Peptide refers to polypeptides and proteins.

[0075] "Pharmaceutical composition" means a mixture of substances suitable for administering to an individual. For example, a pharmaceutical composition may comprise one or more active pharmaceutical agents and a sterile aqueous solution.

[0076] "Pharmaceutically acceptable salts" means physiologically and pharmaceutically acceptable salts of antisense compounds, i.e., salts that retain the desired biological activity of the parent oligonucleotide and do not impart undesired toxicological effects thereto.

[0077] "Phosphorothioate linkage" means a linkage between nucleosides where the phosphodiester bond is modified by replacing one of the non-bridging oxygen atoms with a sulfur atom. A phosphorothioate linkage (P.dbd.S) is a modified internucleoside linkage.

[0078] "Portion" means a defined number of contiguous (i.e. linked) nucleobases of a nucleic acid. In certain embodiments, a portion is a defined number of contiguous nucleobases of a target nucleic acid. In certain embodiments, a portion is a defined number of contiguous nucleobases of an antisense compound.

[0079] "Prevent" refers to delaying or forestalling the onset or development of a disease, disorder, or condition for a period of time from minutes to indefinitely. Prevent also means reducing risk of developing a disease, disorder, or condition.

[0080] "Prodrug" means a therapeutic agent that is prepared in an inactive form that is converted to an active form within the body or cells thereof by the action of endogenous enzymes or other chemicals or conditions.

[0081] "Selectively reducing expression of an allelic variant" means reducing expression of one allele more than the other, differing allele among a set of alleles. For example, a mutant allele containing a single nucleotide polymorphism (SNP) may be reduced more than a wild type allele not containing the SNP.

[0082] "Side effects" means physiological responses attributable to a treatment other than the desired effects. In certain embodiments, side effects include injection site reactions, liver function test abnormalities, renal function abnormalities, liver toxicity, renal toxicity, central nervous system abnormalities, myopathies, and malaise. For example, increased aminotransferase levels in serum may indicate liver toxicity or liver function abnormality. For example, increased bilirubin may indicate liver toxicity or liver function abnormality.

[0083] "Single nucleotide polymorphism" or "SNP" means a single nucleotide variation between the genomes of individuals of the same species. In some cases, a SNP may be a single nucleotide deletion or insertion. In general, SNPs occur relatively frequently in genomes and thus contribute to genetic diversity. SNPs are thought to be mutationally more stable than other polymorphisms, lending their use in association studies in which linkage disequilibrium between markers and an unknown variant is used to map disease-causing mutations. The location of a SNP is generally flanked by highly conserved sequences. An individual may be homozygous or heterozygous for an allele at each SNP site. A heterozygous SNP allele can be a differentiating polymorphism. A SNP may be targeted with an antisense oligonucleotide, meaning that the SNP anneals to (or aligns with) position 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 of the antisense oligonucleotide. The remainder of the antisense oligonucleotide bases must have sufficient complementarity to the SNP site to facilitate hybridization.

[0084] "Single nucleotide polymorphism position" or "SNP position" refers to the nucleotide position of the SNP on a reference sequence.

[0085] "Single nucleotide polymorphism site" or "SNP site" refers to the nucleotides surrounding a SNP contained in a target nucleic acid to which an antisense compound is targeted.

[0086] "Single-stranded oligonucleotide" means an oligonucleotide which is not hybridized to a complementary strand.

[0087] "Specifically hybridizable" refers to an antisense compound having a sufficient degree of complementarity between an antisense oligonucleotide and a target nucleic acid to induce a desired effect, while exhibiting minimal or no effects on non-target nucleic acids under conditions in which specific binding is desired, i.e. under physiological conditions in the case of in vivo assays and therapeutic treatments.

[0088] "Targeting" or "targeted" means the process of design and selection of an antisense compound that will specifically hybridize to a target nucleic acid and induce a desired effect.

[0089] "Target nucleic acid," "target RNA," and "target RNA transcript" all refer to a nucleic acid capable of being targeted by antisense compounds.

[0090] "Target segment" means the sequence of nucleotides of a target nucleic acid to which an antisense compound is targeted. For example, for the purposes of this patent application, the target segment may be within the SNP site. "5' target site" refers to the 5'-most nucleotide of a target segment. "3' target site" refers to the 3'-most nucleotide of a target segment.

[0091] "Therapeutically effective amount" means an amount of a pharmaceutical agent that provides a therapeutic benefit to an individual.

[0092] "Treat" refers to administering a pharmaceutical composition to effect an alteration or improvement of a disease, disorder, or condition.

[0093] "Unmodified nucleotide" means a nucleotide composed of naturally occurring nucleobases, sugar moieties, and internucleoside linkages. In certain embodiments, an unmodified nucleotide is an RNA nucleotide (i.e. .beta.-D-ribonucleosides) or a DNA nucleotide (i.e. .beta.-D-deoxyribonucleoside).

Certain Embodiments

[0094] Embodiments of the present invention provide methods, compounds, and compositions for selectively inhibiting mRNA and protein expression of an allelic variant of a gene or DNA sequence. In certain embodiments, the allelic variant contains a single nucleotide polymorphism (SNP). In certain embodiments, the SNP is a differentiating polymorphism. In certain embodiments, a SNP is associated with a mutant allele. In certain embodiments, a SNP is in linkage disequilibrium with another polymorphism that is associated with or is causative of disease. In certain embodiments, a mutant allele is associated with disease. In certain embodiments, mRNA and protein expression of a mutant allele is associated with disease.

[0095] In certain embodiments, the expressed gene product of a mutant allele results in aggregation of the mutant proteins causing disease. In certain embodiments, the expressed gene product of a mutant allele results in gain of function causing disease. In certain embodiments, genes with an autosomal dominant mutation resulting in a toxic gain of function of the protein are the APP gene encoding amyloid precursor protein involved in Alzheimer's disease (Gene, 371: 68, 2006); the PrP gene encoding prion protein involved in Creutzfeldt-Jakob disease and in fatal familial insomnia (Nat. Med. 1997, 3: 1009); GFAP gene encoding glial fibrillary acidic protein involved in Alexander disease (J. Neurosci. 2006, 26:111623); alpha-synuclein gene encoding alpha-synuclein protein involved in Parkinson's disease (J. Clin. Invest. 2003, 111: 145); SOD-1 gene encoding the SOD-1 protein involved in amyotrophic lateral sclerosis (Science 1998, 281: 1851); atrophin-1 gene encoding atrophin-1 protein involved in dentato-rubral and pallido-luysian atrophy (DRPA) (Trends Mol. Med. 2001, 7: 479); SCA1 gene encoding ataxin-1 protein involved in spino-cerebellar ataxia-1 (SCA1) (Protein Sci. 2003, 12: 953); PLP gene encoding proteolipid protein involved in Pelizaeus-Merzbacher disease (NeuroMol Med. 2007, 4: 73); DYT1 gene encoding torsinA protein involved in Torsion dystonia (Brain Res. 2000, 877: 379); and alpha-B crystalline gene encoding alpha-B crystalline protein involved in protein aggregation diseases, including cardiomyopathy (Cell 2007, 130: 427); alpha1-antitrypsin gene encoding alpha1-antitrypsin protein involved in chronic obstructive pulmonary disease (COPD), liver disease and hepatocellular carcinoma (New Engl J Med. 2002, 346: 45); Ltk gene encoding leukocyte tyrosine kinase protein involved in systemic lupus erythematosus (Hum. Mol. Gen. 2004, 13: 171); PCSK9 gene encoding PCSK9 protein involved in hypercholesterolemia (Hum Mutat. 2009, 30: 520); prolactin receptor gene encoding prolactin receptor protein involved in breast tumors (Proc. Natl. Assoc. Sci. 2008, 105: 4533); CCL5 gene encoding the chemokine CCL5 involved in COPD and asthma (Eur. Respir. J. 2008, 32: 327); PTPN22 gene encoding PTPN22 protein involved in Type 1 diabetes, Rheumatoid arthritis, Graves disease, and SLE (Proc. Natl. Assoc. Sci. 2007, 104: 19767); androgen receptor gene encoding the androgen receptor protein involved in spinal and bulbar muscular atrophy or Kennedy's disease (J Steroid Biochem. Mol. Biol. 2008, 108: 245); CHMP4B gene encoding chromatin modifying protein-4B involved in progressive childhood posterior subcapsular cataracts (Am. J. Hum. Genet 2007, 81: 596); FXR/NR1H4 gene encoding Farnesoid X receptor protein involved in cholesterol gallstone disease, arthrosclerosis and diabetes (Mol. Endocrinol. 2007, 21: 1769); ABCA1 gene encoding ABCA1 protein involved in cardiovascular disease (Transl. Res. 2007, 149: 205); CaSR gene encoding the calcium sensing receptor protein involved in primary hypercalciuria (Kidney Int. 2007, 71: 1155); alpha-globin gene encoding alpha-globin protein involved in alpha-thallasemia (Science 2006, 312: 1215); httlpr gene encoding HTTLPR protein involved in obsessive compulsive disorder (Am. J. Hum. Genet. 2006, 78: 815); AVP gene encoding arginine vasopressin protein in stress-related disorders such as anxiety disorders and comorbid depression (CNS Neurol. Disord. Drug Targets 2006, 5: 167); GNAS gene encoding G proteins involved in congenital visual defects, hypertension, metabolic syndrome (Trends Pharmacol. Sci. 2006, 27: 260); APAF1 gene encoding APAF1 protein involved in a predisposition to major depression (Mol. Psychiatry 2006, 11: 76); TGF-beta1 gene encoding TGF-beta1 protein involved in breast cancer and prostate cancer (Cancer Epidemiol. Biomarkers Prev. 2004, 13: 759); AChR gene encoding acetylcholine receptor involved in congenital myasthenic syndrome (Neurology 2004, 62: 1090); P2Y12 gene encoding adenosine diphosphate (ADP) receptor protein involved in risk of peripheral arterial disease (Circulation 2003, 108: 2971); LQT1 gene encoding LQT1 protein involved in atrial fibrillation (Cardiology 2003, 100: 109); RET protooncogene encoding RET protein involved in sporadic pheochromocytoma (J. Clin. Endocrinol. Metab. 2003, 88: 4911); filamin A gene encoding filamin A protein involved in various congenital malformations (Nat. Genet. 2003, 33: 487); TARDBP gene encoding TDP-43 protein involved in amyotrophic lateral sclerosis (Hum. Mol. Gene.t 2010, 19: 671); SCA3 gene encoding ataxin-3 protein involved in Machado-Joseph disease (PLoS One 2008, 3: e3341); SCAT gene encoding ataxin-7 protein involved in spino-cerebellar ataxia-7 (PLoS One 2009, 4: e7232); and HTT gene encoding huntingtin protein involved in Huntington's disease (Neurobiol Dis. 1996, 3:183); and the CA4 gene encoding carbonic anhydrase 4 protein, CRX gene encoding cone-rod homeobox transcription factor protein, FSCN2 gene encoding retinal fascin homolog 2 protein, IMPDH1 gene encoding inosine monophosphate dehydrogenase 1 protein, NR2E3 gene encoding nuclear receptor subfamily 2 group E3 protein, NRL gene encoding neural retina leucine zipper protein, PRPF3 (RP18) gene encoding pre-mRNA splicing factor 3 protein, PRPF8 (RP13) gene encoding pre-mRNA splicing factor 8 protein, PRPF31 (RP11) gene encoding pre-mRNA splicing factor 31 protein, RDS gene encoding peripherin 2 protein, ROM1 gene encoding rod outer membrane protein 1 protein, RHO gene encoding rhodopsin protein, RP1 gene encoding RP1 protein, RPGR gene encoding retinitis pigmentosa GTPase regulator protein, all of which are involved in Autosomal Dominant Retinitis Pigmentosa disease (Adv Exp Med Biol. 2008, 613:203)

[0096] In certain embodiments, selective reduction of mRNA and protein expression of a mutant allele is achieved by targeting a SNP located on the mutant allele with an antisense compound. In certain embodiments, the antisense compound is an antisense oligonucleotide. In certain embodiments, the antisense compound is not a ribozyme, a double stranded siRNA, or an shRNA. In certain embodiments, the antisense oligonucleotide may have one or more modified sugar(s), nucleobase(s), or internucleoside linkage(s). In certain embodiments, the antisense oligonucleotide is complementary to the SNP site. In certain embodiments, the antisense oligonucleotide is at least 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% complementary to the SNP site. In certain embodiments, the antisense oligonucleotide is 100% complementary to the SNP site. In certain embodiments, the SNP site is 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length. In certain embodiments, the SNP anneals to position 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 of the antisense oligonucleotide.

[0097] In certain embodiments, antisense compounds designed to selectively reduce an allelic variant of a gene containing a SNP are created based on potency and selectivity of the antisense compound as well as population genetics.

[0098] In certain embodiments, selective reduction of mRNA and protein expression of an allelic variant of a gene containing a SNP occurs in a cell or tissue. In certain embodiments, the cell or tissue is in an animal. In certain embodiments, the animal is a human.

[0099] In certain embodiments, described herein are compounds comprising a modified antisense oligonucleotide consisting of 12 to 30 linked nucleosides targeted to a single nucleotide polymorphism site, wherein the modified oligonucleotide comprises a wing-gap-wing motif with a 5' wing region positioned at the 5' end of a deoxynucleoside gap, and a 3' wing region positioned at the 3' end of the deoxynucleoside gap, wherein position 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, or positions 1, 2, 3, 4, 5, 6, 7, 8, or 9 of the modified oligonucleotide, as counted from the 5' terminus of the gap, aligns with the single nucleotide polymorphism.

[0100] In certain embodiments, the single nucleotide polymorphism site is on a mutant allele that is associated with a disease. In certain embodiments, the single nucleotide polymorphism site contains a differentiating polymorphism.

[0101] In certain embodiments, the modified antisense oligonucleotide consists of 12 to 20 linked nucleosides. In certain embodiments, modified antisense oligonucleotide consists of 15 to 20 linked nucleosides. In certain embodiments, the modified antisense oligonucleotide consists of 15 to 19 linked nucleosides.

[0102] In certain embodiments, position 8, 9, or 10 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, or positions 4, 5, or 6 of the modified oligonucleotide, as counted from the 5' terminus of the gap, aligns with the single nucleotide polymorphism.

[0103] In certain embodiments, the gap region is 7-11 nucleosides in length, the 5' wing region is 1-6 nucleobases in length and the 3' wing region is 1-6 nucleobases in length.

[0104] In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 5-10-5, 2-9-6, 3-9-3, 3-9-4, 3-9-5, 4-7-4, 4-9-3, 4-9-4, 4-9-5, 4-10-5, 4-11-4, 4-11-5, 5-7-5, 5-8-6, 5-9-3, 5-9-5, 5-10-4, 5-10-5, 6-7-6, 6-8-5, and 6-9-2. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 4-9-5, and 4-11-4.

[0105] In certain embodiments, at least one internucleoside linkage is a modified internucleoside linkage. In certain embodiments, each internucleoside linkage is a phosphorothioate internucleoside linkage.

[0106] In certain embodiments, at least one nucleoside comprises a modified nucleobase. In certain embodiments, the modified nucleobase is a 5'-methylcytosine.

[0107] In certain embodiments, at least one nucleoside of at least one of the wing regions comprises a modified sugar or sugar surrogate. In certain embodiments, each of the nucleosides of each wing region comprises a modified sugar or sugar surrogate. In certain embodiments, the sugar or sugar surrogate is a 2'-O-methoxyethyl modified sugar.

[0108] In certain embodiments, at least one of the wing regions comprises a 4' to 2' bicyclic nucleoside and at least one of the remaining wing nucleosides is a non-bicyclic 2'-modified nucleoside.

[0109] In certain embodiments, the non-bicyclic 2'-modified nucleoside is a 2'-O-methoxyethyl nucleoside.

[0110] In certain embodiments, the 4' to 2' bicyclic nucleoside is 4'-CH(CH.sub.3)--O-2' bicyclic nucleoside.

[0111] In certain embodiments, the modified antisense oligonucleotide consisting of 17 linked nucleosides and wherein position 9 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, aligns with the differentiating polymorphism. In certain embodiments, the wing-gap-wing motif is 2-9-6.

[0112] In certain embodiments, described herein are compounds comprising a modified oligonucleotide consisting of 18 linked nucleosides and 90% complementary to a differentiating polymorphism, wherein the modified oligonucleotide comprises a wing-gap-wing motif, wherein position 9 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, aligns with the differentiating polymorphism; wherein each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar; and wherein the wing-gap-wing motif is 4-9-5.

[0113] In certain embodiments, described herein are compounds comprising a modified oligonucleotide consisting of 19 linked nucleosides and 90% complementary to a differentiating polymorphism, wherein the modified oligonucleotide comprises a wing-gap-wing motif, wherein position 10 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, aligns with the differentiating polymorphism; wherein each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar; and wherein the wing-gap-wing motif is 4-11-4.

[0114] In certain embodiments, described herein are compounds comprising a modified oligonucleotide consisting of 15 to 19 linked nucleosides and fully complementary to a differentiating polymorphism, wherein the modified oligonucleotide comprises a wing-gap-wing motif, wherein position 6, 7, 8, 9, 10, 11, 12, 13, or 14 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, aligns with the differentiating polymorphism; and at least one high-affinity sugar modification. In certain embodiments, the modified oligonucleotide is 100% complementary to the single nucleotide polymorphism site.

[0115] In certain embodiments, at least one of the wing regions comprises a high-affinity sugar modification. In certain embodiments, the high-affinity sugar modification is a bicyclic sugar. In certain embodiments, the bicyclic sugar comprises a 4'-CH(CH.sub.3)--O-2' bridge.

[0116] In certain embodiments, at least one of positions 2, 3, 6, 9, 10, 11, 13, or 14 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, comprises the at least one high-affinity sugar modification.

[0117] In certain embodiments, at least one of positions 2, 3, 13, and 14 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, comprises the at least one high-affinity sugar modification.

[0118] In certain embodiments, each of nucleoside positions 2, 3, 13, and 14 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, comprise the at least one high-affinity sugar modification.

[0119] In certain embodiments, the high-affinity sugar modification is a bicyclic sugar. In certain embodiments, the bicyclic sugar comprises a 4'-CH(CH.sub.3)--O-2' bridge.

[0120] In certain embodiments, the wing-gap-wing motif is any of the group consisting of 3-9-3, 4-9-4, and 5-9-5.

[0121] In certain embodiments, described herein are compounds comprising a modified oligonucleotide consisting of 15, 17, or 19 linked nucleosides and fully complementary to a differentiating polymorphism, wherein the modified oligonucleotide comprises a wing-gap-wing motif, wherein position 6, 8, 10, or 14 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, aligns with the differentiating polymorphism; and at least one high-affinity sugar modification.

[0122] In certain embodiments, at least one of positions 2, 3, 6, 9, 10, 11, 13, or 14 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, comprises the at least one high-affinity sugar modification.

[0123] In certain embodiments, the high-affinity sugar modification is a bicyclic sugar. In certain embodiments, the bicyclic sugar comprises a 4'-CH(CH.sub.3)--O-2' bridge.

[0124] In certain embodiments, the wing-gap-wing motif is any of the group consisting of 3-9-3, 4-9-4, and 5-95.

[0125] In certain embodiments, described herein are compounds comprising a modified oligonucleotide consisting of 15 linked nucleosides and 90% complementary to a differentiating polymorphism, wherein the modified oligonucleotide comprises a wing-gap-wing motif, wherein position 8 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, aligns with the differentiating polymorphism; and at least one high-affinity sugar modification. In certain embodiments, the modified oligonucleotide is 100% complementary to the differentiating polymorphism.

[0126] In certain embodiments, each of nucleoside positions 2, 3, 13, and 14 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, comprise the at least one high-affinity sugar modification.

[0127] In certain embodiments, the high-affinity sugar modification is a bicyclic sugar. In certain embodiments, the bicyclic sugar comprises a 4'-CH(CH.sub.3)--O-2' bridge.

[0128] In certain embodiments, the wing-gap-wing motif is 3-9-3.

[0129] In certain embodiments, described herein are methods of selectively reducing expression of an allelic variant of a gene containing a single nucleotide polymorphism in a cell, tissue, or animal, comprising administering to the cell, tissue, or animal a compound comprising a modified oligonucleotide complementary to a differentiating polymorphism, wherein the modified oligonucleotide comprises a wing-gap-wing motif and wherein position 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, aligns with the differentiating polymorphism. In certain embodiments, the modified oligonucleotide is 90% complementary to the single differentiating polymorphism. In certain embodiments, the modified oligonucleotide is 95% complementary to the single nucleotide polymorphism site. In certain embodiments, the modified oligonucleotide is 100% complementary to the single nucleotide polymorphism site.

[0130] In certain embodiments, the single nucleotide polymorphism site is from 12 to 30 nucleobases in length. In certain embodiments, the single nucleotide polymorphism site is from 15 to 25 nucleobases in length. In certain embodiments, the single nucleotide polymorphism site is from 17 to 22 nucleobases in length. In certain embodiments, the single nucleotide polymorphism site is 17 nucleobases in length. In certain embodiments, the single nucleotide polymorphism site is 18 nucleobases in length. In certain embodiments, the single nucleotide polymorphism site is 19 nucleobases in length. In certain embodiments, the single nucleotide polymorphism site is 20 nucleobases in length.

[0131] In certain embodiments, the allelic variant is associated with disease. In certain embodiments, the disease is Huntington's Disease.

[0132] In certain embodiments, the modified oligonucleotide is a single-stranded oligonucleotide.

[0133] In certain embodiments, at least one internucleoside linkage is a modified internucleoside linkage. In certain embodiments, each internucleoside linkage is a phosphorothioate internucleoside linkage.

[0134] In certain embodiments, at least one nucleoside comprises a modified nucleobase. In certain embodiments, the at least one modified nucleobase is a 5'-methylcytosine.

[0135] In certain embodiments, at least one nucleoside comprises a modified sugar. In certain embodiments, the modified sugar is a high-affinity sugar modification. In certain embodiments, the high-affinity sugar is a bicyclic sugar. In certain embodiments, each bicyclic sugar comprises a 4'-CH(CH.sub.3)--O-2' bridge.

[0136] In certain embodiments, at least one of nucleoside positions 2, 3, 13, and 14 of the modified oligonucleotide, counting from the 5' terminus of the modified oligonucleotide, comprises a nucleoside having a bicyclic sugar wherein the bicyclic sugar comprises a 4'-CH(CH.sub.3)--O-2' bridge.

[0137] In certain embodiments, each of nucleoside positions 2, 3, 13, and 14 of the modified oligonucleotide, counting from the 5' terminus of the modified oligonucleotide, comprises a bicyclic sugar wherein the bicyclic sugar comprises a 4'-CH(CH.sub.3)--O-2' bridge.

[0138] In certain embodiments, the at least one modified sugar comprises a 2'-O-methoxyethyl. In certain embodiments, each nucleoside positioned in a wing segment of the modified oligonucleotide comprises a 2'-O-methoxyethyl modification.

[0139] In certain embodiments, the wing-gap-wing motif is any of the group consisting of 2-9-6, 3-9-3, 3-9-4, 3-9-5, 4-7-4, 4-9-4, 4-9-5, 4-10-5, 4-11-4, 4-11-5, 5-7-5, 5-8-6, 5-9-3, 5-9-5, 5-10-4, 5-10-5, 6-7-6, 6-8-5, and 6-9-2.

[0140] In certain embodiments, the modified oligonucleotide is not a ribozyme, a double stranded siRNA, or an shRNA.

[0141] In certain embodiments, the single nucleotide polymorphism site is on a mutant allele that is associated with disease. In certain embodiments, the single nucleotide polymorphism site contains a differentiating polymorphism.

[0142] In certain embodiments, the modified antisense oligonucleotide consists of 12 to 20 linked nucleosides. In certain embodiments, the modified antisense oligonucleotide consists of 15 to 19 linked nucleosides.

[0143] In certain embodiments, the gap region is 7 to 11 nucleosides in length, the 5' wing region is 1 to 6 nucleobases in length and 3' wing region is 1 to 6 nucleobases in length.

[0144] In certain embodiments, wherein at least one nucleoside of at least one of the wing regions comprises a modified sugar or sugar surrogate.

[0145] In certain embodiments, each of the nucleosides of each wing region comprises a modified sugar or sugar surrogate. In certain embodiments, the sugar or sugar surrogate is a 2'-O-methoxyethyl modified sugar.

[0146] In certain embodiments, at least one of the wing regions comprises a 4' to 2' bicyclic nucleoside and at least one of the remaining wing nucleosides is a non-bicyclic 2'-modified nucleoside.

[0147] In certain embodiments, the non-bicyclic 2'-modified nucleoside is a 2'-O-methoxyethyl nucleoside.

[0148] In certain embodiments, 4' to 2' bicyclic nucleoside is a 4'-CH(CH.sub.3)--O-2' bicyclic nucleoside.

[0149] In certain embodiments, described herein are methods of selectively reducing expression of an allelic variant of a gene containing a single nucleotide polymorphism in a cell, tissue, or animal, comprising administering to the cell, tissue, or animal a compound comprising a modified oligonucleotide complementary to a differentiating polymorphism, wherein the modified oligonucleotide comprises a wing-gap-wing motif and wherein position 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, aligns with the differentiating polymorphism.

[0150] In certain embodiments, described herein are methods of selectively reducing expression of an allelic variant of a gene containing a single nucleotide polymorphism in a cell, tissue, or animal, comprising administering to the cell, tissue, or animal a compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and complementary to a differentiating polymorphism, wherein the modified oligonucleotide comprises a wing-gap-wing motif and wherein position 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide aligns with the differentiating polymorphism; and wherein the allelic variant is a mutant allele.

[0151] In certain embodiments, the mutant allele is associated with any disease from the group consisting of Alzheimer's disease, Creutzfeldt-Jakob disease, fatal familial insomnia, Alexander disease, Parkinson's disease, amyotrophic lateral sclerosis, dentato-rubral and pallido-luysian atrophy DRPA, spino-cerebellar ataxia, Torsion dystonia, cardiomyopathy, chronic obstructive pulmonary disease (COPD), liver disease, hepatocellular carcinoma, systemic lupus erythematosus, hypercholesterolemia, breast cancer, asthma, Type 1 diabetes, Rheumatoid arthritis, Graves disease, SLE, spinal and bulbar muscular atrophy, Kennedy's disease, progressive childhood posterior subcapsular cataracts, cholesterol gallstone disease, arthrosclerosis, cardiovascular disease, primary hypercalciuria, alpha-thallasemia, obsessive compulsive disorder, Anxiety, comorbid depression, congenital visual defects, hypertension, metabolic syndrome, prostate cancer, congenital myasthenic syndrome, peripheral arterial disease, atrial fibrillation, sporadic pheochromocytoma, congenital malformations, Machado-Joseph disease, Huntington's disease, and Autosomal Dominant Retinitis Pigmentosa disease.

[0152] In certain embodiments, described herein are methods of treating Huntington's Disease, comprising selectively reducing expression of an allelic variant of a gene containing a single nucleotide polymorphism in a cell, tissue, or animal, comprising administering to the cell, tissue, or animal a compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and complementary to differentiating polymorphism, wherein the modified oligonucleotide comprises a wing-gap-wing motif and wherein position 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, aligns with differentiating polymorphism; and wherein the allelic variant is associated with Huntington's Disease.

[0153] In certain embodiments, position 8, 9, or 10 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, or positions 4, 5, or 6 of the modified oligonucleotide, as counted from the 5' terminus of the gap, aligns with the single nucleotide polymorphism.

Single Nucleotide Polymorphisms (SNPs)

[0154] Single-nucleotide polymorphisms (SNPs) are single base-pair alterations in the DNA sequence that represent a major source of genetic heterogeneity (Gene. 1999, 234:177). SNP genotyping is an important tool with which to investigate these genetic variants (Genome Res. 2000, 10:895; Trends Biotechnol. 2000, 18:77). In certain embodiments, antisense compounds designed to selectively reduce an allelic variant of a gene containing an SNP were selected based on potency, selectivity and population genetics coverage.

Potency

[0155] In certain embodiments, antisense compounds designed to selectively reduce an allelic variant of a gene containing a SNP are created based on potency of the antisense compound. Potency generally refers to how amenable the targeted sequence area is to antisense inhibition. In certain embodiments, specific SNP sites may be particularly amenable to antisense inhibition. Certain such highly amenable SNP sites may be targeted by antisense compounds for selectively reducing an allelic variant of a gene. Potency is demonstrated by the percent inhibition of mutant mRNA achieved by the antisense oligonucleotides targeting a SNP compared to the percent inhibition of mutant mRNA achieved by the benchmark oligonucleotide.

Selectivity

[0156] In certain embodiments, antisense compounds designed to selectively reduce an allelic variant of a gene containing a SNP are created based on selectivity of the antisense compound. Selectivity generally refers to antisense compounds comprising a particular sequence, motif, and chemical modification(s) that preferentially target the one or more differentiating polymorphisms (SNPs) in the RNA encoding a mutant HTT protein compared to the RNA encoding a wild type HTT protein. In certain embodiments, specific sequences, motifs, and chemical modification(s) are particularly selective in reducing an allelic variant of a gene containing a SNP. Certain such sequences, motifs, and chemical modification(s) are utilized to selectively reduce an allelic variant of a gene. Selectivity is demonstrated by the ability of the antisense oligonucleotide targeting a SNP to inhibit expression of the major allele or mutant allele preferentially compared to the minor allele or wild type allele.

Population Genetics

[0157] In certain embodiments, antisense compounds designed to selectively reduce an allelic variant of a gene containing an SNP are created based on the population genetics of a population afflicted with disease. Population genetics means the frequency at which the SNP appears in the disease chromosome of patients afflicted with a particular disease. In certain embodiments, the disease is Huntington disease. Where potency and selectivity amongst antisense compounds is equal, SNP targets that have higher population genetics coverage are favored over SNPs that have a weaker association with disease chromosomes.

Antisense Compounds

[0158] Oligomeric compounds may include, but are not limited to, oligonucleotides, oligonucleosides, oligonucleotide analogs, oligonucleotide mimetics, antisense compounds, antisense oligonucleotides, and siRNAs. An oligomeric compound may be "antisense" to a target nucleic acid, meaning that is is capable of undergoing hybridization to a target nucleic acid through hydrogen bonding.

[0159] In certain embodiments, an antisense compound is an antisense oligonucleotide. In certain embodiments, the antisense compound is not a ribozyme, a double stranded siRNA, or an shRNA.

[0160] In certain embodiments, an antisense compound has a nucleobase sequence that, when written in the 5' to 3' direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is targeted. In certain such embodiments, an antisense oligonucleotide has a nucleobase sequence that, when written in the 5' to 3' direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is targeted.

[0161] In certain embodiments, antisense compounds are 12 to 30 subunits in length. In other words, such antisense compounds are from 12 to 30 linked subunits. In other embodiments, the antisense compound is 8 to 80, 12 to 50, 15 to 30, 18 to 24, 19 to 22, or 20 linked subunits. In certain such embodiments, the antisense compounds are 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or 80 linked subunits in length, or a range defined by any two of the above values. In some embodiments the antisense compound is an antisense oligonucleotide, and the linked subunits are nucleosides.

[0162] In certain embodiments antisense oligonucleotides targeted to a nucleic acid may be shortened or truncated. For example, a single subunit may be deleted from the 5' end (5' truncation), or alternatively from the 3' end (3' truncation). A shortened or truncated antisense compound targeted to a nucleic acid may have two subunits deleted from the 5' end, or alternatively may have two subunits deleted from the 3' end, of the antisense compound. Alternatively, the deleted nucleosides may be dispersed throughout the antisense compound, for example, in an antisense compound having one nucleoside deleted from the 5' end and one nucleoside deleted from the 3' end.

[0163] When a single additional subunit is present in a lengthened antisense compound, the additional subunit may be located at the 5' or 3' end of the antisense compound. When two or more additional subunits are present, the added subunits may be adjacent to each other, for example, in an antisense compound having two subunits added to the 5' end (5' addition), or alternatively to the 3' end (3' addition), of the antisense compound. Alternatively, the added subunits may be dispersed throughout the antisense compound, for example, in an antisense compound having one subunit added to the 5' end and one subunit added to the 3' end.

[0164] It is possible to increase or decrease the length of an antisense compound, such as an antisense oligonucleotide, and/or introduce mismatch bases without eliminating activity. For example, in Woolf et al. (Proc. Natl. Acad. Sci. USA 89:7305-7309, 1992), a series of antisense oligonucleotides 13-25 nucleobases in length were tested for their ability to induce cleavage of a target RNA in an oocyte injection model. Antisense oligonucleotides 25 nucleobases in length with 8 or 11 mismatch bases near the ends of the antisense oligonucleotides were able to direct specific cleavage of the target mRNA, albeit to a lesser extent than the antisense oligonucleotides that contained no mismatches. Similarly, target specific cleavage was achieved using 13 nucleobase antisense oligonucleotides, including those with 1 or 3 mismatches.

[0165] Gautschi et al (J. Natl. Cancer Inst. 93:463-471, March 2001) demonstrated the ability of an oligonucleotide having 100% complementarity to the bc1-2 mRNA and having 3 mismatches to the bc1-xL mRNA to reduce the expression of both bc1-2 and bc1-xL in vitro and in vivo. Furthermore, this oligonucleotide demonstrated potent anti-tumor activity in vivo.

[0166] Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358,1988) tested a series of tandem 14 nucleobase antisense oligonucleotides, and a 28 and 42 nucleobase antisense oligonucleotides comprised of the sequence of two or three of the tandem antisense oligonucleotides, respectively, for their ability to arrest translation of human DHFR in a rabbit reticulocyte assay. Each of the three 14 nucleobase antisense oligonucleotides alone was able to inhibit translation, albeit at a more modest level than the 28 or 42 nucleobase antisense oligonucleotides.

[0167] However, selective reduction of expression of an allelic variant is optimized when the SNP contained in the target nucleic anneals to a complementary base in the antisense compound and not a mismatched base. Moreover, selectivity in general is increased when there are fewer mismatches between the SNP site and the antisense compound. However, a certain number of mismatches may be tolerated.

Antisense Compound Motifs

[0168] In certain embodiments, antisense compounds targeted to a nucleic acid have chemically modified subunits arranged in patterns, or motifs, to confer to the antisense compounds properties such as enhanced the inhibitory activity, increased binding affinity for a target nucleic acid, or resistance to degradation by in vivo nucleases.

[0169] Chimeric antisense compounds typically contain at least one region modified so as to confer increased resistance to nuclease degradation, increased cellular uptake, increased binding affinity for the target nucleic acid, and/or increased inhibitory activity. A second region of a chimeric antisense compound may optionally serve as a substrate for the cellular endonuclease RNase H, which cleaves the RNA strand of an RNA:DNA duplex.

[0170] Antisense compounds having a gapmer motif are considered chimeric antisense compounds. In a gapmer an internal region having a plurality of nucleotides that supports RNaseH cleavage is positioned between external regions having a plurality of nucleotides that are chemically distinct from the nucleosides of the internal region. In the case of an antisense oligonucleotide having a gapmer motif, the gap segment generally serves as the substrate for endonuclease cleavage, while the wing segments comprise modified nucleosides. In the case of an antisense oligonucleotide for selectively reducing expression of an allelic variant of a gene containing a SNP, the SNP anneals to a nucleobase within the gap segment.

[0171] In certain embodiments, the SNP anneals or is complementary to a nucleobase at position 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 of the antisense oligonucleotide, wherein position refers to the orientation of a nucleobase within the antisense oligonucleotide counting from the 5' terminus of the antisense oligonucleotide. For example, the 5' most nucleobase within the antisense oligonucleotide is in the first position of the antisense oligonucleotide. In certain embodiments, the SNP anneals or is complementary to a nucleobase at position 6, 7, 8, 9, or 10 of the antisense oligonucleotide (counting from the 5' terminus). In certain embodiments, the SNP anneals or is complementary to a nucleobase at position 9 or 10 of the antisense oligonucleotide (counting from the 5' terminus).

[0172] In certain embodiments, the SNP anneals to a nucleobase at position 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 of the gap segment, wherein position refers to the orientation of a nucleobase within the gap segment counting from the 5' terminus of the gap segment. For example, the 5' most nucleobase within the gap segment is in the first position of the gap segment. In certain embodiments, the SNP anneals to a nucleobase at position 4, 5, 6, or 7 counting from the 5' terminus of the gap segment. In certain embodiments, the SNP anneals to a nucleobase at position 4 or 5 beginning from the 5' terminus of the gap segment.

[0173] In certain embodiments, the regions of a gapmer are differentiated by the types of sugar moieties comprising each distinct region. The types of sugar moieties that are used to differentiate the regions of a gapmer may in some embodiments include .beta.-D-ribonucleosides, .beta.-D-deoxyribonucleosides, 2'-modified nucleosides (such 2'-modified nucleosides may include 2'-MOE, and 2'-O--CH.sub.3, among others), and bicyclic sugar modified nucleosides (such bicyclic sugar modified nucleosides may include those having a 4'-(CH.sub.2)n-O-2' bridge, where n=1 or n=2). The bicyclic moiety may be a cEt having the formula 4'-CH(CH.sub.3)--O-2.'

[0174] The wing-gap-wing motif is frequently described as "X--Y--Z", where "X" represents the length of the 5' wing region, "Y" represents the length of the gap region, and "Z" represents the length of the 3' wing region. As used herein, a gapmer described as "X--Y--Z" has a configuration such that the gap segment is positioned immediately adjacent to each of the 5' wing segment and the 3' wing segment. Thus, no intervening nucleotides exist between the 5' wing segment and gap segment, or the gap segment and the 3' wing segment. Any of the antisense compounds described herein can have a gapmer motif. In some embodiments, X and Z are the same, in other embodiments they are different. In certain embodiments, Y is between 8 and 15 nucleotides. In certain embodiments, Y is comprised of deoxynucleotides. X, Y or Z can be any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or more nucleotides. Thus, gapmers of the present invention include, but are not limited to, for example 1-10-1, 1-18-1, 2-8-2, 2-9-6, 2-10-2, 2-13-5, 2-16-2, 3-9-3, 3-9-5, 3-10-3, 3-14-3, 4-8-4, 4-9-5, 4-10-5, 4-11-4, 4-12-3, 4-12-4, 5-8-5, 5-9-5, 5-10-4, 5-10-5, or 6-8-6.

[0175] In certain embodiments, the antisense compound has a "wingmer" motif, having a wing-gap or gap-wing configuration, i.e. an X--Y or Y--Z configuration as described above for the gapmer configuration. Thus, wingmer configurations of the present invention include, but are not limited to, for example 5-10, 8-4, 4-12, 12-4, 3-14, 16-2, 18-1, 10-3, 2-10, 1-10, 8-2, 2-13, 5-13, 5-8, or 6-8.

[0176] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 2-9-6 gapmer motif or a 6-9-2 gapmer motif.

[0177] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 3-9-3 gapmer motif.

[0178] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 3-9-5 gapmer motif or 5-9-3 gapmer motif.

[0179] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 4-9-5 gapmer motif or 5-9-4 gapmer motif.

[0180] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 4-10-5 gapmer motif or 5-10-4 gapmer motif.

[0181] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 4-11-4 gapmer motif.

[0182] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 5-9-5 gapmer motif.

[0183] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 5-8-6 gapmer motif or a 6-8-5 gapmer motif.

[0184] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 6-7-6 gapmer motif.

[0185] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 6-8-5 gapmer motif or a 5-8-6 gapmer motif.

[0186] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 3-9-4 gapmer motif or a 4-9-3 gapmer motif.

[0187] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 5-7-5 gapmer motif.

[0188] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 4-7-4 gapmer motif.

[0189] In certain embodiments, antisense compounds targeted to a nucleic acid possess a 5-10-5 gapmer motif.

[0190] In certain embodiments, an antisense compound targeted to a nucleic acid has a gap-widened motif.

Certain Mixed Wings

[0191] In certain embodiments, the invention provides gapmer compounds wherein at least one nucleoside of one wing is differently modified compared to at least one other nucleoside of the same wing. Such antisense compounds are referred to as mixed wing antisense compounds (see WO 2008/049085). In certain embodiments, the modifications (or no modification) of one or more nucleosides of the 3' wing are different from those of one or more other nucleosides of the 3' wing. Such antisense compounds may be referred to as 3' mixed wing gapmers. In certain embodiments, the modifications (or no modification) of one or more nucleosides of the 5' wing are different from those of one or more other nucleosides of the 5' wing. Such antisense compounds may be referred to as 5' mixed wing gapmers. In certain embodiments, the modifications (or no modification) of one or more nucleosides of the 3' wing are different from those of one or more other nucleosides of the 3' wing and the modifications (or no modification) of one or more nucleosides of the 5' wing are different from those of one or more other nucleosides of the 5' wing. Such antisense compounds may be referred to as 3', 5' mixed wing gapmers. In such embodiment, the modifications and combination of modifications at the 3' wing and at the 5' wing may be the same or they may be different.

[0192] In certain embodiments, mixed wing compounds have desirable properties. Certain nucleoside modifications confer on the antisense compound a desirable property, for example increased affinity for a target or nuclease resistance, but also confer an undesirable property, for example increased toxicity. Incorporation of certain other nucleoside modifications results in antisense compounds with different profiles of properties. In certain embodiments, one may combine modifications in one or both wings to optimize desirable characteristics and/or minimize undesirable characteristics. In certain embodiments, the wings of a mixed wing antisense compound comprise one or more nucleoside comprising a first modification that increases affinity of the antisense compound for a target nucleic acid compared to an antisense compound comprising unmodified nucleosides; and one or more nucleoside comprising a second modification that results in reduced toxicity compared to an antisense compound with wings comprising nucleosides that all comprise the first modification.

[0193] In certain embodiments, an antisense compound comprises at least one wing comprising at least one MOE substituted nucleoside and at least one high affinity modification. In certain such embodiments, the at least one MOE substituted nucleoside and the at least one high affinity are in the 3' wing. In certain such embodiments, the at least one MOE substituted nucleoside and the at least one high affinity are in the 5' wing.

[0194] In certain embodiments, an antisense compound comprises 1, 2 or 3 high affinity modifications in the 5' and/or 3' wings.

Target Nucleic Acids, Target Regions and Nucleotide Sequences

[0195] In certain embodiments, an allelic variant of huntingtin is selectively reduced. Nucleotide sequences that encode huntingtin include, without limitation, the following: GENBANK Accession No. NT.sub.--006081.18, truncated from nucleotides 1566000 to 1768000 (replaced by GENBANK Accession No. NT.sub.--006051), incorporated herein as SEQ ID NO: 1, and NM.sub.--002111.6, incorporated herein as SEQ ID NO: 2.

[0196] It is understood that the sequence set forth in each SEQ ID NO in the Examples contained herein is independent of any modification to a sugar moiety, an internucleo side linkage, or a nucleobase. As such, antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an internucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis No) indicate a combination of nucleobase sequence and motif.

[0197] In certain embodiments, a target region is a structurally defined region of the target nucleic acid. For example, a target region may encompass a 3' UTR, a 5' UTR, an exon, an intron, an exon/intron junction, a coding region, a translation initiation region, translation termination region, or other defined nucleic acid region. The structurally defined regions for huntingtin can be obtained by accession number from sequence databases such as NCBI and such information is incorporated herein by reference. In certain embodiments, a target region may encompass the sequence from a 5' target site of one target segment within the target region to a 3' target site of another target segment within the same target region.

[0198] Targeting includes determination of at least one target segment to which an antisense compound hybridizes, such that a desired effect occurs. In certain embodiments, the desired effect is a reduction in mRNA target nucleic acid levels of a particular allelic variant. In certain embodiments, the desired effect is reduction of levels of the protein encoded by the target nucleic acid or a phenotypic change associated with a particular alleleic variant.

[0199] A target region may contain one or more target segments. Multiple target segments within a target region may be overlapping. Alternatively, they may be non-overlapping. In certain embodiments, target segments within a target region are separated by no more than about 300 nucleotides. In certain embodiments, target segments within a target region are separated by a number of nucleotides that is, is about, is no more than, is no more than about, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 nucleotides on the target nucleic acid, or is a range defined by any two of the preceeding values. In certain embodiments, target segments within a target region are separated by no more than, or no more than about, 5 nucleotides on the target nucleic acid. In certain embodiments, target segments are contiguous. Contemplated are target regions defined by a range having a starting nucleic acid that is any of the 5' target sites or 3' target sites listed herein.

[0200] Suitable target segments may be found within a 5' UTR, a coding region, a 3' UTR, an intron, an exon, or an exon/intron junction. Target segments containing a start codon or a stop codon are also suitable target segments. A suitable target segment may specifically exclude a certain structurally defined region such as the start codon or stop codon.

[0201] The determination of suitable target segments may include a comparison of the sequence of a target nucleic acid to other sequences throughout the genome. For example, the BLAST algorithm may be used to identify regions of similarity amongst different nucleic acids. This comparison can prevent the selection of antisense compound sequences that may hybridize in a non-specific manner to sequences other than a selected target nucleic acid (i.e., non-target or off-target sequences).

Cell Lines

[0202] In certain embodiments, the GM04281, GM02171, and GM02173B cell lines are used in experiments described herein below. The GM04281 cell line has a wild-type HTT allele that contains 17 repeats and a mutant HTT allele that contains 69 repeats. The cell line was derived from a patient both of whose parents were also affected by the disease. The GM02171 cell line was chosen as a counter screen control to the GM04281. This cell line was derived from the daughter of parents, only one of whom had the disease. The daughter had not developed HD but was considered to be at risk. The GM02173B cell line was also patient-derived and was used as a haplotype test control.

[0203] Table 1 provides SNPs found in the GM04281, GM02171, and GM02173B cell lines. Also provided are the allelic variants found at each SNP position, the genotype for each of the cell lines, and the percentage of HD patients having a particular allelic variant. For example, the two allelic variants for SNP rs6446723 are T and C. The GM02171 cell line is homozygous CC, the GM02173 cell line is heterozygous TC, and the GM04281 cell line is homozygous TT. Fifty percent of HD patients have a T at SNP position rs6446723.

TABLE-US-00001 TABLE 1 Allelic Variations for SNPs Associated with HD SNP Variation GM02171 GM02173 GM04281 TargetPOP allele rs6446723 T/C CC TC TT 0.50 T rs3856973 A/G AA AG GG 0.50 G rs2285086 A/G GG AG AA 0.50 A rs363092 A/C AA AC CC 0.49 C rs916171 C/G GG GC CC 0.49 C rs6844859 T/C CC TC TT 0.49 T rs7691627 A/G AA AG GG 0.49 G rs4690073 A/G AA AG GG 0.49 G rs2024115 A/G GG AG AA 0.48 A rs11731237 T/C CC TC TT 0.43 T rs362296 A/C AC AC AC 0.42 C rs10015979 A/G AA AG GG 0.42 G rs7659144 C/G CG CG CC 0.41 C rs363096 T/C CC TC TT 0.40 T rs362273 A/G AG AG AA 0.39 A rs16843804 T/C TC TC CC 0.38 C rs362271 A/G AG AG GG 0.38 G rs362275 T/C TC TC CC 0.38 C rs3121419 T/C TC TC CC 0.38 C rs362272 A/G -- AG GG 0.38 G rs3775061 A/G AG AG AA 0.38 A rs34315806 T/C TC TC CC 0.38 C rs363099 T/C TC TC CC 0.38 C rs2298967 T/C TC TC TT 0.38 T rs363088 A/T TA TA AA 0.38 A rs363064 T/C TC TC CC 0.35 C rs363102 A/G AA AA AA 0.23 G rs2798235 A/G GG GG GG 0.21 A rs363080 T/C CC CC CC 0.21 T rs363072 A/T TA AA AA 0.13 A rs363125 A/C AC CC CC 0.12 C rs362303 T/C TC CC CC 0.12 C rs362310 T/C TC CC CC 0.12 C rs10488840 A/G AG GG GG 0.12 G rs362325 T/C TC TT TT 0.11 T rs35892913 A/G GG GG GG 0.10 A rs363102 A/G AA AA AA 0.09 A rs363096 T/C CC TC TT 0.09 C rs11731237 T/C CC TC TT 0.09 C rs10015979 A/G AA AG GG 0.08 A rs363080 T/C CC CC CC 0.07 C rs2798235 A/G GG GG GG 0.07 G rs1936032 C/G CC CC CC 0.06 C rs2276881 A/G GG GG GG 0.06 G rs363070 A/G AA AA AA 0.06 A rs35892913 A/G GG GG GG 0.04 G rs12502045 T/C CC CC CC 0.04 C rs6446723 T/C CC TC TT 0.04 C rs7685686 A/G GG AG AA 0.04 G rs3733217 T/C CC CC CC 0.03 C rs6844859 T/C CC TC TT 0.03 C rs362331 T/C CC TC TT 0.03 C

Hybridization

[0204] In some embodiments, hybridization occurs between an antisense compound disclosed herein and a SNP site. The most common mechanism of hybridization involves hydrogen bonding (e.g., Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding) between complementary nucleobases of the nucleic acid molecules.

[0205] Hybridization can occur under varying conditions. Stringent conditions are sequence-dependent and are determined by the nature and composition of the nucleic acid molecules to be hybridized.

[0206] In certain embodiments, the antisense compounds provided herein are specifically hybridizable with the nucleic acid of a particular allelic variant.

Complementarity

[0207] An antisense compound and a target nucleic acid are complementary to each other when a sufficient number of nucleobases of the antisense compound can hydrogen bond with the corresponding nucleobases of the target nucleic acid, such that a desired effect will occur (e.g., selective reduction of a gene product of an allelic variant).

[0208] Non-complementary nucleobases between an antisense compound and a target nucleic acid may be tolerated provided that the antisense compound remains able to specifically hybridize to a target nucleic acid. Moreover, an antisense compound may hybridize over one or more segments of a target nucleic acid such that intervening or adjacent segments are not involved in the hybridization event (e.g., a loop structure, mismatch or hairpin structure).

[0209] In certain embodiments, the antisense compounds provided herein, or a specified portion thereof, are, or are at least, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to a target nucleic acid, a target region, target segment, SNP site, or specified portion thereof. Percent complementarity of an antisense compound with a target nucleic acid can be determined using routine methods. For example, an antisense compound in which 18 of 20 nucleobases of the antisense compound are complementary to a target region, and would therefore specifically hybridize, would represent 90 percent complementarity. In this example, the remaining noncomplementary nucleobases may be clustered or interspersed with complementary nucleobases and need not be contiguous to each other or to complementary nucleobases. As such, an antisense compound which is 18 nucleobases in length having 4 (four) noncomplementary nucleobases which are flanked by two regions of complete complementarity with the target nucleic acid would have 77.8% overall complementarity with the target nucleic acid and would thus fall within the scope of the present invention. Percent complementarity of an antisense compound with a region of a target nucleic acid can be determined routinely using BLAST programs (basic local alignment search tools) and PowerBLAST programs known in the art (Altschul et al., J. Mol. Biol., 1990, 215, 403 410; Zhang and Madden, Genome Res., 1997, 7, 649 656). Percent homology, sequence identity or complementarity, can be determined by, for example, the Gap program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, Madison Wis.), using default settings, which uses the algorithm of Smith and Waterman (Adv. Appl. Math., 1981, 2, 482 489).

[0210] In certain embodiments, the antisense compounds provided herein, or specified portions thereof, are fully complementary (i.e. 100% complementary) to a target nucleic acid, a SNP site, target region, target segment, or specified portion thereof. As used herein, "fully complementary" means each nucleobase of an antisense compound is capable of precise base pairing with the corresponding nucleobases of a target nucleic acid. For example, a 20 nucleobase antisense compound is fully complementary to a target sequence that is 400 nucleobases long, so long as there is a corresponding 20 nucleobase portion of the target nucleic acid that is fully complementary to the antisense compound. Fully complementary can also be used in reference to a specified portion of the first and/or the second nucleic acid. For example, a 20 nucleobase portion of a 30 nucleobase antisense compound can be "fully complementary" to a target sequence that is 400 nucleobases long. The 20 nucleobase portion of the 30 nucleobase oligonucleotide is fully complementary to the target sequence if the target sequence has a corresponding 20 nucleobase portion wherein each nucleobase is complementary to the 20 nucleobase portion of the antisense compound. At the same time, the entire 30 nucleobase antisense compound may or may not be fully complementary to the target sequence, depending on whether the remaining 10 nucleobases of the antisense compound are also complementary to the target sequence.

[0211] The location of a non-complementary nucleobase may be at the 5' end or 3' end of the antisense compound. Alternatively, the non-complementary nucleobase or nucleobases may be at an internal position of the antisense compound. When two or more non-complementary nucleobases are present, they may be contiguous (i.e. linked) or non-contiguous. In one embodiment, a non-complementary nucleobase is located in the wing segment of a gapmer antisense oligonucleotide.

[0212] In certain embodiments, antisense compounds that are, or are up to 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length comprise no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 non-complementary nucleobase(s) relative to a target nucleic acid, SNP site, or specified portion thereof.

[0213] In certain embodiments, antisense oligonucleotides that are, or are up to 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length comprise no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 non-complementary nucleobase(s) relative to a target nucleic acid, SNP site, or specified portion thereof.

[0214] The antisense compounds provided herein also include those which are complementary to a portion of a target nucleic acid. As used herein, "portion" refers to a defined number of contiguous (i.e. linked) nucleobases within a region or segment of a target nucleic acid. A "portion" can also refer to a defined number of contiguous nucleobases of an antisense compound. In certain embodiments, the antisense compounds, are complementary to at least an 8 nucleobase portion of a target segment. In certain embodiments, the antisense compounds are complementary to at least a 12 nucleobase portion of a target segment. In certain embodiments, the antisense compounds are complementary to at least a 15 nucleobase portion of a target segment. Also contemplated are antisense compounds that are complementary to at least a 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more nucleobase portion of a target segment, or a range defined by any two of these values.

Identity

[0215] The antisense compounds provided herein may also have a defined percent identity to a particular nucleotide sequence, SEQ ID NO, or compound represented by a specific Isis number, or portion thereof. As used herein, an antisense compound is identical to the sequence disclosed herein if it has the same nucleobase pairing ability. For example, a RNA which contains uracil in place of thymidine in a disclosed DNA sequence would be considered identical to the DNA sequence since both uracil and thymidine pair with adenine. Shortened and lengthened versions of the antisense compounds described herein as well as compounds having non-identical bases relative to the antisense compounds provided herein also are contemplated. The non-identical bases may be adjacent to each other or dispersed throughout the antisense compound. Percent identity of an antisense compound is calculated according to the number of bases that have identical base pairing relative to the sequence to which it is being compared.

[0216] In certain embodiments, the antisense compounds, or portions thereof, are at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to one or more of the antisense compounds or SEQ ID NOs, or a portion thereof, disclosed herein.

[0217] In certain embodiments, a portion of the antisense compound is compared to an equal length portion of the target nucleic acid. In certain embodiments, an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleobase portion is compared to an equal length portion of the target nucleic acid.

[0218] In certain embodiments, a portion of the antisense oligonucleotide is compared to an equal length portion of the target nucleic acid. In certain embodiments, an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleobase portion is compared to an equal length portion of the target nucleic acid.

Modifications

[0219] A nucleoside is a base-sugar combination. The nucleobase (also known as base) portion of the nucleoside is normally a heterocyclic base moiety. Nucleotides are nucleosides that further include a phosphate group covalently linked to the sugar portion of the nucleoside. For those nucleosides that include a pentofuranosyl sugar, the phosphate group can be linked to the 2', 3' or 5' hydroxyl moiety of the sugar. Oligonucleotides are formed through the covalent linkage of adjacent nucleosides to one another, to form a linear polymeric oligonucleotide. Within the oligonucleotide structure, the phosphate groups are commonly referred to as forming the internucleoside linkages of the oligonucleotide.

[0220] Modifications to antisense compounds encompass substitutions or changes to internucleoside linkages, sugar moieties, or nucleobases. Modified antisense compounds are often preferred over native forms because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for nucleic acid target, increased stability in the presence of nucleases, or increased inhibitory activity.

[0221] Chemically modified nucleosides may also be employed to increase the binding affinity of a shortened or truncated antisense oligonucleotide for its target nucleic acid. Consequently, comparable results can often be obtained with shorter antisense compounds that have such chemically modified nucleosides.

[0222] Chemically modified nucleosides may also be employed to increase selectivity in reducing expression the gene product of an allelic variant.

Modified Internucleoside Linkages

[0223] The naturally occurring internucleoside linkage of RNA and DNA is a 3' to 5' phosphodiester linkage. Antisense compounds having one or more modified, i.e. non-naturally occurring, internucleoside linkages are often selected over antisense compounds having naturally occurring internucleoside linkages because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for target nucleic acids, and increased stability in the presence of nucleases.

[0224] Oligonucleotides having modified internucleoside linkages include internucleoside linkages that retain a phosphorus atom as well as internucleoside linkages that do not have a phosphorus atom. Representative phosphorus containing internucleoside linkages include, but are not limited to, phosphodiesters, phosphotriesters, methylphosphonates, phosphoramidate, and phosphorothioate. Methods of preparation of phosphorous-containing and non-phosphorous-containing linkages are well known.

[0225] In certain embodiments, antisense compounds comprise one or more modified internucleoside linkages. In certain embodiments, the modified internucleoside linkages are phosphorothioate linkages. In certain embodiments, each internucleoside linkage of an antisense compound is a phosphorothioate internucleoside linkage.

Modified Sugar Moieties

[0226] Antisense compounds of the invention can optionally contain one or more nucleosides wherein the sugar group has been modified. Such sugar modified nucleosides may impart enhanced nuclease stability, increased binding affinity, increased selectivity for an allelic variant, or some other beneficial biological property to the antisense compounds. In certain embodiments, nucleosides comprise a chemically modified ribofuranose ring moieties. Examples of chemically modified ribofuranose rings include without limitation, addition of substitutent groups (including 5' and 2' substituent groups, bridging of non-geminal ring atoms to form bicyclic nucleic acids (BNA), replacement of the ribosyl ring oxygen atom with S, N(R), or C(R1)(R)2 (R.dbd.H, C1-C12 alkyl or a protecting group) and combinations thereof. Examples of chemically modified sugars include 2'-F-5'-methyl substituted nucleoside (see PCT International Application WO 2008/101157 Published on Aug. 21, 2008 for other disclosed 5',2'-bis substituted nucleosides) or replacement of the ribosyl ring oxygen atom with S with further substitution at the 2'-position (see published U.S. Patent Application US2005-0130923, published on Jun. 16, 2005) or alternatively 5'-substitution of a BNA (see PCT International Application WO 2007/134181 Published on Nov. 22, 2007 wherein LNA is substituted with for example a 5'-methyl or a 5'-vinyl group).

[0227] Examples of nucleosides having modified sugar moieties include without limitation nucleosides comprising 5'-vinyl, 5'-methyl (R or S), 4'-S, 2'-F, 2'-OCH.sub.3 and 2'-O(CH.sub.2)2OCH.sub.3 substituent groups. The substituent at the 2' position can also be selected from allyl, amino, azido, thio, O-allyl, O--C1-C10 alkyl, OCF3, O(CH.sub.2)2SCH.sub.3, O(CH.sub.2)2-O--N(Rm)(Rn), and O--CH2-C(.dbd.O)--N(Rm)(Rn), where each Rm and Rn is, independently, H or substituted or unsubstituted C1-C10 alkyl.

[0228] As used herein, "bicyclic nucleosides" refer to modified nucleosides comprising a bicyclic sugar moiety. Examples of bicyclic nucleosides include without limitation nucleosides comprising a bridge between the 4' and the 2' ribosyl ring atoms. In certain embodiments, antisense compounds provided herein include one or more bicyclic nucleosides wherein the bridge comprises a 4' to 2' bicyclic nucleoside. Examples of such 4' to 2' bicyclic nucleosides, include but are not limited to one of the formulae: 4'-(CH.sub.2)--O-2' (LNA); 4'-(CH.sub.2)--S-2'; 4'-(CH.sub.2).sub.2--O-2' (ENA); 4'-CH(CH.sub.3)--O-2' and 4'-CH(CH.sub.2OCH.sub.3)--O-2' (and analogs thereof see U.S. Pat. No. 7,399,845, issued on Jul. 15, 2008); 4'-C(CH.sub.3)(CH.sub.3)--O-2' (and analogs thereof see published International Application WO/2009/006478, published Jan. 8, 2009); 4'-CH.sub.2--N(OCH.sub.3)-2' (and analogs thereof see published International Application WO/2008/150729, published Dec. 11, 2008); 4'-CH.sub.2--O--N(CH.sub.3)-2' (see published U.S. Patent Application US2004-0171570, published Sep. 2, 2004); 4'-CH.sub.2--N(R)--O-2', wherein R is H, C1-C12 alkyl, or a protecting group (see U.S. Pat. No. 7,427,672, issued on Sep. 23, 2008); 4'-CH.sub.2--C(H)(CH.sub.3)-2' (see Chattopadhyaya, et al., J. Org. Chem., 2009, 74, 118-134); and 4'-CH.sub.2--C--(.dbd.CH.sub.2)-2' (and analogs thereof see published International Application WO 2008/154401, published on Dec. 8, 2008). See, for example: Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 3607-3630; Wahlestedt et al., Proc. Natl. Acad. Sci. U.S.A., 2000, 97, 5633-5638; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Srivastava et al., J. Am. Chem. Soc., 129(26) 8362-8379 (Jul. 4, 2007); U.S. Pat. Nos. 7,053,207; 6,268,490; 6,770,748; 6,794,499; 7,034,133; and 6,525,191; Elayadi et al., Curr. Opinion Invens. Drugs, 2001, 2, 558-561; Braasch et al., Chem. Biol., 2001, 8, 1-7; and Orum et al., Curr. Opinion Mol. Ther., 2001, 3, 239-243; and U.S. Pat. No. 6,670,461; International applications WO 2004/106356; WO 94/14226; WO 2005/021570; U.S. Patent Publication Nos. US2004-0171570; US2007-0287831; US2008-0039618; U.S. Pat. No. 7,399,845; U.S. Patent Serial No. 12/129,154; 60/989,574; 61/026,995; 61/026,998; 61/056,564; 61/086,231; 61/097,787; 61/099,844; PCT International Applications Nos. PCT/US2008/064591; PCT/US2008/066154; PCT/US2008/068922; and Published PCT International Applications WO 2007/134181. Each of the foregoing bicyclic nucleosides can be prepared having one or more stereochemical sugar configurations including for example .alpha.-L-ribofuranose and .beta.-D-ribofuranose (see PCT international application PCT/DK98/00393, published on Mar. 25, 1999 as WO 99/14226).

[0229] In certain embodiments, bicyclic sugar moieties of BNA nucleosides include, but are not limited to, compounds having at least one bridge between the 4' and the 2' position of the pentofuranosyl sugar moiety wherein such bridges independently comprises 1 or from 2 to 4 linked groups independently selected from --[C(R.sub.a)(R.sub.b)].sub.n--, --C(R.sub.a).dbd.C(R.sub.b)--, --C(R.sub.a).dbd.N--, --C(.dbd.NR.sub.a)--, --C(.dbd.O)--, --C(.dbd.S)--, --O--, --Si(R.sub.a).sub.2--, --S(.dbd.O).sub.x--, and --N(R.sub.a)--;

[0230] wherein: [0231] x is 0, 1, or 2; [0232] n is 1, 2, 3, or 4; [0233] each R.sub.a and R.sub.b is, independently, H, a protecting group, hydroxyl, C.sub.1-C.sub.12 alkyl, substituted C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl, substituted C.sub.2-C.sub.12 alkenyl, C.sub.2-C.sub.12 alkynyl, substituted C.sub.2-C.sub.12 alkynyl, C.sub.5-C.sub.20 aryl, substituted C.sub.5-C.sub.20 aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C.sub.5-C.sub.7 alicyclic radical, substituted C.sub.5-C.sub.7 alicyclic radical, halogen, OJ.sub.1, NJ.sub.1J.sub.2, SJ.sub.1, N.sub.3, COOJ.sub.1, acyl (C(.dbd.O)--H), substituted acyl, CN, sulfonyl (S(.dbd.O).sub.2-J.sub.1), or sulfoxyl (S(.dbd.O)-J.sub.1); and

[0234] each J.sub.1 and J.sub.2 is, independently, H, C.sub.1-C.sub.12 alkyl, substituted C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl, substituted C.sub.2-C.sub.12 alkenyl, C.sub.2-C.sub.12 alkynyl, substituted C.sub.2-C.sub.12 alkynyl, C.sub.5-C.sub.20 aryl, substituted C.sub.5-C.sub.20 aryl, acyl (C(.dbd.O)--H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, C.sub.1-C.sub.12 aminoalkyl, substituted C.sub.1-C.sub.12 aminoalkyl or a protecting group.

[0235] In certain embodiments, the bridge of a bicyclic sugar moiety is, --[C(R.sub.a)(R.sub.b)].sub.n--, --[C(R.sub.a)(R.sub.b)].sub.n--O--, --C(R.sub.aR.sub.b)--N(R)--O-- or --C(R.sub.aR.sub.b)--O--N(R)--. In certain embodiments, the bridge is 4'-CH.sub.2-2',4'-(CH.sub.2)2-2',4'-(CH.sub.2)3-2',4'-CH.sub.2--O-2',4'-(- CH.sub.2).sub.2--O-2',4'-CH.sub.2--O--N(R)-2' and 4'-CH.sub.2--N(R)--O-2'- wherein each R is, independently, H, a protecting group or C.sub.1-C.sub.12 alkyl.

[0236] In certain embodiments, bicyclic nucleosides are further defined by isomeric configuration. For example, a nucleoside comprising a 4'-2' methylene-oxy bridge, may be in the .alpha.-L configuration or in the .beta.-D configuration. Previously, .alpha.-L-methyleneoxy(4'-CH.sub.2--O-2') BNA's have been incorporated into antisense oligonucleotides that showed antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372).

[0237] In certain embodiments, bicyclic nucleosides include, but are not limited to, (A) .alpha.-L-Methyleneoxy(4'-CH.sub.2--O-2') BNA, (B) .beta.-D-Methyleneoxy(4'-CH.sub.2--O-2') BNA, (C) Ethyleneoxy(4'-(CH.sub.2)2-O-2') BNA, (D) Aminooxy(4'-CH.sub.2--O--N(R)-2') BNA, (E) Oxyamino (4'-CH.sub.2--N(R)--O-2') BNA, and (F) Methyl(methyleneoxy) (4'-CH(CH.sub.3)--O-2') BNA, (G) methylene-thio(4'-CH.sub.2--S-2') BNA, (H) methylene-amino(4'-CH.sub.2--N(R)-2') BNA, (I) methyl carbocyclic (4'-CH.sub.2--CH(CH.sub.3)-2') BNA, (J) propylene carbocyclic (4'-(CH.sub.2)3-2') BNA, and (K) ethylene carbocyclic (4'-CH.sub.2--CH.sub.2-2') (carba LNA or "cLNA") as depicted below.

##STR00001## ##STR00002##

wherein Bx is the base moiety and R is independently H, a protecting group or C.sub.1-C.sub.12 alkyl.

[0238] In certain embodiments, bicyclic nucleoside having Formula I:

##STR00003##

[0239] wherein:

[0240] Bx is a heterocyclic base moiety;

[0241] -Q.sub.a-Q.sub.b-Q.sub.c- is --CH.sub.2--N(R.sub.c)--CH.sub.2--, --C(.dbd.O)--N(R.sub.c)--CH.sub.2--, --CH.sub.2--O--N(R.sub.c)--, --CH.sub.2--N(R.sub.c)--O-- or --N(R.sub.c)--O--CH.sub.2;

[0242] R.sub.c is C.sub.1-C.sub.12 alkyl or an amino protecting group; and

[0243] T.sub.a and T.sub.b are each, independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety or a covalent attachment to a support medium.

[0244] In certain embodiments, bicyclic nucleoside having Formula II:

##STR00004##

wherein:

[0245] Bx is a heterocyclic base moiety;

[0246] T.sub.a and T.sub.b are each, independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety or a covalent attachment to a support medium;

[0247] Z.sub.a is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, substituted C.sub.1-C.sub.6 alkyl, substituted C.sub.2-C.sub.6 alkenyl, substituted C.sub.2-C.sub.6 alkynyl, acyl, substituted acyl, substituted amide, thiol or substituted thio.

[0248] In one embodiment, each of the substituted groups, is, independently, mono or poly substituted with substituent groups independently selected from halogen, oxo, hydroxyl, OJ.sub.c, NJ.sub.cT.sub.d, SJ.sub.c, N.sub.3, OC(.dbd.X)J.sub.c, and NJ.sub.cC(.dbd.X)NJ.sub.cJ.sub.d, wherein each J.sub.c, J.sub.d and J.sub.e is, independently, H, C.sub.1-C.sub.6 alkyl, or substituted C.sub.1-C.sub.6 alkyl and X is O or NJ.sub.c.

[0249] In certain embodiments, bicyclic nucleoside having Formula III:

##STR00005##

wherein:

[0250] Bx is a heterocyclic base moiety;

[0251] T.sub.a and T.sub.b are each, independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety or a covalent attachment to a support medium;

[0252] Z.sub.b is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl, substituted C.sub.1-C.sub.6 alkyl, substituted C.sub.2-C.sub.6 alkenyl, substituted C.sub.2-C.sub.6 alkynyl or substituted acyl (C(.dbd.O)--).

[0253] In certain embodiments, bicyclic nucleoside having Formula IV:

##STR00006##

wherein:

[0254] Bx is a heterocyclic base moiety;

[0255] T.sub.a and T.sub.b are each, independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety or a covalent attachment to a support medium;

[0256] R.sub.d is C.sub.1-C.sub.6 alkyl, substituted C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, substituted C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl or substituted C.sub.2-C.sub.6 alkynyl;

[0257] each q.sub.a, q.sub.b, q.sub.c and q.sub.d is, independently, H, halogen, C.sub.1-C.sub.6 alkyl, substituted C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, substituted C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl or substituted C.sub.2-C.sub.6 alkynyl, C.sub.1-C.sub.6 alkoxyl, substituted C.sub.1-C.sub.6 alkoxyl, acyl, substituted acyl, C.sub.1-C.sub.6 aminoalkyl or substituted C.sub.1-C.sub.6 aminoalkyl;

[0258] In certain embodiments, bicyclic nucleoside having Formula V:

##STR00007##

wherein:

[0259] Bx is a heterocyclic base moiety;

[0260] T.sub.a and T.sub.b are each, independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety or a covalent attachment to a support medium;

[0261] q.sub.a, q.sub.b, q.sub.e and q.sub.f are each, independently, hydrogen, halogen, C.sub.1-C.sub.12 alkyl, substituted C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl, substituted C.sub.2-C.sub.12 alkenyl, C.sub.2-C.sub.12 alkynyl, substituted C.sub.2-C.sub.12 alkynyl, C.sub.1-C.sub.12 alkoxy, substituted C.sub.1-C.sub.12 alkoxy, OJ.sub.j, SJ.sub.j, SO.sub.2J.sub.j, SO.sub.2J.sub.j, NJ.sub.jJ.sub.k, N.sub.3, CN, C(.dbd.O)OJ.sub.j, C(.dbd.O)NJ.sub.jJ.sub.k, C(.dbd.O)J.sub.j, O--C(.dbd.O)NJ.sub.jJ.sub.k, N(H)C(.dbd.NH)NJ.sub.jJ.sub.k, N(H)C(.dbd.O)NJ.sub.jJ.sub.k or N(H)C(.dbd.S)NJ.sub.jJ.sub.k;

[0262] or q.sub.e and q.sub.f together are .dbd.C(q.sub.g)(q.sub.h);

[0263] q.sub.g and q.sub.h are each, independently, H, halogen, C.sub.1-C.sub.12 alkyl or substituted C.sub.1-C.sub.12 alkyl.

[0264] The synthesis and preparation of the methyleneoxy(4'-CH.sub.2--O-2') BNA monomers adenine, cytosine, guanine, 5-methyl-cytosine, thymine and uracil, along with their oligomerization, and nucleic acid recognition properties have been described (Koshkin et al., Tetrahedron, 1998, 54, 3607-3630). BNAs and preparation thereof are also described in WO 98/39352 and WO 99/14226.

[0265] Analogs of methyleneoxy(4'-CH.sub.2--O-2') BNA, methyleneoxy(4'-CH.sub.2--O-2') BNA and 2'-thio-BNAs, have also been prepared (Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222). Preparation of locked nucleoside analogs comprising oligodeoxyribonucleotide duplexes as substrates for nucleic acid polymerases has also been described (Wengel et al., WO 99/14226). Furthermore, synthesis of 2'-amino-BNA, a novel comformationally restricted high-affinity oligonucleotide analog has been described in the art (Singh et al., J. Org. Chem., 1998, 63, 10035-10039). In addition, 2'-Amino- and 2'-methylamino-BNA's have been prepared and the thermal stability of their duplexes with complementary RNA and DNA strands has been previously reported.

[0266] In certain embodiments, bicyclic nucleoside having Formula VI:

##STR00008##

wherein:

[0267] Bx is a heterocyclic base moiety;

[0268] T.sub.a and T.sub.b are each, independently H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety or a covalent attachment to a support medium;

[0269] each q.sub.i, q.sub.j, q.sub.k and q.sub.i is, independently, H, halogen, C.sub.1-C.sub.12 alkyl, substituted C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl, substituted C.sub.2-C.sub.12 alkenyl, C.sub.2-C.sub.12 alkynyl, substituted C.sub.2-C.sub.12 alkynyl, C.sub.1-C.sub.12 alkoxyl, substituted C.sub.1-C.sub.12 alkoxyl, OJ.sub.j, SJ.sub.j, SOJ.sub.j, SO.sub.2J.sub.j, NJ.sub.jJ.sub.k, N.sub.3, CN, C(.dbd.O)OJ.sub.j, C(.dbd.O)NJ.sub.jJ.sub.k, C(.dbd.O)J.sub.j, O--C(.dbd.O)NJ.sub.jJ.sub.k, N(H)C(.dbd.NH)NJ.sub.jJ.sub.k, N(H)C(.dbd.O)NJ.sub.jJ.sub.k or N(H)C(.dbd.S)NJ.sub.jJ.sub.k; and

[0270] q.sub.i and q.sub.j or q.sub.l and q.sub.k together are .dbd.C(q.sub.g)(q.sub.h), wherein q.sub.g and q.sub.h are each, independently, H, halogen, C.sub.1-C.sub.12 alkyl or substituted C.sub.1-C.sub.12 alkyl.

[0271] One carbocyclic bicyclic nucleoside having a 4'-(CH.sub.2)3-2' bridge and the alkenyl analog bridge 4'-CH.dbd.CH--CH.sub.2-2' have been described (Frier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443 and Albaek et al., J. Org. Chem., 2006, 71, 7731-7740). The synthesis and preparation of carbocyclic bicyclic nucleosides along with their oligomerization and biochemical studies have also been described (Srivastava et al., J. Am. Chem. Soc. 2007, 129(26), 8362-8379).

[0272] As used herein, "4'-2' bicyclic nucleoside" or "4' to 2' bicyclic nucleoside" refers to a bicyclic nucleoside comprising a furanose ring comprising a bridge connecting two carbon atoms of the furanose ring connects the 2' carbon atom and the 4' carbon atom of the sugar ring.

[0273] As used herein, "monocylic nucleosides" refer to nucleosides comprising modified sugar moieties that are not bicyclic sugar moieties. In certain embodiments, the sugar moiety, or sugar moiety analogue, of a nucleoside may be modified or substituted at any position.

[0274] As used herein, "2'-modified sugar" means a furanosyl sugar modified at the 2' position. In certain embodiments, such modifications include substituents selected from: a halide, including, but not limited to substituted and unsubstituted alkoxy, substituted and unsubstituted thioalkyl, substituted and unsubstituted amino alkyl, substituted and unsubstituted alkyl, substituted and unsubstituted allyl, and substituted and unsubstituted alkynyl. In certain embodiments, 2' modifications are selected from substituents including, but not limited to: O[(CH.sub.2).sub.nO].sub.mCH.sub.3, O(CH.sub.2).sub.nNH.sub.2, O(CH.sub.2).sub.nCH.sub.3, O(CH.sub.2).sub.nONH.sub.2, OCH.sub.2C(.dbd.O)N(H)CH.sub.3, and O(CH.sub.2).sub.nON[(CH.sub.2).sub.nCH.sub.3].sub.2, where n and m are from 1 to about 10. Other 2'- substituent groups can also be selected from: C.sub.1-C.sub.12 alkyl, substituted alkyl, alkenyl, alkynyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH.sub.3, OCN, Cl, Br, CN, CF.sub.3, OCF.sub.3, SOCH.sub.3, SO.sub.2CH.sub.3, ONO.sub.2, NO.sub.2, N.sub.3, NH.sub.2, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving group, a reporter group, an intercalator, a group for improving pharmacokinetic properties, or a group for improving the pharmacodynamic properties of an antisense compound, and other substituents having similar properties. In certain embodiments, modified nucleosides comprise a 2'-MOE side chain (Baker et al., J. Biol. Chem., 1997, 272, 11944-12000). Such 2'-MOE substitution have been described as having improved binding affinity compared to unmodified nucleosides and to other modified nucleosides, such as 2'-O-methyl, O-propyl, and O-aminopropyl. Oligonucleotides having the 2'-MOE substituent also have been shown to be antisense inhibitors of gene expression with promising features for in vivo use (Martin, P., Helv. Chim. Acta, 1995, 78, 486-504; Altmann et al., Chimia, 1996, 50, 168-176; Altmann et al., Biochem. Soc. Trans., 1996, 24, 630-637; and Altmann et al., Nucleosides Nucleotides, 1997, 16, 917-926).

[0275] As used herein, a "modified tetrahydropyran nucleoside" or "modified THP nucleoside" means a nucleoside having a six-membered tetrahydropyran "sugar" substituted in for the pentofuranosyl residue in normal nucleosides (a sugar surrogate). Modified THP nucleosides include, but are not limited to, what is referred to in the art as hexitol nucleic acid (HNA), anitol nucleic acid (ANA), manitol nucleic acid (MNA) (see Leumann, C J. Bioorg. & Med. Chem. (2002) 10:841-854), fluoro HNA (F-HNA) or those compounds having Formula X:

[0276] Formula X:

##STR00009##

wherein independently for each of said at least one tetrahydropyran nucleoside analog of Formula X:

[0277] Bx is a heterocyclic base moiety;

[0278] T.sub.3 and T.sub.4 are each, independently, an internucleoside linking group linking the tetrahydropyran nucleoside analog to the antisense compound or one of T.sub.3 and T.sub.4 is an internucleoside linking group linking the tetrahydropyran nucleoside analog to the antisense compound and the other of T.sub.3 and T.sub.4 is H, a hydroxyl protecting group, a linked conjugate group or a 5' or 3'-terminal group;

[0279] q.sub.1, q.sub.2, q.sub.3, q.sub.4, q.sub.5, q.sub.6 and q.sub.7 are each independently, H, C.sub.1-C.sub.6 alkyl, substituted C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, substituted C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl or substituted C.sub.2-C.sub.6 alkynyl; and

[0280] one of R.sub.1 and R.sub.2 is hydrogen and the other is selected from halogen, substituted or unsubstituted alkoxy, NJ.sub.1J.sub.2, SJ.sub.1, N.sub.3, OC(.dbd.X)J.sub.1, OC(.dbd.X)NJ.sub.1J.sub.2, NJ.sub.3C(.dbd.X)NJ.sub.1J.sub.2 and CN, wherein X is O, S or NJ.sub.1 and each J.sub.1, J.sub.2 and J.sub.3 is, independently, H or C.sub.1-C.sub.6 alkyl.

[0281] In certain embodiments, the modified THP nucleosides of Formula X are provided wherein q.sub.m, q.sub.n, q.sub.p, q.sub.r, q.sub.s, q.sub.t and q.sub.u are each H. In certain embodiments, at least one of q.sub.m, q.sub.n, q.sub.p, q.sub.r, q.sub.s, q.sub.t and q.sub.u is other than H. In certain embodiments, at least one of q.sub.m, q.sub.n, q.sub.p, q.sub.r, q.sub.s, q.sub.t and q.sub.u is methyl. In certain embodiments, THP nucleosides of Formula X are provided wherein one of R.sub.1 and R.sub.2 is F. In certain embodiments, R.sub.1 is fluoro and R.sub.2 is H; R.sub.1 is methoxy and R.sub.2 is H, and R.sub.1 is methoxyethoxy and R.sub.2 is H.

[0282] As used herein, "2'-modified" or "2'-substituted" refers to a nucleoside comprising a sugar comprising a substituent at the 2' position other than H or OH. 2'-modified nucleosides, include, but are not limited to, bicyclic nucleosides wherein the bridge connecting two carbon atoms of the sugar ring connects the 2' carbon and another carbon of the sugar ring; and nucleosides with non-bridging 2' substituents, such as allyl, amino, azido, thio, 0-allyl, O--C.sub.1-C.sub.10 alkyl, --OCF.sub.3, O--(CH.sub.2).sub.2--O--CH.sub.3, 2'-O(CH.sub.2).sub.2SCH.sub.3, O--(CH.sub.2).sub.2--O--N(R.sub.m)(R.sub.n), or O--CH.sub.2--C(.dbd.O)--N(R.sub.m)(R.sub.n), where each R.sub.m and R.sub.n is, independently, H or substituted or unsubstituted C.sub.1-C.sub.10 alkyl. 2'-modified nucleosides may further comprise other modifications, for example at other positions of the sugar and/or at the nucleobase.

[0283] As used herein, "2'-F" refers to a nucleoside comprising a sugar comprising a fluoro group at the 2' position.

[0284] As used herein, "2'-OMe" or "2'-OCH.sub.3" or "2'-O-methyl" each refers to a nucleoside comprising a sugar comprising an --OCH.sub.3 group at the 2' position of the sugar ring.

[0285] As used herein, "MOE" or "2'-MOE" or "2'-OCH.sub.2CH.sub.2OCH.sub.3" or "2'-O-methoxyethyl" each refers to a nucleoside comprising a sugar comprising a --OCH.sub.2CH.sub.2OCH.sub.3 group at the 2' position of the sugar ring.

[0286] As used herein, "oligonucleotide" refers to a compound comprising a plurality of linked nucleosides. In certain embodiments, one or more of the plurality of nucleosides is modified. In certain embodiments, an oligonucleotide comprises one or more ribonucleosides (RNA) and/or deoxyribonucleosides (DNA).

[0287] Many other bicyclo and tricyclo sugar surrogate ring systems are also know in the art that can be used to modify nucleosides for incorporation into antisense compounds (see for example review article: Leumann, J. C, Bioorganic & Medicinal Chemistry, 2002, 10, 841-854). Such ring systems can undergo various additional substitutions to enhance activity.

[0288] Methods for the preparations of modified sugars are well known to those skilled in the art.

[0289] In nucleotides having modified sugar moieties, the nucleobase moieties (natural, modified or a combination thereof) are maintained for hybridization with an appropriate nucleic acid target.

[0290] In certain embodiments, antisense compounds comprise one or more nucleotides having modified sugar moieties. In certain embodiments, the modified sugar moiety is 2'-MOE. In certain embodiments, the 2'-MOE modified nucleotides are arranged in a gapmer motif. In certain embodiments, the modified sugar moiety is a cEt. In certain embodiments, the cEt modified nucleotides are arranged throughout the wings of a gapmer motif.

Modified Nucleobases

[0291] Nucleobase (or base) modifications or substitutions are structurally distinguishable from, yet functionally interchangeable with, naturally occurring or synthetic unmodified nucleobases. Both natural and modified nucleobases are capable of participating in hydrogen bonding. Such nucleobase modifications may impart nuclease stability, binding affinity, increased selectivity for an allelic variant, or some other beneficial biological property to antisense compounds. Modified nucleobases include synthetic and natural nucleobases such as, for example, 5-methylcytosine (5-me-C). Certain nucleobase substitutions, including 5-methylcytosine substitutions, are particularly useful for increasing the binding affinity of an antisense compound for a target nucleic acid. For example, 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2.degree. C. (Sanghvi, Y. S., Crooke, S. T. and Lebleu, B., eds., Antisense Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278).

[0292] Additional modified nucleobases include 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl (--C.ident.C--CH.sub.3) uracil and cytosine and other alkynyl derivatives of pyrimidine bases, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 2-F-adenine, 2-amino-adenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3-deazaadenine.

[0293] Heterocyclic base moieties may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2-pyridone. Nucleobases that are particularly useful for increasing the binding affinity of antisense compounds include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and 0-6 substituted purines, including 2 aminopropyladenine, 5-propynyluracil and 5-propynylcytosine.

[0294] In certain embodiments, antisense compounds comprise one or more modified nucleobases. In certain embodiments, gap-widened antisense oligonucleotides comprise one or more modified nucleobases. In certain embodiments, the modified nucleobase is 5-methylcytosine. In certain embodiments, each cytosine is a 5-methylcytosine.

Compositions and Methods for Formulating Pharmaceutical Compositions

[0295] Antisense oligonucleotides may be admixed with pharmaceutically acceptable active or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions are dependent upon a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

[0296] An antisense compound can be utilized in pharmaceutical compositions by combining the antisense compound with a suitable pharmaceutically acceptable diluent or carrier. A pharmaceutically acceptable diluent includes phosphate-buffered saline (PBS). PBS is a diluent suitable for use in compositions to be delivered parenterally. Accordingly, in one embodiment, employed in the methods described herein is a pharmaceutical composition comprising an antisense compound and a pharmaceutically acceptable diluent. In certain embodiments, the pharmaceutically acceptable diluent is PBS. In certain embodiments, the antisense compound is an antisense oligonucleotide.

[0297] Pharmaceutical compositions comprising antisense compounds encompass any pharmaceutically acceptable salts, esters, or salts of such esters, or any other oligonucleotide which, upon administration to an animal, including a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. Accordingly, for example, the disclosure is also drawn to pharmaceutically acceptable salts of antisense compounds, prodrugs, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

[0298] A prodrug can include the incorporation of additional nucleosides at one or both ends of an antisense compound which are cleaved by endogenous nucleases within the body, to form the active antisense compound.

Conjugated Antisense Compounds

[0299] Antisense compounds may be covalently linked to one or more moieties or conjugates which enhance the activity, cellular distribution, increased selectivity for an allelic variant, or cellular uptake of the resulting antisense oligonucleotides. Typical conjugate groups include cholesterol moieties and lipid moieties. Additional conjugate groups include carbohydrates, phospholipids, biotin, phenazine, folate, phenanthridine, anthraquinone, acridine, fluoresceins, rhodamines, coumarins, and dyes.

[0300] Antisense compounds can also be modified to have one or more stabilizing groups that are generally attached to one or both termini of antisense compounds to enhance properties such as, for example, nuclease stability. Included in stabilizing groups are cap structures. These terminal modifications protect the antisense compound having terminal nucleic acid from exonuclease degradation, and can help in delivery and/or localization within a cell. The cap can be present at the 5'-terminus (5'-cap), or at the 3'-terminus (3'-cap), or can be present on both termini. Cap structures are well known in the art and include, for example, inverted deoxy abasic caps. Further 3' and 5'-stabilizing groups that can be used to cap one or both ends of an antisense compound to impart nuclease stability include those disclosed in WO 03/004602 published on Jan. 16, 2003.

Cell Culture and Antisense Compounds Treatment

[0301] The effects of antisense compounds on the level, activity or expression target nucleic acids can be tested in vitro in a variety of cell types. Cell types used for such analyses are available from commercial vendors (e.g. American Type Culture Collection, Manassas, Va.; Zen-Bio, Inc., Research Triangle Park, N.C.; Clonetics Corporation, Walkersville, Md.) and are cultured according to the vendor's instructions using commercially available reagents (e.g. Invitrogen Life Technologies, Carlsbad, Calif.). Illustrative cell types include, but are not limited to, HepG2 cells, Hep3B cells, and primary hepatocytes. Illustrative cell lines include GM04281, GM02171, and GM02173B cells.

In Vitro Testing of Antisense Oligonucleotides

[0302] Described herein are methods for treatment of cells with antisense oligonucleotides, which can be modified appropriately for treatment with other antisense compounds.

[0303] In general, cells are treated with antisense oligonucleotides when the cells reach approximately 60-80% confluency in culture.

[0304] One reagent commonly used to introduce antisense oligonucleotides into cultured cells includes the cationic lipid transfection reagent LIPOFECTIN (Invitrogen, Carlsbad, Calif.). Antisense oligonucleotides are mixed with LIPOFECTIN in OPTI-MEM 1 (Invitrogen, Carlsbad, Calif.) to achieve the desired final concentration of antisense oligonucleotide and a LIPOFECTIN concentration that typically ranges 2 to 12 ug/mL per 100 nM antisense oligonucleotide.

[0305] Another reagent used to introduce antisense oligonucleotides into cultured cells includes LIPOFECTAMINE (Invitrogen, Carlsbad, Calif.). Antisense oligonucleotide is mixed with LIPOFECTAMINE in OPTI-MEM 1 reduced serum medium (Invitrogen, Carlsbad, Calif.) to achieve the desired concentration of antisense oligonucleotide and a LIPOFECTAMINE concentration that typically ranges 2 to 12 ug/mL per 100 nM antisense oligonucleotide.

[0306] Another technique used to introduce antisense oligonucleotides into cultured cells includes electroporation.

[0307] Cells are treated with antisense oligonucleotides by routine methods. Cells are typically harvested 16-24 hours after antisense oligonucleotide treatment, at which time RNA or protein levels of target nucleic acids are measured by methods known in the art and described herein. In general, when treatments are performed in multiple replicates, the data are presented as the average of the replicate treatments.

[0308] The concentration of antisense oligonucleotide used varies from cell line to cell line. Methods to determine the optimal antisense oligonucleotide concentration for a particular cell line are well known in the art. Antisense oligonucleotides are typically used at concentrations ranging from 1 nM to 300 nM when transfected with LIPOFECTAMINE. Antisense oligonucleotides are used at higher concentrations ranging from 625 to 20,000 nM when transfected using electroporation.

RNA Isolation

[0309] RNA analysis can be performed on total cellular RNA or poly(A)+mRNA. Methods of RNA isolation are well known in the art. RNA is prepared using methods well known in the art, for example, using the TRIZOL Reagent (Invitrogen, Carlsbad, Calif.) according to the manufacturer's recommended protocols.

Analysis of Inhibition of Target Levels or Expression

[0310] Reduction, inhibition, or expression of a target nucleic acid can be assayed in a variety of ways known in the art. For example, target nucleic acid levels can be quantitated by, e.g., Northern blot analysis, competitive polymerase chain reaction (PCR), or quantitative real-time PCR. RNA analysis can be performed on total cellular RNA or poly(A)+mRNA. Methods of RNA isolation are well known in the art. Northern blot analysis is also routine in the art. Quantitative real-time PCR can be conveniently accomplished using the commercially available ABI PRISM 7600, 7700, or 7900 Sequence Detection System, available from PE-Applied Biosystems, Foster City, Calif. and used according to manufacturer's instructions.

Quantitative Real-Time PCR Analysis of Target RNA Levels

[0311] Quantitation of target RNA levels may be accomplished by quantitative real-time PCR using the ABI PRISM 7600, 7700, or 7900 Sequence Detection System (PE-Applied Biosystems, Foster City, Calif.) according to manufacturer's instructions. Methods of quantitative real-time PCR are well known in the art.

[0312] Prior to real-time PCR, the isolated RNA is subjected to a reverse transcriptase (RT) reaction, which produces complementary DNA (cDNA) that is then used as the substrate for the real-time PCR amplification. The RT and real-time PCR reactions are performed sequentially in the same sample well. RT and real-time PCR reagents are obtained from Invitrogen (Carlsbad, Calif.). RT real-time-PCR reactions are carried out by methods well known to those skilled in the art.

[0313] Gene (or RNA) target quantities obtained by real time PCR are normalized using either the expression level of a gene whose expression is constant, such as cyclophilin A, or by quantifying total RNA using RIBOGREEN (Invitrogen, Inc. Carlsbad, Calif.). Cyclophilin A expression is quantified by real time PCR, by being run simultaneously with the target, multiplexing, or separately. Total RNA is quantified using RIBOGREEN RNA quantification reagent (Invetrogen, Inc. Eugene, Oreg.). Methods of RNA quantification by RIBOGREEN are taught in Jones, L. J., et al, (Analytical Biochemistry, 1998, 265, 368-374). A CYTOFLUOR 4000 instrument (PE Applied Biosystems) is used to measure RIBOGREEN fluorescence.

[0314] Probes and primers are designed to hybridize to target nucleic acids. Methods for designing real-time PCR probes and primers are well known in the art, and may include the use of software such as PRIMER EXPRESS Software (Applied Biosystems, Foster City, Calif.).

Analysis of Protein Levels

[0315] Reduction, inhibition, or expression of target nucleic acids can be assessed by measuring target protein levels. Target protein levels can be evaluated or quantitated in a variety of ways well known in the art, such as immunoprecipitation, Western blot analysis (immunoblotting), enzyme-linked immunosorbent assay (ELISA), quantitative protein assays, protein activity assays (for example, caspase activity assays), immunohistochemistry, immunocytochemistry or fluorescence-activated cell sorting (FACS). Antibodies directed to a target can be identified and obtained from a variety of sources, such as the MSRS catalog of antibodies (Aerie Corporation, Birmingham, Mich.), or can be prepared via conventional monoclonal or polyclonal antibody generation methods well known in the art. Antibodies useful for the detection of mouse, rat, monkey, and human proteins are commercially available.

In Vivo Testing of Antisense Compounds

[0316] Antisense compounds, for example, antisense oligonucleotides, are tested in animals to assess their ability to selectively reduce or inhibit expression of target gene product and produce phenotypic changes, such as, amelioration of a disease symptom. Testing may be performed in normal animals, or in experimental disease models. For administration to animals, antisense oligonucleotides are formulated in a pharmaceutically acceptable diluent, such as phosphate-buffered saline. Administration includes parenteral routes of administration, such as intraperitoneal, intravenous, and subcutaneous. Calculation of antisense oligonucleotide dosage and dosing frequency is within the abilities of those skilled in the art, and depends upon factors such as route of administration and animal body weight. Following a period of treatment with antisense oligonucleotides, RNA or protein is isolated from tissue and changes in target nucleic acid or protein expression are measured.

Administration

[0317] In certain embodiments, the compounds and compositions described herein may be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration may be topical (including ophthalmic, vaginal, rectal, intranasal), oral, pulmonary (including by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal, epidermal and transdermal) or parenteral, for example, by intravenous drip, intravenous injection or subcutaneous, intraperitoneal, intraocular, intravitreal, or intramuscular injection.

[0318] In certain embodiments, the compounds and compositions as described herein are administered parenterally.

[0319] In certain embodiments, parenteral administration is by infusion. Infusion can be chronic or continuous or short or intermittent. In certain embodiments, infused pharmaceutical agents are delivered with a pump. In certain embodiments, parenteral administration is by injection.

[0320] In certain embodiments, compounds and compositions are delivered to the CNS. In certain embodiments, compounds and compositions are delivered to the cerebrospinal fluid. In certain embodiments, compounds and compositions are administered to the brain parenchyma. In certain embodiments, compounds and compositions are delivered to an animal by intrathecal administration, or intracerebroventricular administration. Broad distribution of compounds and compositions, described herein, within the central nervous system may be achieved with intraparenchymal administration, intrathecal administration, or intracerebroventricular administration.

[0321] In certain embodiments, parenteral administration is by injection. The injection may be delivered with a syringe or a pump. In certain embodiments, the injection is a bolus injection. In certain embodiments, the injection is administered directly to a tissue, such as striatum, caudate, cortex, hippocampus and cerebellum.

[0322] In certain embodiments, methods of specifically localizing a pharmaceutical agent, such as by bolus injection, decreases median effective concentration (EC50) by a factor of 20, 25, 30, 35, 40, 45 or 50. In certain embodiments, the pharmaceutical agent in an antisense compound as further described herein. In certain embodiments, the targeted tissue is brain tissue. In certain embodiments the targeted tissue is striatal tissue. In certain embodiments, decreasing EC50 is desirable because it reduces the dose required to achieve a pharmacological result in a patient in need thereof.

[0323] In certain embodiments, an antisense oligonucleotide is delivered by injection or infusion once every month, every two months, every 90 days, every 3 months, every 6 months, twice a year or once a year.

Certain Compounds and Indications

[0324] Provided herein are compounds and methods that provide potent inhibition and increased selectivity for a mutant allele. Potency is demonstrated by the percent inhibition of mutant mRNA achieved by the antisense oligonucleotides targeting a SNP compared to the percent inhibition of mutant mRNA achieved by the benchmark oligonucleotide. Selectivity is demonstrated by the ability of the antisense oligonucleotide targeting a SNP to inhibit expression of the major allele or mutant allele preferentially compared to the minor allele or wild type allele. The usage of three cell lines with different genotypes at each SNP position have facilitated the determination of design rules that provide for potent and selective SNP targeting antisense oligonucleotides.

[0325] In certain embodiments, the compounds are antisense oligonucleotides as further described herein. The antisense oligonucleotides preferentially target a SNP or differentiating polymorphism. Oligonucleotides of various lengths were tested and certain lengths were determined to be beneficial for the targeting of SNPs.

[0326] In certain embodiments, the antisense oligonucleotides have a sequence that is 12-30 nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 12-25 nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 12-21 nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 12-20 nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 13-20 nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 14-20 nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 15-20 nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 12-19 nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 13-19 nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 14-19 nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 15-19, nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 16-19 nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 17-19 nucleobases in length. In certain embodiments, the antisense oligonucleotides have a sequence that is 12, 13, 14, 15, 16, 17, 18, 19, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 nucleobases in length.

[0327] For oligonucleotides of various lengths, the position of the nucleoside complementary to the SNP position was shifted within the gap and the wings and the effect was tested. Certain positions within the antisense oligonucleotide are shown to be beneficial for targeting SNPs.

[0328] In certain embodiments, the antisense oligonucleotide is at least 12, at least 13, at least 14, at least 15, at least 16, at least 17 at least 18 or at least 19 nucleobases in length and the SNP is complementary to positions 6-15 counting from the 5' terminus of the antisense oligonucleotide and/or positions 1-9 counting from the 5' end of the gap. In certain embodiments, the antisense oligonucleotide is at least 12, at least 13, at least 14, at least 15, at least 16, at least 17 at least 18 or at least 19 nucleobases in length and the SNP is complementary to positions 8-14 counting from the 5' terminus of the antisense oligonucleotide and/or positions 1-9 counting from the 5' end of the gap. In certain embodiments, the antisense oligonucleotide is at least 12, at least 13, at least 14, at least 15, at least 16, at least 17 at least 18 or at least 19 nucleobases in length and the SNP is complementary to positions 8-14 counting from the 5' terminus of the antisense oligonucleotide and/or positions 4-7 counting from the 5' end of the gap. In certain embodiments, the antisense oligonucleotide is at least 12, at least 13, at least 14, at least 15, at least 16, at least 17 at least 18 or at least 19 nucleobases in length and the SNP is complementary to positions 8-10 counting from the 5' terminus of the antisense oligonucleotide and/or positions 4-6 counting from the 5' end of the gap.

[0329] In certain embodiments, the SNP is complementary to position 8, 9, or 10 counting from the 5' terminus of the oligonucleotide or position 4, 5, or 6, counting from the 5' end of the gap. For oligonucleotides of various lengths, the effect of the length of the gap, 5' wing, and 3' wing was tested.

[0330] Certain wing-gap-wing combinations were shown to be beneficial for a SNP targeting antisense oligonucleotide. In certain embodiments the gap is 7-11 nucleobases in length and each wing is independently 1-6 nucleobases in length. In certain embodiments the gap is 7-11 nucleobases in length and each wing is independently 2-6 nucleobases in length. In certain embodiments the gap is 8-11 nucleobases in length and each wing is independently 2-6 nucleobases in length. In certain embodiments the gap is 9-11 nucleobases in length and each wing is independently 2-6 nucleobases in length. In certain embodiments the gap is 9 nucleobases in length and each wing is independently 2-6 nucleobases in length. In certain embodiments the gap is 10 nucleobases in length and each wing is independently 2-6 or 4-5 nucleobases in length. In certain embodiments the gap is 11 nucleobases in length and each wing is independently 2-6, or 4-5 nucleobases in length. In certain embodiments, the wing-gap-wing configuration is one of 4-7-4, 5-8-6, 6-8-5, 6-7-6, 5-7-5. 6-8-5, 5-8-6, 3-9-4, 4-9-3, 2-9-6, 6,9,2,3-9-3, 3-9-5, 5-9-3, 5-9-4, 4-9-5, 5-9-5, 4-11-4, 4-10-5 and 5-10-4.

[0331] For oligonucleotides of various lengths, the effect of certain chemistries was tested. Certain chemistry modifications were shown to be beneficial for a SNP targeting antisense oligonucleotide. In certain embodiments, each nucleoside of each wing of the modified antisense oligonucleotide has a 2'-MOE modification. In certain embodiments, each nucleoside of each wing of the modified antisense oligonucleotide has a high affinity modification. In certain embodiments, the antisense oligonucleotide is a mixed wing gapmer. In such embodiment, the modifications and combination of modifications at the 3' wing and at the 5' wing may be the same or they may be different. In certain embodiments, the antisense oligonucleotide has one or more 2'-MOE modifications in the wings and/or one or more high affinity modifications in the wings. In certain embodiments, the high affinity modification is a cEt modification. In certain embodiments, the antisense oligonucleotide has a high affinity modification at positions 2, 3, 13, and 14 of the antisense oligonucleotide (counting from the 5' terminus). In certain embodiments, the antisense oligonucleotide has one, two, three, or four high affinity modifications in at least one of the wings. In certain embodiments, the antisense oligonucleotide has one, two, three, or four high affinity modifications in each of the 5' and 3' wings independently. In certain embodiments, the antisense oligonucleotide has a high affinity modification at positions 2 and 3 in one or both of the 5' and 3' wings (counting from the 5' terminus of the 5' wing and the 3' terminus of the 3' wing). In certain embodiments, the antisense oligonucleotide has a high affinity modification at positions 2, 3 and 4 in one or both of the 5' and 3' wings (counting from the 5' terminus of the 5' wing and the 3' terminus of the 3' wing,). In certain embodiments, the antisense oligonucleotide has a high affinity modification at positions 1 of the 5' and/or 3' wings (counting from the 5' terminus of the 5' wing and the 3' terminus of the 3' wing,). In certain embodiments, the antisense oligonucleotide has a high affinity modification at positions 1 of the 5' and 3' wings (counting from the 5' terminus of the 5' wing and the 3' terminus of the 3' wing,) and at least one other position in the wing. In certain embodiments, the antisense oligonucleotide has alternating 2'-MOE and high affinity modification in at least one of the 5' and 3' wings.

[0332] In certain embodiments, the compound comprises an antisense oligonucleotide incorporating one or more of the design rules provided above.

[0333] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 12 to 30 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein the single nucleotide polymorphism aligns with any one of positions 6-15 beginning from the 5' terminus of the antisense oligonucleotide or positions 1-9 beginning from the 5' end of the gap of the modified antisense oligonucleotide; and wherein each nucleoside of each wing has a modified sugar or sugar surrogate. In certain embodiments the single nucleotide polymorphism site contains a differentiating polymorphism. In certain embodiments, the single nucleotide polymorphism site is on a mutant allele. In certain embodiments, the mutant allele is associated with disease. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 4-7-4, 5-8-6, 6-8-5, 6-7-6, 5-7-5. 6-8-5, 5-8-6, 3-9-4, 4-9-3, 2-9-6, 6,9,2,3-9-3, 3-9-5, 5-9-3, 5-9-4, 4-9-5, 5-9-5, 4-11-4, 4-10-5 and 5-10-4.

[0334] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 12 to 20 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein the single nucleotide polymorphism aligns with any one of positions 6-15 beginning from the 5' terminus of the antisense oligonucleotide or positions 1-9 beginning from the 5' end of the gap of the modified antisense oligonucleotide; and wherein each nucleoside of each wing has a modified sugar or sugar surrogate. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 4-7-4, 5-8-6, 6-8-5, 6-7-6, 5-7-5. 6-8-5, 5-8-6, 3-9-4, 4-9-3, 2-9-6, 6,9,2,3-9-3, 3-9-5,5-9-3, 5-9-4, 4-9-5, 5-9-5, 4-11-4, 4-10-5 and 5-10-4.

[0335] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 12 to 20 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein the single nucleotide polymorphism aligns with any one of positions 8-14 beginning from the 5' terminus of the antisense oligonucleotide or positions 1-9 beginning from the 5' end of the gap of the modified antisense oligonucleotide; and wherein each nucleoside of each wing has a modified sugar or sugar surrogate. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 4-7-4, 5-8-6, 6-8-5, 6-7-6, 5-7-5. 6-8-5, 5-8-6, 3-9-4, 4-9-3, 2-9-6, 6,9,2,3-9-3, 3-9-5,5-9-3, 5-9-4, 4-9-5, 5-9-5, 4-11-4,4-10-5 and 5-10-4.

[0336] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 12 to 20 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein the single nucleotide polymorphism aligns with any one of positions 8-14 beginning from the 5' terminus of the antisense oligonucleotide or positions 4-7 beginning from the 5' end of the gap of the modified antisense oligonucleotide; and wherein each nucleoside of each wing has a modified sugar or sugar surrogate. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 4-7-4, 5-8-6, 6-8-5, 6-7-6, 5-7-5. 6-8-5, 5-8-6, 3-9-4, 4-9-3, 2-9-6, 6,9,2,3-9-3, 3-9-5,5-9-3, 5-9-4, 4-9-5, 5-9-5, 4-11-4,4-10-5 and 5-10-4.

[0337] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 12 to 20 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein the single nucleotide polymorphism aligns with any one of positions 8-10 beginning from the 5' terminus of the antisense oligonucleotide or positions 4-6 beginning from the 5' end of the gap of the modified antisense oligonucleotide; and wherein each nucleoside of each wing has a modified sugar or sugar surrogate. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 4-7-4, 5-8-6, 6-8-5, 6-7-6, 5-7-5. 6-8-5, 5-8-6, 3-9-4, 4-9-3, 2-9-6, 6,9,2,3-9-3, 3-9-5,5-9-3, 5-9-4, 4-9-5, 5-9-5, 4-11-4,4-10-5 and 5-10-4.

[0338] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 12 to 19 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein the single nucleotide polymorphism aligns with any one of positions 8-10 beginning from the 5' terminus of the antisense oligonucleotide or positions 4-6 beginning from the 5' end of the gap of the modified antisense oligonucleotide; and wherein each nucleoside of each wing has a modified sugar or sugar surrogate. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 4-7-4, 5-8-6, 6-8-5, 6-7-6, 5-7-5. 6-8-5, 5-8-6, 3-9-4, 4-9-3, 2-9-6, 6,9,2,3-9-3, 3-9-5,5-9-3, 5-9-4, 4-9-5, 5-9-5, 4-11-4,4-10-5 and 5-10-4.

[0339] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 13 to 19 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein the single nucleotide polymorphism aligns with any one of positions 8-10 beginning from the 5' terminus of the antisense oligonucleotide or positions 4-6 beginning from the 5' end of the gap of the modified antisense oligonucleotide; and wherein each nucleoside of each wing has a modified sugar or sugar surrogate. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 4-7-4, 5-8-6, 6-8-5, 6-7-6, 5-7-5. 6-8-5, 5-8-6, 3-9-4, 4-9-3, 2-9-6, 6,9,2,3-9-3, 3-9-5,5-9-3, 5-9-4, 4-9-5, 5-9-5, 4-11-4,4-10-5 and 5-10-4.

[0340] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 14 to 19 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein the single nucleotide polymorphism aligns with any one of positions 8-10 beginning from the 5' terminus of the antisense oligonucleotide or positions 4-6 beginning from the 5' end of the gap of the modified antisense oligonucleotide; and wherein each nucleoside of each wing has a modified sugar or sugar surrogate. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 4-7-4, 5-8-6, 6-8-5, 6-7-6, 5-7-5. 6-8-5, 5-8-6, 3-9-4, 4-9-3, 2-9-6, 6,9,2,3-9-3, 3-9-5,5-9-3, 5-9-4, 4-9-5, 5-9-5, 4-11-4,4-10-5 and 5-10-4.

[0341] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 15 to 19 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein the single nucleotide polymorphism aligns with any one of positions 6-15 beginning from the 5' terminus of the antisense oligonucleotide or positions 1-9 beginning from the 5' end of the gap of the modified antisense oligonucleotide; and wherein each nucleoside of each wing has a modified sugar or sugar surrogate. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 4-7-4, 5-8-6, 6-8-5, 6-7-6, 5-7-5. 6-8-5, 5-8-6, 3-9-4, 4-9-3, 2-9-6, 6,9,2,3-9-3, 3-9-5,5-9-3, 5-9-4, 4-9-5, 5-9-5, 4-11-4,4-10-5 and 5-10-4.

[0342] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 15 to 19 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein the single nucleotide polymorphism aligns with any one of positions 8-10 beginning from the 5' terminus of the antisense oligonucleotide or positions 4-6 beginning from the 5' end of the gap of the modified antisense oligonucleotide; and wherein each nucleoside of each wing has a modified sugar or sugar surrogate. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 4-7-4, 5-8-6, 6-8-5, 6-7-6, 5-7-5. 6-8-5, 5-8-6, 3-9-4, 4-9-3, 2-9-6, 6,9,2,3-9-3, 3-9-5,5-9-3, 5-9-4, 4-9-5, 5-9-5, 4-11-4,4-10-5 and 5-10-4.

[0343] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 15 to 19 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein position 6, 8, 9, 10, 11, or 14 beginning from the 5' terminus of the modified antisense oligonucleotide aligns with the single nucleotide polymorphism; and wherein each nucleoside of each wing segment modified sugar or sugar surrogate. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 3-9-3, 3-9-5, 4-9-5, 4-11-4, and 5-10-4.

[0344] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 15 to 19 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein position 1, 4, 5, 6, 7, or 9 of the gap segment aligns with the single nucleotide polymorphism; and wherein each nucleoside of each wing segment has a modified sugar or sugar surrogate. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 3-9-3, 3-9-5, 4-9-5, 4-11-4, and 5-10-4.

[0345] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 15 to 19 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein position 6, 7, 8, 9, 10, 11, or 12 of the modified antisense oligonucleotide aligns with the single nucleotide polymorphism; and positions 2 and 3 of the 5' and 3' wing segments comprise a 4'-CH(CH.sub.3)--O-2' bridge. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 3-9-3, 3-9-5, 4-9-5, 4-11-4, and 5-10-4.

[0346] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 15 to 19 linked nucleosides and fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein position 3, 4, 5, 6, 7, 8 or 9 of the gap segment aligns with the single nucleotide polymorphism; and positions 2 and 3 of the 5' and 3' wing segments comprise a 4'-CH(CH.sub.3)--O-2' bridge. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 3-9-3, 3-9-5, 4-9-5, 4-11-4, and 5-10-4.

[0347] A compound comprising a modified antisense oligonucleotide consisting of 15 to 19 linked nucleosides and fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein position 6, 7, 8, 9, 10, 11, or 12 of the modified antisense oligonucleotide aligns with the single nucleotide polymorphism; and positions 2, 3, 13, and 14 of the antisense oligonucleotide comprise a 4'-CH(CH.sub.3)--O-2' bridge. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 3-9-3, 3-9-5, 4-9-5, 4-11-4, and 5-10-4.

[0348] A compound comprising a modified antisense oligonucleotide consisting of 15 to 19 linked nucleosides and fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein position 3, 4, 5, 6, 7, 8, or 9 of the gap segment aligns with the single nucleotide polymorphism; and positions 2, 3, 13, and 14 of the antisense antisense oligonucleotide comprise a 4'-CH(CH.sub.3)--O-2' bridge. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 3-9-3, 3-9-5, 4-9-5, 4-11-4, and 5-10-4.

[0349] In certain embodiments, the compound comprise a modified antisense oligonucleotide consisting of 17 to 19 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein position 8, 9, or 10 of the modified antisense oligonucleotide aligns with the single nucleotide polymorphism; and wherein each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 3-9-3, 3-9-5, 4-9-5, 4-11-4, and 5-10-4.

[0350] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 17 to 19 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein position 5, 6, or 7 of the gap segment aligns with the single nucleotide polymorphism; and wherein each nucleoside of each wing segment comprises a 2'-O-methoxyethyl sugar. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 3-9-3, 3-9-5, 4-9-5, 4-11-4, and 5-10-4.

[0351] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 17 to 19 linked nucleosides, fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein position 8, 9, or 10 of the modified antisense oligonucleotide aligns with the single nucleotide polymorphism; and positions 2 and 3 of the 5' and 3' wing segments comprise a 4'-CH(CH.sub.3)--O-2' bridge. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 3-9-3, 3-9-5, 4-9-5, 4-11-4, and 5-10-4.

[0352] In certain embodiments, the compound comprises a modified antisense oligonucleotide consisting of 17 to 19 linked nucleosides and fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein position 5, 6, or 7 of the gap segment aligns with the single nucleotide polymorphism; and positions 2 and 3 of the 5' and 3' wing segments comprise a 4'-CH(CH.sub.3)--O-2' bridge. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 3-9-3, 3-9-5, 4-9-5, 4-11-4, and 5-10-4.

[0353] A compound comprising a modified antisense oligonucleotide consisting of 17 to 19 linked nucleosides and fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein position 8, 9, or 10 of the modified oligonucleotide aligns with the single nucleotide polymorphism; and positions 2, 3, 13, and 14 of the antisense antisense oligonucleotide comprise a 4'-CH(CH.sub.3)--O-2' bridge. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 3-9-3, 3-9-5, 4-9-5, 4-11-4, and 5-10-4.

[0354] A compound comprising a modified antisense oligonucleotide consisting of 17 to 19 linked nucleosides and fully complementary to a single nucleotide polymorphism site, wherein the modified antisense oligonucleotide comprises a wing-gap-wing motif, wherein position 5, 6, or 7 of the gap segment aligns with the single nucleotide polymorphism; and positions 2, 3, 13, and 14 of the antisense oligonucleotide comprise a 4'-CH(CH.sub.3)--O-2' bridge. In certain embodiments, the wing-gap-wing motif is any one of the group consisting of 2-9-6, 3-9-3, 3-9-5, 4-9-5, 4-11-4, and 5-10-4.

[0355] In a certain embodiment, the antisense oligonucleotide is 11 to 20 linked nucleosides in length and has, independently, 2 to 5 linked nucleosides in the 5' and 3' wings and 7 to 11 linked nucleosides in the gap. The SNP is complementary to position 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 of the antisense oligonucleotide (counting from the 5' terminus of the antisense oligonucleotide) or position 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 counting from the 5' terminus of the gap segment.

[0356] In a certain embodiment, the antisense oligonucleotide is 15 to 19 linked nucleosides in length and has, independently, 2 to 5 linked nucleosides in the 5' and 3' wings and 7 to 11 linked nucleosides in the gap. The SNP is complementary to position 6, 7, 8, 9, or 10 of the antisense oligonucleotide (counting from the 5' terminus of the antisense oligonucleotide) or position 4, 5, 6, or 7 counting from the 5' terminus of the gap segment.

[0357] In a certain embodiment, the antisense oligonucleotide is 17 linked nucleosides in length and has, independently, 2 to 5 linked nucleosides in the 5' and 3' wing segments and 9 to 11 linked nucleosides in the gap segment. The SNP is complementary to position 8, 9, or 10 of the antisense oligonucleotide (counting from the 5' terminus of the antisense oligonucleotide) or position 5, 6, or 7 (counting from the 5' terminus of the gap segment).

[0358] In a certain embodiment, the antisense oligonucleotide is 18 linked nucleosides in length and has, independently, 2 to 5 linked nucleosides in the 5' and 3' wing segments and 9 to 11 linked nucleosides in the gap segment. The SNP is complementary to position 8, 9, or 10 of the antisense oligonucleotide (counting from the 5' terminus of the antisense oligonucleotide) or position 5, 6, or 7 (counting from the 5' terminus of the gap segment).

[0359] In a certain embodiment, the antisense oligonucleotide is 19 linked nucleosides in length and has, independently, 2 to 5 linked nucleosides in the 5' and 3' wing segments and 9 to 11 linked nucleosides in the gap segment. The SNP is complementary to position 8, 9, or 10 of the antisense oligonucleotide (counting from the 5' terminus of the antisense oligonucleotide) or position 5, 6, or 7 (counting from the 5' terminus of the gap segment).

[0360] In certain embodiments, the invention provides methods of treating an individual comprising administering one or more pharmaceutical compositions described herein. In certain embodiments, the individual has an allelic variant associated with a disease or disorder. The pharmaceutical compositions provided herein preferentially target a SNP. In certain embodiments, the SNP is a differentiating polymorphism.

[0361] Methods have been described for determining whether a SNP is specific to a disease associated allele and more specifically whether a SNP variant of an allele of a heterozygous patient is on the same allele as a disease-causing mutation that is at a remote region of the gene's mRNA (WO 2008/147930 and WO 2008/143774).

[0362] Diseases associated with SNPs have been described for certain genes. In certain embodiments, the gene and associated disease are any of the following: APP gene encoding amyloid precursor protein involved in Alzheimer's disease (Gene, 371: 68, 2006); the PrP gene encoding prion protein involved in Creutzfeldt-Jakob disease and in fatal familial insomnia (Nat. Med. 1997, 3: 1009); GFAP gene encoding glial fibrillary acidic protein involved in Alexander disease (J. Neurosci. 2006, 26:111623); alpha-synuclein gene encoding alpha-synuclein protein involved in Parkinson's disease (J. Clin. Invest. 2003, 111: 145); SOD-1 gene encoding the SOD-1 protein involved in amyotrophic lateral sclerosis (Science 1998, 281: 1851); atrophin-1 gene encoding atrophin-1 protein involved in dentato-rubral and pallido-luysian atrophy (DRPA) (Trends Mol. Med. 2001, 7: 479); SCA1 gene encoding ataxin-1 protein involved in spino-cerebellar ataxia-1 (SCA1) (Protein Sci. 2003, 12: 953); PLP gene encoding proteolipid protein involved in Pelizaeus-Merzbacher disease (Neuro Mol Med. 2007, 4: 73); DYT1 gene encoding torsinA protein involved in Torsion dystonia (Brain Res. 2000, 877: 379); and alpha-B crystalline gene encoding alpha-B crystalline protein involved in protein aggregation diseases, including cardiomyopathy (Cell 2007, 130: 427); alpha1-antitrypsin gene encoding alpha1-antitrypsin protein involved in chronic obstructive pulmonary disease (COPD), liver disease and hepatocellular carcinoma (New Engl J Med. 2002, 346: 45); Ltk gene encoding leukocyte tyrosine kinase protein involved in systemic lupus erythematosus (Hum. Mol. Gen. 2004, 13: 171); PCSK9 gene encoding PCSK9 protein involved in hypercholesterolemia (Hum Mutat. 2009, 30: 520); prolactin receptor gene encoding prolactin receptor protein involved in breast tumors (Proc. Natl. Assoc. Sci. 2008, 105: 4533); CCL5 gene encoding the chemokine CCL5 involved in COPD and asthma (Eur. Respir. J. 2008, 32: 327); PTPN22 gene encoding PTPN22 protein involved in Type 1 diabetes, Rheumatoid arthritis, Graves disease, and SLE (Proc. Natl. Assoc. Sci. 2007, 104: 19767); androgen receptor gene encoding the androgen receptor protein involved in spinal and bulbar muscular atrophy or Kennedy's disease (J Steroid Biochem. Mol. Biol. 2008, 108: 245); CHMP4B gene encoding chromatin modifying protein-4B involved in progressive childhood posterior subcapsular cataracts (Am. J. Hum. Genet 2007, 81: 596); FXR/NR1H4 gene encoding Farnesoid X receptor protein involved in cholesterol gallstone disease, arthrosclerosis and diabetes (Mol. Endocrinol. 2007, 21: 1769); ABCA1 gene encoding ABCA1 protein involved in cardiovascular disease (Transl. Res. 2007, 149: 205); CaSR gene encoding the calcium sensing receptor protein involved in primary hypercalciuria (Kidney Int. 2007, 71: 1155); alpha-globin gene encoding alpha-globin protein involved in alpha-thallasemia (Science 2006, 312: 1215); httlpr gene encoding HTTLPR protein involved in obsessive compulsive disorder (Am. J. Hum. Genet. 2006, 78: 815); AVP gene encoding arginine vasopressin protein in stress-related disorders such as anxiety disorders and comorbid depression (CNS Neurol. Disord. Drug Targets 2006, 5: 167); GNAS gene encoding G proteins involved in congenital visual defects, hypertension, metabolic syndrome (Trends Pharmacol. Sci. 2006, 27: 260); APAF1 gene encoding APAF1 protein involved in a predisposition to major depression (Mol. Psychiatry 2006, 11: 76); TGF-beta1 gene encoding TGF-beta1 protein involved in breast cancer and prostate cancer (Cancer Epidemiol. Biomarkers Prev. 2004, 13: 759); AChR gene encoding acetylcholine receptor involved in congenital myasthenic syndrome (Neurology 2004, 62: 1090); P2Y12 gene encoding adenosine diphosphate (ADP) receptor protein involved in risk of peripheral arterial disease (Circulation 2003, 108: 2971); LQT1 gene encoding LQT1 protein involved in atrial fibrillation (Cardiology 2003, 100: 109); RET protooncogene encoding RET protein involved in sporadic pheochromocytoma (J. Clin. Endocrinol. Metab. 2003, 88: 4911); filamin A gene encoding filamin A protein involved in various congenital malformations (Nat. Genet. 2003, 33: 487); TARDBP gene encoding TDP-43 protein involved in amyotrophic lateral sclerosis (Hum. Mol. Gene.t 2010, 19: 671); SCA3 gene encoding ataxin-3 protein involved in Machado-Joseph disease (PLoS One 2008, 3: e3341); SCAT gene encoding ataxin-7 protein involved in spino-cerebellar ataxia-7 (PLoS One 2009, 4: e7232); HTT gene encoding huntingtin protein involved in Huntington's disease (Neurobiol Dis. 1996, 3:183); and the CA4 gene encoding carbonic anhydrase 4 protein, CRX gene encoding cone-rod homeobox transcription factor protein, FSCN2 gene encoding retinal fascin homolog 2 protein, IMPDH1 gene encoding inosine monophosphate dehydrogenase 1 protein, NR2E3 gene encoding nuclear receptor subfamily 2 group E3 protein, NRL gene encoding neural retina leucine zipper protein, PRPF3 (RP18) gene encoding pre-mRNA splicing factor 3 protein, PRPF8 (RP13) gene encoding pre-mRNA splicing factor 8 protein, PRPF31 (RP11) gene encoding pre-mRNA splicing factor 31 protein, RDS gene encoding peripherin 2 protein, ROM1 gene encoding rod outer membrane protein 1 protein, RHO gene encoding rhodopsin protein, RP1 gene encoding RP1 protein, RPGR gene encoding retinitis pigmentosa GTPase regulator protein, all of which are involved in Autosomal Dominant Retinitis Pigmentosa disease (Adv Exp Med Biol. 2008, 613:203)

[0363] In certain embodiments, the disease is a neurodegenerative disorder. In certain embodiments, the neurodegenerative disorder is Huntington's Disease. In certain embodiments, the targeted SNP is one or more of: rs6446723, rs3856973, rs2285086, rs363092, rs916171, rs6844859, rs7691627, rs4690073, rs2024115, rs11731237, rs362296, rs10015979, rs7659144, rs363096, rs362273, rs16843804, rs362271, rs362275, rs3121419, rs362272, rs3775061, rs34315806, rs363099, rs2298967, rs363088, rs363064, rs363102, rs2798235, rs363080, rs363072, rs363125, rs362303, rs362310, rs10488840, rs362325, rs35892913, rs363102, rs363096, rs11731237, rs10015979, rs363080, rs2798235, rs1936032, rs2276881, rs363070, rs35892913, rs12502045, rs6446723, rs7685686, rs3733217, rs6844859, rs362331, rs1143646, rs2285086, rs2298969, rs4690072, rs916171, rs3025849, rs7691627, rs4690073, rs3856973, rs363092, rs362310, rs362325, rs363144, rs362303, rs34315806, rs363099, rs363081, rs3775061, rs2024115, rs10488840, rs363125, rs362296, rs2298967, rs363088, rs363064, rs362275, rs3121419, rs3025849, rs363070, rs362273, rs362272, rs362306, rs362271, rs363072, rs16843804, rs7659144, rs363120, and rs12502045. In certain embodiments the compounds are ISIS460065, ISIS 459978, ISIS 460028, ISIS 460209, ISIS 460208, and ISIS 460206.

Therapeutically Effective Dosages

[0364] In certain embodiments, administration of a therapeutically effective amount of an antisense compound targeted to the mutant huntingtin allele is accompanied by monitoring of expression of a gene product in an individual, to determine an individual's response to administration of the antisense compound. In certain embodiments, the gene product is huntingtin mRNA or protein. An individual's response to administration of the antisense compound is used by a physician to determine the amount and duration of therapeutic intervention.

[0365] In certain embodiments, administration of an antisense compound targeted to a mutant nucleic acid results in reduction of mRNA or protein expression by at least 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 99%, or a range defined by any two of these values. In certain embodiments, the mutant nucleic acid is huntingtin nucleic acid, the mRNA is huntingtin mRNA, and the protein is huntingtin protein.

[0366] In certain embodiments, pharmaceutical compositions comprising an antisense compound targeted to a mutant allele are used for the preparation of a medicament for treating a patient suffering or susceptible to any of Huntington's Disease, Alzheimer's Disease, Crutzfeldt-Jakob Disease, Fatal Familial Insomnia, Huntington's Disease, Alexander Disease, Parkinson's Disease, Amyotrophic Lateral Sclerosis (ALS), Dentato-Rubral and Pallido-Luysian Atrophy, Spino-Cerebellar Ataxia 1, Pelizaeus-Merzbacher Disease, Torsion Dystonia, Cardiomyopathy, Chronic Obstructive Pulmonary Disease (COPD), liver disease and hepatocellular carcinoma, SLE, Hypercholesterolemia, breast tumors, Asthma, Type 1 Diabetes, Rheumatoid Arthritis, Graves Disease, Spinal and Bulbar Muscular Atrophy, Kennedy's Disease, progressive childhood posterior subcapsular cataracts, Cholesterol Gallstone Disease, Arthrosclerosis, cardiovascular disease, primary hypercalciuria, alpha-thallasemia, OCD, stress-related disorders (including anxiety disorders and comorbid depression), congenital visual defects, hypertension, metabolic syndrome, major depression, breast cancer, prostate cancer, congenital myasthenic syndrome, peripheral arterial syndrome, atrial fibrillation, sporadic pheochromocytoma, congenital malformations, NJD, SCA7, and autosomal dominant retinitis pigmentosa adRP.

Certain Combination Therapies

[0367] In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with one or more other pharmaceutical agents. In certain embodiments, such one or more other pharmaceutical agents are designed to treat the same disease, disorder, or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat a different disease, disorder, or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat an undesired side effect of one or more pharmaceutical compositions of the present invention. In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with another pharmaceutical agent to treat an undesired effect of that other pharmaceutical agent. In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with another pharmaceutical agent to produce a combinational effect. In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with another pharmaceutical agent to produce a synergistic effect.

[0368] In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at the same time. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at different times. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared together in a single formulation. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared separately.

EXAMPLES

Non-Limiting Disclosure and Incorporation by Reference

[0369] While certain compounds, compositions and methods described herein have been described with specificity in accordance with certain embodiments, the following examples serve only to illustrate the compounds described herein and are not intended to limit the same. Each of the patents, applications, printed publications, and other published documents mentioned or referred to in this specification are herein incorporated by reference in their entirety.

Example 1

Single Nucleotide Polymorphisms (SNPs) in the Huntingtin (HTT) Gene Sequence

[0370] The HTT genomic sequence, designated herein as SEQ ID NO: 1 (NT.sub.--006081.18 truncated from nucleotides 1566000 to 1768000) was aligned with the HTT mRNA, designated herein as SEQ ID NO: 2 (NM.sub.--002111.6), using the EMBL-EBI sequence database (ClustalW2, http://www.ebi.ac.uk/Tools/clustalw2/index.html), and the output is presented in FIG. 1. SNP positions (identified by Hayden et al, WO/2009/135322) associated with the HTT gene were mapped to the two sequences and have been demarcated in FIG. 1 by their reference SNP ID number from the Entrez SNP database at the National Center for Biotechnology Information (NCBI, http://www.ncbi.nlm.nih.gov/sites/entrez?db=snp), incorporated herein by reference. Table 2 furnishes further details on each SNP. The `Reference SNP ID number` or `RS number` is the number designated to each SNP from the Entrez SNP database at NCBI, incorporated herein by reference. `SNP position` refers to the nucleotide position of the SNP on SEQ ID NO: 1. `Polymorphism` indicates the nucleotide variants at that SNP position. `Major allele` indicates the nucleotide associated with the major allele, or the nucleotide present in a statistically significant proportion of individuals in the human population. `Minor allele` indicates the nucleotide associated with the minor allele, or the nucleotide present in a relatively small proportion of individuals in the human population.

TABLE-US-00002 TABLE 2 Single Nuclear Polymorphisms (SNPs) and their positions on SEQ ID NO: 1 SNP Major Minor RS No. position Polymorphism allele allele rs2857936 1963 C/T C T rs12506200 3707 A/G G A rs762855 14449 A/G G A rs3856973 19826 G/A G A rs2285086 28912 G/A A G rs7659144 37974 C/G C G rs16843804 44043 C/T C T rs2024115 44221 G/A A G rs10015979 49095 A/G A G rs7691627 51063 A/G G A rs2798235 54485 G/A G A rs4690072 62160 G/T T G rs6446723 66466 C/T T C rs363081 73280 G/A G A rs363080 73564 T/C C T rs363075 77327 G/A G A rs363064 81063 T/C C T rs3025849 83420 A/G A G rs6855981 87929 A/G G A rs363102 88669 G/A A G rs11731237 91466 C/T C T rs4690073 99803 A/G G A rs363144 100948 T/G T G rs3025838 101099 C/T C T rs34315806 101687 A/G G A rs363099 101709 T/C C T rs363096 119674 T/C T C rs2298967 125400 C/T T C rs2298969 125897 A/G G A rs6844859 130139 C/T T C rs363092 135682 C/A C A rs7685686 146795 A/G A G rs363088 149983 A/T A T rs362331 155488 C/T T C rs916171 156468 G/C C G rs362322 161018 A/G A G rs362275 164255 T/C C T rs362273 167080 A/G A G rs2276881 171314 G/A G A rs3121419 171910 T/C C T rs362272 174633 G/A G A rs362271 175171 G/A G A rs3775061 178407 C/T C T rs362310 179429 A/G G A rs362307 181498 T/C C T rs362306 181753 G/A G A rs362303 181960 T/C C T rs362296 186660 C/A C A rs1006798 198026 A/G A G

Example 2

Design of Antisense Oligonucleotides Targeting Huntingtin Gene SNPs and Inhibition of HTT mRNA in Coriell Fibroblast Cell Lines (GM04281, GM02171, and GM02173B)

[0371] Antisense oligonucleotides targeting nucleotides overlapping SNP positions presented in Table 1 were designed and tested for potency in three huntingtin patient-derived Coriell fibroblast cell lines, GM04281, GM02171, and GM02173B (from the Coriell Institute for Medical Research). Cultured GM04281 cells or GM02171 cells or GM02173B cells at a density of 20,000 cells per well were transfected using electroporation with 10,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and HTT mRNA levels were measured by quantitative real time PCR using primer probe set RTS2617 (forward sequence CTCCGTCCGGTAGACATGCT, designated herein as SEQ ID NO: 3; reverse sequence GGAAATCAGAACCCTCAAAATGG, designated herein as SEQ ID NO: 4; probe sequence TGAGCACTGTTCAACTGTGGATATCGGGAX, designated herein as SEQ ID NO: 5). HTT mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN. Results are presented as percent inhibition of HTT mRNA, relative to untreated control cells.

[0372] ISIS 387916 (TCTCTATTGCACATTCCAAG, 5-10-5 MOE (SEQ ID NO: 6)) and ISIS 388816 (GCCGTAGCCTGGGACCCGCC, 5-10-5 MOE (SEQ ID NO: 7)) were included in each study as benchmark oligonucleotides against which the potency of the antisense oligonucleotides targeting nucleotides overlapping each SNP position could be compared.

[0373] The chimeric antisense oligonucleotides in Tables 3 and 4 were designed as 5-9-5 MOE gapmers. The gapmers are 19 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on both sides (in the 5' and 3' directions) by wings comprising five nucleotides each. Each nucleotide in the 5' wing segment and each nucleotide in the 3' wing segment has a 2'-MOE modification. The internucleoside linkages throughout each gapmer are phosphorothioate (P.dbd.S) linkages. All cytosine nucleobases throughout each gapmer are 5-methylcytosines.

[0374] The oligonucleotides are further described in Table 3. The percent inhibition of HTT mRNA by the antisense oligonucleotides in each cell line is shown in Table 4. `Target allele` indicates whether the gapmer is targeted to the major or the minor allele at the SNP position. The number in parentheses indicates the nucleotide position in the gapmer opposite to the SNP position, starting from the 5'-terminus of the oligonucleotide. `Start site` indicates the 5'-most nucleotide to which the gapmer is targeted. "Stop site" indicates the 3'-most nucleotide to which the gapmer is targeted. Each gapmer listed in Tables 3 and 4 is targeted to human HTT pre-mRNA, which is SEQ ID NO: 1.

TABLE-US-00003 TABLE 3 Chimeric oligonucleotides targeting SNP positions on the HTT gene SEQ ISIS SNP RS Target Start Stop ID No No. allele Sequence Site Site NO 387916 n/a n/a TCTCTATTGCACATTCCAAG 145466 145485 6 388816 n/a n/a GCCGTAGCCTGGGACCCGCC 16501 16520 7 435330 rs3856973 Major (8) TAACACTCGATTAACCCTG 19815 19833 8 435348 rs3856973 Minor (8) TAACACTTGATTAACCCTG 19815 19833 9 435294 rs3856973 Major (10) GTTAACACTCGATTAACCC 19817 19835 10 435312 rs3856973 Minor (10) GTTAACACTTGATTAACCC 19817 19835 11 435864 rs2285086 Major (10) GCTAGTTCATCCCAGTGAG 28903 28921 12 435889 rs2285086 Minor (10) GCTAGTTCACCCCAGTGAG 28903 28921 13 435878 rs7659144 Major (10) TGGAAATGGGTTTTTCCAC 37965 37983 14 435903 rs7659144 Minor (10) TGGAAATGGCTTTTTCCAC 37965 37983 15 435863 rs16843804 Major (10) TTTAACCGTGGCATGGGCA 44034 44052 16 435888 rs16843804 Minor (10) TTTAACCGTAGCATGGGCA 44034 44052 17 435331 rs2024115 Major (8) TTCAAGCTAGTAACGATGC 44210 44228 18 435349 rs2024115 Minor (8) TTCAAGCCAGTAACGATGC 44210 44228 19 435295 rs2024115 Major (10) ACTTCAAGCTAGTAACGAT 44212 44230 20 435313 rs2024115 Minor (10) ACTTCAAGCCAGTAACGAT 44212 44230 21 435862 rs10015979 Major (10) GCAGCTAGGTTAAAGAGTC 49086 49104 22 435887 rs10015979 Minor (10) GCAGCTAGGCTAAAGAGTC 49086 49104 23 435880 rs7691627 Major (10) AATAAGAAACACAATCAAA 51054 51072 24 435905 rs7691627 Minor (10) AATAAGAAATACAATCAAA 51054 51072 25 435885 rs2798235 Major (10) CAGAGGAGGCATACTGTAT 54476 54494 26 435910 rs2798235 Minor (10) CAGAGGAGGTATACTGTAT 54476 54494 27 435874 rs4690072 Major (10) CACAGTGCTACCCAACCTT 62151 62169 28 435899 rs4690072 Minor (10) CACAGTGCTCCCCAACCTT 62151 62169 29 435875 rs6446723 Major (10) TAATTTTCTAGACTTTATG 66457 66475 30 435900 rs6446723 Minor (10) TAATTTTCTGGACTTTATG 66457 66475 31 435332 rs363081 Major (8) GCTACAACGCAGGTCAAAT 73269 73287 32 435350 rs363081 Minor (8) GCTACAATGCAGGTCAAAT 73269 73287 33 435296 rs363081 Major (10) GAGCTACAACGCAGGTCAA 73271 73289 34 435314 rs363081 Minor (10) GAGCTACAATGCAGGTCAA 73271 73289 35 435886 rs363080 Major (10) AGAGAGAACGAGAAGGCTC 73555 73573 36 435911 rs363080 Minor (10) AGAGAGAACAAGAAGGCTC 73555 73573 37 435914 rs363075 Major (6) AGCCCCTCTGTGTAAGTTT 77314 77332 38 435926 rs363075 Minor (6) AGCCCTTCTGTGTAAGTTT 77314 77332 39 435916 rs363075 Major (7) GAGCCCCTCTGTGTAAGTT 77315 77333 40 435928 rs363075 Minor (7) GAGCCCTTCTGTGTAAGTT 77315 77333 41 435333 rs363075 Major (8) TGAGCCCCTCTGTGTAAGT 77316 77334 42 435351 rs363075 Minor (8) TGAGCCCTTCTGTGTAAGT 77316 77334 43 435918 rs363075 Major (9) ATGAGCCCCTCTGTGTAAG 77317 77335 44 435930 rs363075 Minor (9) ATGAGCCCTTCTGTGTAAG 77317 77335 45 435297 rs363075 Major (10) GATGAGCCCCTCTGTGTAA 77318 77336 46 435315 rs363075 Minor (10) GATGAGCCCTTCTGTGTAA 77318 77336 47 435920 rs363075 Major (11) TGATGAGCCCCTCTGTGTA 77319 77337 48 435932 rs363075 Minor (11) TGATGAGCCCTTCTGTGTA 77319 77337 49 435366 rs363075 Major (12) ATGATGAGCCCCTCTGTGT 77320 77338 50 435924 rs363075 Minor (12) ATGATGAGCCCTTCTGTGT 77320 77338 51 435922 rs363075 Major (14) TAATGATGAGCCCCTCTGT 77322 77340 52 435934 rs363075 Minor (14) TAATGATGAGCCCTTCTGT 77322 77340 53 435334 rs363064 Major (8) AGAATACGGGTAACATTTT 81052 81070 54 435352 rs363064 Minor (8) AGAATACAGGTAACATTTT 81052 81070 55 435298 rs363064 Major (10) GGAGAATACGGGTAACATT 81054 81072 56 435316 rs363064 Minor (10) GGAGAATACAGGTAACATT 81054 81072 57 435335 rs3025849 Major (8) TTAGTAATCAATTTTAATG 83409 83427 58 435353 rs3025849 Minor (8) TTAGTAACCAATTTTAATG 83409 83427 59 435299 rs3025849 Major (10) AGTTAGTAATCAATTTTAA 83411 83429 60 435317 rs3025849 Minor (10) AGTTAGTAACCAATTTTAA 83411 83429 61 435877 rs6855981 Major (10) GAAGGAATGCTTTTACTAG 87920 87938 62 435902 IS6855981 Minor (10) GAAGGAATGTTTTTACTAG 87920 87938 63 435336 rs363102 Major (8) CTAAAACTAACTTGAGAAT 88658 88676 64 435354 rs363l02 Minor (8) CTAAAACCAACTTGAGAAT 88658 88676 65 435300 rs363102 Major (10) ATCTAAAACTAACTTGAGA 88660 88678 66 435318 rs363102 Minor (10) ATCTAAAACCAACTTGAGA 88660 88678 67 435884 rs11731237 Major (10) GGTGGGCAGGAAGGACTGA 91457 91475 68 435909 rs11731237 Minor (10) GGTGGGCAGAAAGGACTGA 91457 91475 69 435337 rs4690073 Major (8) CCTAAATCAATCTACAAGT 99792 99810 70 435355 rs4690073 Minor (8) CCTAAATTAATCTACAAGT 99792 99810 71 435301 rs4690073 Major (10) TCCCTAAATCAATCTACAA 99794 99812 72 435319 rs4690073 Minor (10) TCCCTAAATTAATCTACAA 99794 99812 73 435883 rs363144 Major (10) GAAAATGTGAGTGGATCTA 100939 100957 74 435908 rs363144 Minor (10) GAAAATGTGCGTGGATCTA 100939 100957 75 435338 rs3025838 Major (8) GTAAGGCGAGACTGACTAG 101088 101106 76 435356 rs3025838 Minor (8) GTAAGGCAAGACTGACTAG 101088 101106 77 435302 rs3025838 Major (10) AGGTAAGGCGAGACTGACT 101090 101108 78 435320 rs3025838 Minor (10) AGGTAAGGCAAGACTGACT 101090 101108 79 435339 rs363099 Major (8) CTGAGCGGAGAAACCCTCC 101698 101716 80 435357 rs363099 Minor (8) CTGAGCGAAGAAACCCTCC 101698 101716 81 435303 rs363099 Major (10) GGCTGAGCGGAGAAACCCT 101700 101718 82 435321 rs363099 Minor (10) GGCTGAGCGAAGAAACCCT 101700 101718 83 435367 rs363099 Major (12) AAGGCTGAGCGGAGAAACC 101702 101720 84 435340 rs363096 Major (8) TTCCCTAAAAACAAAAACA 119663 119681 85 435358 rs363096 Minor (8) TTCCCTAGAAACAAAAACA 119663 119681 86 435304 rs363096 Major (10) GATTCCCTAAAAACAAAAA 119665 119683 87 435322 rs363096 Minor (10) GATTCCCTAGAAACAAAAA 119665 119683 88 435341 rs2298967 Major (8) CTTTTCTATTGTCTGTCCC 125389 125407 89 435359 rs2298967 Minor (8) CTTTTCTGTTGTCTGTCCC 125389 125407 90 435305 rs2298967 Major (10) TGCTTTTCTATTGTCTGTC 125391 125409 91 435323 rs2298967 Minor (10) TGCTTTTCTGTTGTCTGTC 125391 125409 92 435865 rs2298969 Major (10) AAGGGATGCCGACTTGGGC 125888 125906 93 435890 rs2298969 Minor (10) AAGGGATGCTGACTTGGGC 125888 125906 94 435876 rs6844859 Major (10) ACCTTCCTCACTGAGGATG 130130 130148 95 435901 rs6844859 Minor (10) ACCTTCCTCGCTGAGGATG 130130 130148 96 435872 rs363092 Major (10) CAAACCACTGTGGGATGAA 135673 135691 97 435897 rs363092 Minor (10) CAAACCACTTTGGGATGAA 135673 135691 98 435879 rs7685686 Major (10) AATAAATTGTCATCACCAG 146786 146804 99 435904 rs7685686 Minor (10) AATAAATTGCCATCACCAG 146786 146804 100 435871 rs363088 Major (10) TCACAGCTATCTTCTCATC 149974 149992 101 435896 rs363088 Minor (10) TCACAGCTAACTTCTCATC 149974 149992 102 435870 rs362331 Major (10) GCACACAGTAGATGAGGGA 155479 155497 103 435895 rs362331 Minor (10) GCACACAGTGGATGAGGGA 155479 155497 104 435881 rs916171 Major (10) CAGAACAAAGAGAAGAATT 156459 156477 105 435906 rs916171 Minor (10) CAGAACAAACAGAAGAATT 156459 156477 106 435342 rs362322 Major (8) GCTTACATGCCTTCAGTGA 161007 161025 107 435360 rs362322 Minor (8) GCTTACACGCCTTCAGTGA 161007 161025 108 435306 rs362322 Major (10) CAGCTTACATGCCTTCAGT 161009 161027 109 435324 rs362322 Minor (10) CAGCTTACACGCCTTCAGT 161009 161027 110 435868 rs362275 Major (10) AAGAAGCCTGATAAAATCT 164246 164264 111 435893 rs362275 Minor (10) AAGAAGCCTAATAAAATCT 164246 164264 112 435343 rs2276881 Major (8) CATACATCAGCTCAAACTG 171303 171321 113 435361 rs2276881 Minor (8) CATACATTAGCTCAAACTG 171303 171321 114 435307 rs2276881 Major (10) CACATACATCAGCTCAAAC 171305 171323 115 435325 rs2276881 Minor (10) CACATACATTAGCTCAAAC 171305 171323 116 435368 rs2276881 Major (12) GTCACATACATCAGCTCAA 171307 171325 117 435866 rs3121419 Major (10) GAGACTATAGCACCCAGAT 171901 171919 118 435891 rs3121419 Minor (10) GAGACTATAACACCCAGAT 171901 171919 119 435344 rs362272 Major (8) TAGAGGACGCCGTGCAGGG 174622 174640 120 435362 rs362272 Minor (8) TAGAGGATGCCGTGCAGGG 174622 174640 121 435308 rs362272 Major (10) CATAGAGGACGCCGTGCAG 174624 174642 122 435326 rs362272 Minor (10) CATAGAGGATGCCGTGCAG 174624 174642 123 435369 rs362272 Major (12) CACATAGAGGACGCCGTGC 174626 174644 124 435867 rs362271 Major (10) ACGTGTGTACAGAACCTGC 175162 175180 125 435892 rs362271 Minor (10) ACGTGTGTATAGAACCTGC 175162 175180 126

435873 rs3775061 Major (10) TGTTCAGAATGCCTCATCT 178398 178416 127 435898 rs3775061 Minor (10) TGTTCAGAACGCCTCATCT 178398 178416 128 435345 rs362310 Major (8) AAACGGCGCAGCGGGAAGG 179418 179436 129 435363 rs362310 Minor (8) AAACGGCACAGCGGGAAGG 179418 179436 130 435309 rs362310 Major (10) AGAAACGGCGCAGCGGGAA 179420 179438 131 435327 rs362310 Minor (10) AGAAACGGCACAGCGGGAA 179420 179438 132 435915 rs362307 Major (6) AGGGCGCAGACTTCCAAAG 181485 181503 133 435927 rs362307 Minor (6) AGGGCACAGACTTCCAAAG 181485 181503 134 435917 rs362307 Major (7) AAGGGCGCAGACTTCCAAA 181486 181504 135 435929 rs362307 Minor (7) AAGGGCACAGACTTCCAAA 181486 181504 136 435346 rs362307 Major (8) CAAGGGCGCAGACTTCCAA 181487 181505 137 435364 rs362307 Minor (8) CAAGGGCACAGACTTCCAA 181487 181505 138 435919 rs362307 Major (9) ACAAGGGCGCAGACTTCCA 181488 181506 139 435931 rs362307 Minor (9) ACAAGGGCACAGACTTCCA 181488 181506 140 435310 rs362307 Major (10) CACAAGGGCGCAGACTTCC 181489 181507 141 435328 rs362307 Minor (10) CACAAGGGCACAGACTTCC 181489 181507 142 435921 rs362307 Major (11) GCACAAGGGCGCAGACTTC 181490 181508 143 435933 rs362307 Minor (11) GCACAAGGGCACAGACTTC 181490 181508 144 435370 rs362307 Major (12) GGCACAAGGGCGCAGACTT 181491 181509 145 435925 rs362307 Minor (12) GGCACAAGGGCACAGACTT 181491 181509 146 435923 rs362307 Major (14) AGGGCACAAGGGCGCAGAC 181493 181511 147 435935 rs362307 Minor (14) AGGGCACAAGGGCACAGAC 181493 181511 148 435869 rs362306 Major (10) GAGCAGCTGCAACCTGGCA 181744 181762 149 435894 rs362306 Minor (10) GAGCAGCTGTAACCTGGCA 181744 181762 150 435347 rs362303 Major (8) TGGTGCCGGGTGTCTAGCA 181949 181967 151 435365 rs362303 Minor (8) TGGTGCCAGGTGTCTAGCA 181949 181967 152 435311 rs362303 Major (10) AATGGTGCCGGGTGTCTAG 181951 181969 153 435329 rs362303 Minor (10) AATGGTGCCAGGTGTCTAG 181951 181969 154 435882 rs362296 Major (10) GGGGACAGGGTGTGCTCTC 186651 186669 155 435907 rs362296 Minor (10) GGGGACAGGTTGTGCTCTC 186651 186669 156

TABLE-US-00004 TABLE 4 Comparison of inhibition of HTT mRNA levels by ISIS 387916 and ISIS 388816 with that by chimeric oligonucleotides targeting SNP positions on the HTT gene (SEQ ID NO: 1) SEQ SNP RS Target % inhibition ID ISIS No No. allele GM04281 GM02171 GM02173B NO 387916 n/a n/a 96 96 98 6 388816 n/a n/a 76 88 85 7 435330 rs3856973 Major (8) 64 51 36 8 435348 rs3856973 Minor (8) 50 88 80 9 435294 rs3856973 Major (10) 54 46 54 10 435312 rs3856973 Minor (10) 20 82 58 11 435864 rs2285086 Major (10) 54 28 26 12 435889 rs2285086 Minor (10) 17 43 41 13 435878 rs7659144 Major (10) 43 32 39 14 435903 rs7659144 Minor (10) 16 37 29 15 435863 rs16843804 Major (10) 63 78 81 16 435888 rs16843804 Minor (10) 58 75 77 17 435331 rs2024115 Major (8) 56 27 56 18 435349 rs2024115 Minor (8) 26 91 66 19 435295 rs2024115 Major (10) 53 57 62 20 435313 rs2024115 Minor (10) 25 87 53 21 435862 rs10015979 Major (10) 8 51 40 22 435887 rs10015979 Minor (10) 40 22 28 23 435880 rs7691627 Major (10) 43 17 21 24 435905 rs7691627 Minor (10) 13 27 15 25 435885 rs2798235 Major (10) 38 39 30 26 435910 rs2798235 Minor (10) 17 30 16 27 435874 rs4690072 Major (10) 61 34 48 28 435899 rs4690072 Minor (10) 50 41 45 29 435875 rs6446723 Major (10) 28 13 35 30 435900 rs6446723 Minor (10) 24 56 37 31 435332 rs363081 Major (8) 76 95 88 32 435350 rs363081 Minor (8) 27 61 43 33 435296 rs363081 Major (10) 59 77 66 34 435314 rs363081 Minor (10) 38 66 40 35 435886 rs363080 Major (10) 74 72 79 36 435911 rs363080 Minor (10) 57 58 54 37 435914 rs363075 Major (6) 95 92 95 38 435926 rs363075 Minor (6) 88 81 79 39 435916 rs363075 Major (7) 90 92 94 40 435928 rs363075 Minor (7) 83 79 85 41 435333 rs363075 Major (8) 86 97 91 42 435351 rs363075 Minor (8) 59 80 58 43 435918 rs363075 Major (9) 83 90 91 44 435930 rs363075 Minor (9) 29 49 49 45 435297 rs363075 Major (10) 74 84 83 46 435315 rs363075 Minor (10) 47 63 45 47 435920 rs363075 Major (11) 78 66 83 48 435932 rs363075 Minor (11) 39 20 19 49 435366 rs363075 Major (12) 80 91 85 50 435924 rs363075 Minor (12) 37 49 58 51 435922 rs363075 Major (14) 80 90 91 52 435934 rs363075 Minor (14) 63 70 80 53 435334 rs363064 Major (8) 50 59 44 54 435352 rs363064 Minor (8) 12 37 48 55 435298 rs363064 Major (10) 81 92 87 56 435316 rs363064 Minor (10) 69 90 80 57 435335 rs3025849 Major (8) 0 40 37 58 435353 rs3025849 Minor (8) 0 29 18 59 435299 rs3025849 Major (10) 0 34 67 60 435317 rs3025849 Minor (10) 0 38 34 61 435877 rs6855981 Major (10) 31 59 58 62 435902 rs6855981 Minor (10) 0 43 27 63 435336 rs363102 Major (8) 0 21 19 64 435354 rs363102 Minor (8) 0 36 33 65 435300 rs363102 Major (10) 0 34 24 66 435318 rs363102 Minor (10) 0 30 20 67 435884 rs11731237 Major (10) 7 46 51 68 435909 rs11731237 Minor (10) 30 47 41 69 435337 rs4690073 Major (8) 12 0 12 70 435355 rs4690073 Minor (8) 0 26 33 71 435301 rs4690073 Major (10) 23 0 10 72 435319 rs4690073 Minor (10) 0 45 53 73 435883 rs363144 Major (10) 24 23 39 74 435908 rs363144 Minor (10) 27 20 22 75 435338 rs3025838 Major (8) 31 46 69 76 435356 rs3025838 Minor (8) 3 25 17 77 435302 rs3025838 Major (10) 39 73 67 78 435320 rs3025838 Minor (10) 21 49 32 79 435339 rs363099 Major (8) 84 87 76 80 435357 rs363099 Minor (8) 71 91 90 81 435303 rs363099 Major (10) 83 92 85 82 435321 rs363099 Minor (10) 84 95 89 83 435367 rs363099 Major (12) 76 82 72 84 435340 rs363096 Major (8) 0 47 52 85 435358 rs363096 Minor (8) 0 25 35 86 435304 rs363096 Major (10) 5 33 36 87 435322 rs363096 Minor (10) 2 30 32 88 435341 rs2298967 Major (8) 54 72 56 89 435359 rs2298967 Minor (8) 25 59 63 90 435305 rs2298967 Major (10) 66 80 78 91 435323 rs2298967 Minor (10) 36 79 66 92 435865 rs2298969 Major (10) 53 72 79 93 435890 rs2298969 Minor (10) 65 46 54 94 435876 rs6844859 Major (10) 70 67 77 95 435901 rs6844859 Minor (10) 39 83 80 96 435872 rs363092 Major (10) 46 41 54 97 435897 rs363092 Minor (10) 37 69 57 98 435879 rs7685686 Major (10) 83 31 70 99 435904 rs7685686 Minor (10) 30 92 72 100 435871 rs363088 Major (10) 70 55 70 101 435896 rs363088 Minor (10) 66 74 80 102 435870 rs362331 Major (10) 88 74 88 103 435895 rs362331 Minor (10) 78 92 86 104 435881 rs916171 Major (10) 0 57 51 105 435906 rs916171 Minor (10) 14 26 17 106 435342 rs362322 Major (8) 47 74 67 107 435360 rs362322 Minor (8) 17 58 52 108 435306 rs362322 Major (10) 50 77 65 109 435324 rs362322 Minor (10) 42 61 64 110 435868 rs362275 Major (10) 54 35 43 111 435893 rs362275 Minor (10) 3 27 33 112 435343 rs2276881 Major (8) 59 76 65 113 435361 rs2276881 Minor (8) 58 44 20 114 435307 rs2276881 Major (10) 69 82 81 115 435325 rs2276881 Minor (10) 17 47 43 116 435368 rs2276881 Major (12) 84 96 92 117 435866 rs3121419 Major (10) 67 61 64 118 435891 rs3121419 Minor (10) 53 76 73 119 435344 rs362272 Major (8) 35 46 36 120 435362 rs362272 Minor (8) 34 68 57 121 435308 rs362272 Major (10) 26 30 35 122 435326 rs362272 Minor (10) 29 50 39 123 435369 rs362272 Major (12) 66 74 65 124 435867 rs362271 Major (10) 73 74 75 125 435892 rs362271 Minor (10) 52 74 79 126 435873 rs3775061 Major (10) 40 32 47 127 435898 rs3775061 Minor (10) 13 20 24 128 435345 rs362310 Major (8) 38 55 52 129 435363 rs362310 Minor (8) 45 67 60 130 435309 rs362310 Major (10) 33 44 56 131 435327 rs362310 Minor (10) 33 71 61 132 435915 rs362307 Major (6) 61 54 58 133 435927 rs362307 Minor (6) 31 35 44 134 435917 rs362307 Major (7) 67 76 66 135 435929 rs362307 Minor (7) 33 34 55 136 435346 rs362307 Major (8) 67 89 66 137 435364 rs362307 Minor (8) 46 72 66 138 435919 rs362307 Major (9) 84 79 70 139 435931 rs362307 Minor (9) 74 74 86 140 435310 rs362307 Major (10) 74 81 71 141 435328 rs362307 Minor (10) 47 69 75 142 435921 rs362307 Major (11) 74 77 69 143 435933 rs362307 Minor (11) 38 47 74 144 435370 rs362307 Major (12) 64 74 38 145 435925 rs362307 Minor (12) 60 66 80 146 435923 rs362307 Major (14) 73 66 71 147 435935 rs362307 Minor (14) 68 75 87 148 435869 rs362306 Major (10) 82 77 81 149 435894 rs362306 Minor (10) 28 79 72 150 435347 rs362303 Major (8) 68 74 71 151 435365 rs362303 Minor (8) 69 83 76 152 435311 rs362303 Major (10) 46 56 72 153 435329 rs362303 Minor (10) 49 62 39 154 435882 rs362296 Major (10) 29 48 56 155 435907 rs362296 Minor (10) 42 56 52 156

Example 3

Dose-Dependent Antisense Inhibition of Human Huntingtin mRNA Levels in Coriell Fibroblast Cell Lines

[0375] Gapmers from the study described in Example 2 were selected and tested at various doses in GM04281, GM02171, and GM02173B cell lines. Each cell line was plated at a density of 25,000 cells per well and transfected using electroporation with 750 nM, 1,500 nM, 3,000 nM, 6,000 nM, and 12,000 nM concentrations of antisense oligonucleotide, as specified in Table 5, 6, and 7. After a treatment period of approximately 16 hours, RNA was isolated from the cells and HTT mRNA levels were measured by quantitative real-time PCR. Human HTT primer probe set RTS2617 was used to measure mRNA levels. HTT mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN. Results are presented as percent inhibition of HTT mRNA, relative to untreated control cells. IC.sub.50 values are also provided in Tables 5, 6, and 7.

TABLE-US-00005 TABLE 5 Dose-dependent antisense inhibition of human HTT in GM04281 cells ISIS 12,000 IC.sub.50 No. 750 nM 1,500 nM 3,000 nM 6,000 nM nM (.mu.M) 387916 51 81 80 91 97 0.6 435330 24 49 50 73 85 2.5 435331 23 38 64 72 74 2.4 435868 3 17 7 29 63 6.7 435870 53 73 77 86 93 0.6 435871 28 51 52 78 89 1.7 435874 14 21 28 64 82 3.3 435879 42 57 57 81 91 1.1 435890 48 56 62 76 91 0.9 435929 10 0 5 12 48 13.8 435931 20 17 53 62 81 2.9 435933 0 7 24 43 49 10.7 435935 0 38 38 62 29 4.2

TABLE-US-00006 TABLE 6 Dose-dependent antisense inhibition of human HTT in GM02171 cells ISIS 12,000 IC.sub.50 No. 750 nM 1,500 nM 3,000 nM 6,000 nM nM (.mu.M) 387916 57 73 81 93 98 0.4 435330 27 37 0 44 63 4.4 435331 35 34 19 41 63 3.5 435868 21 21 39 24 12 >12.0 435870 50 53 57 70 79 0.9 435871 32 46 45 58 62 3.9 435874 1 0 4 11 6 >12.0 435879 32 14 17 45 38 >12.0 435890 34 33 40 51 62 5.4 435929 25 22 31 5 29 >12.0 435931 15 28 27 60 79 3.7 435933 13 36 21 43 48 12.2 435935 25 42 27 61 68 3.2

TABLE-US-00007 TABLE 7 Dose-dependent antisense inhibition of human HTT in GM02173B cells ISIS 12,000 IC.sub.50 No. 750 nM 1,500 nM 3,000 nM 6,000 nM nM (.mu.M) 387916 43 67 80 86 97 1.1 435330 22 21 0 52 62 5.3 435331 19 17 32 50 55 9.4 435868 17 25 41 13 26 >12.0 435870 24 57 70 78 75 1.8 435871 8 30 42 50 48 5.0 435874 31 35 28 35 42 >12.0 435879 39 44 42 60 64 2.5 435890 38 36 50 65 73 3.1 435929 19 17 19 42 35 7.7 435931 40 19 31 48 71 5.8 435933 35 24 47 52 59 4.4 435935 25 23 40 73 77 3.7

Example 4

Dose-Dependent Antisense Inhibition of Human Huntingtin mRNA Levels in Coriell Fibroblast Cell Lines

[0376] Gapmers from the study described in Example 2 were selected and tested at various doses in GM04281, GM02171, and GM02173B cell lines. Each cell line was plated at a density of 25,000 cells per well and transfected using electroporation with 750 nM, 1,500 nM, 3,000 nM, 6,000 nM, and 12,000 nM concentrations of antisense oligonucleotide, as specified in Table 8, 9, and 10. After a treatment period of approximately 16 hours, RNA was isolated from the cells and HTT mRNA levels were measured by quantitative real-time PCR. Human HTT primer probe set RTS2617 was used to measure mRNA levels. HTT mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN. Results are presented as percent inhibition of HTT mRNA relative to untreated control cells. IC.sub.50 values are also provided in Tables 8, 9, and 10.

TABLE-US-00008 TABLE 8 Dose-dependent antisense inhibition of human HTT in GM04281 cells 12,000 IC.sub.50 ISIS No. 750 nM 1,500 nM 3,000 nM 6,000 nM nM (.mu.M) 387916 61 78 90 94 97 <0.8 435303 33 39 69 79 91 1.5 435328 0 12 16 51 75 5.3 435331 27 48 48 70 82 2.1 435339 46 37 61 73 89 2.3 435869 17 35 44 66 80 3.3 435870 44 60 64 84 84 1.1 435871 41 50 71 78 87 1.2 435874 24 36 35 65 73 3.1 435879 46 52 78 81 92 0.9 435890 41 53 63 80 86 1.3 435925 0 14 39 60 87 4.2 435926 20 28 67 81 89 2.0 435928 32 49 73 86 86 1.8 435931 22 24 40 59 90 3.8

TABLE-US-00009 TABLE 9 Dose-dependent antisense inhibition of human HTT in GM02171 cells 12,000 IC.sub.50 ISIS No. 750 nM 1,500 nM 3,000 nM 6,000 nM nM (.mu.M) 387916 50 64 90 95 96 0.7 435303 14 32 68 79 85 2.8 435328 0 12 20 38 55 10.3 435331 0 13 5 30 36 >12.0 435339 30 40 58 63 49 2.5 435869 13 25 31 47 87 4.0 435870 18 31 44 66 74 3.5 435871 1 20 29 49 64 6.5 435874 3 6 12 17 31 >12.0 435879 0 2 12 35 44 >12.0 435890 15 16 30 48 72 5.8 435925 0 0 22 48 29 6.3 435926 25 28 58 74 85 2.3 435928 18 53 61 86 83 2.5 435931 0 4 25 46 68 6.7

TABLE-US-00010 TABLE 10 Dose-dependent antisense inhibition of human HTT in GM02173B cells 12,000 IC.sub.50 ISIS No. 750 nM 1,500 nM 3,000 nM 6,000 nM nM (.mu.M) 387916 27 65 84 81 96 1.9 435303 23 48 52 76 76 2.9 435328 8 14 19 34 50 15.7 435331 10 17 16 27 32 >12.0 435339 28 26 38 67 82 3.8 435869 12 24 37 45 79 4.2 435870 20 26 58 53 78 2.7 435871 15 16 32 45 71 6.0 435874 13 8 28 36 31 >12.0 435879 22 20 36 53 60 6.0 435890 21 28 34 54 71 4.3 435925 2 10 28 43 78 5.9 435926 7 25 37 73 79 3.5 435928 15 39 60 73 87 2.5 435931 13 13 32 61 62 6.7

Example 5

Antisense Inhibition of Human HTT in GM04281 Cells

[0377] Additional antisense oligonucleotides were designed based on the gapmers selected from studies described in Example 4. These oligonucleotides were designed by creating gapmers shifted slightly upstream and downstream (i.e. "microwalk") of the original gapmers from Tables 8, 9, and 10. Antisense oligonucleotides were also created with uniform MOE, as well as with various motifs, 2-9-6 MOE, 3-9-3 MOE, 3-9-4 MOE, 3-9-5 MOE, 4-10-5 MOE, 4-11-4 MOE, 4-7-4 MOE, 4-9-4 MOE, 4-9-5 MOE, 5-10-4 MOE, 5-7-5 MOE, 5-8-6 MOE, 5-9-3 MOE, 5-9-5 MOE, 6-7-6 MOE, 6-9-2 MOE, and 6-8-5 MOE.

[0378] In addition, antisense oligonucleotides were designed targeting SNP RS Nos. rs2857936, rs12506200, rs762855, and rs1006798 (refer to Table 2). The oligonucleotides were designed targeting either the major allele or the minor allele, and with the SNP position opposite either position 8 or position 10 of the gapmer.

[0379] These gapmers were tested in vitro. Cultured GM04281 cells at a density of 25,000 cells per well were transfected using electroporation with 10,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and HTT mRNA levels were measured by quantitative real-time PCR. HTT mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN. Results are presented in Tables 11-19 as percent inhibition of HTT mRNA, relative to untreated control cells.

[0380] The gapmers, ISIS 435869, ISIS 435870, ISIS 435874, ISIS 435879, and ISIS 435890, from which some of the newly designed gapmers were derived are marked with an asterisk (*) in the table. ISIS 387916 was included in the study as a benchmark oligonucleotide against which the potency of the antisense oligonucleotides targeting nucleotides overlapping each SNP position could be compared.

[0381] The uniform MOE oligonucleotides are 15 nucleotides in length.

[0382] The 2-9-6 gapmers are 17 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on the 5' direction by a wing comprising 2 nucleotides and on the 3' direction by a wing comprising 6 nucleotides.

[0383] The 3-9-3 gapmers are 15 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on both 5' and 3' directions by wings comprising 3 nucleotides each.

[0384] The 3-9-4 gapmers are 16 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on the 5' direction by a wing comprising 3 nucleotides and on the 3' direction by a wing comprising 4 nucleotides.

[0385] The 3-9-5 gapmers are 17 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on the 5' direction by a wing comprising 3 nucleotides and on the 3' direction by a wing comprising 5 nucleotides.

[0386] The 4-10-5 gapmers are 19 nucleotides in length, wherein the central gap segment is comprised of ten 2'-deoxynucleotides and is flanked on the 5' direction by a wing comprising 4 nucleotides and on the 3' direction by a wing comprising 5 nucleotides.

[0387] The 4-11-4 gapmers are 19 nucleotides in length, wherein the central gap segment is comprised of eleven 2'-deoxynucleotides and is flanked on both 5' and 3' directions by wings comprising 4 nucleotides each.

[0388] The 4-7-4 gapmers are 15 nucleotides in length, wherein the central gap segment is comprised of seven 2'-deoxynucleotides and is flanked on both 5' and 3' directions by wings comprising 4 nucleotides each.

[0389] The 4-9-4 gapmers are 17 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on both 5' and 3' directions by wings comprising 4 nucleotides each.

[0390] The 4-9-5 gapmers are 18 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on the 5' direction by a wing comprising 4 nucleotides and on the 3' direction by a wing comprising 5 nucleotides.

[0391] The 5-10-4 gapmers are 19 nucleotides in length, wherein the central gap segment is comprised of ten 2'-deoxynucleotides and is flanked on the 5' direction by a wing comprising 5 nucleotides and on the 3' direction by a wing comprising 4 nucleotides.

[0392] The 5-7-5 gapmers are 17 nucleotides in length, wherein the central gap segment is comprised of seven 2'-deoxynucleotides and is flanked on both 5' and 3' directions by wings comprising 5 nucleotides each.

[0393] The 5-8-6 gapmers are 19 nucleotides in length, wherein the central gap segment is comprised of eight 2'-deoxynucleotides and is flanked on the 5' direction by a wing comprising 5 nucleotides and on the 3' direction by a wing comprising 6 nucleotides.

[0394] The 5-9-3 gapmers are 17 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on the 5' direction by a wing comprising 5 nucleotides and on the 3' direction by a wing comprising 3 nucleotides.

[0395] The 5-9-5 gapmers are 19 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on both 5' and 3' directions by wings comprising 5 nucleotides each.

[0396] The 6-7-6 gapmers are 19 nucleotides in length, wherein the central gap segment is comprised of seven 2'-deoxynucleotides and is flanked on both 5' and 3' directions by wings comprising 6 nucleotides each.

[0397] The 6-9-2 gapmers are 17 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on the 5' direction by a wing comprising 6 nucleotides and on the 3' direction by a wing comprising 2 nucleotides.

[0398] The 6-8-5 gapmers are 19 nucleotides in length, wherein the central gap segment is comprised of eight 2'-deoxynucleotides and is flanked on the 5' direction by a wing comprising 6 nucleotides and on the 3' direction by a wing comprising 5 nucleotides.

[0399] For each of the motifs, each nucleotide in the 5' wing segment and each nucleotide in the 3' wing segment has a 2'-MOE modification. The internucleoside linkages throughout each gapmer are phosphorothioate (P.dbd.S) linkages. All cytosine nucleobases throughout each gapmer are 5-methylcytosines.

[0400] The oligonucleotides are organized in tables according to the SNP they target. "Start site" indicates the 5'-most nucleotide to which the gapmer is targeted. "Stop site" indicates the 3'-most nucleotide to which the gapmer is targeted. `Target allele` indicates whether the gapmer is targeted to the major or the minor allele. The number in parentheses indicates the position on the oligonucleotide opposite to the SNP position.

TABLE-US-00011 TABLE 11 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs2857936 (nucleobases 1952 to 1972 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAA 5-10-5 98 6 G 1952 1970 Minor (8) 459908 GCTTTTCATTGAAAAGAAA 5-9-5 26 157 1952 1970 Major (8) 459916 GCTTTTCGTTGAAAAGAAA 5-9-5 8 158 1954 1972 Minor (10) 459904 CTGCTTTTCATTGAAAAGA 5-9-5 23 159 1954 1972 Major (10) 459912 CTGCTTTTCGTTGAAAAGA 5-9-5 8 160

TABLE-US-00012 TABLE 12 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs12506200 (nucleobases 3695 to 3715 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAA 5-10-5 98 6 G 3695 3713 Major (8) 459909 ACTAGGCCGGGCATGCTGG 5-9-5 48 161 3695 3713 Minor (8) 459917 ACTAGGCTGGGCATGCTGG 5-9-5 35 162 3697 3715 Major (10) 459905 AGACTAGGCCGGGCATGCT 5-9-5 33 163 3697 3715 Minor (10) 459913 AGACTAGGCTGGGCATGCT 5-9-5 45 164

TABLE-US-00013 TABLE 13 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs762855 (nucleobases 14437 to 14457 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAA 5-10-5 98 6 G 14437 14455 Minor (8) 459910 AAACAGCTGTTAGTTCCCA 5-9-5 27 165 14437 14455 Major (8) 459918 AAACAGCCGTTAGTTCCCA 5-9-5 39 166 14439 14457 Minor (10) 459906 AGAAACAGCTGTTAGTTCC 5-9-5 24 167 14439 14457 Major (10) 459914 AGAAACAGCCGTTAGTTCC 5-9-5 28 168

TABLE-US-00014 TABLE 14 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs4690072 (nucleobases 62147 to 62173 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAAG 5-10-5 98 6 62147 62165 Major (6) 460145 GTGCTACCCAACCTTTCTG 5-9-5 62 169 62148 62166 Major (7) 460144 AGTGCTACCCAACCTTTCT 5-9-5 61 170 62149 62167 Major (8) 460143 CAGTGCTACCCAACCTTTC 5-9-5 65 171 62150 62168 Major (9) 460142 ACAGTGCTACCCAACCTTT 5-9-5 83 172 62151 62169 Major (10) *435874 CACAGTGCTACCCAACCTT 5-9-5 76 28 62151 62169 Major (10) 460022 CACAGTGCTACCCAACCTT 4-10-5 75 28 62151 62169 Major (10) 460033 CACAGTGCTACCCAACCTT 4-11-4 89 28 62151 62168 Major (9) 460063 ACAGTGCTACCCAACCTT 4-9-5 77 173 62151 62169 Major (10) 460073 CACAGTGCTACCCAACCTT 5-10-4 86 28 62151 62169 Major (10) 460093 CACAGTGCTACCCAACCTT 5-8-6 61 28 62151 62169 Major (10) 460169 CACAGTGCTACCCAACCTT 6-7-6 16 28 62151 62169 Major (10) 460188 CACAGTGCTACCCAACCTT 6-8-5 53 28 62152 62168 Major (9) 459978 ACAGTGCTACCCAACCT 2-9-6 87 174 62152 62167 Major (8) 459999 CAGTGCTACCCAACCT 3-9-4 48 175 62152 62168 Major (9) 460012 ACAGTGCTACCCAACCT 3-9-5 84 174 62152 62168 Major (9) 460052 ACAGTGCTACCCAACCT 4-9-4 51 174 62152 62168 Major (9) 460083 ACAGTGCTACCCAACCT 5-7-5 37 174 62152 62168 Major (9) 460103 ACAGTGCTACCCAACCT 5-9-3 80 174 62152 62170 Major (11) 460137 TCACAGTGCTACCCAACCT 5-9-5 65 176 62152 62168 Major (9) 460179 ACAGTGCTACCCAACCT 6-9-2 67 174 62153 62167 Major (8) 459989 CAGTGCTACCCAACC 3-9-3 60 177 62153 62167 Major (8) 460043 CAGTGCTACCCAACC 4-7-4 24 177 62153 62171 Major (12) 460138 ATCACAGTGCTACCCAACC 5-9-5 76 178 62154 62172 Major (13) 460139 TATCACAGTGCTACCCAAC 5-9-5 68 179 62155 62173 Major (14) 460140 ATATCACAGTGCTACCCAA 5-9-5 79 180

TABLE-US-00015 TABLE 15 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs2298969 (nucleobases 125883 to 125911 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAAG 5-10-5 98 6 125883 125901 Minor (5) 460166 ATGCTGACTTGGGCCATTC 5-9-5 83 181 125884 125902 Minor (6) 460165 GATGCTGACTTGGGCCATT 5-9-5 88 182 125885 125903 Minor (7) 460164 GGATGCTGACTTGGGCCAT 5-9-5 68 183 125886 125904 Minor (8) 460163 GGGATGCTGACTTGGGCCA 5-9-5 73 184 125887 125905 Minor (9) 460162 AGGGATGCTGACTTGGGCC 5-9-5 88 185 125888 125906 Minor (10) *435890 AAGGGATGCTGACTTGGGC 5-9-5 83 94 125888 125906 Minor (10) 460026 AAGGGATGCTGACTTGGGC 4-10-5 90 94 125888 125906 Minor (10) 460037 AAGGGATGCTGACTTGGGC 4-11-4 86 94 125888 125905 Minor (9) 460068 AGGGATGCTGACTTGGGC 4-9-5 90 186 125888 125906 Minor (10) 460076 AAGGGATGCTGACTTGGGC 5-10-4 90 94 125888 125906 Minor (10) 460096 AAGGGATGCTGACTTGGGC 5-8-6 88 94 125888 125906 Minor (10) 460171 AAGGGATGCTGACTTGGGC 6-7-6 87 94 125888 125906 Minor (10) 460190 AAGGGATGCTGACTTGGGC 6-8-5 69 94 125889 125905 Minor (9) 459983 AGGGATGCTGACTTGGG 2-9-6 80 187 125889 125904 Minor (8) 460005 GGGATGCTGACTTGGG 3-9-4 80 284 125889 125905 Minor (9) 460016 AGGGATGCTGACTTGGG 3-9-5 90 187 125889 125905 Minor (9) 460057 AGGGATGCTGACTTGGG 4-9-4 86 187 125889 125905 Minor (9) 460087 AGGGATGCTGACTTGGG 5-7-5 86 187 125889 125905 Minor (9) 460107 AGGGATGCTGACTTGGG 5-9-3 79 187 125889 125907 Major (11) 460157 CAAGGGATGCTGACTTGGG 5-9-5 88 188 125889 125905 Minor (9) 460181 AGGGATGCTGACTTGGG 6-9-2 62 187 125890 125904 Minor (8) 459972 GGGATGCTGACTTGG Uniform 18 189 125890 125904 Minor (8) 459992 GGGATGCTGACTTGG 3-9-3 90 189 125890 125904 Minor (8) 460046 GGGATGCTGACTTGG 4-7-4 59 189 125890 125908 Major (12) 460158 CCAAGGGATGCTGACTTGG 5-9-5 79 190 125891 125909 Major (13) 460159 GCCAAGGGATGCTGACTTG 5-9-5 82 191 125892 125910 Major (14) 460160 TGCCAAGGGATGCTGACTT 5-9-5 87 192 125893 125911 Major (15) 460161 CTGCCAAGGGATGCTGACT 5-9-5 78 193

TABLE-US-00016 TABLE 16 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs7685686 (nucleobases 146781 to 146809 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAAG 5-10-5 98 6 146781 146799 Major (5) 460156 ATTGTCATCACCAGAAAAA 5-9-5 88 194 146782 146800 Major (6) 460155 AATTGTCATCACCAGAAAA 5-9-5 89 195 146783 146801 Major (7) 460154 AAATTGTCATCACCAGAAA 5-9-5 89 196 146784 146802 Major (8) 460153 TAAATTGTCATCACCAGAA 5-9-5 93 197 146785 146803 Major (9) 460152 ATAAATTGTCATCACCAGA 5-9-5 95 198 146786 146804 Major (10) *435879 AATAAATTGTCATCACCAG 5-9-5 94 99 146786 146804 Major (10) 460024 AATAAATTGTCATCACCAG 4-10-5 88 99 146786 146804 Major (10) 460035 AATAAATTGTCATCACCAG 4-11-4 91 99 146786 146803 Major (9) 460065 ATAAATTGTCATCACCAG 4-9-5 96 199 146786 146804 Major (10) 460074 AATAAATTGTCATCACCAG 5-10-4 94 99 146786 146804 Major (10) 460095 AATAAATTGTCATCACCAG 5-8-6 92 99 146786 146804 Major (10) 460170 AATAAATTGTCATCACCAG 6-7-6 91 99 146786 146804 Major (10) 460189 AATAAATTGTCATCACCAG 6-8-5 94 99 146787 146803 Major (9) 459981 ATAAATTGTCATCACCA 2-9-6 85 200 146787 146802 Major (8) 460002 TAAATTGTCATCACCA 3-9-4 86 201 146787 146803 Major (9) 460014 ATAAATTGTCATCACCA 3-9-5 91 200 146787 146803 Major (9) 460055 ATAAATTGTCATCACCA 4-9-4 90 200 146787 146803 Major (9) 460085 ATAAATTGTCATCACCA 5-7-5 94 200 146787 146803 Major (9) 460104 ATAAATTGTCATCACCA 5-9-3 93 200 146787 146805 Major (11) 460147 TAATAAATTGTCATCACCA 5-9-5 91 202 146787 146803 Major (9) 460180 ATAAATTGTCATCACCA 6-9-2 91 200 146788 146802 Major (8) 459970 TAAATTGTCATCACC Uniform 9 203 146788 146802 Major (8) 459990 TAAATTGTCATCACC 3-9-3 67 203 146788 146802 Major (8) 460045 TAAATTGTCATCACC 4-7-4 84 203 146788 146806 Major (12) 460148 TTAATAAATTGTCATCACC 5-9-5 88 204 146789 146807 Major (13) 460149 ATTAATAAATTGTCATCAC 5-9-5 32 205 146790 146808 Major (14) 460150 TATTAATAAATTGTCATCA 5-9-5 29 206 146791 146809 Major (15) 460151 CTATTAATAAATTGTCATC 5-9-5 33 207

TABLE-US-00017 TABLE 17 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs362331 (nucleobases 155474 to 155502 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAAG 5-10-5 98 6 155474 155492 Major (5) 460136 CAGTAGATGAGGGAGCAGG 5-9-5 81 208 155475 155493 Major (6) 460135 ACAGTAGATGAGGGAGCAG 5-9-5 84 209 155476 155494 Major (7) 460134 CACAGTAGATGAGGGAGCA 5-9-5 87 210 155477 155495 Major (8) 460133 ACACAGTAGATGAGGGAGC 5-9-5 85 211 155478 155496 Major (9) 460132 CACACAGTAGATGAGGGAG 5-9-5 86 212 155479 155497 Major (10) *435870 GCACACAGTAGATGAGGGA 5-9-5 91 103 155479 155497 Major (10) 460019 GCACACAGTAGATGAGGGA 4-10-5 92 103 155479 155497 Major (10) 460031 GCACACAGTAGATGAGGGA 4-11-4 95 103 155479 155496 Major (9) 460061 CACACAGTAGATGAGGGA 4-9-5 87 213 155479 155497 Major (10) 460071 GCACACAGTAGATGAGGGA 5-10-4 94 103 155479 155497 Major (10) 460090 GCACACAGTAGATGAGGGA 5-8-6 86 103 155479 155497 Major (10) 460168 GCACACAGTAGATGAGGGA 6-7-6 84 103 155479 155497 Major (10) 460187 GCACACAGTAGATGAGGGA 6-8-5 89 103 155480 155496 Major (9) 459977 CACACAGTAGATGAGGG 2-9-6 90 214 155480 155495 Major (8) 459996 ACACAGTAGATGAGGG 3-9-4 37 215 155480 155496 Major (9) 460009 CACACAGTAGATGAGGG 3-9-5 90 214 155480 155496 Major (9) 460051 CACACAGTAGATGAGGG 4-9-4 73 214 155480 155496 Major (9) 460081 CACACAGTAGATGAGGG 5-7-5 77 214 155480 155496 Major (9) 460101 CACACAGTAGATGAGGG 5-9-3 84 214 155480 155498 Major (11) 460127 TGCACACAGTAGATGAGGG 5-9-5 89 216 155480 155496 Major (9) 460178 CACACAGTAGATGAGGG 6-9-2 92 214 155481 155495 Major (8) 459967 ACACAGTAGATGAGG Uniform 81 217 155481 155495 Major (8) 459987 ACACAGTAGATGAGG 3-9-3 18 217 155481 155495 Major (8) 460041 ACACAGTAGATGAGG 4-7-4 54 217 155481 155499 Major (12) 460128 GTGCACACAGTAGATGAGG 5-9-5 73 218 155482 155500 Major (13) 460129 AGTGCACACAGTAGATGAG 5-9-5 86 219 155483 155501 Major (14) 460130 AAGTGCACACAGTAGATGA 5-9-5 60 220 155484 155502 Major (15) 460131 GAAGTGCACACAGTAGATG 5-9-5 73 221

TABLE-US-00018 TABLE 18 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs362306 (nucleobases 181739 to 181767 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAAG 5-10-5 98 6 181739 181757 Major (5) 460126 GCTGCAACCTGGCAACAAC 5-9-5 87 222 181740 181758 Major (6) 460125 AGCTGCAACCTGGCAACAA 5-9-5 70 223 181741 181759 Major (7) 460123 CAGCTGCAACCTGGCAACA 5-9-5 83 224 181742 181760 Major (8) 460121 GCAGCTGCAACCTGGCAAC 5-9-5 47 225 181743 181761 Major (9) 460118 AGCAGCTGCAACCTGGCAA 5-9-5 75 226 181744 181762 Major (10) *435869 GAGCAGCTGCAACCTGGCA 5-9-5 91 149 181744 181762 Major (10) 460018 GAGCAGCTGCAACCTGGCA 4-10-5 86 149 181744 181762 Major (10) 460028 GAGCAGCTGCAACCTGGCA 4-11-4 89 149 181744 181761 Major (9) 460058 AGCAGCTGCAACCTGGCA 4-9-5 85 227 181744 181762 Major (10) 460069 GAGCAGCTGCAACCTGGCA 5-10-4 91 149 181744 181762 Major (10) 460089 GAGCAGCTGCAACCTGGCA 5-8-6 54 149 181744 181762 Major (10) 460167 GAGCAGCTGCAACCTGGCA 6-7-6 85 149 181744 181762 Major (10) 460186 GAGCAGCTGCAACCTGGCA 6-8-5 84 149 181745 181761 Major (9) 459975 AGCAGCTGCAACCTGGC 2-9-6 86 228 181745 181760 Major (8) 459995 GCAGCTGCAACCTGGC 3-9-4 87 229 181745 181761 Major (9) 460008 AGCAGCTGCAACCTGGC 3-9-5 83 228 181745 181761 Major (9) 460049 AGCAGCTGCAACCTGGC 4-9-4 88 228 181745 181761 Major (9) 460079 AGCAGCTGCAACCTGGC 5-7-5 46 228 181745 181761 Major (9) 460099 AGCAGCTGCAACCTGGC 5-9-3 44 228 181745 181763 Major (11) 460108 AGAGCAGCTGCAACCTGGC 5-9-5 50 230 181745 181761 Major (9) 460177 AGCAGCTGCAACCTGGC 6-9-2 67 228 181746 181760 Major (8) 459966 GCAGCTGCAACCTGG Uniform 26 231 181746 181760 Major (8) 459985 GCAGCTGCAACCTGG 3-9-3 69 231 181746 181760 Major (8) 460039 GCAGCTGCAACCTGG 4-7-4 56 231 181746 181764 Major (12) 460110 AAGAGCAGCTGCAACCTGG 5-9-5 75 232 181747 181765 Major (13) 460113 CAAGAGCAGCTGCAACCTG 5-9-5 36 233 181748 181766 Major (14) 460115 GCAAGAGCAGCTGCAACCT 5-9-5 78 234 181749 181767 Major (15) 460117 TGCAAGAGCAGCTGCAACC 5-9-5 73 235

TABLE-US-00019 TABLE 19 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs1006798 (nucleobases 198015 to 198035 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAAG 5-10-5 98 6 198015 198033 Minor (8) 459911 ACCATGATATCTCCAGCAC 5-9-5 33 236 198015 198033 Minor (8) 459919 ACCATGACATCTCCAGCAC 5-9-5 26 237 198017 198035 Major (10) 459907 CCACCATGATATCTCCAGC 5-9-5 32 238 198017 198035 Minor (10) 459915 CCACCATGACATCTCCAGC 5-9-5 51 239

Example 6

Dose-Dependent Antisense Inhibition of Human Huntingtin mRNA Levels in Coriell Fibroblast Cell Lines

[0401] Gapmers from the studies described in Example 5 were selected and tested at various doses in GM04281, GM02171, and GM02173B cell lines. Each cell line was plated at a density of 25,000 cells per well and transfected using electroporation with 750 nM, 1,500 nM, 3,000 nM, 6,000 nM, and 12,000 nM concentrations of antisense oligonucleotide, as specified in Tables 20, 21, and 22. After a treatment period of approximately 16 hours, RNA was isolated from the cells and HTT mRNA levels were measured by quantitative real-time PCR. Human HTT primer probe set RTS2617 was used to measure mRNA levels. HTT mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN. Results are presented as percent inhibition of HTT mRNA, relative to untreated control cells. IC.sub.50 values are also provided in Tables 20, 21, and 22.

TABLE-US-00020 TABLE 20 Dose-dependent antisense inhibition of human HTT in GM04281 cells ISIS 12,000 IC.sub.50 No. 750 nM 1,500 nM 3,000 nM 6,000 nM nM (.mu.M) 387916 56 81 89 96 98 0.6 435869 38 49 66 86 91 1.4 435874 33 27 37 49 62 8.4 435879 42 55 73 86 96 1.1 435890 39 51 74 83 89 1.3 459978 29 33 51 69 86 2.5 459992 14 27 51 54 84 3.2 460012 15 24 54 70 81 3.1 460016 3 36 48 71 77 3.3 460019 54 59 74 87 94 0.7 460026 48 47 71 79 88 0.8 460028 39 38 73 77 87 1.4 460031 44 62 72 87 92 0.9 460033 11 38 52 64 87 3.0 460065 43 54 74 89 96 1.1 460068 47 28 63 76 90 2.6 460069 38 50 65 77 91 1.4 460071 53 61 80 89 93 0.6 460073 16 39 42 58 75 4.0 460076 26 47 54 70 86 2.1 460085 48 60 79 89 94 0.8 460140 6 24 44 44 64 6.6 460142 2 38 46 46 68 4.8 460152 35 61 76 92 94 1.2 460157 51 36 53 74 89 2.6 460162 64 41 71 76 85 2.1 460165 41 50 56 76 84 1.5

TABLE-US-00021 TABLE 21 Dose-dependent antisense inhibition of human HTT in GM02171 cells ISIS 12,000 IC.sub.50 No. 750 nM 1,500 nM 3,000 nM 6,000 nM nM (.mu.M) 387916 53 66 88 96 98 0.7 435869 4 20 36 63 86 3.9 435870 25 39 48 62 83 2.8 435874 12 20 18 27 37 >12.0 435879 10 7 11 42 51 10.6 435890 10 23 29 29 55 9.2 459978 15 7 6 29 52 12.7 459992 11 19 26 39 62 8.7 460012 3 3 10 19 41 >12.0 460016 0 14 12 22 48 >12.0 460019 27 21 41 60 73 4.4 460026 9 25 30 46 58 7.8 460028 24 8 32 54 77 5.3 460031 8 25 42 60 83 3.8 460033 11 25 30 40 75 4.1 460065 11 16 11 31 53 10.3 460068 15 13 39 44 53 8.8 460069 17 28 37 60 79 3.9 460071 16 36 58 70 88 2.6 460073 5 19 24 33 56 8.7 460076 19 29 44 54 83 3.3 460085 10 15 17 28 31 >12.0 460140 8 22 22 28 47 >12.0 460142 11 24 28 36 38 >12.0 460152 14 21 8 25 44 22 460157 22 21 29 44 66 6.7 460162 24 55 52 62 82 2.8 460165 14 34 50 69 81 3.1

TABLE-US-00022 TABLE 22 Dose-dependent antisense inhibition of human HTT in GM02173B cells ISIS 12,000 IC.sub.50 No. 750 nM 1,500 nM 3,000 nM 6,000 nM nM (.mu.M) 387916 37 63 86 88 98 1.0 435869 10 20 43 70 85 3.5 435870 24 24 56 72 87 2.3 435874 0 11 12 30 44 >12.0 435879 4 17 43 64 74 4.3 435890 31 29 54 57 69 4.4 459978 7 13 17 35 64 8.4 459992 18 15 30 51 71 5.7 460012 0 10 24 37 72 7.1 460016 15 5 30 38 59 9.5 460019 10 32 51 65 87 3.1 460026 0 34 21 55 65 6.4 460028 0 14 31 51 77 5.2 460031 0 31 53 71 88 3.2 460033 11 8 6 52 84 5.0 460065 19 37 53 58 74 3.6 460068 17 11 31 59 69 5.5 460069 11 21 37 55 75 4.6 460071 6 42 61 83 88 2.6 460073 7 13 19 49 66 6.3 460076 27 31 49 43 81 2.9 460085 17 34 51 54 68 4.4 460140 0 2 28 18 46 >12.0 460142 2 32 37 42 59 7.6 460152 17 32 35 51 66 5.5 460157 9 34 38 52 74 4.5 460162 22 45 57 65 79 2.5 460165 5 45 52 72 84 3.2

Example 7

Antisense Inhibition of Human HTT in GM04281 Cells and GM02171 Cells

[0402] Additional antisense oligonucleotides were designed based on the gapmers selected from studies described in Example 2. These oligonucleotides were designed by creating gapmers shifted slightly upstream and downstream (i.e. "microwalk") of the original gapmers from Table 4.

[0403] The gapmers were tested in the GM04281 and the GM02171 cell lines. Cultured GM04281 or GM02171 cells at a density of 25,000 cells per well were transfected using electroporation with 10,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and HTT mRNA levels were measured by quantitative real-time PCR using primer probe set RTS2617. HTT mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN. Results are presented as percent inhibition of HTT mRNA, relative to untreated control cells.

[0404] The gapmers, from which the newly designed oligonucleotides were derived, were also included in the assay. These parent gapmers, ISIS 435294, ISIS 435295, ISIS 435301, ISIS 435303, ISIS 435304, ISIS 435305, ISIS 435308, ISIS 435330, ISIS 435331, ISIS 435337, ISIS 435339, ISIS 435340, ISIS 435341, ISIS 435344, ISIS 435862, ISIS 435863, ISIS 435864, ISIS 435866, ISIS 435867, ISIS 435868, ISIS 435871, ISIS 435873, ISIS 435875, ISIS 435876, ISIS 435878, ISIS 435880, ISIS 435881, ISIS 435882, ISIS 435884, ISIS 435890, and ISIS 435897 are marked with an asterisk (*) in the table. ISIS 387916 was included in the study as a benchmark oligonucleotide against which the potency of the antisense oligonucleotides targeting nucleotides overlapping each SNP position could be compared.

[0405] The chimeric antisense oligonucleotides in Tables 23-48 were designed as 5-9-5 MOE gapmers. The 5-9-5 gapmers are 19 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on both 5' and 3' directions by wings comprising 5 nucleotides each. Each nucleotide in the 5' wing segment and each nucleotide in the 3' wing segment has a 2'-MOE modification. The internucleoside linkages throughout each gapmer are phosphorothioate (P.dbd.S) linkages. All cytosine nucleobases throughout each gapmer are 5-methylcytosines.

[0406] The gapmers are organized in Tables 23-48, according to the SNP site they target. "Start site" indicates the 5'-most nucleotide to which the gapmer is targeted. "Stop site" indicates the 3'-most nucleotide to which the gapmer is targeted. `Target allele` indicates whether the gapmer is targeted to the major or the minor allele. The number in parentheses indicates the position on the oligonucleotide opposite to the SNP position.

TABLE-US-00023 TABLE 23 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs3856973 (nucleobases 19815 to 19835 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 19815 19833 *435330 Major (8) TAACACTCGATTAACCCTG 88 31 8 19816 19834 476441 Major (9) TTAACACTCGATTAACCCT 88 0 240 19817 19835 *435294 Major (10) GTTAACACTCGATTAACCC 72 30 10

TABLE-US-00024 TABLE 24 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs2285086 (nucleobases 28901 to 28921 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 28901 28919 463570 Major (8) TAGTTCATCCCAGTGAGAA 66 12 241 28902 28920 463573 Major (9) CTAGTTCATCCCAGTGAGA 66 36 242 28903 28921 *435864 Major (10) GCTAGTTCATCCCAGTGAG 40 18 12

TABLE-US-00025 TABLE 25 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs7659144 (nucleobases 37963 to 37983 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 37963 37981 476462 Major (8) GAAATGGGTTTTTCCACAT 38 0 243 37964 37982 476439 Major (9) GGAAATGGGTTTTTCCACA 80 45 244 37965 37983 *435878 Major (10) TGGAAATGGGTTTTTCCAC 76 3 14

TABLE-US-00026 TABLE 26 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs16843804 (nucleobases 44032 to 44052 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 44032 44050 476471 Major (8) TAACCGTGGCATGGGCAGT 82 53 245 44033 44051 476452 Major (9) TTAACCGTGGCATGGGCAG 84 44 246 44034 44052 *435863 Major (10) TTTAACCGTGGCATGGGCA 89 89 16

TABLE-US-00027 TABLE 27 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs2024115 (nucleobases 44210 to 44230 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 44210 44228 *435331 Major (8) TTCAAGCTAGTAACGATGC 84 20 18 44211 44229 476447 Major (9) CTTCAAGCTAGTAACGATG 87 57 247 44212 44230 *435295 Major (10) ACTTCAAGCTAGTAACGAT 85 67 20

TABLE-US-00028 TABLE 28 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs10015979 (nucleobases 49084 to 49104 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 49084 49102 476470 Major (8) AGCTAGGTTAAAGAGTCAC 55 74 248 49085 49103 476450 Major (9) CAGCTAGGTTAAAGAGTCA 44 5 249 49086 49104 *435862 Major (10) GCAGCTAGGTTAAAGAGTC 56 49 22

TABLE-US-00029 TABLE 29 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs7691627 (nucleobases 51052 to 51072 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 51052 51070 476467 Major (8) TAAGAAACACAATCAAAGA 45 21 250 51053 51071 476445 Major (9) ATAAGAAACACAATCAAAG 34 1 251 51054 51072 *435880 Major (10) AATAAGAAACACAATCAAA 68 7 24

TABLE-US-00030 TABLE 30 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs6446723 (nucleobases 66455 to 66475 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 66455 66473 476463 Major (8) ATTTTCTAGACTTTATGAT 37 7 252 66456 66474 476440 Major (9) AATTTTCTAGACTTTATGA 57 0 253 66457 66475 *435875 Major (10) TAATTTTCTAGACTTTATG 42 0 30

TABLE-US-00031 TABLE 31 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and a chimeric antisense oligonucleotide targeted to SNP rs363064 (nucleobases 81053 to 81071 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 81053 81071 476461 Major (9) GAGAATACGGGTAACATTT 87 62 254

TABLE-US-00032 TABLE 32 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs11731237 (nucleobases 91455 to 91475 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 91455 91473 476468 Major (8) TGGGCAGGAAGGACTGAAC 58 56 255 91456 91474 476448 Major (9) GTGGGCAGGAAGGACTGAA 61 69 256 91457 91475 *435884 Major (10) GGTGGGCAGGAAGGACTGA 59 49 68

TABLE-US-00033 TABLE 33 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs4690073 (nucleobases 99792 to 99812 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 99792 99810 *435337 Major (8) CCTAAATCAATCTACAAGT 69 7 70 99793 99811 476446 Major (9) CCCTAAATCAATCTACAAG 61 0 257 99794 99812 *435301 Major (10) TCCCTAAATCAATCTACAA 63 1 72

TABLE-US-00034 TABLE 34 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs34315806 (nucleobases 101676 to 101696 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 101676 101694 463569 Major (8) CTTTTCCGTGCTGTTCTGA 96 95 258 101677 101695 463572 Major (9) ACTTTTCCGTGCTGTTCTG 93 91 259 101678 101696 463567 Major (10) AACTTTTCCGTGCTGTTCT 98 97 260

TABLE-US-00035 TABLE 35 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs363099 (nucleobases 101698 to 101718 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 101698 101716 *435339 Major (8) CTGAGCGGAGAAACCCTCC 94 85 80 101699 101717 476458 Major (9) GCTGAGCGGAGAAACCCTC 92 79 261 101700 101718 *435303 Major (10) GGCTGAGCGGAGAAACCCT 96 93 82

TABLE-US-00036 TABLE 36 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs363096 (nucleobases 119663 to 119683 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 119663 119681 *435340 Major (8) TTCCCTAAAAACAAAAACA 42 21 85 119664 119682 476451 Major (9) ATTCCCTAAAAACAAAAAC 0 0 262 119665 119683 *435304 Major (10) GATTCCCTAAAAACAAAAA 41 27 87

TABLE-US-00037 TABLE 37 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs2298967 (nucleobases 125389 to 125409 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 125389 125407 *435341 Major (8) CTTTTCTATTGTCTGTCCC 83 65 89 125390 125408 476459 Major (9) GCTTTTCTATTGTCTGTCC 89 82 263 125391 125409 *435305 Major (10) TGCTTTTCTATTGTCTGTC 92 85 91

TABLE-US-00038 TABLE 38 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and a chimeric antisense oligonucleotide targeted to SNP rs2298969 (nucleobases 125888 to 125906 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 125888 125906 *435890 Minor (10) AAGGGATGCTGACTTGGGC 91 64 94

TABLE-US-00039 TABLE 39 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs6844859 (nucleobases 130128 to 130148 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 130128 130146 476466 Major (8) CTTCCTCACTGAGGATGAA 87 64 264 130129 130147 476444 Major (9) CCTTCCTCACTGAGGATGA 92 77 265 130130 130148 *435876 Major (10) ACCTTCCTCACTGAGGATG 94 87 95

TABLE-US-00040 TABLE 40 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs363092 (nucleobases 135671 to 135691 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 135671 135689 476464 Major (8) AACCACTTTGGGATGAATA 51 71 266 135672 135690 476442 Major (9) AAACCACTTTGGGATGAAT 58 59 267 135673 135691 *435897 Minor (10) CAAACCACTTTGGGATGAA 48 78 98

TABLE-US-00041 TABLE 41 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs363088 (nucleobases 149972 to 149992 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 149972 149990 476476 Major (8) ACAGCTATCTTCTCATCAA 90 65 268 149973 149991 476460 Major (9) CACAGCTATCTTCTCATCA 86 39 269 149974 149992 *435871 Major (10) TCACAGCTATCTTCTCATC 91 54 101

TABLE-US-00042 TABLE 42 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs916171 (nucleobases 156457 to 156477 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 156457 156475 476465 Major (8) GAACAAAGAGAAGAATTTC 38 0 270 156458 156476 476443 Major (9) AGAACAAAGAGAAGAATTT 58 0 271 156459 156477 *435881 Major (10) CAGAACAAAGAGAAGAATT 59 16 105

TABLE-US-00043 TABLE 43 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs362275 (nucleobases 164244 to 164264 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site NO allele Sequence GM04281 GM02171 No 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 164244 164262 476473 Major (8) GAAGCCTGATAAAATCTCT 83 51 272 164245 164263 476454 Major (9) AGAAGCCTGATAAAATCTC 79 61 273 164246 164264 *435868 Major (10) AAGAAGCCTGATAAAATCT 69 56 111

TABLE-US-00044 TABLE 44 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs362273 (nucleobases 167061 to 167081 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 167061 167079 463568 Major (8) TGATCTGTAGCAGCAGCTT 96 78 274 167062 167080 463571 Major (9) TTGATCTGTAGCAGCAGCT 95 86 275 167063 167081 463566 Major (10) GTTGATCTGTAGCAGCAGC 94 78 276

TABLE-US-00045 TABLE 45 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs362272 (nucleobases 174622 to 174642 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 174622 174640 *435344 Major (8) TAGAGGACGCCGTGCAGGG 78 63 120 174623 174641 476456 Major (9) ATAGAGGACGCCGTGCAGG 87 60 277 174624 174642 *435308 Major (10) CATAGAGGACGCCGTGCAG 76 48 122

TABLE-US-00046 TABLE 46 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs362271 (nucleobases 175160 to 175180 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 175160 175178 476472 Major (8) GTGTGTACAGAACCTGCCG 85 52 278 175161 175179 476453 Major (9) CGTGTGTACAGAACCTGCC 88 69 279 175162 175180 *435867 Major (10) ACGTGTGTACAGAACCTGC 91 80 125

TABLE-US-00047 TABLE 47 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs3775061 (nucleobases 178396 to 178416 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 178396 178414 476475 Major (8) TTCAGAATGCCTCATCTGG 61 1 280 178397 178415 476457 Major (9) GTTCAGAATGCCTCATCTG 80 50 281 178398 178416 *435873 Major (10) TGTTCAGAATGCCTCATCT 80 43 127

TABLE-US-00048 TABLE 48 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs362296 (nucleobases 186649 to 1786669 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Target in in ID Site Site No allele Sequence GM04281 GM02171 NO 145466 145485 387916 n/a TCTCTATTGCACATTCCAAG 100 99 6 186649 186667 476469 Major (8) GGACAGGGTGTGCTCTCCG 80 58 282 186650 186668 476449 Major (9) GGGACAGGGTGTGCTCTCC 80 64 283 186651 186669 *435882 Major (10) GGGGACAGGGTGTGCTCTC 61 61 155

Example 8

Dose-Dependent Antisense Inhibition of Human Huntingtin mRNA Levels in Coriell Fibroblast Cell Lines

[0407] Gapmers from the studies described in Example 7 were selected and tested at various doses in GM04281, GM02171, and GM02173B cell lines. Each cell line was plated at a density of 25,000 cells per well and transfected using electroporation with 750 nM, 1,500 nM, 3,000 nM, 6,000 nM, and 12,000 nM concentrations of antisense oligonucleotide, as specified in Tables 49, 50, and 51. After a treatment period of approximately 16 hours, RNA was isolated from the cells and HTT mRNA levels were measured by quantitative real-time PCR. Human HTT primer probe set RTS2617 was used to measure mRNA levels. HTT mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN. Results are presented as percent inhibition of HTT mRNA, relative to untreated control cells. IC.sub.50 values are also provided in Tables 49, 50, and 51.

TABLE-US-00049 TABLE 49 Dose-dependent antisense inhibition of human HTT in GM04281 cells ISIS IC.sub.50 No. 750 nM 1500 nM 3000 nM 6000 nM 12000 nM (.mu.M) 387916 67 88 95 97 99 <0.8 463566 25 65 79 88 95 1.5 463567 34 73 90 93 98 1.1 463568 33 56 75 87 92 1.3 463571 32 21 70 90 93 1.4 476441 11 27 50 70 87 3.1 476444 20 31 68 49 93 2.3 476449 4 28 34 47 77 4.9 476453 21 21 48 73 85 2.7 476455 5 19 34 56 80 4.6 476458 36 72 83 93 96 1.1 476459 23 59 75 85 91 1.5 476469 17 27 47 47 67 5.5 476473 0 6 32 50 68 6.2 476476 3 7 32 53 86 4.9

TABLE-US-00050 TABLE 50 Dose-dependent antisense inhibition of human HTT in GM02171 cells ISIS 750 1500 3000 6000 12000 IC.sub.50 No. nM nM nM nM nM (.mu.M) 387916 59 79 93 98 98 <0.8 463566 4 33 42 62 79 3.8 463567 38 41 69 85 94 1.5 463568 21 26 41 58 64 4.8 463571 8 23 56 63 75 3.7 476441 0 13 7 0 12 >12.0 476444 11 0 0 67 59 8.8 476449 4 27 37 51 63 5.8 476453 6 40 40 51 73 4.9 476455 32 15 18 47 61 7.8 476458 42 54 71 86 84 1.2 476459 22 38 70 44 73 4.3 476469 7 24 30 56 58 7.8 476473 4 10 15 33 43 >12.0 476476 5 16 18 23 41 >12.0

TABLE-US-00051 TABLE 51 Dose-dependent antisense inhibition of human HTT in GM02171 cells ISIS 750 1500 3000 6000 12000 IC.sub.50 No. nM nM nM nM nM (.mu.M) 387916 66 89 95 97 99 <0.8 463566 32 55 76 77 93 1.3 463567 51 61 87 94 97 0.7 463568 26 23 72 87 94 1.6 463571 32 34 60 86 94 1.9 476441 18 18 27 47 44 >12.0 476444 15 0 31 51 58 7.1 476449 27 33 56 80 81 2.6 476453 24 28 55 75 83 2.7 476455 24 26 52 55 73 3.7 476458 63 77 87 89 94 0.2 476459 37 55 56 62 86 1.5 476469 22 41 40 63 76 2.9 476473 7 28 33 51 73 5.0 476476 11 29 26 55 69 4.6

Example 9

Antisense Inhibition of Human HTT in GM04281 Cells by Oligonucleotides Designed by Microwalk

[0408] Additional gapmers were designed based on the gapmers selected from studies described in Example 4. These gapmers were designed by creating gapmers shifted slightly upstream and downstream (i.e. "microwalk") of the original gapmers from Tables 8, 9, and 10. Gapmers were also created with 3-9-3 or 5-9-5 motifs, and with constrained 6(S)--CH.sub.3-bicyclic nucleic acid (BNA) molecules at various nucleoside positions.

[0409] These gapmers were tested in vitro. Cultured GM04281 cells at a density of 25,000 cells per well were transfected using electroporation with 5,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and HTT mRNA levels were measured by quantitative real-time PCR. HTT mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN. Results are presented as percent inhibition of HTT mRNA, relative to untreated control cells.

[0410] The chimeric antisense oligonucleotides in Tables 52-56 were designed as 3-9-3 or 5-9-5 gapmers. The parent gapmers, ISIS 435869, ISIS 435870, ISIS 435874, ISIS 435879, and ISIS 435890, from which the newly designed gapmers were derived are marked with an asterisk (*) in the table. ISIS 387916 was included in the study as a benchmark oligonucleotide against which the potency of the antisense oligonucleotides targeting nucleotides overlapping each SNP position could be compared.

[0411] The 3-9-3 gapmers are 15 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleosides and is flanked on both 5' and 3' directions by wings comprising 3 sugar modified nucleosides each.

[0412] The 5-9-5 gapmers are 19 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleosides and is flanked on both 5' and 3' directions by wings comprising 5 sugar modified nucleosides each.

[0413] The internucleoside linkages throughout each gapmer are phosphorothioate (P.dbd.S) linkages. All cytosine nucleobases throughout each gapmer are 5-methylcytosines. Bolded and underlined nucleotides in Tables 52-56 indicate the positions of the 6(S)--CH.sub.3-BNA molecules (e.g. cEt molecules) in each gapmer. Italicized nucleotides are MOE subunits.

[0414] "Start site" indicates the 5'-most nucleotide to which the gapmer is targeted. "Stop site" indicates the 3'-most nucleotide to which the gapmer is targeted. `Target allele` indicates whether the gapmer is targeted to the major or the minor allele. The number in parentheses indicates the position on the oligonucleotide opposite to the SNP position.

TABLE-US-00052 TABLE 52 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs4690072 (nucleobases 62147 to 62173 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAAG 5-10-5 97 6 62147 62165 Major (6) 460266 GTGCTACCCAACCTTTCTG 5-9-5 63 169 62151 62169 Major (10) *435874 CACAGTGCTACCCAACCTT 5-9-5 50 28 62151 62169 Major (10) 460213 CACAGTGCTACCCAACCTT 5-9-5 22 28 62151 62169 Major (10) 460220 CACAGTGCTACCCAACCTT 5-9-5 24 28 62151 62169 Major (10) 460221 CACAGTGCTACCCAACCTT 5-9-5 28 28 62153 62167 Major (8) 460208 CAGTGCTACCCAACC 3-9-3 81 177 62155 62173 Major (14) 460267 ATATCACAGTGCTACCCAA 5-9-5 37 180

TABLE-US-00053 TABLE 53 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs2298969 (nucleobases 125884 to 125910 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAAG 5-10-5 97 6 125884 125902 Minor (6) 460233 GATGCTGACTTGGGCCATT 5-9-5 76 182 125888 125906 Minor (10) *435890 AAGGGATGCTGACTTGGGC 5-9-5 75 94 125888 125906 Minor (10) 460215 AAGGGATGCTGACTTGGGC 5-9-5 26 94 125888 125906 Minor (10) 460224 AAGGGATGCTGACTTGGGC 5-9-5 38 94 125888 125906 Minor (10) 460225 AAGGGATGCTGACTTGGGC 5-9-5 49 94 125890 125904 Minor (8) 460210 GGGATGCTGACTTGG 3-9-3 97 189 125892 125910 Minor (14) 460229 TGCCAAGGGATGCTGACTT 5-9-5 60 192

TABLE-US-00054 TABLE 54 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs7685686 (nucleobases 146782 to 146808 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAAG 5-10-5 97 6 146782 146800 Major (6) 460232 AATTGTCATCACCAGAAAA 5-9-5 82 195 146786 146804 Major (10) *435879 AATAAATTGTCATCACCAG 5-9-5 84 99 146786 146804 Major (10) 460214 AATAAATTGTCATCACCAG 5-9-5 33 99 146786 146804 Major (10) 460222 AATAAATTGTCATCACCAG 5-9-5 87 99 146786 146804 Major (10) 460223 AATAAATTGTCATCACCAG 5-9-5 75 99 146788 146802 Major (8) 460209 TAAATTGTCATCACC 3-9-3 96 203 146790 146808 Major (14) 460228 TATTAATAAATTGTCATCA 5-9-5 0 206

TABLE-US-00055 TABLE 55 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs362331 (nucleobases 155475 to 155501 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAAG 5-10-5 97 6 155475 155493 Major (6) 460231 ACAGTAGATGAGGGAGCAG 5-9-5 88 209 155479 155497 Major (10) *435870 GCACACAGTAGATGAGGGA 5-9-5 86 103 155479 155497 Major (10) 460212 GCACACAGTAGATGAGGGA 5-9-5 89 103 155479 155497 Major (10) 460218 GCACACAGTAGATGAGGGA 5-9-5 90 103 155479 155497 Major (10) 460219 GCACACAGTAGATGAGGGA 5-9-5 88 103 155481 155495 Major (8) 460207 ACACAGTAGATGAGG 3-9-3 89 217 155483 155501 Major (14) 460227 AAGTGCACACAGTAGATGA 5-9-5 45 220

TABLE-US-00056 TABLE 56 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs362306 (nucleobases 181740 to 181766 of SEQ ID NO: 1) SEQ Start Stop Target ISIS % ID Site Site allele No. Sequence Motif inhibition NO 145466 145485 n/a 387916 TCTCTATTGCACATTCCAAG 5-10-5 97 6 181740 181758 Major (6) 460230 AGCTGCAACCTGGCAACAA 5-9-5 66 223 181744 181762 Major (10) *435869 GAGCAGCTGCAACCTGGCA 5-9-5 69 149 181744 181762 Major (10) 460211 GAGCAGCTGCAACCTGGCA 5-9-5 22 149 181744 181762 Major (10) 460216 GAGCAGCTGCAACCTGGCA 5-9-5 18 149 181744 181762 Major (10) 460217 GAGCAGCTGCAACCTGGCA 5-9-5 56 149 181746 181760 Major (8) 460206 GCAGCTGCAACCTGG 3-9-3 83 231 181748 181766 Major (14) 460226 GCAAGAGCAGCTGCAACCT 5-9-5 51 234

Example 10

Dose-Dependent Antisense Inhibition of Human Huntingtin mRNA Levels in Coriell Fibroblast Cell Lines

[0415] Gapmers from studies described in Example 9 were selected and tested at various doses in GM04281, GM02171 and GM02173B cell lines. Each cell line was plated at a density of 25,000 cells per well and transfected using electroporation with 312.5 nM, 625 nM, 1,250 nM, 2,500 nM, 5,000 nM and 10,000 nM concentrations of antisense oligonucleotide, as specified in Tables 75, 58, and 59. After a treatment period of approximately 16 hours, RNA was isolated from the cells and HTT mRNA levels were measured by quantitative real-time PCR. Human HTT primer probe set RTS2617 was used to measure mRNA levels. HTT mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN. Results are presented as percent inhibition of HTT mRNA, relative to untreated control cells. IC.sub.50 values are also provided in Tables 57, 58, and 59.

TABLE-US-00057 TABLE 57 Dose-dependent antisense inhibition of human HTT in GM04281 cells ISIS 312.5 625 1,250 2,500 5,000 10,000 IC.sub.50 No. nM nM nM nM nM nM (.mu.M) 387916 26 49 68 86 94 97 0.7 435869 0 0 23 48 62 82 3.2 435870 15 38 50 65 85 88 1.3 435874 14 22 32 49 65 73 2.7 435879 0 17 40 61 83 94 1.8 435890 5 13 37 56 70 82 2.3 460206 10 18 37 52 66 85 2.3 460207 20 27 50 65 80 91 1.4 460208 21 34 51 63 70 79 1.5 460209 52 74 89 94 94 95 0.2 460210 34 61 84 91 97 98 0.5 460212 13 31 50 62 75 82 1.6 460218 14 27 50 63 78 86 1.8 460219 9 32 42 64 77 87 1.6 460222 19 21 42 57 73 78 1.7 460231 12 24 41 57 71 84 1.9 460233 16 28 59 66 72 74 1.8 460266 4 17 32 48 60 75 2.9

TABLE-US-00058 TABLE 58 Dose-dependent antisense inhibition of human HTT in GM02171 cells ISIS 312.5 625 1,250 2,500 5,000 10,000 IC.sub.50 No. nM nM nM nM nM nM (.mu.M) 387916 32 56 77 89 95 97 0.7 435869 0 6 22 40 69 84 2.9 435870 15 19 32 51 68 77 2.4 435874 0 5 1 17 17 30 >10.0 435879 0 8 0 16 36 47 15.3 435890 14 16 19 19 39 57 9.3 460206 5 13 33 41 68 80 2.7 460207 13 10 22 22 33 39 45.6 460208 13 15 11 11 15 53 10.8 460209 8 27 46 70 80 86 1.6 460210 19 37 55 75 88 96 1.1 460212 8 23 30 43 57 74 2.2 460218 15 26 27 36 52 78 3.2 460219 16 17 32 44 69 76 2.5 460222 14 3 0 0 13 0 >10.0 460231 6 8 13 16 33 56 10.4 460233 27 30 39 46 61 73 2.4 460266 0 15 20 15 18 34 >10.0

TABLE-US-00059 TABLE 59 Dose-dependent antisense inhibition of human HTT in GM02173B cells ISIS 312.5 625 1,250 2,500 5,000 10,000 IC.sub.50 No. nM nM nM nM nM nM (.mu.M) 387916 22 47 76 88 96 98 0.7 435869 10 0 16 38 59 76 3.9 435870 22 36 44 58 69 81 2.0 435874 11 6 25 23 32 42 >10.0 435879 0 9 21 30 52 68 4.8 435890 12 16 30 31 48 66 4.5 460206 11 13 18 35 59 74 3.5 460207 15 25 30 37 42 66 4.3 460208 5 14 27 32 52 51 9.0 460209 27 49 61 79 81 74 0.8 460210 19 40 61 77 89 95 1.0 460212 0 19 32 32 61 78 2.9 460218 4 17 26 38 64 82 3.0 460219 5 6 26 47 68 84 2.9 460222 13 19 23 30 35 50 16.1 460231 7 33 25 35 54 77 3.7 460233 11 20 37 52 68 69 2.3 460266 12 6 10 21 25 47 >10.0

Example 11

Dose-Dependent Antisense Inhibition of Human HTT in GM04281 and GM02171 Cells by Oligonucleotides Designed by Microwalk

[0416] Additional gapmers were designed based on the gapmers selected from studies described in Example 10. These gapmers were designed by creating gapmers shifted slightly upstream and downstream (i.e. "microwalk") of the original gapmers from Tables 57, 58, and 59. Gapmers were also created with 4-9-4 MOE or 5-9-5 MOE motifs, and with constrained 6(S)--CH.sub.3-bicyclic nucleic acid (BNA) molecules at various nucleotide positions.

[0417] These gapmers were tested in the GM04281 and GM02171 cell lines. Cultured GM04281 or GM02171 cells at a density of 25,000 cells per well were transfected using electroporation with 2,500 nM or 5,000 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and HTT mRNA levels were measured by quantitative real-time PCR. HTT mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN. Results are presented as percent inhibition of HTT mRNA, relative to untreated control cells.

[0418] The chimeric antisense oligonucleotides in Tables 60, 61, and 62 were designed as 3-9-3, 4-9-4, or 5-9-5 MOE gapmers. The parent gapmers, ISIS 435890, ISIS 460210, ISIS 435879, ISIS 460209, ISIS 435870, and ISIS 460207, from which the newly designed gapmers were derived are marked with an asterisk (*) in the table. ISIS 387916 was included in the study as a benchmark oligonucleotide against which the potency of the antisense oligonucleotides targeting nucleotides overlapping each SNP position could be compared.

[0419] The 3-9-3 gapmers are 15 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on both 5' and 3' directions by wings comprising 3 nucleotides each.

[0420] The 4-9-4 gapmers are 17 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on both 5' and 3' directions by wings comprising 4 nucleotides each.

[0421] The 5-9-5 gapmers are 19 nucleotides in length, wherein the central gap segment is comprised of nine 2'-deoxynucleotides and is flanked on both 5' and 3' directions by wings comprising 5 nucleotides each.

[0422] The internucleoside linkages throughout each gapmer are phosphorothioate (P.dbd.S) linkages. All cytosine nucleobases throughout each gapmer are 5-methylcytosines. Bolded and underlined nucleotides in Tables 60, 61, and 62 indicate the positions of the 6(S)--CH.sub.3-BNA (e.g. cEt molecules) molecules in each gapmer. Italicized nucleotides are MOE subunits.

[0423] The gapmers are organized in Tables 60, 61, and 62, according to the SNP site they target. "Start site" indicates the 5'-most nucleotide to which the gapmer is targeted. "Stop site" indicates the 3'-most nucleotide to which the gapmer is targeted. `Target allele` indicates whether the gapmer is targeted to the major or the minor allele. The number in parentheses indicates the position on the oligonucleotide opposite to the SNP position.

TABLE-US-00060 TABLE 60 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs2298969 (nucleobases 125888 to 125907 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Concentration in in ID position position No. Sequence Motif (nM) GM04281 GM02171 NO 145466 145485 387916 TCTCTATTGCA 5-10-5 5000 57 24 6 CATTCCAAG 125888 125907 *435890 AAGGGATGCTG 5-9-5 2500 22 0 94 ACTTGGGC 5000 41 23 125890 125904 *460210 GGGATGCTGAC 3-9-3 2500 59 24 189 TTGG 5000 81 33 125889 125905 474870 AGGGATGCTG 4-9-4 2500 23 3 187 ACTTGGG 5000 44 34 125889 125905 474890 AGGGATGCTG 4-9-4 2500 38 6 187 ACTTGGG 5000 49 25 125889 125905 474910 AGGGATGCTGA 4-9-4 2500 34 8 187 CTTGGG 5000 49 41 125889 125905 474914 AGGGATGCTGA 4-9-4 2500 44 14 187 CTTGGG 5000 44 21 125888 125907 474918 AAGGGATGCT 5-9-5 2500 31 0 94 GACTTGGGC 5000 26 25 125888 125907 474922 AAGGGATGCT 5-9-5 2500 33 14 94 GACTTGGGC 5000 65 24 125889 125905 476332 AGGGATGCTG 4-9-4 2500 23 13 187 ACTTGGG 5000 51 42 125888 125907 476336 AAGGGATGCTG 5-9-5 2500 5 0 94 ACTTGGGC 5000 43 9

TABLE-US-00061 TABLE 61 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs7685686 (nucleobases 146786 to 146805 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Concentration in in ID position position No. Sequence Motif (nM) GM04281 GM02171 NO 145466 145485 387916 TCTCTATTGCA 5-10-5 5000 57 24 6 CATTCCAAG 146786 146805 *435879 AATAAATTGTC 5-9-5 2500 39 0 99 ATCACCAG 5000 59 19 146788 146802 *460209 TAAATTGTCAT 3-9-3 2500 3 0 203 CACC 5000 13 5 146787 146803 474871 ATAAATTGTCA 4-9-4 2500 82 32 200 TCACCA 5000 83 58 146787 146803 474891 ATAAATTGTCA 4-9-4 2500 84 29 200 TCACCA 5000 89 56 146787 146803 474911 ATAAATTGTCA 4-9-4 2500 70 18 200 TCACCA 5000 83 40 146787 146803 474915 ATAAATTGTCA 4-9-4 2500 38 9 200 TCACCA 5000 74 14 146786 146805 474919 AATAAATTGTC 5-9-5 2500 80 7 99 ATCACCAG 5000 84 37 146786 146805 474923 AATAAATTGTC 5-9-5 2500 74 32 99 ATCACCAG 5000 83 51 146787 146803 476333 ATAAATTGTCA 4-9-4 2500 75 28 200 TCACCA 5000 86 21 146786 146805 476337 AATAAATTGTC 5-9-5 2500 71 6 99 ATCACCAG 5000 83 31

TABLE-US-00062 TABLE 62 Comparison of inhibition of human HTT mRNA levels by ISIS 387916 and chimeric antisense oligonucleotides targeted to SNP rs362331 (nucleobases 155478 to 155498 of SEQ ID NO: 1) % % inhibition inhibition SEQ Start Stop ISIS Concentration in in ID position position No. Sequence Motif (nM) GM04281 GM02171 NO 145466 145485 387916 TCTCTATTGCAC 5-10-5 5000 57 24 6 ATTCCAAG 155479 155498 *435870 GCACACAGTAG 5-9-5 2500 19 1 103 ATGAGGGA 5000 49 34 155481 155495 *460207 ACACAGTAGAT 3-9-3 2500 0 0 217 GAGG 5000 7 8 155480 155496 474872 CACACAGTAGA 4-9-4 2500 35 9 214 TGAGGG 5000 63 37 155480 155496 474892 CACACAGTAGA 4-9-4 2500 43 16 214 TGAGGG 5000 69 31 155480 155496 474912 CACACAGTAGA 4-9-4 2500 16 9 214 TGAGGG 5000 36 6 155480 155496 474916 CACACAGTAGA 4-9-4 2500 22 5 214 TGAGGG 5000 47 7 155479 155498 474920 GCACACAGTAG 5-9-5 2500 19 0 103 ATGAGGGA 5000 43 23 155479 155498 474924 GCACACAGTAG 5-9-5 2500 29 8 103 ATGAGGGA 5000 48 22 155480 155496 476334 CACACAGTAGA 4-9-4 2500 35 7 214 TGAGGG 5000 62 32 155479 155498 476338 GCACACAGTAG 5-9-5 2500 26 9 103 ATGAGGGA 5000 40 4 155479 155495 474873 ACACAGTAGAT 4-9-4 2500 53 9 285 GAGGGA 5000 61 29 155479 155495 474893 ACACAGTAGAT 4-9-4 2500 47 5 285 GAGGGA 5000 59 30 155479 155495 474913 ACACAGTAGAT 4-9-4 2500 30 16 285 GAGGGA 5000 29 17 155479 155495 474917 ACACAGTAGAT 4-9-4 2500 23 12 285 GAGGGA 5000 40 5 155478 155497 474921 CACACAGTAGA 5-9-5 2500 28 0 212 TGAGGGAG 5000 43 23 155478 155497 474925 CACACAGTAGA 5-9-5 2500 30 9 212 TGAGGGAG 5000 61 34 155479 155495 476335 ACACAGTAGAT 4-9-4 2500 35 2 285 GAGGGA 5000 53 31 155478 155497 476339 CACACAGTAGA 5-9-5 2500 15 0 212 TGAGGGAG 5000 34 13

Example 12

Dose-Dependent Antisense Inhibition of Human Huntingtin mRNA Levels in Coriell Fibroblast Cell Lines

[0424] Gapmers from the studies described in Example 11 were selected and tested at various doses in GM04281, GM02171 and GM02173B cell lines. Each cell line was plated at a density of 25,000 cells per well and transfected using electroporation with 625 nM, 1,250 nM, 2,500 nM, 5,000 nM and 10,000 nM concentrations of antisense oligonucleotide, as specified in Tables 63, 64, and 65. After a treatment period of approximately 16 hours, RNA was isolated from the cells and HTT mRNA levels were measured by quantitative real-time PCR. Human HTT primer probe set RTS2617 was used to measure mRNA levels. HTT mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN. Results are presented as percent inhibition of HTT mRNA, relative to untreated control cells. IC.sub.50 values are also provided in Tables 63, 64, and 65.

TABLE-US-00063 TABLE 63 Dose-dependent antisense inhibition of human HTT in GM04281 cells ISIS 625 1250 2500 5000 10000 IC.sub.50 No nM nM nM nM nM (.mu.M) 387916 70 83 94 96 98 <0.6 460207 51 63 83 91 93 0.5 460209 83 93 96 97 97 <0.6 460210 70 89 94 97 98 0.6 474871 94 97 96 96 95 <0.6 474873 51 73 89 94 95 0.5 474891 93 95 97 96 95 <0.6 474892 48 72 89 93 95 0.6 474911 85 92 96 95 94 <0.6 474919 89 94 95 94 96 <0.6 474922 21 47 73 86 96 1.5 474923 86 94 96 95 94 <0.6 476333 92 94 95 95 96 <0.6 476334 45 70 87 92 95 0.6 476337 83 92 95 96 96 <0.6

TABLE-US-00064 TABLE 64 Dose-dependent antisense inhibition of human HTT in GM02171 cells ISIS 625 1250 2500 5000 10000 IC.sub.50 No nM nM nM nM nM (.mu.M) 387916 28 38 63 82 99 1.6 460207 16 0 20 22 55 10.0 460209 27 50 61 87 94 9.9 460210 34 60 80 86 97 0.9 474871 62 74 84 87 90 0.1 474873 13 29 61 77 89 2.2 474891 57 72 80 83 88 0.2 474892 23 26 51 68 81 2.5 474911 47 58 68 72 82 0.7 474919 44 48 65 71 83 1.1 474922 15 27 49 74 79 2.6 474923 27 53 74 79 84 1.5 476333 42 53 75 76 84 1.0 476334 20 23 58 71 87 2.3 476337 23 34 60 62 75 2.7

TABLE-US-00065 TABLE 65 Dose-dependent antisense inhibition of human HTT in GM02173B cells ISIS 625 1250 2500 5000 10000 IC.sub.50 No nM nM nM nM nM (.mu.M) 387916 38 75 89 95 99 0.9 460207 13 27 52 46 63 6.5 460209 79 68 84 90 92 <0.6 460210 37 62 79 92 97 0.9 474871 74 83 87 92 89 <0.6 474873 22 32 67 72 92 1.9 474891 69 78 84 89 89 <0.6 474892 26 50 75 83 91 1.3 474911 50 66 76 86 86 0.6 474919 57 67 74 87 82 <0.6 474922 15 32 61 71 90 2.2 474923 49 67 78 83 85 0.5 476333 58 71 78 87 89 <0.6 476334 20 42 63 76 91 1.8 476337 48 63 71 79 80 0.6

Example 13

Strategy for Selection of Antisense Oligonucleotides Based on Potency and Selectivity

[0425] Gapmers from each of the studies described above were selected for further analysis based on potency and selectivity.

[0426] Potency was based on the percent inhibition of HTT mRNA achieved by the antisense oligonucleotides targeting a SNP compared to the percent inhibition of HTT mRNA achieved by the benchmark oligonucleotide, ISIS 387916.

[0427] Selectivity was based on the ability of the antisense oligonucleotides targeting a SNP to inhibit expression of the major allele and not of the minor allele. The usage of the three cell lines with different genotypes at each SNP position facilitated this process.

[0428] ISIS 460065 (5'-ATAAATTGTCATCACCAG-3' (SEQ ID NO: 199)) is a 4-9-5 MOE gapmer targeted to SNP rs7685686 (major allele A, minor allele G) at position 9 of the oligonucleotide. The GM04281 cell line is homozygous AA at SNP position rs7685686. The GM02173B cell line is heterozygous AG at SNP position rs7685686. The GM02171 cell line is homozygous GG at SNP position rs7685686. Therefore, selectivity is shown if ISIS 460065 causes potent inhibition of HTT mRNA in GM04281, less potent inhibition of HTT mRNA in GM02173, and little to no significant inhibition of HTT mRNA in GM02171. IC50 values taken from Table 20, 21, and 22, and presented below in Table 66, confirm varying degrees of inhibition in the three cell lines, wherein expression was most reduced in the homozygous AA cell line, moderately reduced in the heterozygous AG cell line, and less reduced in the homozygous GG cell line. IC50 is the concentration of antisense oligonucleotide required for 50 percent inhibition HTT mRNA. IC50 values are in .mu.M.

TABLE-US-00066 TABLE 66 Genotype of the Coriell cell lines for SNP rs7685686 and comparison of inhibition of HTT mRNA by ISIS 460065 in each cell line GM04281 GM02173B GM02171 Genotype AA AG GG IC.sub.50 with 1.1 3.6 10.3 ISIS 460065

[0429] ISIS 459978 (5'-ACAGTGCTACCCAACCT-3' (SEQ ID NO: 174)) is a 2-9-6 MOE gapmer targeted to SNP rs4690072 (major allele T, minor allele G) at position 9 of the oligonucleotide. The GM04281 cell line is homozygous TT at SNP position rs4690072. The GM02173B cell line is heterozygous TG at SNP position rs4690072. The GM02171 cell line is homozygous GG at SNP position rs4690072. Therefore, selectivity is shown if ISIS 459978 causes potent inhibition of HTT mRNA in GM04281, less potent inhibition of HTT mRNA in GM02173, and little to no significant inhibition of HTT mRNA in GM02171. IC50 values taken from Table 20, 21, and 22, and presented below in Table 67, confirm varying degrees of inhibition in the three cell lines, wherein expression was most reduced in the homozygous TT cell line, moderately reduced in the heterozygous TG cell line, and less reduced in the homozygous GG cell line. IC50 is the concentration of antisense oligonucleotide required for 50 percent inhibition HTT mRNA. IC50 values are in .mu.M.

TABLE-US-00067 TABLE 67 Genotype of the Coriell cell lines for SNP rs4690072 and comparison of inhibition of HTT mRNA by ISIS 459978 in each cell line GM04281 GM02173B GM02171 Genotype TT TG GG IC.sub.50 with 2.5 8.4 12.7 ISIS 459978

[0430] ISIS 460028 (5'-GAGCAGCTGCAACCTGGCA-3' (SEQ ID NO: 149)) is a 4-11-4 MOE gapmer targeted to SNP rs362306 (major allele G, minor allele A) at position 10 of the oligonucleotide. The GM04281 cell line is homozygous GG at SNP position rs362306. The GM02173B and GM02171 cell lines are heterozygous GA at SNP position rs362306. Therefore, selectivity is shown if ISIS 460028 causes potent inhibition of HTT mRNA in GM04281 and less potent inhibition of HTT mRNA in GM02173 and GM02171. IC50 values taken from Table 20, 21, and 22, and presented below in Table 68, confirm varying degrees of inhibition between the GM04281 cell line and the GM02173B and GM02171 cell lines, wherein expression was most reduced in the homozygous GG cell line and less reduced in the heterozygous AG cell line. IC50 is the concentration of antisense oligonucleotide required for 50 percent inhibition HTT mRNA. IC50 values are in .mu.M.

TABLE-US-00068 TABLE 68 Genotype of the Coriell cell lines for SNP rs362306 and comparison of inhibition of HTT mRNA by ISIS 460028 in each cell line GM04281 GM02173B GM02171 Genotype GG AG AG IC.sub.50 with 1.4 5.2 5.3 ISIS 460028

Example 14

Strategy for Selection of Antisense Oligonucleotides with cEt Motifs Based on Potency and Selectivity

[0431] Gapmers from each of the studies described above were selected for further analysis based on potency and selectivity.

[0432] Potency was based on the percent inhibition of HTT mRNA achieved by the antisense oligonucleotides targeting a SNP compared to the percent inhibition of HTT mRNA achieved by the benchmark oligonucleotide, ISIS 387916.

[0433] Selectivity was based on the ability of the antisense oligonucleotides targeting a SNP to inhibit expression of the major allele and not of the minor allele. The usage of the three cell lines with different genotypes at each SNP position facilitated this process.

[0434] ISIS 460209 (5'-TAAATTGTCATCACC-3' (SEQ ID NO: 203)) is a 3-9-3 gapmer with cEt subunits at positions 2, 3, 13, and 14, targeted to SNP rs7685686 (major allele A, minor allele G) at position 8 of the oligonucleotide. The GM04281 cell line is homozygous AA at SNP position rs7685686. The GM02173B cell line is heterozygous AG at SNP position rs7685686. The GM02171 cell line is homozygous GG at SNP position rs7685686. Therefore, selectivity is shown if ISIS 460209 causes potent inhibition of HTT mRNA in GM04281, less potent inhibition of HTT mRNA in GM02173, and little to no significant inhibition of HTT mRNA in GM02171. IC.sub.50 values taken from Table 57, 58, and 59, and presented below in Table 69, confirm varying degrees of inhibition in the three cell lines, wherein expression was most reduced in the homozygous AA cell line, moderately reduced in the heterozygous AG cell line, and less reduced in the homozygous GG cell line. IC.sub.50 is the concentration of antisense oligonucleotide required for 50 percent inhibition HTT mRNA. IC.sub.50 values are in .mu.M.

TABLE-US-00069 TABLE 69 Genotype of the Coriell cell lines for SNP rs7685686 and comparison of inhibition of HTT mRNA by ISIS 460209 in each cell line GM04281 GM02173B GM02171 Genotype AA AG GG IC.sub.50 with 0.2 0.8 1.6 ISIS 460209

[0435] ISIS 460208 (5'-CAGTGCTACCCAACC-3' (SEQ ID NO: 177)) is a 3-9-3 gapmer with cEt subunits at positions 2, 3, 13, and 14, targeted to SNP rs4690072 (major allele T, minor allele G) at position 8 of the oligonucleotide. The GM04281 cell line is homozygous TT at SNP position rs4690072. The GM02173B cell line is heterozygous TG at SNP position rs4690072. The GM02171 cell line is homozygous GG at SNP position rs4690072. Therefore, selectivity is shown if ISIS 460208 causes potent inhibition of HTT mRNA in GM04281, less potent inhibition of HTT mRNA in GM02173, and little to no significant inhibition of HTT mRNA in GM02171. IC.sub.50 values taken from Table 57, 58, and 59, and presented below in Table 70, confirm varying degrees of inhibition in the three cell lines, wherein expression was most reduced in the homozygous TT cell line, moderately reduced in the heterozygous TG cell line, and less reduced in the homozygous GG cell line. IC.sub.50 is the concentration of antisense oligonucleotide required for 50 percent inhibition HTT mRNA. IC.sub.50 values are in .mu.M.

TABLE-US-00070 TABLE 70 Genotype of the Coriell cell lines for SNP rs4690072 and comparison of inhibition of HTT mRNA by ISIS 460208 in each cell line GM04281 GM02173B GM02171 Genotype TT TG GG IC.sub.50 with 1.5 9.0 10.8 ISIS 460208

[0436] ISIS 460206 (5'-GCAGCTGCAACCTGG-3' (SEQ ID NO: 231)) is a 3-9-3 gapmer with cEt subunits at positions 2, 3, 13, and 14, targeted to SNP rs362306 (major allele G, minor allele A) at position 8 of the oligonucleotide. The GM04281 cell line is homozygous GG at SNP position rs362306. The GM02173B and GM02171 cell lines are heterozygous GA at SNP position rs362306. Therefore, selectivity is shown if ISIS 460206 causes potent inhibition of HTT mRNA in GM04281 and less potent inhibition of HTT mRNA in GM02173 and GM02171. IC.sub.50 values taken from Table 57, 58, and 59, and presented below in Table 71, confirm varying degrees of inhibition between the GM04281 cell line and the GM02173B and GM02171 cell lines, wherein expression was most reduced in the homozygous GG cell line and less reduced in the heterozygous AG cell line. IC.sub.50 is the concentration of antisense oligonucleotide required for 50 percent inhibition HTT mRNA. IC.sub.50 values are in .mu.M.

TABLE-US-00071 TABLE 71 Genotype of the Coriell cell lines for SNP rs362306 and comparison of inhibition of HTT mRNA by ISIS 460206 in each cell line GM04281 GM02173B GM02171 Genotype GG AG AG IC.sub.50 with 2.3 2.7 2.7 ISIS 460206

Example 15

Comparison of SNPs in Various Cell Lines and Mouse Models Associated with Huntington's Disease

[0437] The genotype at various SNP positions associated with Huntington's disease was compared amongst the three Cornell cell lines, used in the above Examples, as well as with the GM04022 fibroblast, the BACHD mouse model and the YAC18 mouse model.

[0438] The donor patient of the GM04022 fibroblast cell line was heterozygous at SNP position rs363125 (NCBI Entrez SNP database), harboring an A allele (adenine) and a C allele (cytosine) at nucleotide 5310 of SEQ ID NO: 2 (van Bilsen, P. H. J. et al., Human Gene Therapy. 19: 710-718, 2008). YAC18 mice were developed with a YAC transgene containing human huntingtin gene (Hodgson, et al. Hum. Mol. Genet. 5: 1875-85, 1996). BACHD mice were developed expressing a full-length mutant huntingtin gene with 97 glutamine repeats under the control of a bacterial artificial chromosome (Gray, M. et al., J. Neurosc. 28: 6182-95, 2008). The comparative genotype at the indicated SNP positions in all four cell lines and mouse models is presented in Table 72.

TABLE-US-00072 TABLE 72 Genotypes of the Coriell cell lines and Huntington mouse models SNP GM02171 GM02173 GM04281 GM04022 BACHD YAC18 rs3856973 AA AG GG AG GG AA rs2285086 GG AG AA AG AA GG rs7659144 CG CG CC CG CC GG rs16843804 TC TC CC CC CC TT rs2024115 GG AG AA AG AA GG rs3733217 CC CC CC CC CC CC rs10015979 AA AG GG AA AA AA rs7691627 AA AG GG AG GG AA rs2798235 GG GG GG AG GG GG rs4690072 GG TG TT TG TT GG rs6446723 CC TC TT TC TT CC rs363081 GG GG GG GG GG GG rs363080 CC CC CC TC CC CC rs363075 GG GG GG GG GG GG rs363064 TC TC CC CC CC TT rs3025849 AA AA AA AA AA AA rs363102 AA AA AA AG AA AA rs11731237 CC TC TT CC CC CC rs4690073 AA AG GG AG GG AA rs363144 TT TT TT TT TT TT rs3025838 CC CC CC CC CC CC rs34315806 TC TC CC CC CC TT rs363099 TC TC CC CC CC TT rs363096 CC TC TT CC TT CC rs2298967 TC TC TT TT TT CC rs2298969 GG AG AA AG AA GG rs6844859 CC TC TT TC TT CC rs363092 AA AC CC AC AA AA rs7685686 GG AG AA AG AA GG rs363088 TA TA AA AA AA TT rs362331 CC TC TT TC TT CC rs916171 GG GC CC GC CC GG rs362322 AA AA AA AA AA AA rs362275 TC TC CC CC CC TT rs362273 AG AG AA AA AA GG rs2276881 GG GG GG GG GG GG rs3121419 TC TC CC CC CC TT rs362272 -- AG GG GG GG AA rs362271 AG AG GG GG GG AA rs3775061 AG AG AA AA AA GG rs362310 TC CC CC TC CC CC rs362307 CC TC CC CC CC CC rs362306 AG AG GG GG GG AA rs362303 TC CC CC TC CC CC rs362296 AC AC AC CC CC AA

Example 16

Allele-Specific Inhibition Measured in BacHD Cortical Neurons

[0439] Antisense oligonucleotides, ISIS 460209 (5'-TAAATTGTCATCACC-3' (SEQ ID NO: 203)), targeting SNP rs7685686 of human HTT, and ISIS 387916 (TCTCTATTGCACATTCCAAG (SEQ ID NO: 6)), and with no human or murine SNP target site, were tested for their effect on Htt protein levels in vitro. ISIS 387916 is cross-reactive with murine Htt mRNA (GENBANK Accession No. NM.sub.--010414.1, designated herein as SEQ ID NO: 286) at target start site 5763 with one mismatch. ISIS 460209 is cross-reactive with murine Htt mRNA at target start site 6866 with three mismatches.

[0440] Primary BacHD cortical neurons, which express human Htt and murine Htt, were isolated in the following way: Embryos were dissected from E15.5-E17.5 pregnant females. Cortices were dissected into ice-cold divalent-free Hank's Balanced Salt Solution (Invitrogen, 14025-134). The cortices were chopped into pieces and digested with 0.05% Trypsin-EDTA (Invitrogen, 25300-120) at 37.degree. C. for 8 minutes. The digestion was halted by addition of complete neurobasal media (Invitrogen, 10888-022). Cells were resuspended in media and treated with DNAse I (Invitrogen, 18047-019). After titration through a 100 ul pipette tip, cells are resuspended in neurobasal media with B27 supplement (Invitrogen, 17504-044), and counted. 1.7.times.10.sup.5 cells/well were plated in 24-well plates precoated with poly-D-lysine (BD Biosciences, 354210). Neurons were fed with 200 .mu.l neurobasal media with B27 on the second day in vitro.

[0441] ISIS 460209 or ISIS 387916 was added to the supplementary media fed to neurons on division 2 at 0.7 .mu.M, 1.4 .mu.M or 1.5 .mu.M final concentrations. Cells were harvested after 8 days with into 1 mL of media using a cell scraper. Cells were centrifuged at 2,500 rpm for 5 min at 4.degree. C. and the pellets were resuspended in a buffer of 50 mM Tris, pH=8.0, 150 mM NaCl, 1% Igepal, 40 mM .beta.-glycerophosphate, 10 mM NaF, 1.times. Roche complete protease inhibitor, 1 mM Sodium Orthovanadate and 800 .mu.M PMSF. The lysates were centrifuged after 15 min incubation and protein concentration was measured with the DC assay (BioRad).

[0442] Protein lysates were run on low-bis gels to separate huntingtin alleles (resolving gel--2001:Acrylamide:BIS (10% acrylamide, 0.5% BIS, 375mM Tris pH 8.8; stacking gel--4% Acrylamide-BIS(29:1), 156 mM Tris pH6.8; Running buffer--25 mM Tris, 190 mM Glycine, 0.1% SDS+10 .mu.M beta-mercaptoethanol added fresh). After electrophoresis, proteins in the gel were transferred to a nitrocellulose membrane (Hybond-C Extra; GE Healthcare Bio-Sciences) at 90V for 40' to allow samples to penetrate the stacking gel and then at 190V for 2.5 h to resolve proteins.

[0443] Primary antibodies specific for human Htt and murine calnexin protein were used at 1:10,000 dilutions. HRP-conjugated anti-mouse secondary antibody (1:10,000, Jackson ImmunoResearch Laboratories) was used for visualizing proteins using SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific). Protein bands were quantified using ImageJ software and normalized to calnexin levels. Protein bands were quantified using ImageJ software. Table 73 provides an estimate of the percentage inhibition relative to the negative control sample. The comparative percent inhibitions of the human Htt protein and the murine Htt protein are presented.

TABLE-US-00073 TABLE 73 Effect of antisense inhibition on mutant human and wild-type murine Htt protein (percent inhibition normalized to PBS control) Dose (.mu.M) Human Murine ISIS 387916 0.7 54 38 1.4 75 58 1.5 92 88 ISIS 460209 0.2 71 35 0.4 82 41 1.5 94 56

Example 17

Dose-Dependent Antisense Inhibition of Human Huntingtin mRNA Levels in Coriell Fibroblast Cell Lines

[0444] Gapmers from the studies described in Examples, 3, 4, 10, and 12 were selected and tested at various doses in GM04281, GM02171 and GM02173B cell lines. Each cell line was plated at a density of 25,000 cells per well and transfected using electroporation with 0.4747 nM, 1.5011 nM, 4.7463 nM, 15.0079 nM 45.455 nM, 150.0527 nM, 474.4673 nM, 1,500.27 nM, 4,743.833 nM, and 15,000 nM concentrations of antisense oligonucleotide, as specified in Tables 72, 73, and 74. After a treatment period of approximately 16 hours, RNA was isolated from the cells and HTT mRNA levels were measured by quantitative real-time PCR. Human HTT primer probe set RTS2617 was used to measure mRNA levels. HTT mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN. Results are presented as percent inhibition of HTT mRNA, relative to untreated control cells. IC.sub.50 values are also provided in Tables 72, 73, and 74.

TABLE-US-00074 TABLE 74 Dose-dependent antisense inhibition of human HTT in GM04281 cells ISIS 0.4747 1.5011 4.7463 15.0079 47.455 150.0527 474.4673 1500.27 4743.833 15000.0 IC.sub.50 No nM nM nM nM nM nM nM nM nM nM (.mu.M) 387916 15 12 4 5 7 26 70 89 98 99 0.33 435879 0 8 19 13 24 23 45 53 84 93 0.25 435890 16 1 8 12 25 23 32 52 61 91 0.82 460209 2 9 21 17 36 46 80 89 94 93 0.09 460210 4 7 5 19 20 35 69 85 98 98 0.21 476333 7 10 8 11 42 65 86 93 93 95 0.05

TABLE-US-00075 TABLE 75 Dose-dependent antisense inhibition of human HTT in GM02171 cells ISIS 0.4747 1.5011 4.7463 15.0079 47.455 150.0527 474.4673 1500.27 4743.833 15000.0 IC.sub.50 No nM nM nM nM nM nM nM nM nM nM (.mu.M) 387916 22 8 0 9 0 32 60 90 96 97 0.27 435879 0 1 6 2 0 0 8 9 46 57 7.62 435890 0 0 0 6 0 0 0 31 27 71 4.37 460209 11 5 15 0 0 7 30 69 82 88 0.96 460210 0 0 0 2 17 18 38 70 93 95 0.56 476333 0 0 0 0 13 18 44 69 72 91 0.75

TABLE-US-00076 TABLE 76 Dose-dependent antisense inhibition of human HTT in GM02173B cells ISIS 0.4747 1.5011 4.7463 15.0079 47.455 150.0527 474.4673 1500.27 4743.833 15000.0 IC.sub.50 No nM nM nM nM nM nM nM nM nM nM (.mu.M) 387916 3 17 7 25 27 33 65 88 98 99 0.19 435879 0 6 0 8 3 10 16 24 50 68 3.72 435890 0 13 0 1 2 12 16 23 49 82 4.60 460209 0 7 29 2 9 32 52 71 82 86 0.27 460210 0 13 0 5 16 18 49 74 93 97 0.27 476333 11 13 20 7 23 36 63 75 83 90 0.13

Example 18

Validation of the Specificity of ISIS Oligonucleotides Targeting SNPs of Human Huntingtin by the Molecular Beacon Assay

[0445] Some of the gapmers from the study described in Example 17 were tested in GM04022 fibroblasts (from the Coriell Institute for Medical Research).

[0446] To verify allele-specific suppression of HTT mRNA in GM04022 fibroblasts by ISIS 435879, ISIS 460209, and ISIS 476333, the Molecular Beacon assay, as described in the van Bilsen at el publication (van Bilsen, P. H. J. et al., Human Gene Therapy. 19: 710-718, 2008), was conducted using `molecular beacon` synthetic oligonucleotides linked with a fluorophore and quencher. GM04022 fibroblasts were transfected by electroporation with ISIS 435879, ISIS 460209, or ISIS 476333 at 0.06 .mu.M, 0.19 .mu.M, 0.56 .mu.M, 1.67 .mu.M, 5 .mu.M and 15 .mu.M concentrations of antisense oligonucleotide, as specified in Tables 75-77. ISIS 387916 was included in the assay as a benchmark oligonucleotide. The qRT-PCR assay for molecular beacon for the A allele was conducted with the annealing temperature at 56.5.degree. C. The qRT-PCR assay for molecular beacon for the C allele was conducted with the annealing temperature at 62.0.degree. C. Primer probe set RTS2617 was used to measure the total HTT mRNA reduction. The results of the assay are presented in Tables 77-79 as percent inhibition over the PBS control. The results demonstrate that the SNP-specific ISIS oligonucleotides specifically target the C allele of rs7685686 compared to the A allele (Table 80).

TABLE-US-00077 TABLE 77 Dose-dependent antisense inhibition of the A allele of rs7685686 in GM04022 fibroblasts ISIS 0.06 0.19 0.56 1.67 5.00 15.00 IC.sub.50 No .mu.M .mu.M .mu.M .mu.M .mu.M .mu.M (.mu.M) 387916 33 40 53 90 99 98 0.56 435879 0 0 50 29 38 47 10.8 460209 14 4 54 73 81 95 0.53 476333 2 44 41 77 91 86 0.64

TABLE-US-00078 TABLE 78 Dose-dependent antisense inhibition of the C allele of rs7685686 in GM04022 fibroblasts ISIS 0.06 0.19 0.56 1.67 5.00 15.00 IC.sub.50 No .mu.M .mu.M .mu.M .mu.M .mu.M .mu.M (.mu.M) 387916 41 42 46 86 95 92 0.54 435879 0 0 75 60 68 81 2.9 460209 35 48 76 84 88 92 0.19 476333 22 60 75 84 90 93 0.15

TABLE-US-00079 TABLE 79 Dose-dependent antisense inhibition of total HTT mRNA in GM04022 fibroblasts ISIS 0.06 0.19 0.56 1.67 5.00 15.00 No .mu.M .mu.M .mu.M .mu.M .mu.M .mu.M 387916 32 59 49 89 98 99 435879 0 0 42 25 41 62 460209 26 27 54 75 84 96 476333 25 51 58 82 92 90

TABLE-US-00080 TABLE 80 IC.sub.50 ratio (A/C) in GM04022 fibroblasts ISIS No Ratio 387916 1.0 435879 4.2 460209 2.8 476333 4.3

Example 19

Allele-Specific Inhibition Measured in Cortical Neurons from BACHD and YAC18 Mice

[0447] In order to identify potential SNPs for screening of human allele-specific ISIS oligonucleotides, the HTT mRNA of YAC18 and BACHD mice were sequenced by the Goldengate 96SNP assay. It was determined that the BAC and YAC mice carried different alleles at several key SNP positions (Table 72) and could therefore be used as a screening tool for allele-specific knockdown. Each of the SNP positions chosen for targeting in the mouse strains were also compared to human HD chromosomes. For each target, approximately 50% of the human HD population is heterozygous for the target expressed in the BACHD mice, but not the YAC18 mice.

[0448] In order to verify the allele-specificity of the ISIS oligonucleotides (described in Examples 2, 9, 17 and 18), the antisense oligonucleotides, ISIS 460207, targeting SNP rs362331; ISIS 460209, targeting SNP rs7685686; ISIS 435879, targeting SNP rs7685686; ISIS 476333, targeting SNP rs7685686; ISIS 460210, targeting SNP rs2298969; ISIS 435874, targeting SNP rs4690072; ISIS 460208, targeting SNP rs4690072; ISIS 435331, targeting SNP rs2024115; and ISIS 435871, targeting SNP rs363088, were tested for their effect on HTT protein levels in BACHD and YAC18 cortical neurons. ISIS 387916, which has no human or murine SNP target site, was used as the benchmark. ISIS 387916 is cross-reactive with murine HTT mRNA (GENBANK Accession No. NM.sub.--010414.1, designated herein as SEQ ID NO: 286) at target start site 5763 with one mismatch. It was expected that treatment with the allele-specific antisense oligonucleotides would cause significant inhibition of HTT mRNA in the BACHD neurons and not in the YAC18 neurons. It was also expected that treatment with ISIS 387916 would cause inhibition of HTT mRNA in both sets of neurons.

[0449] YAC18 cultures were prepared from E16.5 pregnant female YAC18 (line 60, +/+) mice who had been bred with YAC18 (line 60, +/+) males. All progeny are thus homozygous YAC18 (line 60), facilitating pooled cortical cultures. BACHD E16.5 embryos were isolated from pregnant BACHD (+/-) mice who had been bred with pregnant BACHD (+/-) male mice, necessitating single pup cultures and genotyping. Single cortices were isolated, using caution to prevent cross-contamination of samples. Each dissociated cortex was used to seed 5 wells of a 6-well plate. After genotyping, only BACHD (+/-) cultures were used for ASO treatment. The antisense oligonucleotides were added to the supplementary media fed to the neurons on division 2. Cells were harvested after 8 days with into 1 mL of media using a cell scraper. Cells were centrifuged at 2,500 rpm for 5 min at 4.degree. C. and the pellets were resuspended in a buffer of 50 mM Tris, pH=8.0, 150 mM NaCl, 1% Igepal, 40 mM .beta.-glycerophosphate, 10 mM NaF, 1.times. Roche complete protease inhibitor, 1 mM Sodium Orthovanadate and 800 .mu.M PMSF. The lysates were centrifuged after 15 min incubation and protein concentration was measured with the DC assay (BioRad).

[0450] Protein lysates were run on low-bis gels to separate huntingtin alleles (resolving gel--2001:Acrylamide:BIS (10% acrylamide, 0.5% BIS, 375 mM Tris pH 8.8; stacking gel--4% Acrylamide-BIS(29:1), 156 mM Tris pH6.8; Running buffer--25 mM Tris, 190 mM Glycine, 0.1% SDS+10 .mu.M beta-mercaptoethanol added fresh). After electrophoresis, proteins in the gel were transferred to a nitrocellulose membrane (Hybond-C Extra; GE Healthcare Bio-Sciences) at 90V for 40' to allow samples to penetrate the stacking gel and then at 190V for 2.5 h to resolve proteins.

[0451] Primary antibodies specific for human HTT and murine calnexin protein were used at 1:10,000 dilutions. HRP-conjugated anti-mouse secondary antibody (1:10,000, Jackson ImmunoResearch Laboratories) was used for visualizing proteins using SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific). Protein bands were quantified using ImageJ software and normalized to calnexin levels. Tables 81-91 provide the percentage inhibition relative to the untreated control sample. The percentage inhibition of human HTT protein levels in BACHD and YAC18 neurons are presented.

TABLE-US-00081 TABLE 81 HTT SNPs in BACHD and YAC18 mice and correlation with human HTT SNPs Allele % of human Allele Allele present in patients present in present in human patients heterozgous YAC18 BACHD with high at the SNP SNP Mice Mice CAG repeats position rs2024115 G A A 48 rs2298969 G A A 52 rs362331 C T T 49 rs363088 G T T 38 rs4690072 T A A 49 rs7685686 G A A 49

TABLE-US-00082 TABLE 82 Effect of antisense inhibition by ISIS 387916 in BACHD and YAC18 neurons 500 nM 1500 nM YAC18 69 81 BACHD 84 90

TABLE-US-00083 TABLE 83 Effect of antisense inhibition by ISIS 435331, targeting rs2024115 in BACHD and YAC18 neurons 500 nM 1500 nM YAC18 0 0 BACHD 39 43

TABLE-US-00084 TABLE 84 Effect of antisense inhibition by ISIS 460210, targeting rs2298969 in BACHD and YAC18 neurons 500 nM 1500 nM YAC18 31 51 BACHD 79 89

TABLE-US-00085 TABLE 85 Effect of antisense inhibition by ISIS 460207, targeting rs362331 in BACHD and YAC18 neurons 500 nM 1500 nM YAC18 0 0 BACHD 29 44

TABLE-US-00086 TABLE 86 Effect of antisense inhibition by ISIS 435871, targeting rs363088 in BACHD and YAC18 neurons 500 nM 1500 nM YAC18 0 0 BACHD 51 68

TABLE-US-00087 TABLE 87 Effect of antisense inhibition by ISIS 435874, targeting rs4690072 in BACHD and YAC18 neurons 500 nM 1500 nM YAC18 9 5 BACHD 30 44

TABLE-US-00088 TABLE 88 Effect of antisense inhibition by ISIS 460208, targeting rs4690072 in BACHD and YAC18 neurons 500 nM 1500 nM YAC18 1 8 BACHD 54 68

TABLE-US-00089 TABLE 89 Effect of antisense inhibition by ISIS 460209, targeting rs7685686 in BACHD and YAC18 neurons 500 nM 1500 nM YAC18 12 32 BACHD 72 83

TABLE-US-00090 TABLE 90 Effect of antisense inhibition by ISIS 435879, targeting rs7685686 in BACHD and YAC18 neurons 500 nM 1500 nM YAC18 0 7 BACHD 36 58

TABLE-US-00091 TABLE 91 Effect of antisense inhibition by ISIS 476333, targeting rs7685686 in BACHD and YAC18 neurons 500 nM 1500 nM YAC18 46 61 BACHD 89 91

Sequence CWU 1

1

2881202001DNAHomo sapiens 1gcccagcagg tgtcagcctc attttacccc gcccctattc aagatgaagt tgttctggtt 60ccaacgcctc tgacatatta gctgcatcat tttacatttc tttttttttt ttccttttaa 120atggggtctt gctctgtcac ccaggctgga gtgctgtggt atgatctcgg ctcactgcaa 180tctccacctc cgaggttcca gcgattctct tgcctcagcc tcccgagtag ctgggactac 240aggcacccac catcatactg ggctaatttt tgtgttttta gtagagatgg ggtttcccca 300tgttgcccag gctgatctca aactcctggg cttaagcaat acagccgcgt tggcctccca 360aagtgttggg attacaagca tgagctaccc cacccagctc attttacatt tccacttgtt 420aaactgaaaa ctggcccgag aaagcttctg tactgccatc cttgcgtcct tgcagatgaa 480tcgtaaccta gcatagtagg taggcagact gaaaacctaa cttagcagta ggcttctgta 540acaacagctg tgtctcagcc agttcctgca gccagacttc aaccactcac aggccgcaaa 600ctgttcaaac tgtgttcgga gaaggcgaat tcatctggct gttaacgtgc ctcacttctg 660ctttctgtgg ccactttccc ttttctgtcc ataaatttgc tttgaccaca cagcatccct 720agagtctccc tgaatctgct gtgattctgg gacctgcacc atttgtgaat tgtttttttt 780ttccttgatc agctaaactc tgttcaattc aatttgttgg aagtttttaa cataccaatg 840gtgcaccaag gttccaattt ctccacttcc tcataaataa gtcattttaa atggcttttc 900agtattccaa tatttggaag tattaatgtt tctaccaatt ttctattttt ggacattgag 960gttgtttcat tttttttttc tttttttgag acagagtctc gctccgtcac ccaggctgga 1020gtgcagtggc ctgatcccgg cccactgcaa cctccacctc cctcctcagc ctcctgagta 1080gctgggatta caggtgcatg caccaccaca cccagctaat ttttgtattt ttagtagaga 1140tggggtttca ccatgttggt caggctggtc tcaaactcct gacctcaggt ggtccacctg 1200ccttggcctc ccaaaatgct gggattacag gcctgagcca ctgcgcctgg cctcatcttc 1260ttgatattaa tgttgcttta acatctttgt ccctgtgttt tttgtttttt tttttgagac 1320ggagtctcat tcattctgtc acccaggctg gagttcagtg gcgtgatctc agctcactgc 1380aacctctgtc tcctgggttc cagtgattct cctgcgtcgg tctcctgagt agctgtgttc 1440ctgggtcttt cgatggttat ttaatacttc cctacagtaa tgccctgtgc gtacatgcta 1500agtgtgatga aatggttggc acagttaaat cttttgaaag acattgccaa gtcactcttc 1560agaaaagtga taggaggtca tagcaatttt aagaagtcct catttctaca tttccttact 1620aatctcggtt ggtgtctctt caatctttcc tcacactttt cttgggtttt tcctgaatca 1680tgagtctact acatttacac attttaaagc atctttagaa acaggatctc attttgttgc 1740ccaggctaga gtttggtggc atgattatag ctcctcatac tcctgggctc aagtgatcct 1800tccacctctg aaaccccaaa atttgagaaa ggtctcattt aatttagaaa gtttattttg 1860ccaaggttga gggtgcacac ctgtgatgat atacgagtta aaaagaaatt atttaggcag 1920atactgaggg taagaaagtc ctcggtaagg ttttcttttc aatgaaaagc agcccccaag 1980cattttcttt tctaacaaag agcagcctgt aaaatcgagc tgcagacata cacaagcaag 2040ctggaagctt gcacaggtga atgctggcag ctgtgccaat aagaaaaggc tacctggggc 2100caggcagatc caacatggcg gctccatctt ccctttcctt gtcaaccatg tgcacagtaa 2160ggagcaggca acatagtgtc ccccgagtag agaccaattt gcataataaa aggtgagggt 2220agggtgggca gcttctttgc atgctatgta aacattatgc ctggtccaac caatctttgg 2280gccctgtgta aattagacac cacctcctca agcctgtcta taaaaccctg tccattctgc 2340cgcaggctgg aagacccact ggggcacccc tctctctcta taggagacag ctattcattt 2400ttctctttct ttcacctatt aaagctccac tcttaacccc actccgtgtg tatctatgtt 2460cttgatttcc ttggcatgag gcaatgaacc ttgggtatta ccccagaacc ttgggtatta 2520tgccacttca gtgacacagc ctcaggaaat cctgatgaca tgttcccaag atggtcgggg 2580cacagcttgg ttttatacat tttagggaga catgagacgt caattcatat atgtaagaag 2640tacattggtt ccgtccagaa aggcggggac aacttgaggc agggagagag cttctaggtc 2700acaggtagac aaatggttgc attcttttga atctccgata agcctttcca aaggaggcaa 2760tcagaatatg cgtctattga ctgggcgcag tggctcatgc ctgtaatgcc agcactttgg 2820gaggcggagg tgggtggatc acctgaggtc aggagtttga gagcagcccg gccaacatgg 2880tgaaaccctg tctctactaa aaatacaaaa aattagctgg gcgtggtggc gggcgcctgt 2940aatcccagct actcgggagg ctgaggcagg agaatagctt gaacccagaa ggaagaggtt 3000gcagtgagct gagatggtgc cattgcactc cagcctgggc aacaagagtg aaactccatc 3060tcagaaaaaa aaaaaaaagg cctgggcaaa gtggctcacg cctgtaatcc cagcactttg 3120ggaagccgag gcgggcaggt cacaaagtca ggagattgag accatcctgg ctaacatgat 3180gaaaccccat ctctactaaa aaatacaaaa aactagctgg gtgtggtggc gagcacctgt 3240agtcccagct actcggcagg ctgaggcagg agaatggcgt gaaccgggga ggcggagctt 3300gcagtgagcc gagatcacac cactgcactc cagcccggac gacagggcaa gactctatct 3360caaattaaaa aaaaaaaaaa aaaaaaaaaa aaagagagag agaatatgca tctatctcag 3420tgagcagaag gatgactttg aatggaatgg gagcagttcc tagcttgaac ttccccttta 3480gcttcagtga tttgggggct caaggtatgt tcctttcaca tacctcagcc tcccaagtag 3540ctgggaccac aagtgcatgc caccacacgt ggctaatgtt ttattttttt tgtaggaata 3600gggtctcact atgtgtccag gctggtctaa aacccctgag ctcaaatggt cctcccgcct 3660cagcctcccg aaatgctggg attacaggca tgagccagca tgcccggcct agtctacatt 3720tttataaatt gctaattcaa agttccctct ccaaaacctc atggttttcc ctgttctcat 3780cccctgcacc ctcccttccc ctggagtact cacctggcct tggaggtctg gtgtgagccc 3840ggacttcgat tctaggcaca gcatgtgatg agcgccccca ggtcaaacac ctcccctctg 3900cggcctgtgc ttcaccgcct tgacagtgag aaaggtctcc cttcggctca ttctcgaagt 3960ctcaaacttc acttctcctg tgcgctgatt ctgaattcag cccccgtcca aggtcctggc 4020ccctttctct tctgcttggc gtgttgttca tcaccactgt gcactgctga gggtaagtgc 4080ggttctctgg acctctgctt tatcattaga acagactctt gcggtttccc acgacattcc 4140tttcacttct cacttggaag atgagccgtg aggaaatcct gtgttgtgtg gtatgtgggc 4200tgtgcttctg cttgacttga gggccaagca gcattgcaag ccatggtttt aaataagaaa 4260gaacatttct aaccttcatc ttctagtaag gaaacaagtg ggctttagag ttcttgctca 4320ggaaagacct atgtcccagt ccaaccggac cttttactaa agagatcttc ctgatcctcc 4380tccccaggcc aggggagggg tcctccctgg ggttggagcc tttagtaggg ggtcggagac 4440acgacgtagc cttcatgaca ttcatagtct agttacacga tccctgtaag ggtcagttga 4500agtaagtgct acaaaggaag ggaggtgctc agtggagagg gctctctttt atgtattata 4560tttctttcat ggggagggat atggatcagg gatcagcaga ggtgtttcag tcccgaggga 4620aagaaagtca gcgtggcttg ggagttggga gcagcaagac agtggctcaa gatatcttaa 4680gactagtgga gtacaccttg catgttaaaa gccttgctca gggctgcctg gttcttgtag 4740gacgacagag atggcctagc tctgcatact gcacccccag gggctcagaa cagtgcaaat 4800gtcagtctat ctgtcagtgg cagagccagc cttggagcag gggtgcaagg aggtctctgc 4860actggccagg catgcagaac attctgttca gtagcactgg acagaaggcc ccatctagat 4920gagacagagc tggtggggca ggacaaagac tcctggcagc tcaaacggcc tggcagatgc 4980ttggagagag ggggcttctt gagacagcac catttctggg aagagagtca cctgggaggg 5040atgaggccac gctccggctt ggaggtgaag agaggggctg ctgcaagaaa gaattagaga 5100catgccagcc tttgctgtgt tgcccaggct ggtcatgaac tcttggcctc aagcaatctt 5160cccacctcag cctccccaag cgctgggatt atagacatga gcccccatgc tggccaataa 5220aagatgattt tatggagggg atggtggtga aggttgtggg tggtatgaaa tagtaagaaa 5280tatatattgg tctgcaccca gttcctgcca cagagctcct aaaatcctga gaacttcctg 5340ggtgagcatc ttttgttcta atgaggtgac tcttggtggc tcctggatag gagtgaatca 5400ccagaaagat caagccagag ttagaagcag aaagtgctgg ctataacaca ggaaagctgt 5460aacacaaata ataaagtttt tttttttttt tttgagatgg agcctcactc tgttgcccag 5520gctggagtgc aatggtgcaa tctcagctca ctacaagctc tgcctcccag gttcaagtga 5580ttctcctgcc tcagcctcct gagcagttgg gactacaggt gtgtgccacc acatctggct 5640aatttttgta tttttagcag agacggggtt tcaccatatt aaccaggctg gcctcaaact 5700ccttaccttg tgatccgcct gcctcagcct cccaaagtgc tgggattaca ggcatgagcc 5760accgtgcctg gccaaaagac attgttctta aaagaatcaa ctaactaacc aaataaataa 5820aaatctaacc taattaagaa actaaaaata cacaaaaatt aatttcaagg ggagaaaaat 5880catgtaaaga gagaaagata atgaatactt tgcagaaatt tatgaacata aacataaaac 5940ttggatgaaa tgcatttcta ggaaaacata atttatcaaa actaaccaca agtaaaatag 6000aagcctaaat aggatatttt caagagaaga agtaaagttg tcaaagtgct acccttcaaa 6060aaaacaccag gctcaaacaa tctgacatgg gaatgttagc acaccttaga gagcaaataa 6120aactttgaat gggcttgaaa tattccagac tctagaaaaa caaaacttcc caattctttt 6180tataaagcaa gtataaattg ataccaaaat cttataaaga ccttatacaa aacttcatac 6240caatctcttt tatgaataca aaacccttaa taaagtatta ccagacagaa cccaacaata 6300cataaaaatg tcacatcata acatagtggg gtttatttca ataatgcatg gatggttcaa 6360tacaaggaaa ttcagtaaca caatataata gatcatgtga atatacccaa agaaaaaata 6420gattattttc atagatgctg taaaggcatt tgaccaaatt caacacctac tttttaggtg 6480gtcaataaaa taaattagtt actccttctt tagcatgata aaatatattt atcagcccag 6540aaggcatcat tttacccgat aagggcacac gctggaggga ataatgttaa aattaggaat 6600aagaggatag ctagtttctt tcttcttttt tttttttgag acggagtctt gctctgttgc 6660caggctggag tgcagtggtg caatgttggc tcactgcacg ccccccgcct cccaggttca 6720agcgattctc ctgcctcagc ctcccgagta gctgggacta caggcgcgca ccaccatgcc 6780cggctaattt ttttttgtat tttagtagag atggggtttc accatgttgg tcaggctggt 6840cttgaactcc caacctcacg tactgggatt accggtgtga gccaccacgc cagcccaact 6900actttcaaca ttatccttaa tactgatgct tattgactta ctatggggtt acctctagat 6960aaatccataa taagttgaaa atataagtaa aaaatgccct taatacacct aacctaccaa 7020acatcatagc tgagcccagc ctgccttagc tatgctcaga cactgacgtc agcctacaat 7080tggcaaaatc acacagcagc acagtctact gcagagcatc tgctgtttgc ccttgtgact 7140gcgtggctgc ctgggagctt cccagcttca caagacagta ttacgtagca catcactagc 7200ctggggaaag atcaaagttg aaaatttgaa gtgtggtttc cattgaatgt gtactgcttt 7260tgcaccatca tcaagtcaaa aaattttagt tgaaccagcc taagtttggg accatcttta 7320ttttcaggag gaacttccat gtacattgat gacggacgat agaatccgtt tctatcatcc 7380taatgaacat aatgaataaa tccagacaaa cataaacatt aacagagtaa gcagctttcg 7440gggctggaag ccagaagagg gtgggagcgc agagagagag gccaaacacc agggctgctt 7500ctgctttgcg ggtatttgct gatctggaca aggtatctgg aaggctgagc taagcctcct 7560ttttttttga ggtggcgtct cactctgttg ccaggctgga gtgcaatggt gcgatctcag 7620ctcactgcaa cctccacctc cctggttcaa gcgattctcc tgcctcagcc tcccgagtag 7680ctgggattac aggctcccgc cactacaccc agctgatttt tgtaatttta gtagagacgg 7740ggtttcacca tgttggccag gatggtctcg atctcttgac gtcatgatct gtccacctcg 7800gcctcccaaa gtgctgggat tataggcgtg acccaccgtg ccccgtctga gctaagcctc 7860ttgagcatag gggactaaaa atgaaatcta gcgcatgcca agtttagggt cccaggcaat 7920tcctttccac tttggggtcc actttggggt ccaccccacc caagaagaag gatgacttgg 7980aagtaaacca gctctgaaat atggatggtc ctctgggacc ataccaatcc cttcatatca 8040accacatcca gttcctcaaa actggaactt ggattaagat ggcctaggac ttctagtgtc 8100ccaggagcct ggcattgcaa acaaaaatcc tctccggaag aagataatac cttaagcttc 8160aaatgactct ctaataaatt tcaaatacaa tgtccagcac acaaacacaa attaccagga 8220acgtgatatg aggcctgatg gatgggaatt agcagaaact tcaggcatga gaaacatacc 8280ctcagaggcc tagaatctat ctagtgtcta gataatggag atatgaaata cagacactta 8340aacaactatg tttcccatgt tcaaagagga aatttgcaaa acttgaaagt gttggcagga 8400aatcagaaac tataaaatgt gacaacagca tactttagag tcagtataaa ttacggtccc 8460gaaaactgca gaattccaga acttaatggt aaagcaaggg tttaacagca gaatagaaat 8520agccagagag aactaggaag taagtcagat gacactaccc agaataaggc actgagaggc 8580caaggaatgg aaaatgcaga agaaaggata tggtgagagg atctaatata catttatttg 8640gagtaccagg gagagagaga aggagaagaa cagaagccgt gtttcaagga cggtgactga 8700gaggcttcga aactgatgaa agccatcagt tcacaaattc aaagcccagt gaattccaag 8760gagaaaaaaa gaaatccata ctgtgaaagc aagtccagac aatgacaaac accatcaaca 8820atacacagga caggcataag atgcatttaa tggggacact cagaggcaga gggttatcag 8880aaggaggcac ttctctccca agttctcatc atcccagggc cagggacagc tggtcacacc 8940ttagggagtt cactaggaga gggatctggc ttcttgtcat tctgggtatt tgtagggaaa 9000ttggaaggga accgagagca cctagccaat cgcatagcaa tgggagattt caggctgtgg 9060ggaatgtctt tgctggtgaa aagaacatcc tgaccttaga aatctttcac cgagggggat 9120ctgcgttcca gaacttctgg agctggtata ggtaaggctt tgagctttcc tactgagcca 9180gcctgttgct aggttaccaa aggggacctc gagggccatc tggccaacaa gcagacttgt 9240ctctccttac acccccagac gtatcactgc aaaactacag aaaaccaaag acagagaaaa 9300tcttaaaagc agccagattt aaaaaatggc atattagttt caaagcagca gccatgaaat 9360tgacagctga tgtctcaaca gcaagaatga aaagtggaag acaggccagg tgtggtggct 9420caggcctgta atcccagcac tttgggaggc cgaggcgggt ggatcacgag gtcaggagac 9480caagaccatc ctggctaaca tggtgaaacc ccgtctctac taaaaataca aaaaaattag 9540tcgggcatgg tggtgggtgc ctgtagtccc agctactcgg gaggctgagg caggagaatg 9600gcgtgaaccc gggaggcgga gcttgcagtg agccgagatt gtgccactgc actccagcct 9660gggtgacaga gcaagactct gtctcaaaaa aaaaaaaaaa aaaaaaaaaa aaagggtgac 9720gaagcttcaa tctcctgaaa ggaagcaact gccgcctttg attcgatacc caccaaaatc 9780cgtgaagaag gaaggcaaaa taaaaacact tcctgattga actggaaaga tttccgcaat 9840agaagaccca ctgtccaagg aattctaaag gatgctttcc aggcagaaga aaatgacccc 9900agaggaagat cagagattca ggaaagaaat ggagagtgat aaaaatggaa aattcggggg 9960ccaatttaaa caaaagctga ctgctctaca actgttgtgt ctctatcttt tgtaacatat 10020atgtgtgtgt agcttttttt tttttttttg tcaagatgga ttctcactct gtcgcccagg 10080ctacagtgaa atggcacggt ctcggctcac tgcaacctct gccccttggg ctcaaatgat 10140tctcttgcct cagcctcctg agtagctgag attacaggtg cctggcacaa tgcctggcta 10200atttttgtat ttttactaga gatgggattt ctccatgttg gccaggctgg tcttgaacac 10260ctgacctcag gtgatccacc tgcctgggcc tcccaaagtg ctaggattac aggcgcgagc 10320cactgcatct ggcctatgtg tgtgtttata tggaattaaa acacatggca ataataccct 10380ccaaattggg agaaaccaaa aatagcattt aaatgttgta agctccctgc ataatcaaga 10440agagaataga tttacgttag attttgatac ctggaggatg aatgttgtaa tttctagggt 10500gaccatgaaa agaggagaca acggtgtatg tttttttttt tttgagatgg agtctcactt 10560tgtcacccag gctggagtgt tgtggtgtga tcttggctca ctgcaacctc ctcctcttgg 10620gttcaggcca tcctcccacc taggcctcca gagtaggtgg gatcacaggc acctgccacc 10680acacctggct aatttttttt tttttttaaa tatttagtag agatggggtt tcaccatgtt 10740ggccaggctg gtcttgaact cctgacctca ggcgatctgc ctacctctgc ctctcaaagt 10800gctgggatta caggtgtgag ccatcgcgcc cggccaacag tgatcacttt caaactaaca 10860gaggttcaaa aataaaatca gacttaacca aaaaccaggt aacagagctg gtaggatata 10920cagaaagact gacctcacgt atatcaacga ttacagttaa tattaatgaa ggaaatgctc 10980tagtttaaaa acgagggttg tcaaagaccc cacataagaa gctccttacc agcggtgcac 11040ctagaaccta aggaaacagg acagatgaag gaggacgcgc ccccgccgct gtcctgcgcc 11100tcagccatcc tatgagacgg gaaaggtttc tgtctgcagc tgggcccgtg ctctttacca 11160gctcctggct ttcttctctg gaaggttcct gcctgttttg ccctcacacc tgctcctctc 11220tcagccctct caggggtggg gctggaggcc accaaagagc ctcctctgct ctccagttgc 11280tcgactgctc ctcatttccc cctggggtct gcgtcagggt ttccttcttt tccagcccca 11340ccccgcgtgc atcccacctg gtctcgggtc ggggctgctc ccgcttactg ccccctgccc 11400aggctggtgt gcaccccctc tggctgcttt caaggcctct tctctcttct cggcaggaca 11460ggcacaggca ggtggccagg tgtcatgctt agctccccgc ccagtgagat tctttcattt 11520aacaatcttc ccctgaatag ttcatgttca ttgctgaaaa tttgaaaaat atggaaaagc 11580acaaagatta agatataaac cgccctcaat tcccctgccc agagagagtc actgctatga 11640cttggtgact aggaacctta tttctctctc gctctttttt ttttttttga gacagagtct 11700tgctctgtca cccaggctgg agtgcagtgg ctcgatctca gctcactgca acctccgcct 11760cctgggttca agcgattctc ctgcctcagc ctcttgagta gctgggatta caggcacctg 11820ccaccatgcc cggctaattt ttgtattttt agttgagaga gggtttcatc ttgttggtca 11880ggcggacttg aactcctgac ctcaggtgat cagcccacct cggcctccca aagtgctggg 11940attacaggtg tgagccactg cgccttcatc tctcttctgt gtatgtgtac gctgtttttt 12000ctttagaatg ggggacgtta tcaggctcta catggtgtgt agtcggctag catgttgtaa 12060gcctttccct gtgtcacaag tgctcatctg gaacaggatt ctaatgactg cctgtggcta 12120tgttgggatt cctttaactc agctccttct gcccagcatc tatctttttt ccatcttttg 12180tcctaagtgt tgctataata aatcattgat cacacatgcc tgactgtttg cataggataa 12240attacgggaa atgtttttgc tgttcaggga ctgtgcccat ttttaggcct cagagacacc 12300atgccagact gcccagtatt gatctttact ctttttagat gatgccaaac ttttctgtga 12360actttaaaaa cctgtgtctt gacagtccat ttctgtaagt ctttcacatt agatttcctg 12420tcaggatgat agtcaattct aggcagatga tgttttctca gccatggctg aagcagttgt 12480gatttgttgt ggccatgtaa agtcccgatg atccattgcc tccctggatg ggttggaata 12540atttggtttg ggagcatata acagaatgac ctggagtcac agcagctcag acggaagtgt 12600atttctccct tacagatgaa agaattccag gccaggctgg aatgacaact gcacacagtc 12660atctgggccc cctccttcca gctcccatca ccccaggatg tggcttttat gcagatgatc 12720caaaatggct gctcaagtcc cagccaacac atcccattcc agggagcagg aaaaaggtgt 12780gtctttccct tcattttatg tgattccttt ctagaagtac tactcattac ttctgcttgc 12840atctccctgg ctagcactta cttagttata tggccatagc tagctgaagg aaggacaggg 12900actgtcatac actagctaag aggcaaactg cttagataaa aaggtctcta aagaaggtca 12960gagcggctgc tagggtgcaa ctctattact tattgttatg ggacgaactg tgtccctcat 13020tcaggttgat gtcctaagcc ccagaacctc agaatgggat tgtatttgga gacaggttct 13080ttaaggaggt aaggaggcta aaatgagatc attagggtgg gccataatcc gactgatgtc 13140ttacaagaag agattaggac acggacatgc tcagagggac ggccacgtga ggacaccaag 13200aaaggcagct gtctgcaagt caaggacagg gctcagggga aaccaacctt gccaacacct 13260tcatctcgga cttctagcct ctaggaccat gagaagatac atttctgttg tttaagctgc 13320ccggtctgtg gtactttgtt atggcagccc aagtaaacaa atacagtcat ctgctgctgg 13380aacaaatcac cccagcactg tggcttggca gcacacatgt ctagtcatag agttatatgt 13440agttacgtgt agagccatat gtatcgtcac acgttctgtg ggtcaggaat ttggacccag 13500cttaaccagc tccacttctc gccagggttc agtcaaatac cagctgcctc ccacctgaga 13560gctcagccgg ggaagggtcc ctttccaatc tcacgtggtg ttggcaggat ccagttcctc 13620atggcctgct ggactgagaa cctcagttct cactgcctgt tggccagagg ccgcctttat 13680gtcctcgcca tgtgggcctc tccaacatgg cagctgactt catcagagca tccatgccaa 13740gaaggcaaca gagagggcca gggagactga agtcataccc ttttgcgacc tagtcatggg 13800gtgacattcc atcacctttg cccattggtt agaagcaggc caccaggtac agcccaagct 13860cacggggagg ggtcatacaa gggtgtcaat accaggaggt gaggggtgct ggggccatct 13920tatgagtctg cccactgagg taactaacaa ccttgaggcc tgacacagtg gacaaaggcc 13980cttattaaca gcagagaact gggaacttta tttatttatt tatttttgag acagagtctc 14040actcttgtca cccaggctgg agtgcaatgg catgatcttg gctcactgca acctccacct 14100cccaggttca agcaattctg cctcagcctc cggaatagct gggactacag gcatgcacca 14160ctacacccgg ctaatttttg tatttttagt agagacaggg tttcgccatg ttggccaggc 14220tggtctcgaa ctcctgacct ctggtgatct gcctgccttg gcctcccaaa gtgctgggat 14280tacaggcgtg agccaccgca cctcgctgga acttaatttt tttagagaca gtgtcgctct 14340atcacccaag ctggagtgca gtggtgcaat cctagctcac ttgcagcctc aaattcctgg 14400gttcaggtga tcctcccaca tcagcctccc aagaactggg aactaacagc tgtttctctg 14460ctgtccttct caagaaaagg gaggctactg ctaccccact ggggacaatg ctgggtttcc 14520ctttaggaca ggctctgaga caaggcggag gtgctgtttg tggccacaga gcaggggact 14580ctgggttgca ggtgtggcct ggctaaagta ggctttactg ggctcctctc tgcctgcatc 14640accccccggc tgggcggttg tctctgaggc caaccttact ccctgctggg caggctggac 14700agctgccctc tccgtttgcc cctctaccac ccaaaaggca ggaggctctg gagaccagga 14760ccctgcccgc cacggcctgt gtcccaggcg tgagggggtg ccccacagac ctctgctgag 14820ctgctgctga atgacgcccc ttgggggtcc tgccggaagg tcagagcagg ggtgcactcc 14880cataaagaaa cgcccccagg tcgggactca ttcctgtggg cggcatcttg tggccatagc 14940tgcttctcgc tgcactaatc acagtgcctc tgtgggcagc aggcgctgac cacccaggcc 15000tgccccagac cctctcctcc cttccggggc gctgcgctgg

gaccgatggg gggcgccagg 15060cctgtggaca ccgccctgca ggggcctctc cagctcactg ggggtggggt gggggtcaca 15120cttggggtcc tcaggtcgtg ccgaccacgc gcattctctg cgctctgcgc aggagctcgc 15180ccaccctctc cccgtgcaga gagccccgca gctggctccc cgcagggctg tccgggtgag 15240tatggctctg gccacgggcc agtgtggcgg gagggcaaac cccaaggcca cctcggctca 15300gagtccacgg ccggctgtcg ccccgctcca ggcgtcggcg ggggatcctt tccgcatggg 15360cctgcgcccg cgctcggcgc cccctccacg gccccgcccc gtccatggcc ccgtccttca 15420tgggcgagcc cctccatggc cctgcccctc cgcgccccac ccctccctcg ccccacctct 15480caccttcctg ccccgccccc agcctcccca cccctcaccg gccagtcccc tcccctatcc 15540cgctccgccc ctcagccgcc ccgcccctca gccggcctgc ctaatgtccc cgtccccagc 15600atcgccccgc cccgcccccg tctcgccccg cccctcaggc ggcctccctg ctgtgccccg 15660ccccggcctc gccacgcccc tacctcacca cgccccccgc atcgccacgc cccccgcatc 15720gccacgcctc ccttaccatg cagtcccgcc ccgtcccttc ctcgtcccgc ctcgccgcga 15780cacttcacac acagcttcgc ctcaccccat tacagtctca ccacgccccg tcccctctcc 15840gttgagcccc gcgccttcgc ccgggtgggg cgctgcgctg tcagcggcct tgctgtgtga 15900ggcagaacct gcgggggcag gggcgggctg gttccctggc cagccattgg cagagtccgc 15960aggctagggc tgtcaatcat gctggccggc gtggccccgc ctccgccggc gcggccccgc 16020ctccgccggc gcagcgtctg ggacgcaagg cgccgtgggg gctgccggga cgggtccaag 16080atggacggcc gctcaggttc tgcttttacc tgcggcccag agccccattc attgccccgg 16140tgctgagcgg cgccgcgagt cggcccgagg cctccgggga ctgccgtgcc gggcgggaga 16200ccgccatggc gaccctggaa aagctgatga aggccttcga gtccctcaag tccttccagc 16260agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcaacagc 16320cgccaccgcc gccgccgccg ccgccgcctc ctcagcttcc tcagccgccg ccgcaggcac 16380agccgctgct gcctcagccg cagccgcccc cgccgccgcc cccgccgcca cccggcccgg 16440ctgtggctga ggagccgctg caccgaccgt gagtttgggc ccgctgcagc tccctgtccc 16500ggcgggtccc aggctacggc ggggatggcg gtaaccctgc agcctgcggg ccggcgacac 16560gaacccccgg ccccgcagag acagagtgac ccagcaaccc agagcccatg agggacaccc 16620gccccctcct ggggcgaggc cttcccccac ttcagccccg ctccctcact tgggtcttcc 16680cttgtcctct cgcgagggga ggcagagcct tgttggggcc tgtcctgaat tcaccgaggg 16740gagtcacggc ctcagccctc tcgcccttcg caggatgcga agagttgggg cgagaacttg 16800tttcttttta tttgcgagaa accagggcgg gggttctttt aactgcgttg tgaagagaac 16860ttggaggagc cgagatttgc tcagtgccac ttccctcttc tagtctgaga gggaagaggg 16920ctgggggcgc gggacacttc gagaggaggc ggggtttgga gctggagaga tgtgggggca 16980gtggatgaca taatgctttt aggacgcctc ggcgggagtg gcggggcagg gggggggcgg 17040ggagtgaggg cgcgtccaat gggagatttc ttttcctagt ggcacttaaa acagcctgag 17100atttgaggct cttcctacat tgtcaggaca tttcatttag ttcatgatca cggtggtagt 17160aacacgattt taagcaccac ctaagagatc tgctcatcta agcctaagtt ggtctgcagg 17220cgtttgaatg agttgtggtt gccaagtaaa gtggtgaact tacgtggtga ttaatgaaat 17280tatcttaaat attaggaaga gttgattgaa gttttttgcc tatgtgtgtt gggaataaaa 17340ccaacacgtt gctgatgggg aggttaattg ccgagggatg aatgaggtgt acattttacc 17400agtattccag tcaggcttgc cagaatacgg ggggtccgca gactccgtgg gcatctcaga 17460tgtgccagtg aaagggtttc tgtttgcttc attgctgaca gcttgttact ttttggaagc 17520taggggtttc tgttgcttgt tcttggggag aatttttgaa acaggaaaag agagaccatt 17580aaaacatcta gcggaacccc aggactttcc ctggaagtct gtgtgtcgag tgtacagtag 17640gagttaggaa gtactctggt gcagttcagg cctttctctt acctctcagt attctatttc 17700cgatctggat gtgtcccaga tggcatttgg taagaatatc tctgttaaga ctgattaatt 17760tttagtaata tttcttgttc tttgtttctg ttatgatcct tgtctcgtct tcaaagttta 17820attagaaaat gattcggaga gcagtgttag cttatttgtt ggaataaaat ttaggaataa 17880attattctaa aggatggaaa aactttttgg atatttggag aaattttaaa acaatttggc 17940ttatctcttc agtaagtaat ttctcatcca gaaatttact gtagtgcttt tctaggaggt 18000aggtgtcata aaagttcaca cattgcatgt atcttgtgta aacactaaac agggctcctg 18060atgggaagga agacctttct gctgggctgc ttcagacact tgatcattct aaaaatatgc 18120cttctctttc ttatgctgat ttgacagaac ctgcatttgc ttatcttcaa aatatgggta 18180tcaagaaatt tcctttgctg ccttgacaaa ggagatagat tttgtttcat tactttaagg 18240taatatatga ttaccttatt taaaaaattt aatcaggact ggcaaggtgg cttacacctt 18300taatccgagc actttgggag gcctaggtgg acgaatcacc tgaggtcagg agtttgagac 18360cagcctggct aacatggtga aaccctgtct ctactaaaaa tacaaaaatt agctggtcat 18420ggtggcacgt gcctgtaatc caagctacct gggaggctga ggcaggaaaa tcgcttgaac 18480ccgggaggca gagtctgcag tgagttgaga tcacgccact gcactccagc ctgggtgaca 18540gagcgagact ctatctcaaa aaaaattttt tttaatgtat tatttttgca taagtaatac 18600attgacatga tacaaattct gtaattacaa aagggcaata attaaaatat cttccttcca 18660cccctttcct ctgagtacct aactttgtcc ccaagaacaa gcactatttc agttcctcat 18720gtatcctgcc agatataacc tgttcatatt gtaagataga tttaaaatgc tctaaaaaca 18780aaagtagttt agaataatat atatctatat attttttgag atgtagtctc acattgtcac 18840ccaggctgga gtgcagtgat acaatctcgg ctcactgcag tctctgcctc ccaggttcaa 18900atgcttctcc tgcctcagcc ttctgagtag ctgggattac aggcgcccac caccatgtcc 18960agctaatttt tgtattttta gtagagatgg ggtttcacca tgttggccag gctggtcttg 19020aactcctgac cttgtgatct gtccacctcg gcctcccaaa gtgctgggat tacaggtgtg 19080agccaccatg cctggctaga ataataactt ttaaaggttc ttagcatgct ctgaaatcaa 19140ctgcattagg tttatttata gttttatagt tattttaaat aaaatgcata tttgtcatat 19200ttctctgtat tttgctgttg agaaaggagg tattcactaa ttttgagtaa caaacactgc 19260tcacaaagtt tggattttgg cagttctgtt cacgtgcttc agccaaaaaa tcctcttctc 19320aaagtaagat tgatgaaagc aatttagaaa gtatctgttc tgtttttatg gctcttgctc 19380tttggtgtgg aactgtggtg tcacgccatg catgggcctc agtttatgag tgtttgtgct 19440ctgctcagca tacaggatgc aggagttcct tatggggctg gctgcaggct cagcaaatct 19500agcatgcttg ggagggtcct cacagtaatt aggaggcaat taatacttgc ttctggcagt 19560ttcttattct ccttcagatt cctatctggt gtttccctga ctttattcat tcatcagtaa 19620atatttacta aacatgtact atgtgcctgg cactgttata ggtgcagggc tcagcagtga 19680gcagacaaag ctctgccctc gtgaagcttt cattctaatg aaggacatag acagtaagca 19740agatagataa gtaaaatata cagtacgtta atacgtggag gaacttcaaa gcagggaagg 19800ggatagggaa atgtcagggt taatcgagtg ttaacttatt tttattttta aaaaaattgt 19860taagggcttt ccagcaaaac ccagaaagcc tgctagacaa attccaaaag agctgtagca 19920ctaagtgttg acatttttat tttattttgt tttgttttgt tttttttgag acagttcttg 19980ctctatcagc caggctggag tgcactagtg tgatcttggc tcactgcaac ctctgcctct 20040tgggttcaag tgattctcat gcctcagcct cctgtttagc tgggattata gacatgcact 20100gccatgcctg ggtaattttt tttttttccc ccgagacgga gtcttgctct gtcgcccagg 20160ctggagtgca gtggcgcgat ctcagctcac tgcaagctcc gcttcccgag ttcacgccat 20220tctcctgcct cagtctccca agtagctggg actacaggcg cctgccacca cgtccagcta 20280atttttttgt atttttaata gagacggggt ttcaccgtgt tagccaggat gatcttgatc 20340tcctgacctc gtcatccgcc gaccttgtga tccgcccacc tcggcctccc aaagtgctgg 20400gattacaggc atgagccact gtgcccggcc acgcctgggt aatttttgta tttttagtag 20460agatggggtt ttgccatgat gagcaggctg gtctcgaact cccggcctca tgtgatctgc 20520ctgccttggc ctcccaaagt gctaggatta caggcatgag ccaccatacc tggccagtgt 20580tgatatttta aatacggtgt tcagggaagg tccactgaga agacagcttt tttttttttt 20640ttttttgggg ttggggggca aggtcttgct ctttaaccca ggctggaatg cagtatcact 20700atcgtagctc acttcagcct tgaactcctg ggctcaagtg atcctcccac ctcaacctca 20760caatgtgttg ggactatagg tgtgagccat cacacctggc cagatgatgg cttttgagta 20820aagacctcaa gcgagttaag agtctagtgt aagggtgtat gaagtagtgg tattccagat 20880ggggggaaca ggtccaaaat cttcctgttt caggaatagc aaggatgtca ttttagttgg 20940gtgaattgag tgagggggac atttgtagta agaagtaagg tccaagaggt caagggagtg 21000ccatatcaga ccaatactac ttgccttgta gatggaataa agatattggc atttatgtga 21060gtgagatggg atgtcactgg aggattagag cagaggagta gcatgatctg aatttcaatc 21120ttaagtgaac tctggctgac aacagagtga aggggaacac cggcaaaagc agaaaccagt 21180taggaagcca ctgcagtgct cagataagca tggtgggttc tgtcagggta ccggctgtcg 21240gctgtgggca gtgtgaggaa tgactgactg gattttgaat gcggaaccaa ctgcacttgt 21300tgaactctgc taagtataac aatttagcag tagcttgcgt tatcaggttt gtattcagct 21360gcaagtaaca gaaaatcctg ctgcaatagc ttaaactggt aacaagcaag agcttatcag 21420aagacaaaaa taagtctggg gaaattcaac aataagttaa ggaacccagg ctctttcttt 21480tttttttttt tgaaacggag tttcgctctt gtcacccggg ctggagtgca atgatgtgat 21540ctcagctcac taaaacctct acctcctggg ttcaagtgat tcttctgcct cagcctccca 21600agtaactggg attacaggcg tataccacca tgcccagcta atttttgtgt ttttagtaga 21660gatggggttt caccatgttg gccaggctgg tctcgaactt ctgacctcag gtgatccact 21720cgcctcagcc tgccaaagtg ctgggattac aggtttgggc cactgcaccc ggtcagaacc 21780caggctcttt cttatactta ccttgcaaac ccttgttctc attttttccc tttgtatttt 21840tattgttgaa ttgtaatagt tctttatata ttctggatac tggattctta tcagatagat 21900gatttgtaaa aactctccct tcctttggat tgtcttttta ctttcttgat agtgtctttt 21960gaagtgtaaa agtttttaat tttgatgaag tcgagtttat ctattttgtc tttggttgct 22020gtgcttcaag tgtcatatct aagaaatcat tgtctaatcc aaagtcaaaa aggtttactc 22080ctatgttttc ttctaagaat tttagagttt tacatttaag tctgatccat tttgagttaa 22140tttttatata tggttcaggt agaagtccaa ctttattctt ttccatgtgg ttattcagtt 22200gtcccagcac tgtttgttga agagactatt ctttccccat ggaattatct tagtaccctt 22260gttgaaaatt aatcgtcctt aattgtataa atttatttct agactgtcag ttctacctgt 22320tggtctttat gtcgatcctg tgccagtacc atacagtctt gattactgaa gtttgtgtca 22380cagtttaaat tcatgaaatg tgagttctcc aactttgttc cttttcaaga ttgatttggc 22440catgctgggt cccttgcatt tccgtacgaa ttgtaggatc agcttgtcag tttcaacaaa 22500gaagccaagt aggattctga gagggattgt gttgaatctg tagatcaact tggggagtat 22560tcgcatctta acaatattgt cttccaccta tgaacatggg caaactttgt gtaaatggtc 22620agattgtaag tatttcgggc tgtgtgggca cagtgtctct gtcacagcta cgcggctctg 22680ccattgtagc atgaaagtag ccataagcaa tatgtatgag tgtctgtgtt ccaatagaat 22740tttattaatg acaaggaagt ttgaatttca tataattttc acctgtcatg agatagtatt 22800tgattatttt ggtcaaccat ttaaaaatgt aaaaacattt cttagcttgt gaactagcca 22860aaaatatgca ggttatagtt ttcccactcc taggttaaaa tatgatagga ccacatttgg 22920aaagcatttc tttttttttt tttttttttt tttttgagac ggagtttcac tcttgttgcc 22980caggctggag tgcagtggcg cgatctcggc tcactgcaac ctctgcctcc caggttcaag 23040acattctcct gcacggcctc cctagtagct gggattacag gcatgcgcca ccacacccag 23100ctaattttgt atttttagta gagacggggt ttctccatgt tggtcaggct ggtcttgaac 23160tcctgacctc aggtgatcca cccgcctcag cctcccaaag tgctgggatt acagggtgtg 23220agccaccaca ccctgctgga aagcatttct tttttggctg tttttgtttt ttttttaaac 23280tagttttgaa aattataaaa gttacacata tacattataa aaatatcttc aagcagcaca 23340gatgaaaaac aaagcccttc ttgcaagtct gtcatctttg tctaacttcc taagaacaaa 23400agtgtttctt gtgtcttctt cccagatttt aatatgcata tacaagcatt taaatgtgtc 23460attttttgtt tgcttgactg agatcacatt acatatgtat ttttttactt aacaatgtgt 23520catagatatt gttccatagc agtacctgta attcttatta attgctatgt aatattttag 23580aatttctttt taaaagagga cttttggaga tgtaaaggca aaggtctcac atttttgtgg 23640ctgtagaatg tgctggtgac atattctctc taccttgaga agtccccatc cccatcacct 23700ccatttcctg taaataagtc aaccacttga taaactacct ttgaatggat ccacactcaa 23760aacatttagt cttattcaga caacaaggag gaaaaataaa ataccttata aagcactgtt 23820taatattgta ttaaattgga tcaatttggg ggctagaatg tatgttagag acatgatatg 23880tccataggtc cttgctatca cagtgaggtc tcagggacag tcgtttggta tcatttggga 23940tctcataagc agactctctc tgcttgacct gacaaatcag agtctgtgtt ttaacaggtt 24000cagtgagtga cttacatgca cattggagtt tgggaagctc cactgtaggt gcttagacct 24060tacctttgtt gttgctaata acaatgcaag catttgggag gaagacctgt gttgctcata 24120tgtgtccagg tgtagctgag gtggccttgc ttatctgctg tagggccgtt gagcatttct 24180gtagctgtga tgagtgagct gaggtgagcc tgcggagagc tcccagccat tggtagtggg 24240actcgcttag atgaactgga aggacccttt catctgagca gccactatgg agaaaaacaa 24300ccgaatgagg ggagagacaa tgtgcaattt tatttagggc acaaaggaga gctgtggtta 24360gaaggtgaca tttgagtgga aagggggcaa gccatgtgta tagcgggaga agagaggtcc 24420aggcagagtt aacagaaggc agaaatgctt tccatgtttg agaaccagta aggaggccag 24480tggctgaagt aaggtgaagg gcagaaataa ggatgaggct gcgagagatg agaggttaga 24540gacgagcgtc ttgtgcacca agataagctt gtgtggtcaa aacaagtagt ttaatttatg 24600tttttaaaag atcattttgg ctgggcacaa tggttcatgc ctgtaatacc agtagtttga 24660gacggtgtgg tgggaggatt gcctgaggcc agacgaccag catagccaac atagcagcac 24720ctataaggtc tctacaaaaa actttaaaaa attagctggg catagtggtg tgtgcctgta 24780gtcccagcta ctcaggaggc tgaggaggct ggaggattgc ttgagtccag gagtttgagg 24840ctgcagtgag ctatgattat gccactacac tacaacctgg gcaagagagt gagaccctgt 24900ctctaaatat acacacacac acacacacac acacacacac acacacacac acacacacac 24960acacacatat atatgtatat atatgcattt agatgaaaag atcactttga caataccaca 25020tgctggtgag gatttagaaa aactaggtca cttattgctg gtgggaatat aatatagtac 25080ggccactctg gaaaacagtt tggcagtttg tcataaaact gaacataccg ttagtataca 25140gcccagcagc aactacaatc ctgggcatta atcctagaga aatgaaacct taatgttcac 25200ataaaaacct atactcaagt atgcatagca gctttaccca taatatctaa gaactggaat 25260cagctcagat gtccttcaac aggtgaatgg ttaaactact cagtaataaa aaggaatgag 25320ctactgatag catgcaacag tttaggtgaa gttatgctaa tgaaaaaagc caatcccaaa 25380aggttataca tactgtatga ttctatgttt ttttgcaatg gcacagtttt agggatggag 25440aatagattag tggttgcctg gggttagaga tggggtagta gagtaggtta gtggtggcag 25500aggagagaaa agagagggag gtgaatgtgg ttataaaagg acaacacagg ggaatacttg 25560taatggaaat gctttgtctt tttttttttt tttttttttt tggcgacaga gtcttgctct 25620gttgcccagg ctggagtgca gtggcatgat cttttctcac tgcaacctct gcctcctggg 25680ttcaagtgat acttgtgtct cagtctccca tgttcagagt gaaacaaacc agaggtaatg 25740ttcatccaaa taatccaaca cacatgacat taaaacatca agatcaggtc ggacgtggtg 25800gctcatgcct gtaatcccag cacttttggg aggccaaggt gggcagatca cttgaggtca 25860ggagttcgag accagccggg ccaacatgat gaaaccccat cttgactaaa aatacaaaaa 25920ttagccgggc atggtggtgt gcacctgtag tcccagctac ttgggaggct gaggcaagag 25980aactgcttga acccgagggg cagaggttgc agtgagctga gagtgcgcca ttgcacttca 26040gcctgtgtga cagagtaaga ctccatctcc aaaaaaaaaa aaccaagatc aattaaaata 26100cagcattact gggccgggtg tggtggctca cacctgtaat cccagcactt tgggaggccg 26160agatgggcag atcacgaggt caggagatcc agaccatccc ggctaacacg gtgaaacccc 26220gtctctacta aaaaatacaa aaaattagcc gggtatagtg gtgggtgcct gtagtcccag 26280ctacttggga ggctgaagca ggagaatggt gtgaacccgg gaggcagagc tggcagtgag 26340ctgagatcgc gccactgcac tccagcctgg gcgacagagc aagactccgt ctcgggggaa 26400aaaaaaaaat aaataaatag aatgctgtag tgtccttgag tttacatgcc cctccttacg 26460cttgtgtgcc cgtgcagatt gcttgattac acaattagag gaggctggcg gaggattgtt 26520ttaatttttt tttttttgag acagtctggc tctgttcccc aggctagagt gcaatggcgc 26580aatcttggtg cactgcaacc tctgcctcct gggttcaagc agttcttctg ccgcagcctc 26640ccgagtagct gggattatag gcgcccgcca ccacgcccaa ctattttttg tatttttagt 26700agagcagcgt ttcaccatgc tggccaggct ggtctcgaac tcctgacctc agatgatctg 26760ctgccccagc ctcccaaagt gctgggatta caggcgtgag ccacacctgg ccgtttgttt 26820taattttgaa ggtgaagtga aagtgactac atttaccaaa agtgattgaa aagccaggac 26880tgttcttacc ctgtttttcc agttcttgct cagagcaagg tggtttcttt ttcacttaat 26940caccatactt acttttcatg tagaacaagt cagtttgagt tatcagttca tcatcttaac 27000taaattccat gggggaagga attagtttta gtttcttaaa cttccaggtt tgcttattgg 27060acaaaatgag atagcaaggc agtgttttta agttagattt tttatttctt tggtaataca 27120attttctcag aaacttagta gtcttttagt ttagttgttt ttagttggtc ctatgttttg 27180gatcacccct ctctacttta ttttgatagt gccaactgtg aagacatctg aagccatagg 27240tttggatggg aaggaggcat ctttagcctg atcatcttcg ccaggctgtt tatctccttt 27300tgcttggctg agaagtctta ataggaggct tattcccagc tatttgggga catagaagca 27360gttagccatt gcttatattt tactgaggtc tgtgtggtat gttgattgta gtcagttaac 27420gattttgaga actgaaggca gcctggtata tatagagtag gtattagact gtgtttcttc 27480taattgaatt tcccatctct tgtaatctat gccatcatct tctgtactgc tgagaaagaa 27540agaaagtttc taatcaaact ataccactgg ttgtaagatg cagtttggct ttagtgatgt 27600taacacatga ttcaaacgtg aaattgattg agtattggtg aaatacagag gagatttaaa 27660gccagaagac ctgggtttaa atgctggctg tatgacttca tatctgtgtg atcttgggca 27720tgtcatggtt ggcacttcaa tttcttctct ctataatggg ggaagtgagg ccagtcatgg 27780tggctcatac ctataatccc agtgctttgg gaggccaaga tgggaagatc gcttgaggcc 27840aggagtttga gcaattgggc aacatcgtga ggccccgtct ctacaaaata ttttgaaaaa 27900attagccagg cccagtggtg cgtgcctgtg gtccgcgcca ctcaggaggc tgagacggga 27960ggatcctttc agcctaggag tttaaggcta aagtgagcca tgattgtgct atcgtactcc 28020agcctgggca gcagagcaag atcctgactc taaaaaaaag taaaataaag taaaatgggg 28080gaaatgaact gctttagtaa catcatctgt tttttctgtg agcagcgtag cttgacagcc 28140attggtgaac tcgtgccctg tgcttccctg tccagatccc cattctgccc gcaacatgga 28200gtataacggt ttattcatag tagtcgagaa acactcactg aatgaatgaa tgaggtgtag 28260aactaagtgg agtgggtaat tcaacacata ttaatttcct tctttttttt atttttagaa 28320agaaagaact ttcagctacc aagaaagacc gtgtgaatca ttgtctgaca atatgtgaaa 28380acatagtggc acagtctgtc aggtaattgc actttgaact gtctagagaa aataagaact 28440ttgtatattt tcagtcttaa tgggctagaa tattctttgt gtcccagcta ttttaaatgg 28500attcagaaat ccatttaaga tgaagaagga cccttttccc atatttctgg ctatatacaa 28560ggatatccag acactgaaat gaataatgtt ccctttttgt aatcttttat gcaaaaatta 28620aaaccattat ggtaattgaa caacatgttt atgtttagtt aacaccctta gcaactatag 28680ttattttaaa accatctatg gtttgatatt tttgcatttg ttgcaatagt aggaacagca 28740caagacagtt cagtttgtct ctcttatttg ctttttcttg gcagtttgct gtcctattgt 28800acctctgctc ctagcagtgg ctggagccca ctcctctgtg cttcgggatt agtggggatc 28860gtggggcatt gactgtaggt cagctttcct tgcttgatct ttctcactgg gatgaactag 28920cagcaccttc ttttgtagct gctttgcttt tgactatctt tctgaccgtt gttcctagta 28980gctgtagatg gtaaatatat ttaggcctgt ttccaatggc tcagtaggag acatattcac 29040ctatgatatc tgaattctgt tacccacatg ggcatgcgtg aaatagttgc cttgccttac 29100tttcccttgg aataaataat tcatgttatt ctcctggtag aagctagaaa aagcctttat 29160agtcagtcag aaaaaaattt ttagacaaat aatcttgatt ttagtactga caaaaacgtg 29220tggtgattct ttttttaatt tttttttgag acggagtttc actcttgttg cccaggctgg 29280agtgcaatgg cgtgatctcg gctcactgca acctctgcct cctgggttca agtgattctc 29340ctgcctcagc ctcccaagta gctggagtta caggcatgtg ctactgtgcc cagctaattt 29400tgtattttta gtagagatgt tggtcaggct gatctcgaac tcccaacctt aggtgatctg 29460cccgcctcag cctcccaaag tgctgggatt acaggcgtga gccagggcgc ccggtgattc 29520atttgttttt tcaaaaaatt tcctcttggc cattgctttt cacttttgtt tttttttttt 29580ttttgagacg gagtcacgat ctgtcaccca ggctggagtg cagtggcatg atcttggctt 29640actgcaagct ctgcctccca ggttcacgcc attctcctgc ttcagcctgg cgagtagctg 29700ggactacagg tgctcgccac cacacccggc taattttttg tatttttagt agagatgggg 29760tttcaccgtg gtcttgatct cctgacctca tgacccgctc aactcagcct cccaaagtgc 29820tgggattaca ggcgtgagcc accgcgcccg gccctctctt gtctttttat tgtggtaaaa 29880tgcacataaa attgactgtc ttaaccattt ttaggggtac agttcagtat atatattcgt 29940aatgttgtac agccatcact gccatctact tcataagttt ttcttctgtc aaaactgaac 30000atctgtcttc attaaactcc ctatcatcca ttctttcctg tagtcccttt ctactttctg 30060tctgtatgag tgtaactgct ctggagacct catgtaagtg

gattcctaca ggatttgtgt 30120tttttttttg gtgatctgct tatttttaat gcctctgtgc atttgtatta tatactttca 30180aagtgatttc acaaaaccgt ttcattttag gttaactcat ttctgttgtt tgtgaaatac 30240tgtgtatgat tctgttctgt ttctgtctaa tttgtggaaa tgttgtggga agaaaatgaa 30300ataacaaatg agcatatgtc ctgaaaataa aaatataaaa attctaagtt agcatgctat 30360tgtagaatac aacgctatga taaaagtagg aaaaaaaaag gtttgaattc tatctctgct 30420acctgtgtaa gctgggtgac tttagataag ctgtaacgtg tttgagcctt actggctcat 30480ttttgaaatg taatccctag ttacacagtt cttgtgggat cagatggtac atgtgaaaca 30540ctgtgaaaaa gcaactgcat agatatgttc attagccacc tgagcgggaa gcgtatccca 30600ttgcgatgcc catcatccaa agctatatgt tatctttact tttttttttt tgagacagag 30660tcttgctctg ttgcccaggc tagagtgcag tggtgcaatc tcagctcact gcaagctcca 30720cctcccgggt tcacgctatt ctcctgcccc agcctcccaa gtagctggga ctacaggcac 30780ccgccaccat gcctggctaa atttttgtat ttttagtaga gatggggttt caccgtgtta 30840gccaggatgg tcttgatctc ctgacctcgt gatccgcccg cctcggcctc ccaaagtgct 30900gggattacag gcgtgagcca ctgcccctgg ccatctttac tttttttgtg aaatgacttt 30960aaatacttgg caaacatttg gtcattgttc atctgatctc caccatccag gtctcagaga 31020acataatttc tctctgaaag cttattgacc caggaaataa gatctctttc aatctgagtg 31080cgtcaggctt tattcttgtc attttgtctt ttgataattt tcaaatggaa ttcatggaat 31140gttggcttat attcatatat tagtaaagta tgttgagaca tcttaagatt gatttgtggt 31200tctatatgcc atattaaatc aaaataatag ctgttaatgg ttttcacatt agtctgtctc 31260ttgtttttat ggagtaatgc tgagagttca ttatgcttgt tctacagaag agcatgttaa 31320aaggagtttt tggagtcaga gaggttattc ttggtttcat aggatacact ctatactttt 31380tagggatttc agagtatata gctgaaggtg atattttatg taaatatgtt ttatggaaac 31440ttattgctca tcgctgtttc ctgttaactc tcctaaaata taattaaact tttggaactt 31500ttttatagct tttgtgctag actaattttt gtctctaatg aggttatata aatggcagct 31560tctgacgttt tcaatgtagg aagtcattta aaacttcatg tatattgtga aaatgtagtc 31620tgctttaagc tctctaaagt ggtctaagtt actggttcct aagtatggat gagcatcaaa 31680atcatctgga aaatttgtta aaaatacagt aatgaaggca cctcactgtc ctttttccca 31740aacatacttc tgcattctgt ttgagtaggt agggactaca catttttcac aagtatcctc 31800ttgggaatac ccaggaatgc ttacttgagc aacctcttac taatatgtac cttgataagg 31860tggctaggta aacataaata tacaaaaatc catagatctc ccatatatta gcataaatca 31920gctagaaaat ataacgttta aagatctagt tcacagtagc accaatatat cgaactctaa 31980ggaatcgata aatatgcaaa aactttataa aaacttctgt taatgtttct gaaagatata 32040ggtgaccact ttctagatag gaagatttta tattactaag ttgaattttc tctaaattaa 32100cacagaaatt taaaataatc ttgatcaaaa ttctagtaga ggtatttttg aacttgttca 32160ctgcaagaat aaatacataa ttgcaaagaa tatctcaaaa tcatcaccag gcctggtgtg 32220gtggcccatg cctgtaatcc cagcactttg ggaggctgag gcaggcagat cacctgaggt 32280caagagtttg agaccagctg gaccagtgcg gtgaaacact gcctctacta aaaatacaaa 32340aattagctgg gtgtggtggt gcatgcctgt agtcccagct acttgggagg ctgaggcagg 32400agaattgctt gaacccagga ggtacaggtt gcggtgagcc tagatcgcac cactgcattc 32460cagcctgggc gacaagagca aaattctgtc tcaagaaaaa agagaaaaaa gaaaaagaaa 32520tcaacactaa tatggtgaga cttaatgtat gtgacattaa aatagtgatt ggatgttaaa 32580acaggtatag aacagaaaga agagtgtatg tgtgtatctg tatgaattta tgatgggtgt 32640aacatatatg tattagggaa atgagggaaa tgatacattt ctctgacttt gggagaacat 32700tatatctcta cctcatattg caaacaaaca taaagttcag attaattacc taaatgtgaa 32760aaaatgaaat aatttcttta aaaaatgtaa tcttagtttg aggaaggtta acattataaa 32820ggaaaaaact gttttgagtg gaatatagtt caatatgtca aaatccacct tcaacaaaat 32880tgaaagtaaa ttgaacttgg ggaaagtatt gacagcatat agatcaaagg ttactagcct 32940gtgtaaagag cagttataaa tatcgttaag aaaaacactg tcgacctgtc ggcaccttgt 33000tctccgactc ccagcctcca gaactgtgac gagtaagtgc ttattgttta aaccacccag 33060tctgtatgtg gtattttgtt atagaaactc aagctgatta ggacactagt aatcagtaga 33120ctgaaactga aacaaaaata agaacctttt ttacctgtca aattggcaaa cattaagaat 33180attcagattt ttgtcagagg tgatacaacc ttctaagaag gcaatttggg aaaatataaa 33240gctttagatt attatatgtc tgacctagca gttttacctc tagggtgctt acccctagga 33300aagtgtgtaa tgatattggt gcagtgccct tcatcccatt agaaaattaa aaataacctt 33360aatggcctac cactaaaagg ggattgaaaa tttaagatat atttatttat gtgtttattg 33420agatggagtc ttgcactgtc cgcctgggcc agagtgcaat ggtgcgatct cggctcactg 33480caacctctgc ttcccgggtt catgtgattc tcctgcctca gcctcctgag tagctgggat 33540tacaggctca caccaccgca cccggctaat tttttgtatt tttagtagag atggggtttc 33600actgtgttgg ccagactggt ctcgaactcc tgacctcatg atccgcgccc ctcggcctcc 33660cagtgttggg attacaggtg tgagccactg cgcctggcca gatacattta tacaagagaa 33720tgttagttaa cattcataga tatttatatt ttgtttactt tttattaaaa aaattttttt 33780tagagacagg atcttactct gtcacccagg caggatgcag ttgcacaatc atagcccact 33840gcagcctgaa ctcctgggct taagtgatcc ttctgcctca gccttttgag tacctggggg 33900actttaggca gtgctactat acctggctaa tttttaaatg ttttatagat gagatcttgc 33960tgtattgccc aggctggtct agaattcctg ggcccaagtg atcctcccac cttggcctcc 34020caaagcgctg agattacagg catgagccac cacttctgac caatagatat ttatatttgt 34080gactggaaaa tatattaaca atgtgttaaa aaattcagtt aaaaaataat gaaagatttt 34140tgcttctggc taagatagaa taacaaggac agcatttatc ttcttgcctt gaaatagttg 34200aaaacggaag aaatatatgt aacagtggtt ttcaagttat tgggcatcag gcaaagaaga 34260atagttatcc caggaaaatg aatgtggaga gccctacaat ttccttacat tactgcctgg 34320tcatggcaag aggaaaaact gagaggagac tgaggctgag ccagtggttt gctgggttga 34380ggaggcagag ctgggagtgc agagatgcaa ggtggtgaga gcccatatgg aagaatacca 34440gggaagagag ctgcagaggg agctccggag acctgcaccc tgccctctca gtaccctgtc 34500atgtgtgtag ctgagtactg acgagcactt gcttgtgcgg aaatgaccca gggctggagg 34560tagagccacc tgaaaggatt agaaggaaca gttgctgaaa gtcacacagg gccaggaaga 34620atttctaatc acaccagttg gagtggaaaa cctcagctct catagagcag gtagggtact 34680cagaagggtt tgcccaccta gccccagact aagtttcgtt actctgaccc tacctaatat 34740taaaaagaga ttaattaaat tgttcgcaac aaaaataata tatttcagtg tttgtaacac 34800gtagaagtga attgtatgac aatagcataa aggctggaag agcagaaatt gacatgtatt 34860tgcgctgggc agaataatgc tcccctcttt ccccaaaaga tatcaagtcc taatccctgg 34920agcctgtaaa tattacttta tatggaaaat tgttttatga tgtgattaaa ttcaggatct 34980tgagatgagg gggctatctt ggatgatctg ggtaggcact aaatgcaatc acatatatat 35040aaaaaggagg cagagggaga ttttacacac agagagaagg ccctgtgaag atggaacaga 35100aagatttgaa ggtgctggcc ttgaaaattg gagtgatgaa gctataagcc aaggaatgca 35160gcagccacca aagctggaag aggcacggag cagttctcat ttagagccta ctccagaggg 35220aatgtggtgc tgccaattcc tttttttttt ttttttttaa gatatcattt acccctttaa 35280gttggttttt tttttttttt ttttttttta gtatttattg atcattcttg ggtgtttctt 35340ggagaggggg atttggcagg gtcataggac aatagtggag ggaaggtcag cagataaaca 35400tgtaaacaaa ggtctctggt tttcctaggc agagggccct gccacgttct gcagtgtttg 35460tgtccctggg tacttgagat tagggagtgg tgatgactct taacgagtat gctgccttca 35520agcatctgtt taacaaagca catcttgcac cgcccttaat ccatttaacc cttagtggac 35580acagcacatg tttcagagag cacggggttg ggggtaaggt tatagattaa cagcatccca 35640aggcagaaga atttttctta gtacagaaca aaatggagtg tcctatgtct acttctttct 35700acgcagacac agtaacaatc tgatctctct ttcttttccc acatttcctc cttttctatt 35760cgacaaaact gccaccgtca tcatggactg ttctcaatga gctattgggt acacctccca 35820gatggggtgg cggccgggca gaggggctcc tcacttccca gatggggcgg ccgggcagag 35880gcgcccccca acctcccaga cggggcggcg gctgggcggg ggctgccccc cacctcccgg 35940acggggcggg tggccgggcg ggggctgccc accacctccc ggacggggcg gctggccggg 36000cgggggctgc cccccacctc ccggacgggg cgggtggccg ggcgggggct gccccccacc 36060tcccggacgg ggcggctggc cgggcggggg ctgcccccca cctcccggac ggagcggctg 36120ccgggcggag gggctcctca cttcccggac ggggcggctg ctgggcggag gggctcctca 36180cttctcagac ggggcggctg gtcagagacg ctcctcacct cccagacggg gtggcagtgg 36240ggcagagaca ttcttaagtt cccagacgga gtcacggccg ggcagaggtg ctcttcacat 36300ctcagacggg gcggcggggc agaggtgctc cccacttccc agacgatggg cggccgggca 36360gagatgctcc tcacttccta gatgggatga cagccgggaa gaggcgctcc tcacttccca 36420gactgggcag ccaggcagag gggctcctca catcccagac gatgggcggc caggcagaaa 36480cgctcctcac ttcctagacg gggtggcggc tgggcagagg ccgcaatctt ggcactttgg 36540gaggccaagg caggcggctg ggaggtgaag gttgtagtga cccgagatca cgccactgca 36600ctccagcctg ggcaacactg agcactgagt gagcgagact ccgtctgcaa tcccggcacc 36660tcgggaggcc gaggctggca gatcacttgc agtcaggagc tggagaccag cccggccaac 36720acggcgaaac cccgtctcca ccaaaaaaca cgaaaaccag tcagacatgg cggtgcgtgc 36780ctgcaatccc aggcacttgg caggctgagg caggagaatc aggtagggag gttgcagtga 36840gtagagatgg tggcagtaca gtccagcctt ggctcggcat cagagggaga ctgtgcgagg 36900gcgagggcga gggcgaggga attccttaat ttcagtttag tgatactaat tttggactct 36960ggcctctaaa actgtgaaag aaaaaatttt ttgtttgttt gtttctttta agccacatag 37020tttgtggtaa tttgttacag cagctgcagg aaactaattt atgctgcatg tgaaatggtg 37080taataaggta gattgtgatg aagatacata gtataaacaa ttaagcaaca actaaaagca 37140caacaaggaa ttatagctaa tgaaccaaaa aaggagatta gaataataaa aatggtgaat 37200cccaaagaag ccagaaatag gggaagaggc aaataaagga aagaaagagc ttgatggtag 37260atttcaacct aactatgtca aaaaggacat tacatgtaaa aggcagcgat ttttcagatt 37320gaatggaaaa gtaagactcg gtatatgctg ctgcctgcaa gaaacacatt ctaaatataa 37380aggcaaaaat aacctacagg taacagaacg gaaagaagtt cactgtgctt acaagaatta 37440gatgcaagct agactggttc tgttaatatc agacaaagtg gatttcaaag caaaggctct 37500tgcccaggat gagatggtca tttcataatg atgaagggga ttcgttcatc agcctggcat 37560agcaagctga aatgtttatg caccggacta cagagctaaa atacatgaag caaagcctga 37620cagaactaca agtagaaaca gacaaatcca cagtgataga gatttcagta gccgctctca 37680atgatttgta gaacacgtag ccataatatc tggatctaga acacttgacc aacactgtcc 37740cctgtgcaac ctcattggca tttacaggac actccaccca gcaccagcag aagagacact 37800ctctcaagtg ctcacagaat gtttgccaag atagagcaga tgctgggcca taaaacaagt 37860ctctaaatta aaagcattca aattattcag agtatgtttt ctgacctcag tatcattaag 37920ttggaatata ttataggaag ataacctgga aaagcctcag atatgtggaa aaacccattt 37980ccacatggcc catgggtcag aagtgaagtc aaaagggaaa tttgaaagtc ttttggattg 38040actgatataa aaacaataga tttctaaact tgtggggtgc tgttacagca tagtaaatgg 38100aaatttctag cattaaatgc ctgttttagg aaagaaagat ttcaaatcaa tgacctcagc 38160ttctaccttt ggaaacttga aaatgacaag caaatggaat ccagagttac cagaagggcc 38220aggtacggtg gcttatgcct gcagttctgc cactttggga ggccgaggca ggtggattgt 38280ttgagactgg cagttgaaga ccagcctggg cagcctaggg agaccccata tctacaaaaa 38340acaaaaaaat tagccaggtg tggtggcatg tgcctgtagt cccagctaac caggagtcta 38400aggtgggagg attgcttgag tctgggaggt tgaggctgca gtgaactgtg attgtgccac 38460tgtgttccat cctgggcaac agaatgagac cctgtctcaa aaacaaaaac agttactaga 38520agaatggaca tcataaagat aggagcagaa gtcagtaaaa tagaaaacaa aaatacatag 38580gaaatcaata aaaccaaaag ctggttcatc aagaacatca ataaattggt aaagctgata 38640ggaaaaacag tgaagtcaca aattagcaat atcaggaatg agggagatga cagtagtata 38700gattatatag atattaaaag gactgtatga ggcaggtgtg gtggttcacg cctgtaatcc 38760cagcaccttg ggaggccgag gtggacagat cacctgaggt caggagtttg ggaccagcct 38820ggccaacatg gtgaaactct gtctctacta aaaatacaaa aattagttgg tcgtggtgct 38880gtgtgcctgt aatcccagct acttgggagg ctgaggcagg agaattgctt gaacctggga 38940ggcggaggtt gcagtgagct gagattgtgc cgttgcactc cagcctgggt gacagagcaa 39000gactccatct caaaacaaat aaataaataa aaaggactat atggtaatat tatgaacaac 39060tttatgccaa taaatttgac aacttataga tgaaatggat gagttccttg aaagacacag 39120aaactattaa agctctctca agaagatata gataagctga ttagccctat atctatttta 39180ttgaatttaa atgtaaaaat caatatttag ttactggaaa acttttaagt gtggttggaa 39240atggtatacg aactttttca actgaatttt atgaagtcta atcacaggta aaggttttct 39300gatgaaaatt tagtgtctga attgagatat actgtaaaaa atgttatata tcttaattat 39360ttcttcacat taattacatg ttgaaataat actttgggtg tattgggtta aattaaatat 39420tatgaaaatc ttgcctgttt tctttttact tttgatgcgt cagctaggaa atataaaagt 39480gtagctcaca ttctgtttct gttgacagta ctgctttgga gcacagtgtt tgaatgatct 39540atcatttcaa agacctttcc tcagttcgtt attcatggct gtctgtattc cacatagata 39600aggtctgaaa tactgctaag tggcatgttt tgttttatgc ttttataagt ttgttgatca 39660ttactgatgt ggacttttgg tgcctcttag gctcattgct atcttccaac cattgtttgc 39720aatttttacc tagagataaa gagaaagaga catttggttt cagagtagtt agattgggat 39780catgaaagag caacctcatt ttgatgcttc aaaaatagca catcccccgt attactggga 39840tttgctattc ttgggattac ttcaagaaca tccttgtgtt actggtttgg atgcttctga 39900atgctgtgaa gtcagtttca tgtacatggc tcatcagttt agctctctct tggctttgtt 39960tagacagttg gagcatgatg gcctaaacag cttctttcaa ttaaacattt taaaatagtt 40020tacaaatagt aaacaaactc cagtttttgt gactctttgt ctcgcacaac aaaaacacaa 40080tctgaccatg atcatctggc atcttagggt gaaatatggt tatactttgg cccataccga 40140aagcaagatt aaaaaggggc aggagagata gactgctgaa ctgattttca aggttccaag 40200aatattgtag gttaagagta aaagtaaact tttggtagaa agcagtgggt tgtctaggat 40260tgaagtatct gaagttttta aacgaaaatt taaaaagaaa aatgagaatt gccttacaag 40320tacaatctct tcttttttaa aaaataaact ttattttgaa atagttttag atttatagaa 40380aaaaattaga tagggtagga agttttcata taccctacat ccagttaccc cagttattat 40440catcctaatt tagtgtgaga cattttcatg tttaatgaat caatattgat atgctattaa 40500cttaagtcca gactttattc agattttctt aatttctatg taatgtcctt tttctgttcc 40560agaattccat gcaggacacc ggatacctca ttacatttca ttgtcatgtc accttaggct 40620cctcttgaca gtttctcttc tttttttgct tagaaattct ccagaatttc agaaacttct 40680gggcatcgct atggaacttt ttctgctgtg cagtgatgac gcagagtcag atgtcaggat 40740ggtggctgac gaatgcctca acaaagttat caaagtaaga accgtgtgga tgatgttctc 40800ctcagagcta tcattgttgt aggctgagag aagaagcgat cattgagtgt tcttctgttt 40860tgagtccctg aggatgtctg cacttttttc ctttctgatg tatggtttgg aggtgctctg 40920ttgtatggtt tggaggtgct ctgttgtatg gtttggaggt gctctattgt atggtttgga 40980ggtgctctgt tgtatggttt ggaggtgctc ttgtatggtt tggaggtgct cttgtatggt 41040ttggaggtgc tctgttgtat ggtttggagg tggtcttgta tggtttgcag gtgctctatt 41100gcatggtttg caggtgctct attgtatggt ttggaagtgc tcttgtatgg tttggaggtg 41160ctcttgtatg gtttggagat gctctattgt atggtttgca ggtgctctat tgtatggttt 41220ggaagtgctc ttgtatggtt tggaggtgct cttgtatggt ttggaggtgc tctgttgtat 41280ggtttggagg tgctctgttg tatggtttgg aggtgctctt gtatggtttg gaggtgctct 41340attgtatggt ttggagatgc tctggtatct gcctgcattg cttgccacac ctgcccggtc 41400agaaggcgct atgttgacaa ttgtgcctgc acggtgccta ggtcaatgaa gggaaccgat 41460ggtagccact ggatgctcct gggaaaatgt cactacaggc accagagaag ccagagctat 41520gcccaaattt ctatgagtct cagttttctt aaccataaaa tgggatcaat gtttttgtgg 41580catgtgtatg agtgtgtgtc tgtgtatgtg tgaggattaa attgtgtatg tgtgaggact 41640aattgccact actggatcct caaagtggta agaagtgttc ttattaataa tgacatcctt 41700acactcttac ccagcaagat tgatgggtgt ggcactgctt ctctttttcc atcacatggt 41760ttccatggta tccttttgcc cagggaatct ttgctttgtg gctagcactt tgttgtttgg 41820ctaatcacgc tttctgtggt caggacgctg gcttctctgg agccatggga ttctagctcc 41880ctgtcttgtc cctagagtgg tcactgtctt ctctctccgc ttgcaattcc tgctttgctc 41940gcatctcact tatgcagtga cgtatatcag tttcaccttg ttctccgtgc ctgctgatca 42000ttggcaccac ttgcatggtg ccatttaggg cctgcttcca gttaagcttg cttctccaca 42060ggcctaaata tccttgcttg cttcttttat tctcactggc aggaccaggg cggtctgtct 42120ttgcatgaga cagggtctcg ctcagtcacc caggctggag tgcagtggct gatcacggct 42180cattgcagcc ttgagctacc gggctcaagc tatcctcctg gcttggcccc ttgagtagct 42240gggactacag gcgtgcacca ccatgcccag ctaattttta aaattatttg tagagatggg 42300atctcgccag gttgcccagg ctggtcttga acgcctgggc tcaagtgatc ctccctcctt 42360ggtttcccaa agtgctggga tcacaggtgt gagccactgt gcctggccct tgatgtttca 42420gttcttgata tttgatcctc agagtcagaa aatctaaaaa gagggctatc ccaggttgcc 42480ttggttcatg gcaaatggga cgttaagagg gcagagagaa tatgaacaga aactgttcta 42540atattggtca tttaatgtgt aagtattgtt cttttttaaa cctccttcat tttttttcca 42600ggaattgctg gacacagtgg cttggtgtgt gtctgaggac tgtaggccat ggccctaggt 42660tgtggtttta ggtctcaggt gctcttcctg gctgtctcct tgcttctttc ccatgtcctc 42720ttctttgttt ccagccattt ctcccttatg cttaagtttg gtgcagcagg gtttggctgc 42780tctcagattc ctgcttcctc agatgctgta gttgtcaggc ccagcgggct ggcagcggga 42840tcaggatctg gctaggtttg ctctcactgt ggcagagtag ggggaggcgt gggagagcac 42900gtgtgacccc aggccagctg tagggagcat aggcatggtc acgtagcctt caggtcctag 42960actttgtctt ctcatgagta tggctgtgtg tgtatggtga aaactaggtt ctacttagcc 43020caagaaaatg ggcacatttt gcatgtggtt tctgtagaga aatgcactgg gtatctgaca 43080tagcctggca gcatgcctcc ctcaggtagg ttagtctcag gcggtgaagc acgtgtgtcc 43140agcaagaact tcatatgtgg cataaagtct ccgttctgtg aggtgctggc aaatcaccac 43200caccgtcaag aggctgaagt gatttttgtc tagggaggca ggaaaggctt cctggagtca 43260gcagccagta ggtgaaagag tagattggag accttcttaa tcatcaccgc ctcttgtctc 43320aaggggtgcc aggaagctgt ggaggctgaa cccatcttat gctgccagag agtgggacac 43380catgagggtc aggtcaaggg gttgtacctt gtttggtaga gaattagggg ctcttgaaga 43440ctttggatgt ggtcagggga gtgtatcatt taggaagagt gacccggtga ggacgtgggg 43500tagaggagga caggtgggag ggagtccagg tgggagtgag tagacccagc aggagtgcag 43560ggcctcgagc caggatggtg gcagggctgt gaggagaggc agccacctgt gtgtctgcgg 43620aagcaggggc aagagggaag aggccagcag cgtgctgcca tcacccagcg actggcgtag 43680attgtgagag accattccct gctcttagga ggggctgagt tttagttttc tcttgttata 43740caataagctt ggtatttgtt tacaaaacat ttgtaaagct aaatcaaggt ttgataaggc 43800ttctagtttt atttaagaag taatgttgaa ataaatgttt gtccaattcg ctttgctcat 43860ttaaggactt tcagtacaaa ctgcaacaac aggattagga tttaaacgtt tctgagatgt 43920ttttactcct cagaatttcc cagaatgtga tctggttttg attttcaagc ttgctgaccc 43980aataggttaa cccacaagtt ttacgaagac catctcagtc cacttacatc aactgcccat 44040gccacggtta aagagatcat cgactgatgt ttggcacagc ttcctccctc ttgggtgggc 44100aagcatttgg aagagaaggc tcctatgggt gagagtgggg caccaaagtc ttccctgtcc 44160catcccctag cttgagaagc ccttctctaa tgtggacttt gtgccgttag catcgttact 44220agcttgaagt tgaccatctg gacgtacttt ctggtttagc ctcacaagtg agcaaggagg 44280gttgagagat gtgctgtgag gaatgtgggg ccccagctgg cagcaggctc tgggtcaggg 44340gggcagggac cacgggcata cctgacagtg aggaggggcc acacctgcag aaaaggatgc 44400aggactccgc cttgggaagt gttctaggcc agagcgaggg tctgtggttt ataagtacac 44460ccacagtgct cgggaccctg cagatgtcca gggtgccgtc tgagcccgta tcatccaaca 44520gaatgttctg ctagtgaaga ttaaagattt actccagggg ctttaggatt tattatatat 44580atataaatcc tatatatata attttttttt tttttttttt tgagatggag tttcgctctt 44640gttgcccagg ctggagtgca atggcgtgat cttggctcac tgcaacctcc gcctcccggg 44700ttcaaactat tctcctgcct cagcctctcg agtagctggg attacaggcg cccaccacca 44760cacccggcta atttttgtat tttttagtag agacggagtt tctccatgtt ggtcaggctg 44820gtcttgaact cctgacctca ggtgatctgc ccgccttggc ctcccaaagt gctgggatta 44880caggcatgag ccaccccacc tggccaggat ttattgtatt tgaaccatct accattttaa 44940ttttgatgtt atgtagtatt tgatgataat gaaagttaaa ttgtttttct ttccattttt 45000ctgtttaagt gaatgacctg tatctagttt attcagtaac ttcctgcata tatttgtttc 45060tttcattctt aatgaatata ttcttaattt agttgctatt atgttttgct ttgccccaaa 45120attgaaatct tagtttcctt ttagctcgtt ttagaactag

tgatgggatg tgtcttccat 45180aaatctcttg tgatttgttg taggctttga tggattctaa tcttccaagg ttacagctcg 45240agctctataa ggaaattaaa aaggtgggcc ttgcttttct tttttaaaaa tgttttaaat 45300tttaaatttt tataggtaca cgtattttgt aggtacatgt aaatgtatat atttatgggg 45360tacatgagat attttgatac aggtatacaa tacataataa tcacaccatg gaaagttgga 45420tatccatgcc ctcaagcatt tatcctttgt gttacaaaca atccagttac atgctttact 45480tattttattt tatttttgag acagagtctt gctttcaccc atgctagagt acagtggcat 45540gaccttggct cactgcaacc tccgcctccc gggttcaacc gaactttggg ctggtctcaa 45600actcctgacc tcaggtgatc cgcccgcctc ggcctcccaa agtgttggga ttacaggcgt 45660gagccactgt gccgggcctg attgtacatt ttaaaataac taaaacagtc agggcacagt 45720ggctcatgcc tgtaatccca gcattttggg aggctgaggc aggtgatcac ctgagatcag 45780gagttcgaga ccagcctggc caacatggag aaaccctgtc tctactaaaa atacaaaaat 45840tagccaagtg tggtggcggg cgcctgtaat cctggctact cgggaggctg aggtagggga 45900atcgcttgaa cctgggggtg gaggttgcag tgagccgaga tcacgccact gcattccagc 45960ctgagcgaca gagtgagact ttgtctcaaa aaataaaaat gaaataaaat tgggccgggt 46020gtggtggctc acaccttagt cccagcactt tgggaacctg aggcaggtgg atgcttgaga 46080ccaggagttt gagaccagca tgggcaacat ggcaaaacgc tgtctgtaca gaaattagct 46140gggtgtggtg gtgcacaact atagtctcag ctacttggga gattgaggtg ggaggattaa 46200ttgagcctgg aaggttgaat ctataggtag ctgagattgt gccactgccc ttcagcctgg 46260gcgaccaagt gagaccctgt ctcaaaagaa aaacaaaaaa acaaaaaaca aaccactatt 46320atcgactata tattattgtc tatgatccct ctgctgtgct gtcgaatacc aggtcttggg 46380cccttatttc catcactgag caaacttcac tctgttaagc agcaggtgtg ggatttcatc 46440gttattcagt aattcacaat gttagaagga aatgctgttt ggtagacgat tgctttactt 46500ttcttcaaaa ggttactctt tattagatga gatgagaatt aaaaatggta acttacttta 46560tatctttata attgaagccc actagacctt aaagtagtta ccagatgttt tatgcattta 46620aatggccttt tctctaaaat tagaaagtaa caaggaaaga aaatgcttcg tttctatgca 46680accctcttgg tgactagtat gtgactctta atgcaaccct cattgcaccc cctcagaatg 46740gtgcccctcg gagtttgcgt gctgccctgt ggaggtttgc tgagctggct cacctggttc 46800ggcctcagaa atgcaggtaa gttgtacact ctggatgttg gtttttgtcg ggggccagct 46860gctactgatc ctttatgtct cagctcagat gtcatttcaa aagtctgctc tgccctctcc 46920aaattgcagt cgaccttgcc ctgtttatgt ttccctcata gcactaatcc atgtcagaaa 46980ttgtcacgta cagtctatct gtgtgcttgt ttattttcta tcccaccctt ccgcaagaga 47040cttatgggat gtgtgcccca ggacagcagg ggtcttactg tcttatgctc tgttgcagcc 47100cagcagcgat aacagtgtct gcacatagta cttgcttaaa agatacttgc caaattgttg 47160aaggttgagg taccaatttc attattgctg actataggag ttatagcaaa atatccattt 47220gtctgttaca tgagttaaaa atatggttgt tgcactgtga atagtttggt ttagtcaaaa 47280cagttgtatc ttaacggatt gagaaacaaa agcaggacca cttttcatca gctccctcct 47340tctccttaac cagcaataca tgctgatgct gatatcccat agaccctcag ctccatcctg 47400agtcactggg aatgtggtct aaaccctcac tattaatatg aactgagttt caataagaat 47460cttatatggg tcgggcatag tggctcatac ctttgatccc agcacttcag gaggccaagg 47520caggtggatt gcttgaccca gactaggcaa catggtgaaa cgccgcctct acaaaaaata 47580caaaacttag ccaggcatgg tggtgcgtgc ctgtggtcac agccactcga gaggctgagg 47640tgggaggatc acttgagcct gggaggtgga ggtcgtgttg agccaagatc gcaccactgc 47700actccagcct gggcaacaga gtgagacctg tctcaaaaaa accaaaatcc agaaaagaac 47760ttatatggct gcagaggtat aatcactaag gaaatttcct tttgtataat cttttttctt 47820ttactatcat ttaaaaaaat gtgttatatt tctgaagcaa cacatccagg ttctgcacat 47880agcagccaaa gtgaccttaa agaatataac tgggtcttgt cattccctta tttaaactct 47940tgtacccatt tcccagtgcc gtttagatag agattccaga ctcgtcaatg gctctgtcac 48000ctcagacacc ctgcattgac tcattagtct gattagagtc aggtttttct tcctcctgat 48060ggtttttttt tcccccttag ttctcagcgg aacagtcact tccttaggga ggtttcccca 48120gccaccctct gaggccgtgc ttgttgccag actctgccac tagagggcag ggctgcacca 48180ctcctggcac ctcgcacccg gcctgccctg tcactctgtg tgttgggtga attcctgtga 48240tctgtgactc actgctctgt gtcctacaca ttcggctttt cttctctccc cacaacccca 48300ttttataatt ctcctttttc aggaaagctt tattcccatt taaaaatttt tgtttttaaa 48360atggtatttt cttacactta ttttctaatt aaaaatgagt gttttaagaa gtattatgat 48420ttactgcaaa taatttttaa acccagcctt ttagatcctc tgtgatcata agagaaatga 48480aggatgtctc ccaacacttg agcttcatcc acatttcatc ctcctgttct ttcagctgag 48540ttttccccat cccattaggg actgttggaa tataaaactg gcttttccct aacagggaat 48600gaattgcttc tgtttctcct gaaggagagc tggaagaatg acttgcgttc ttttgcatac 48660acaggcctta cctggtgaac cttctgccgt gcctgactcg aacaagcaag agacccgaag 48720aatcagtcca ggagaccttg gctgcagctg ttcccaaaat tatggcttct tttggcaatt 48780ttgcaaatga caatgaaatt aaggtatgat tgttgcctca ggtcacaaac atgcgagtga 48840tgctgtgagt gagtctgtgg agggtgaggg cttctgaaca gggagtcctg tgggagtgct 48900tcttggggta tgttgtatgt cgtaatttag actaccatca tttgtgttat ttttgaggca 48960cctaaggact tctttccact tctcatttct tactgtgggg tgaagagttg aattgggaga 49020tggtttctag atgcaaattg aaaaggcatt tttccagagc agatttgttt tcggcgtact 49080agagtgactc tttaacctag ctgcgggaag atgactgtgc caagactgca ggtaggagaa 49140agctcactga cgaggccttg tgggtctgaa cgtcctgcag ctatcagagc ctgttggctt 49200cctgttgtgc attccaacaa atcatcttca aacccacttt agtgttttgt ttataatgtc 49260cagaaatagt gaccctgtca catgctctac agattacagg attcttagcc tcttcctttt 49320tggtaggtca gtcctgggtt tgagcccaag tgaccctcct gggaggtgat gatacacact 49380gggtagagtg gaatcagatg gacttggatt agaattctgt cctctttact agttattttc 49440ctctaggcaa actgcccaac agctctaagc tatttccttc gtattctgaa aaataagcct 49500taatgggacc catatagggc aactctgaga gtaaaataaa ggaatatgtg ttagagtgta 49560gcatagtcac ccacgggaag ggcttagatg ttagctgcta ctgctcttat tagctgaatg 49620atttggaata aactgttagc ctctctcatg ttttttctct tgagcttcga agttttcttg 49680ttaatactaa ggagatattc aaactagtca tggggttttg gaatgacgaa gggagatgat 49740gaatctaaag aatttagtgt aatatttctt catgctcagt aaatggtagt ttctgctgct 49800gttattttta ttaccatctc tttggaatgg gagtaggtgc tcctttgtgg tcagaggctg 49860tgagagctcc acagcgccag tttgcccatc tgtacactgg ggtctgttga aggcagtccc 49920ctctgtgata tctctggctg tcagagctca gatgatagat ggtatttttg tactcttagt 49980tctcatcatt ttcatgattt cgatcaccat ttgagtatga tgatgctaac actttgttga 50040acgtagaatc cgttaattac ttccttcctg aacctttggc attaaaaaaa atctattctg 50100ctacctctct gctcatttat ggttattcaa atttattatc aagagcctgg tacagtggct 50160tgtgcctata attgtagcta cttgggaggc tgaggtagga ggattgcttg aggccaggag 50220tttgagacca gcctgggcaa gatagtgaga ccctatctct aaaaaaactg aaaaaaaatt 50280agctggacat gatggcatgt gcctgtggtc ctagctactc aggaggctga gacaggaggc 50340tcggttgagc ccaggagttg gagttcgagg ctacactgag ctgtgattgt gccaccacac 50400tccagcatgg gtggtaaaac aagatgccat ttcttaaaaa aaaaaaatat atatatatat 50460attatcaatg aaattcagta gtaccaacag gattataaac aaagatagta gttcccttcc 50520tactttttct cttaatcctt gtgtctcaca ggcaaacata actcttagta tttcttccaa 50580tatttacttt catgtttctt tctttctttc tttttttttc tttgagatgg agttttgctc 50640ttgttgccaa ggctggagtg caatgacgca atcttggctc accacaacct ctgtctcccg 50700ggttcaagcg attctcctgc ctcagcctcc tagtagctgg gattacaggc atgcatcacc 50760acgctcggct aattttgtac ttttagtaga gatggggttt ctccgggttg gtcaggctgg 50820tctcgaactc ctgacctcag gtgatcctcc cacctcagcc tcccaaagtg ctgggattac 50880aggcgtgagc cactgcgccc agcaacttcc acatttctaa ataacatgct tctactgcta 50940tttttttttt caattttaga cattttttta ctttcactat agttctatca gaattcagtg 51000tgtacgttat tatgcctaag taaatagtca tggttgctta cgtattatat ttctttgatt 51060gtgtttctta tttgatgaga aagctgtgtt ttttgctctg ggttgaaact ggagagagga 51120cctggggagg aggaggagga cagatgaagt tggtgactgt accttcatgg ccatagctgg 51180gttctcagca cccggggatc tgctgatcac ctactcatag gccaggcccc tatcgaagtt 51240ctaggtgacc cagtgctggg gacggggggg ccacctgcaa ggtctaatca tggaggtggg 51300ggctacagtg ttggcttgtg ctggggccag catccttagg aaggcatctt ggaggtggag 51360gagacagccg cccacttctt gattggggcc ttcagcagca ccagcttctt gggcaggctg 51420gtgctggctt tcatcaccat gtcgtgttca atcttcttcc agatcctgac ttctaggttc 51480agctttcctc agaccctggt tcctttcaga ggccattgct gctgccttgc tctttgctgg 51540cttgtgcctt gattatatgt ctttgtacaa ctttttgttt tcctggagtt aatcttcaca 51600tctgttttct tggagttaat cgttacctct atatcgcttg cttattattc tttggccttt 51660ttgtcttctc acaccttcca acttctttgt aatatgtgtt tagtacaatt tttcatgaca 51720ggtagtttac tgaatcagtt tttccccagt gtggtcatcc aacttgagtt atccagctct 51780ctgccccagt ctgggcaggt tgatcttcag gtctgtagta cacttgtatc ctaggacttc 51840tctttgccat tagcctggaa tttcctttgc agttctcccg ttggatgccc agttcctaga 51900tgccatatgt ttttctatcg tctagtagct tcctgagaga agatgaatgg gagggaaatt 51960gtatgaggtt ttgcattcat aaaaatgcca ttttttttcc tgtacacttg gctgggtatg 52020gtgttctggg gtagaaatca ttttccctca gaaatgcaaa gtctttgccc tgttgtctta 52080aaatctccaa cgtgacccga ttccttaacc tatgaatgta cttttctttg gaagctttcc 52140atttttgggg aggtgaagtg ctaggtactt agtaggcctt ttaatttgga aacttacatc 52200ccttcagttc tgggaaaatt ttcttaacat ttctctgaga agttcttgcc ttttattttc 52260tgtgttctct cctgaaattg gttagttgga tgttggtcct cctagattga ctcacatctt 52320acctttttct tttctttttc tggtactttt tagatatcca tctcaaactc ttctattcat 52380tgttatgttt ttaacttctt tcttttcttt gtctcttgat ggggtcttgc cctgttgccc 52440aggttgtggt gcagtggtgc gatcatagct cactgcagcc tcaaattcct gggctcaagc 52500agctgttctg cctcaccctc ccaagtagtt gggactacag gtatgcacca ccacgtccag 52560ctattttctt tacttttttt tttttttttt tgagatggag tcctactctg tcgcccaggc 52620tagagtgcgg tggtgggatt ttggctcact taagcctctg cctcccaggt tcaagcagtt 52680ctcctgcctc agcctctcaa gtagctggga ttacaggtgt gcaccaccat gcccggctaa 52740tttttgtatt tttagtagag ccagagtttc accatgttgg ccaggctggt ctcgaacgcc 52800tgacctcagg tgatccgcct gccttggcct ccgaaagtgc cgggattaca ggcgtgagcc 52860catcattaga tctttaaata ccagtatcta taagtctttt cctcttgagt cagctagtat 52920ccctggaagg aaattactca ttttcctgct tggaggctat aagcttggct atgtttatcc 52980tgcaaccggg gactggaagg gaggggactg acagtgttgc tggtcagggt gccctcttac 53040tttttgtttt ctgtgtgcat ctcacgtctg tcctcagcct atgtaaacac ctcttgagat 53100tatccctctc aatctttgcc ggaggtgggg gaggggctgc ttcctgggct gccttggatt 53160ggagggaaga cctcaggtga gtgggtggga atttgcccaa ggagccatga gaccagccac 53220tatttcaccc tctccatccc tccactttca gatgtatgtg gcgcctccaa agcccgagct 53280cttcttggcg tctgtggctt caataagctt gctttttgct ggtatccctc ctaccctccc 53340ctgtccccag caaagcttgc atttgaactt cttcctacgg gctaacaaat cagtcagtta 53400tgtagctctt gttacttttt agcttccgaa gttttgttga cacccgtagt ctgctaatgt 53460ccctgttctg ttctttctgt tcgtgtaaat atatgcttta tacaacttct ttacatgatt 53520tttgtggggt ttctgggtag cagagcttca caagttcaat ccagcgtgtt ggattagaaa 53580tctcccaccc tctggtttat tcttattctc aaaattacct gccaaacact gatactccct 53640tgtttttcct tttcctgaca ggaaatgtac ataccataca ggacagaaat cattagtgta 53700tcccttggtg aataaccaca aagtgaactt aacccttgta accgccaccc aggtcaagac 53760agaatattac caagcactca gaagcctctc ccctattccc ccgtcactgc tcctgccttc 53820ctccccaagg tcatgactgc tggcttctaa ttccagagtc tgtttttaaa ttctgtgtac 53880atagaccatg gattaagtgt tctttttgtc tggtttattt tggtcgacat taagttcatg 53940agagtcttct atattatcgt gtgtattagt attcctgtag ttttaggagc ttcatagcat 54000tccattgtag ggatatacca cagtttattc attgtattat cactgggttg tttctagttc 54060ttggctattg cgagcagtgc tactgtgacc actcttaggt gtgtcttttg gagtacatgt 54120gcaggtttcc atcttgcaca gctagaggtg gagttgttgg gtgatagggt gtgtgcatct 54180cagctgcagt agaaactgcc aaatagcttt ccttgagtgc ttgtaccagc tcaccctttt 54240gccactgtgt atggggattc caggagctct ggtcctcgct agcacttgga attgctgatg 54300cttttactct tagccttcct gatgggtgtt ttctggaatc acattatgat tttaatttcc 54360attccttaaa gtacccttgg ctctgaagtt taatgattca tgcatctctt cccttttgaa 54420gtactcttac aggtatgttg tgcatgtgtt gaaaagtggc actatctatt ctaaaataca 54480gtatgcctcc tctgtgtttg aacagttgta gcgtggcctt ggggcctcct gttagctggc 54540ttggagaagg gattcttggg attgtagaga ttagacctga ggaggcccct tggagctctc 54600tgactaaatt ttattcttta ttattccaaa ctatttaagc tcaccgtgtg ctgactcatc 54660ataataatga gtagctctca ttgtgcttgt ctatttggac tcatacaatg attttttttt 54720tttctttgag acagagtctt gctctgttgc ctaggctgga gtgcagtggc acaatctcgg 54780ctcactgcag cctccacctc ccaggttcaa gtgattcttg tgcctcagct tctcaagtag 54840ctgagactgc aggtgcgtac caccatgcct ggctaatgtt tgtattttta gtagagacgg 54900ggtttcacca tgttggccag gttggtctca aactcctgac ctcaagtgat ctgccttctt 54960cagcctccca aagtgctggg attacaggtg tgagccactg agcttggcca aagtagtttt 55020ttaagatgtt agtatctttt cttgcagcta aaaaagtttg tcagagatga ttctactttg 55080ttctccaggt gttttctcag ggagaaattg gaggcagtaa gccactgggg gagtcctgtg 55140gctggggggt ggggtagtcc tgtggctcct tgtcagggag tcctgtggct ggcaaggaga 55200gaagtcctgt ggctgggttg ggagggagtc ctgtggctgg ggtctcatcc tgtgcctaac 55260agtgtccaga ggtgccgaga ccagctcagt cggggagacc ctaacccagc agcgctagag 55320gaattaaaga cacacacaca gaaatataga ggtgtgaagt gggaaatcag gggtctcaca 55380gcctttagag ctgagagccc tgaacagaga tttacccaca tatttattaa tagcaaacca 55440gtcattagca ttgtttctat agatgttaaa ttaactaaaa gtatccctta tgggaaacga 55500ggggatgggc cgaattaaaa gaagaggttg ggctagttaa ccgcagcagg agcatgtcct 55560taaggcacag atcgctcatg ctattgtttg tggcttaaga atgcctttaa gcggttttcc 55620accctgggtg ggccaggtgt tccttgccct cattcctgtc aacccacaac cttccagtgt 55680gggcattagg gccattatga acatgttaca gtgcttcaga gattttgttt atggccagtt 55740ttggggccag tttatggcca gattttgggg ggcctgctcc caatacagag gtctcgtgta 55800aattccctgg gaggcgataa gcctctgaga aacagactat gctaaccacg ccatgaaaga 55860gaaacttatt tataaatcag atgccagtta ctagtttact gcttatttgc ccaggcgtag 55920ctctgacaga gtccccgact catagtgctt gctcagtgca tgctgaacaa tgattggaat 55980caagtcatgg ctcagagcat agttttgaat aatgggaaat ggatgttctt aagtaacata 56040gtcaccaaga taatgcgact agctgggtca ccccttttca attttaggat atttttatca 56100agatttaaat ggccatcatt agagttatag cactttctcc tttggattgt cctagaggcc 56160catgagaaag tattccctaa tttcttagga gaacagtttg tgggtagtat gcggtcatgt 56220ccagttaaat tgcagatatt tccgatcgaa gatgttccag tcctgagaac ttcgtgacat 56280tagcaggact tctacaagcc atctcttagg gtggggcatt tactgcagtt ggctagtact 56340cttttctcct taactttgtc atttgttgat ttttttttaa ctgtccccaa atactgtggg 56400cagagtgtat ctagaattga ggcctccacc attgcggaga ggacatggat gctgagcagt 56460cccctgagtg aaggttataa agaagcaaat agactacaca tgtctgtaaa ctgctcttga 56520gtgtcccaaa tttggggtac ttcagttcag ctgtaggaaa agcctcaaac tgtttatact 56580ttgcaagaat tggaaacttc taattcacgt taagttttat gtaatacatg ataagcttca 56640taggagcttc atcttttatc tacttggact tttgcttccg taggttttgt taaaggcctt 56700catagcgaac ctgaagtcaa gctcccccac cattcggcgg acagcggctg gatcagcagt 56760gagcatctgc cagcactcaa gaaggacaca atatttctat agttggctac taaatgtgct 56820cttaggtaag gtggaggcat atgagtggaa gagtctccag catgtactca agatagacct 56880ttgaaataaa taaaaccaga tgatccctca gcttctagac caggctattt ggcactggtt 56940gattgaatgt gaactgcact ggggctgctg tgagcccgca tgggtctctg tgaccctgca 57000gatgcagccg tgcccaggga ctgggcagtg ggtgtgggct ggtgtgagcc ctgtctgcca 57060cccagggcct ggccctctgt ctgtgtcggc catgactatg gtgagtcttg taggcttgag 57120actgtgcctc gggttcctgc gggttctctg taggtcagtt gacagtttct cctgttgttt 57180gggtaactgt ggaaacgaac actggcaagt gctgaagcga gcatgtggac gtgcgatatg 57240aaataacgac ctggctttca aaggcagtga ggctctctgg aaaggacctt gctgagctag 57300ggatgtgggt gtgtagccat tcccagtggg cctcatggcg tactcgttca tgatcatgtt 57360tgtgccatct tgatctctca ggatctcttc ttttttaaca gattaagccg ggaatctcca 57420aacagtgagt cagatgttaa gatgtcttgc ttccaccccc acaggcttac tcgttcctgt 57480cgaggatgaa cactccactc tgctgattct tggcgtgctg ctcaccctga ggtatttggt 57540gcccttgctg cagcagcagg tcaaggacac aagcctgaaa ggcagcttcg gagtgacaag 57600gaaagaaatg gaagtctctc cttctgcaga gcagcttgtc caggtaggag cacagggttt 57660actctaggcc ctgcatgtga atgactgaca ttcaaagaac cgattaattt ggaagagaag 57720cggcagaacc gagagttaga ggtgtggact ctggagctgc gctgctcgtt tccaacccta 57780ggtgctgacc tctagctgtc ttccctctgt atgtccctgt caccgtgagt caaatgcggg 57840tgatgcctcc tcaggtgccg tgttacctaa gcctctcaga gaccactgct accctgtttc 57900taaaaccaga ggtcacgata tgtgttcatc cacccagtaa atactgattg agcacccact 57960gtgtgctagg ctctgggata ggggctgggt atacaatggt gagtatttca gctgcagctt 58020ctgccccgtg gaggctgtgg cctagcacac tggtctaggc acggtggtat atgctcactc 58080aaggagatag ggacgtggtc gtttggggtg tcggaacaaa atgtcggaac ttctctttcc 58140aatgcagaga aaccttgcag taattctaat gtactgtgat tggcagttga cttcagttct 58200ttgtagcacg cttactcagg ttatttcact aactatgtaa ccatgcagcc tcattttaag 58260caattggatt ttttgaactt tacttaaaat gttatgtcag ggtttttatt gtgcttaatg 58320tgtgccattt agctaagttt tgtaggatac gaaattgtaa gtggcttaaa atgattctta 58380atagaatcat gaattgaaga taatgctaat aatttaagca ctgagttagg tagtgtttgt 58440aaaatgctta gaatgcttcc tggcacatgt taaggccatg taagtgctgc gtgttgataa 58500acagctgagc aaaagtggac tcttaagaaa gtattggggc tgagagttct gttccaacca 58560gctgcccttt ggttattttt cagaataaaa gcagagtctc atgggatatg acatttatat 58620ttccttcaca aaaaacactg ctgagtgttt tgttgagtaa aaagggtgta gccatggtaa 58680taatacattt aaaatatagt ttatttcatc tttaccttgc cttgtttttt ttttaagcta 58740gctttttatt gagaattcca cacatacaaa agtatcaact catgaccagt tatatttcat 58800ttataatcct acttctccct ttttttatta tttgaaagca aaccccaatt atcctcttat 58860ttcatctata agtatttcag tatctctata gatgaggact cttctttatt tttaaaactt 58920tatttttaaa atgatggtca gatgcagtgt tcatgcctgt aatcccagaa ctttgggagg 58980ccaagctggg cggatcactt gaacctggga gtttgagacc agcccgggaa acatggcgaa 59040accccatgtc ttaaagaaaa aaatcagcca agtgtggtga tgcatgcctg tagtcccagc 59100tacttgggag gctgagatgg gagggtcaca tgagcctgga agatcaaggc tgcagtgatc 59160catgattgta ccactgcact ccatcctggg tgatggagca agattctgtc tcaaaaaaac 59220aaaactgcaa aacaacgtca caaaacagtg ccattgttag acctgaaaat attaaacatt 59280tcctacatca aatacccacc aactcattat caatttttct ctctactctt ttggaatcag 59340catctaaata aaattggtcg ataaggattg taaatctctt tgatgaactg gttcccctcc 59400atcccagttt ttttccctta gagttcattt attgagaaac cagattgttt gtcttctaag 59460ttttcctgtg gtctgatata ctgcttccat ctccactgtg taaattaaca cctttttctc 59520ttctctgtat ttcctgtaaa tcaataattg gaggaaaagc cttgtcagat ttagtgtata 59580ttttatatct gagtccagta tttcttatat aatattttaa gataagtgta ctcttttaaa 59640aagtattgaa actatatgct caattttttt taactgatgc ttttaagaag gctgcttgat 59700cataaaagtt tagagatcat tggtctgatg ggaaaagcaa ataattacta aaccgtttag 59760caaggttgag gtgcacatgg tggggcctgg agaagttcag tcatgagccg tcacttatgg 59820gcacgtggaa tctgacccgg cacagagttg ggagaagaca ggagctttat agacagaaaa 59880tgtggtcttt gctaagtccc aggagtgaaa gggtgagaca gtgctcacag cacacgagtg 59940tgggtgcgta gacagagcaa gggtgggtcc tgaaaaggcc tgcaggcttt ctcatagatt 60000agcaagagtg ctggttacgg aggtttctaa catttgtgaa cagatcgaaa ctgtgttaaa 60060ttgggattgc agtaatcctg gaaggacagg gatagagggt gaaggggaaa aaagggtatg 60120gatgtgagac ttaattgctg attttcttaa gacctttctc caaagtaaat aaatgatgtg 60180gcacattttt gaactggcaa attctaaact ctagatatga

ttatctctat aacatatctt 60240actccatctt cttttgacta aaaactgttc ttaattaaat taccatgaga cgttcaattc 60300agcaaatgta gtttggctaa ccatatttaa ttagaattta atataatcct aggcctggcc 60360aaactattaa gcaagtgtgg gcaaaatatt gataatttta gatatgcagg aacttagttt 60420gctttccatg tgtgcttttc gaaaaaggaa taaattgaaa aatagaggaa gccctgaaat 60480ccaagaagca aactctctca cctaggcatg cagtaaaagc aattctagga tgattgctgt 60540ttggcgcgta gttcgtatta gaaaccattc ttcttgaata aatagtatgt ttaagaagct 60600gggcagaggg aaggcatatg catatattat caacaaggag ggagaaaaag gcaattagta 60660accatccata ggagggtcag caagatttat aaaggaaatt tgtgatccaa gtatgaagca 60720aaataaggtg cagaataaat tttaagcaag taatagatta gagtaagaga acccatttga 60780ccattaacct tgggacattc tctttcaaat gacatggagt agtactgaaa tctttctttc 60840tttctgagtc taggttattg tgactggact cagaaagaaa tatttcatta ttgcagtgaa 60900taacatttgt gaacattatt gttcataaat tatgcagtga ataacattta tgaacacgtg 60960atgtgtaaga tacatactgt ttatttttag ttaagttttt tggctcaact tctaggcaga 61020gaacattaaa tgtaaatagt gttacctagg agcatgtaaa tggaaatctc catagtatga 61080aagcagtgct gttgctaaca gaatttagga gggggcagat gaggtgaagg aaatgtgggt 61140gctgatttcc ttattacatt gagaggagcc aggagattct ttgttcaaaa tggatggctt 61200aagaagtcaa agtataagct gattacgtag agcaggtacc caaaaatgtt ttgtgtaagg 61260ggccagatag taaatatttt cagtcttgca ggccatccca agtctgtggc agctactcaa 61320cactaccttt gtagcatgaa agcagccaca ggcagcccat aaatgtggct ctgttccggt 61380gaaactttag gtacaaaagc aggtgcaggc cagacctgac ctgtgcactg tggtttgctg 61440acctgggatt caggggtata gaagttacca tcagaagagc taaaagtgag actttttact 61500ttatactctt ctacactgtc tgattttgaa aaaaagaaac atgtatttta taatattaaa 61560gatagggttg gcaaatagca aataaaaata cagaatacca gtgaaatttg aacttcagat 61620acattatgag taattttatg gtgtaagtat attccaaatc atgtgggaca tacttacact 61680acaaaattat ttgttgtttg tttacagttt aaatttgagt gccttgtatt ttatctggca 61740actgtaatta aagggaaaaa gaataaattc attatgttca tataatgtga tatagcaggg 61800gtccccaacc cccaggctgc agagtggtac tggtccatgg gtccccaacc cccaggctgc 61860agagcggtat tggtccatgg cctgttagga accaggctgc ccagcaggaa gtgagcagca 61920ggtgagctgg cattcccacc tgagcaccgc ctcctgtcag atcagtggca gcattagatt 61980cccataggag tgcaaaccct attgtgaact gcacatgtga ggggtctagg ttgtgcgctc 62040cttatgagaa tctaatgcct gatgatctga ggtggaacag tctcgtcttg aaaccatccc 62100ctggccctgt ggaaaaattg tctcccatga aaccagtctc tggtgccaga aaggttgggt 62160agcactgtga tatagtatta aaagtgctaa taaatatggc atactgcctt taaaatgtct 62220ggtagctctt tctcagtggc actcataata gtgttttttg atttttaaat gtgtgtcaag 62280ctgactctcc cctccgtgta tgctgggctt tattttccct ttcctagtca ccagttttgg 62340gaaatagaga tcttcattct catgctgctc ctctagtgca agtgctccat ttatttttaa 62400ggaattaata taacaaaaaa tcatgggaat ttagaaaaca acatggaagc taatgatcac 62460attggtggaa gtgataggga aatatttagg gggagaagtt aaggtataaa ctttgtcaat 62520gaagtcctat taaaaacaac aaaaaagtga agcttaggat gcattttata aactctgacc 62580agaacacctg tgtttctctg tttctaggtt tatgaactga cgttacatca tacacagcac 62640caagaccaca atgttgtgac cggagccctg gagctgttgc agcagctctt cagaacgcct 62700ccacccgagc ttctgcaaac cctgaccgca gtcgggggca ttgggcagct caccgctgct 62760aaggaggagt ctggtggccg aagccgtagt gggagtattg tggaacttat aggcaagtta 62820ttagcaaggt ctactcttac aattaacttt gcagtaatac tagttacact ctattgatta 62880tgggcctgcc ctgtgctaag cagtctgcat tccatcttcc ttgccaaaac ttataataca 62940aatttcatct ttattttata aataggggag ttgggctggg tgtggtggct cacgcctgta 63000atttcagcac tttggaagga tcgcttcagc ccaggagttt gagacaacct ggccaagtga 63060gaccctgtct ctacaaaaaa aaaaaaaaaa aaaaaattag ctgggcatgg tggcacatgc 63120ctgtagtccc agctgctttg gaggctgagg tggtaggatt gcttaagccc aagaggttga 63180ggctgcagtg aatcttgatg gcagctgcac tgagcctggt gacagagcaa gatgctgtct 63240caaaataaat ttaaaaataa aataagagaa ttaaagttta gcaggttggg tggcaaaatg 63300aggccacaca tttaaagccc ctcctcctga ttcttttctc tgccttggct gcctcctgtg 63360gcattttagg tgctgagaaa tgaaaacagt agggaaaata gttccaggat cctcatgtta 63420atttgccaga aatggcatct tcaagtcgtc agagggatct gagagttcct tcctggcctg 63480acttgagaaa atccgtctgt ccccagctct gcgtctgcct ccactgccca gtcacctcct 63540ctccatgctc ttggggctgg gccctacccc accatgcagt gctgccctgg agcagtgagc 63600ttggtgggtc ctgtctggca tgagagctgc ctttgggagc tggatcccag cctctaccac 63660tgggtctggt gcctagcagg ctatggataa acttctgctg actccggcct ctcctaagcc 63720actgcaacgt ggtcggtgta gtgcacagtg tgtgtgcagc gtggccttac tcacagcctc 63780cacattagag agaatctgac tgaagtctta ctgctgcctc gtgtgaacat aaatgtttgc 63840cagaaccatg agcaggaaat gttaatctgc cttgtttcct gtcctttaca cggaagaatt 63900tttttctgta tggaatgcgt gccttacaaa taatgagtgg aaatacccat cgctaatgaa 63960aagttatact tgactgttag tcagctaaat aatctgagat ttctaatact tttaatttgg 64020cttttacaat gcaatttatc ttagcttttt tgatttctta ggtcatatct ttagaactat 64080atatttgaat gttaatgtaa ttttcatatt gaaattaaaa tgttgaactg cgatgttaag 64140tgtttcctgt ggaaaaacgt tcacattttc tctagtttta aagttgaatc aagctgtttg 64200aagattttca catttcttct agattttatc agcttgttac tttatctgtc actttctgtg 64260atttgcagct ggagggggtt cctcatgcag ccctgtcctt tcaagaaaac aaaaaggtga 64320ttatttcaga aatcagagtc ttgtgttgaa tcttactgat tttcttgtat ttctgtaatg 64380taatgtatct tgtatttctt gtaatactgt attggactct gtgtatatct cttctcagat 64440gagtgattat atgtgtgaat gttgctggaa tctgataacc aggcctgaat agttttgtag 64500ggtggctttt aaaaattact ttcatatcag aattgctttg tcataaattt tgaacgcatc 64560ataaatttct aatgttcggg gtcagcagac tttttttgta aagggacaga gtgtaaacat 64620cttagcttta tgggccatat ggtctctttt gcaacattca gctctgccct gtgacaggaa 64680tgcagttgta aagacatgag ctactggcca gctatgttcc agtagaactt tacttacaga 64740aacagacagg ctgtagtttg ccaatacctg ccttagggaa tgtgttgtta tattttgtga 64800gttaccttct cagtaaattt tatttagtat tagtcaggaa tattattaag tagcttcttt 64860tccagcctgg tcaacatagt gagacccggt ctctaccaaa acaaaacaaa acaaaaaaac 64920agccacgcat gtggcatgtg cctgtagcct cagctgctgc tcagggggct gaggcaagag 64980gattgtttga gcccaggagt ttgaggtcac agtgagctgt agtcatgcca ctgcactcca 65040gcctaggcaa cagaatgaga ccttgtgtct taaaaaaaaa aagtttcctt tgttgggtta 65100ttttaatttg gacctggtta tcatttttca gccatattta actttgtaca tatcagaatg 65160ttctgataaa acttaacttt tattaaagtg tttgtgatat aatctgctag ttttggtaca 65220cattatcttt tgcaatgcca gttattttct tttccagtgt gggtttgcat aggaaaagaa 65280ttgctgtcac tttctatttt gaaatcttaa aagactgatc cttttttgtg tcatgatttg 65340agtatttaat tgagagccta atgcctaata ttatttgcag tattaaatgg gatcttaaca 65400ggaatagcat tctagccttc attgaattaa gtaaacattt cttaagagaa cttggaatct 65460ataatatttg cgtcatcata gtatgagata cttaatcaag tttgagattt tagtgaaaca 65520ttgtttagaa gccaaaagga ttctaggaaa aattaatgtc tatattcttg aattaggaga 65580gattttggga cgtgtgacta agttacgctg acacttgttt gtttcttagt cgctttttcc 65640agtggcggtg agaacgaaga tgactgattc acattgctca gatgagttta tcctcttctg 65700gctgggacat gggatatatc ctgtctcttt taagcctttt tggtattttt cccccattga 65760gagctgtgtc ttcaaactct tctgttatag ctggaaaatc ctttttaagt gaaatctgcc 65820caaattataa gacagatgaa ggtagagttg tgttggatat aggattaggg tgaaagtagt 65880gggggtgtcc tggagcctct cttctggtgg cagcctagct cttgtgcctt tgaggaaatt 65940accctgggga cggctctgtg gaacatattt gcaaaccact gatttggaag atagagatgg 66000cttttgttaa gatctgaatt cacctttttg gcattttatt tgatttctca aggtaaagaa 66060cttattttgt aataaagttt cctattattt agtagatagg ccaagttgct gtgttaattc 66120catgtagatt ttgggtttcc tttgctcatt ttttcactct taatctcaca tcattgtaag 66180tttatggaag ttatcatact tctgactttt tctttgaaga gcagaaatta gaaattccca 66240ataattattt tgatagtgtc atttaatgac actcacatgt gatgtagcca caaagattta 66300atgagttcag ttttaaatca tattaagact gttggtttca tttgttctca ttaatgtaat 66360tctgaagatg aacaataaaa tgtattttta gaactttcaa atgaaatatt atttcatcct 66420tccagatcat ataatgctta agttctgatt gttaatcata aagtctagaa aattaaaaga 66480taataaaatg aaagtgactt ttaggtatta gagttttatt ataaattctg gtgtgtcatt 66540ggagctatga catgaatatt tcaaaggcca atagcattgg atctttacag ttataactta 66600ccatttttaa gtttaagtag taatatagat tatttaataa tcaaaatcaa taaatattaa 66660ttattaaaat gttttgtggt atagtttgag aatcattgct tttaactttt tccatatagg 66720tttattgact ttaatagcat tctaaacata acatctctac attctttgtg tttaatactg 66780tggaggtata aaaatactta tatatgatga taaactatat tagagtaaat taaatattct 66840tatgagtttc attttagagt gcatttactt aattttgaag tccttatttt tagcaaacta 66900aaaggaatgt tggtacatta tttactaggc aaagtgctct taggagaaga agaagccttg 66960gaggatgact ctgaatcgag atcggatgtc agcagctctg ccttaacagg tagttctcac 67020tagttagccg ctggtgtgga ccttcactgt ctgccttcca ccccttgccc ttcctgctcg 67080tccccctgca cctggtggac agcacgactg ggggcagcag tggagccagg ttgcttaaat 67140ggggcatatt cgggcttctt ttataatact tactctgaag cttgtgtgtc tgtggtgttt 67200gcatcatata tttgttgttt tccatggttt aggctgtttt aaaattaggt ttatggcttg 67260agcatagggc tttgtgagta ggggatggca ggtcgaaaca tctcatgagt tggatgggtt 67320atgctggggg ttgggaaatg ggatgaaaaa ttatgggatg aaaaattgcc tatggatagt 67380ttaacttgaa agaatctgcc tttgtttaca gatagttatc ttttttcttt tttgagatag 67440agtctcacac tgtcacccag tgcagatacc cagtgtcact ggagtgcagt ggtgtgctct 67500tggtgcactg cagcctccgc cttctgggtt ccagcgattc tcctgcctca gcctcccaag 67560tagctgggac tacaggtgcc cgccaccacg cttggctaat ttttgtattt ttttgtggag 67620acgggttttt gccatgttgg tcaggctggt cttgaactcc tgacctcaag tgatctgcct 67680gcctcagcct cccacagtgc cgggattaca ggagtgagcc actgtgcccg gccagttaca 67740gatacttatc taatgaaatt ctctgtgtac tttataaaag atgaggatta actgaaggta 67800ctaataactg gattatatga gggtggtttt ggttgtataa tcctatctaa aagaatattt 67860tagctataac tgaaagtaag acttaaatat ttagagagga aaatctgaat aattctagta 67920gtaattattt atttacaaaa taaaaataga tttttttttg attacacaaa ttaaacaaca 67980ataaaacatc acagcaatcc ggatactata aagctcacat gcttaccgac ccaactgccc 68040caggagtgac cactgccaac agcttcatgt cgaccttttt gccataattt ttatatagcc 68100ttttttgttt ttaaatggta atttagaaag tcaactagga aaatgtgtta caggtttatc 68160ttccaggaga ataggactgg agtcgagatc ttgaatgtgg cttggaagaa ggcaagccca 68220ccccagagag atgagttgac agttgtttct gaccactgct tgcttagagg gcctgcgtgt 68280ctgtgaccgc ctagctttgc gcccctgact aggctgcccc ttaattacaa atgtctttat 68340atattgctcc agctaaggct tggagtagtc ggttaagaac ttgaacttcg gtttttgcag 68400tgaaacagca tttgagaata tcaccttctg ataagcctta ttttataagg tgggtactgt 68460agtgggaggc agtgtgagag atgcttgaag gatgcactgc tgtcctgcat ttcagcatct 68520tcaggatgct gtgcagctga aacatttgat aacggtggaa ctgttcgtta ttttgcaagc 68580ctgtgattcc ctattgaatg ttttctctcg ccatttgaca aatgagtgtt tctctgtctt 68640cagcctcagt gaaggatgag atcagtggag agctggctgc ttcttcaggg gtttccactc 68700cagggtcagc aggtcatgac atcatcacag aacagccacg gtcacagcac acactgcagg 68760cggactcagt ggatctggcc agctgtgact tgacaagctc tgccactgat ggggatgagg 68820aggatatctt gagccacagc tccagccagg tcagcgccgt cccatctgac cctgccatgg 68880acctgaatga tgggacccag gcctcgtcgc ccatcagcga cagctcccag accaccaccg 68940aagggcctga ttcagctgtt accccttcag acagttctga aattgtaagt gggcagaggg 69000gcctgacatc ttttttttta ttttttattt gagacagagt ctcactccat agtgcagtgg 69060aggccgggca caggggctca tgcctgtaat cccagcactt tgggagactg aggcaggcgg 69120atcacttgag gtcaggagtt cgagaccagc ctggccaaca tggtgaaacc ctgtctctac 69180taaaaataca aaaattagtt gggcgtggtg gcacatgtct gtagtcccag ctgttaggga 69240ggctgaggca ggagaattgc ttgagcctgg gaggcagagg ttgcaatgag ccgagatcgt 69300gacactgcac tccagcccgg gcaacagagc aagactccat ttcaaaaaaa ataaaaaaat 69360aaagtgcagt ggctcgttct cagcccactg caacttctgc ctcccaggct cgagcgattc 69420tcccgcctca gcctcctgag taggtgggat tacaggtggg caccaccaca ctcagctaat 69480gtttgtattt tcagtagaga cagggtttca ccatgttggc caggctggtc tcaaactcct 69540gaccttagat gatccaccca ccttggcctc ctaaagtatt gggattatag ttgtgagcca 69600ccatgcccgg ccctgccacc tgccatcttt tgagttcttc cctggagacc tagacctgaa 69660ccctcctgct tgttctcttg ttatctaata cccctattga cagcgcagct tagatcatta 69720atggagagct tgacctcatc tgataccttc actgaaggaa acaacttagt gtcttttgtg 69780ttgaacactg aggtaaaaaa ttggaatagt tgattatatg aactctgcta aaattgagtg 69840cattttacat tttttaaggc cttgttgggc cctggttaaa taattatttt taaaaatcct 69900taaggagcct attataaaca gatctgtggt cttaatgaaa tgtgattaat actgtgcatt 69960attttaagaa cttttgactt ttcaaaaaac ttttacaaca tttcccattt gatagcggca 70020taggtttaag cacttctcat ctctaagtta gtggacaaaa aaccctcatg gatagtctaa 70080taatgtttgc tacaagtcca tgttgagttt tatactccat tttattttca gttttaaaaa 70140ctgtggttaa atatgtgtaa cataaaattt atgttcttaa ccattttttg cgtatacagt 70200tcgctggtat taaatacatt taaataatgt catggaatca ttgctaccac ccatctctgt 70260aaccttttga tcatgtaaca ctgaagctct gttcccattg aactctattc ctcctttccc 70320gccaagtccc tggcaaccac gattcttctt tctgtcttct gaatttgact actttgggtt 70380ctcatatact ttaggagtca cacagtattt gttttactta gcataatgtc cccaaagctc 70440atgcatgttg tagcctatgt tagaacttcc taatgtttca ggccaaatac tattccattg 70500tatggatagg ccacattttg cttttccatt cctctgtcca tggacacttg tattgcttca 70560tgttttagcc attgtgaatc atgctgttat gaacgtgggt gtacagatag ctcctggaga 70620ctctgctttc catttttttg gctaaatacc cagaaatgga gttgctttta cattccaatt 70680ttaatttaaa acattcatat cattgagtgt tttacttaat agtatagtag ttaacaaact 70740taataaaata gtattttggt aataatttgc tggtagtcca ttgttcagtt tttttaggta 70800aattacacag gacatttcaa gtggacatga aacatcttgt gatgtggaat catgccccaa 70860gctgatggct aaacatatga aataccatac cctaaattta gtagatttag tctttgcaat 70920ttaggagata acctgttata ttgttaggtt tttgtcgaaa agctttgtcc tcatatttcc 70980aacttgctgt aaaatttgtt tgtgaagaca aatatttttg tatgggtttt ttctttttca 71040tattaaaaag aaatgtccac attggaattt ttttggagtt tttagagcta atagagcttt 71100tcataatgta gtgggaatga gtgatcagta agctcttagc agtttccatg cgtgcatttc 71160tgtgccttga aataaatgac agatgagtac atttgtgttc tgtgtgtaaa atgtgctctt 71220tcctcattgc acttccatgt tggagggctt gtctcttggt gatcacactt caaaattctc 71280acagcccccc ttgaaccgtt taggtgttag acggtaccga caaccagtat ttgggcctgc 71340agattggaca gccccaggat gaagatgagg aagccacagg tattcttcct gatgaagcct 71400cggaggcctt caggaactct tccatgggta tgtggactac aggtgatgcg ctacaaagtg 71460gtttgtattc agacctggac atcttaatta tatctttgct tccaagaaga agtcctttga 71520tactgttttc tgagttctga atagctgatg aaaatgacca attgaggaat aatcatactt 71580tttcttgatc taaatcttat acttttgagt tatcttagca taaatgtata attgtatttt 71640aagtggaaat ttgtcactta atcttgattt ctctgttttt aaagcccttc aacaggcaca 71700tttattgaaa aacatgagtc actgcaggca gccttctgac agcagtgttg ataaatttgt 71760gttgagagat gaagctactg aaccgggtga tcaagaaaac aaggtgaggg acataggctt 71820gagacgactt ggtgtttctg agcttgtgtg aggatttaaa atcgccctgg ctactgtcta 71880ctttattgct ttcccatccc tgggccttta aatttcccct ttaaatacca gctcttccca 71940ggcctgttgt tttctgcctt tccaggtact acccacagcc ttgagaattg cctgagttct 72000gcctcctttg agagtgtgcc ccagacaaat ctattctgta ctgaatgttt ccttgtctga 72060tttcttggat cattcatttg atggttgcgt atggcctgca acgtttcttg ttttggttct 72120actgaactgt tctaaaagtc tctcttcata ttatcttttt acatgtaaat gtaactgtct 72180tcacttttaa ttcctcaagg acaaggaata gcgtttcaca gttcgtccca tcaatcagaa 72240ttatagcctt tggcatctcc ctatctacca ggcccacttc ctcttagatt tgggcttccc 72300caggctgttg cctttcccca agtagcttct gcttgtcctg tagaagacct ttcatgcttt 72360gcttctgcag cagccgttcc tgaatgccta gtgtcaactg ccttcttacc acgcccaccc 72420tccctgcatg ctgcatttat cccctgccac agccctgtga ccctgtgtcc tgctgcctct 72480gacttgtctg tttctgcttg gccatggtct ctgtgaggtc aggtgtgcat atgggcacaa 72540accagggcat ctctttatcc ccagcacctg gcttaagtgc tgctctggaa ctatctgttg 72600aatgaactaa tgcatgaatg tattgttgag tatgagacaa acaagtgtca ttgtctcctt 72660tctagccttg ccgcatcaaa ggtgacattg gacagtccac tgatgatgac tctgcacctc 72720ttgtccattg tgtccgcctt ttatctgctt cgtttttgct aacaggggga aaaaatggtg 72780agtacaaaag gggatgtgca cagttgaagg aaataactag gtttcagagg tcagcttggt 72840ggcctgtttt tgccttgcgt gcagcagagg aagtagaatc tgaggatgag tttggttttc 72900actagccgag gggagggagg aaatgatggg agcaggtagg ttattgggtc tggttttgtt 72960catttgaaaa caatctgttg tttgaggctg aaggtggctt gggtgatttc ttggcagtgc 73020tggttccgga cagggatgtg agggtcagcg tgaaggccct ggccctcagc tgtgtgggag 73080cagctgtggc cctccacccg gaatctttct tcagcaaact ctataaagtt cctcttgaca 73140ccacggaata ccctggtatg ttaaaagttc acatcttatt ttctcagatt taatcattat 73200tgtaaaaact atttcagtat tgactatttt agttttagag cagtaagtgt tttgagttca 73260tttgggatat ttgacctgcg ttgtagctct tcagaaaaca catgaatagt gaagttcttt 73320gtttcatggg ttccctttag atgaaaccca tagaggagaa aagtagaaac ctcagcacgt 73380aagagccaac atatatacac atcggattta aacctaaagc acaaattgtg cctggtcgca 73440gtggcgctga gtcgcactca gccaggccag gcattcacac tcagggtgag tgggaaccag 73500gactggctga ggcagcagtg gacccaagtc tccatcgcgc ccatgcttac tatggagcct 73560tctcgttctc tctttttctt tgggtgagag ggtacacttg tgtttttgaa tttatatgag 73620gtaagtgtgt aatagggttt tttctaatct tttttaagtg gaatctggaa ttttaatcag 73680atttattatc tgacaaccta gaattataat ccagaaagtc tgtggtattg aggacatatt 73740ggcaatatga tgaatctcta attcttaaat cctgaaactt tttttttttt aatcacttag 73800ggttattata gtgaagtcat ttctgaattt ggatcttctc ttcacacctc tttttctctt 73860tcctgagaat taagcttttg tttcgagtta gaaagttgat agtagggaat tgttccatgg 73920ctgagcaatt tatctccaca gaggaacagt atgtctcaga catcttgaac tacatcgatc 73980atggagaccc acaggttcga ggagccactg ccattctctg tgggaccctc atctgctcca 74040tcctcagcag gtcccgcttc cacgtgggag attggatggg caccattaga accctcacag 74100gtaacggcca gtttttcagc tgtgtttttt ctagttatgc ttactaaggt ttaagtttag 74160atgatgatgt ttgttgcttg ttcttctggt taggaaatac attttctttg gcggattgca 74220ttcctttgct gcggaaaaca ctgaaggatg agtcttctgt tacttgcaag ttagcttgta 74280cagctgtgag ggtgagcata atcttctgtg gaaccatttc ttcacttagt ggacatttta 74340tcattgctac aattaaaatt ggagcttaat aggaaatatt tccatgcact ctaaagctgt 74400aaccagtaat acccaccatg tatccatctc tcagctttag aaagaaaacg ttgccagtaa 74460agttaatgct tcataaactt cagtttaagt tctaattctc agaatatttg tttgaaatag 74520acctcttcct aaaggatata tttagaaata acctatcatt aagtgtaaag tctgttgaat 74580atgctgggca cggtgactca cacctgtaat ctgaccactt tgggaggcca aggtggaagg 74640attgcttgag cccaggagtt caagactatg ggcaacatag ttgaccctgt ccctacagaa 74700aattaaaaaa aaaaaaaaaa aaagtagctg ggtatggtgg tgcatacctg tagtctcagc 74760tactcgggaa gctgaggtgg aggggggatt gcttgagccc cagagatcaa ggctgcagta 74820aggcgtggtt acaccactgc cctctagcct gggcaacaga gtgagactgt ctcaaaaata 74880atagtaataa taatcagttg aattaaaaaa aaaaaaaaaa aaaccactgt gctaggccca 74940tagtatggta agagttaaag tgagccttag ggattattta ctcaacctct gtttctgtat 75000aaagtggaat aggctcaatt ctttaagtga tagcatgttg aacctttcca taccaactgg 75060ctcataagtc acaactggcc agtcaacaag agtaaaaatt aactggtaaa aatcaaagca 75120aaaaacctac aattgtcaaa tttgtgggat aactccccct tttaaaatgt catgcctgac 75180agtaatttct ctctagtttc caggttttca gtcagttgtg tcttttttga gcagaaggaa 75240gcatgctaag agctcaatct tgtggctagc tgggggtctt

tgtgtcagcc atgcatgtga 75300tggtgcccct gggtgcttgg ggctgcaggg gaggggtaca gcagtagggg cctgttctgt 75360tctctcgtgc tgtggagtac atagtgacat agtggggtgg tccttggtgt aggtcccttg 75420ttcctacccc tgggtctgag atttatttag aagtggtgtt ggggctgtgc ggcaggcccc 75480tctgtaactg atcaatgttt gtgaagttgc tgtttgagag ttgaaaccat gacataagca 75540gaaatggaag gaagaaagaa ccagttatgt gaaagggaca catttacttt taagcttgta 75600tttactgaga taaagtattc ttaatcaatg ttcttgagag gtgtgggaaa aatgcaacat 75660cctggttgca gttaaaccca gaacattgtg tgttgaagag tgacggttct caaaccgtca 75720agacgcgggt actgagtggg actaacctgc tgtcctcttg ccttggacct tgtgttccag 75780aactgtgtca tgagtctctg cagcagcagc tacagtgagt taggactgca gctgatcatc 75840gatgtgctga ctctgaggaa cagttcctat tggctggtga ggacagagct tctggaaacc 75900cttgcagaga ttgacttcag gtaagtgagt cacatccatt agatttcatg aactaagctc 75960aattgaaagt tctgggatca cttgatgcaa ggaatgatgt tatcaagtac cctgtccatc 76020agaaatccga gtggtttagg tagatgacag tgattttctc ctcccagtgg ctttttgctg 76080aactttgccc tatgcttgga attttatttt attttattat ttatttagag acaagatctt 76140gctctgtcgc ccaggcttga atgcagtagc acaatcatag ctcactgaag ctttgaactc 76200taggactcaa gtggtcctcc tgcctcagcc tcccgattag ctaggagaat aggtgtgtgc 76260cgtcacactg gctaatattt tttgtagaaa tggggtcttg ctatgttgcc caggctggtc 76320tcaaactcct gggcttgatt gatcctccat cttggcctcc caaagtgctg ggattacagg 76380catgagccac tgtgcctggc ctagaatttt aaaatataag tagaagagta gatttttttt 76440tttggtagtc ctcgtcattt aagtattctg gatagtggga ataaaagagc ttagaatttt 76500tcatctttgt cttaaacttt taaaaaaatg tagcttatat taattctgct tgtttaaaaa 76560gaatatactc ttcattatac tgaacctagg taagacagct ggtttatatt ttgttgcaat 76620taaaaaacgt gagctgtggt tgcagtgagc caagattgtg gccattgcac ttcagcctgg 76680caacagagtg agacttggcc tcaaaaaaaa aaaaataaca tgagctgtgt tggcactttc 76740attttctaag agtagttttg gctggagaag ttttctttca gtactttctt ttagaaggga 76800aattttcctt tataatttag ggtttgtttt ttttttttcc aagccacctt ttatagagcc 76860cttgtgggtt atttcattta atccttagaa tgtttataaa tctgggcttg ttctcggctc 76920cacccacaga tagggacgct gagcgtgcat gagtgggcag caagatagca ggttatggag 76980ggcccagctc accccttctg tggcttgagc caattttata gggcacttac agagtctttt 77040gaaatagtat ttattttgaa gaaaaagaaa aacagtttac tgagtactgt cttattgagt 77100ctggaattgt gagaggaatg ccacctctat ttatttaaag ccattggcct tttttgttgt 77160tttgagtaag tgctgcccaa ggtccttcca gggcacctgg atgagcctgc tctggagcaa 77220gctggcggta agtgtttact gagtaactaa atgatttcat tgttaaatgt gctcttttgt 77280taggctggtg agctttttgg aggcaaaagc agaaaactta cacagagggg ctcatcatta 77340tacaggggta agcggtttat ttttgtgaga tgctgtttta ccttcaagaa ggtgaaagtg 77400aggctttcct tgtggaattt ctctaaatgc attcgtcatg ttttagatgt ttatttcaca 77460gtttatatca tgaaagttat aatcttgtca tatggattta agtctagtaa tgttgagttc 77520tttctcacta gctttccaaa atatcttacc taaaatttag tcaaatacaa gattatgttt 77580atttttatta tccttctctc taaagctttt aaaactgcaa gaacgagtgc tcaataatgt 77640tgtcatccat ttgcttggag atgaagaccc cagggtgcga catgttgccg cagcatcact 77700aattaggtat ttaccaatat tttatctctt ttcctttttt ggttgaagta ctaaaagata 77760cgagaatgga aagagaggga agaattcaaa ggatgtagag cagtattcct gaatctgagc 77820tcatttcagc cattctattc ttaaactata atgaaaaaaa aatccaaaaa agtctaaaat 77880tataattaaa aaaacaacaa aatactaact gtccattgta aaaagtaatg cactttcatt 77940gtaaaaattt tggactatag agaatagtac taagaagaaa aaaaaaatca ccttcaattc 78000tgctgccacc tggaggtaat cactgttaat attttgctat atactctatg agtttcttgt 78060tcaaaatcag gtcaaaatta catgcaattt tgtaatctga caatttccac ttaatatttt 78120attagcattt tcctgttatg aaacagtaat tttagttatg ggtcgttgtt ttgctatgcg 78180gttgggataa aattttatat actttttttg gcaattactt attatacata aatgtttgtg 78240tatagttttc tttttctgag aattcctgga agttgagtta ccaggcccgg ctttgaattt 78300ttttttttat tttttttttg agacagagtc ctgctctatt gtccaggtgc tatctcggct 78360cactgcaacc tctgtctccc tggttcaagc gattctcctg cctcagcctc ccgagtagct 78420gggattacag gggcacacca ccacgcccaa ttaatttttg tatttttagt agagacaggg 78480tttcacgata ttggccaggc tggtctcgaa cttctgaccc cgtgatccac ctgcattggc 78540ctcccaaagt gctgggatta caggcgtgag ccatggcgcc tggccaggct ttaaatttaa 78600aacaaatctt ctaatagctt tatggaggtt ataatttaca tttcttgaaa tgtactcact 78660ttgagtgtat agtaaactcc aattttatca catttctgtc accccaaatg tatccttgtg 78720cccatttgct gtaacctccg gttcctgccc caactcctag gcagccactc atctattttc 78780tgtcccttaa gatttgtgtt ttcgccaggc gctcatgcct gtaatcccag cactttggga 78840ggccgaggtt ggtggatcac ttgaggtcag gagttcgaga ccagcctggc caacatggtg 78900aaaccttgtc tctactaaaa atacaaaaat tagtcggatg tggtggcaca cgcctgtaat 78960cccagctact cgggaggctg aggcaggaga atcacttgaa cctgggaggc ggaggttgca 79020gtgagcagag atcgcgccac tgccttccaa cctgggcaac agagagagac tgtctcaaaa 79080caaacaaaga tttgtatttt ctggacattt tatagtactg gggtcatagt atagatggac 79140ttttgcattt ggcttctttt acttaattgt gagattggtt cttgttgtag catgtatcag 79200tagtttgttc atttttattg gcgaaagtat tctattatat gaataatacc atattttatc 79260tatccatcag atggatatta tagagttcat gttttggcta atttatgaat tatggtactg 79320tgaacatttg cctgcaagat tttgtgtaga catgtcttca tttctcttga gtagatcacc 79380tagaagtgga tttttaaata attttggtac ttactgtgaa actgctcttc aaaaacatac 79440cattgttcct tccttccttc cttccttcct tccttccttc tttccttcct cccttcctcc 79500ctcccttccc tacttccctc tccctttccc tttcccttcc ccttttccct tccccttccc 79560gcctgcctgc ctgcctgcct tccttccttc cttccttcgt ttctttctac atatacacat 79620ttttttaaat ttcaatggtt tttggggtac aagtggtttt tggttacatg gctgaatttt 79680ggttacatgg tgaagtctga gattttagta cacctgtcac ccgagtagtg taccttgtac 79740ccaatatgta gttttttgtc cctcaccttc cagccttccg ccttgtgagt ctccaatgtc 79800cattatacca cactgtatgc ccttgcgtac ccacagctca gctcccactt ctgagaacat 79860atagcagaaa catgccaaag tatactccca ctaccagaat gtgattgtgc ctgattcttc 79920tcaccagtac aaatatttca aaaaaagtta aatatgtatc agttttttgg gcagaagttg 79980atacttctct ttatttattt attttttttg agatagggtc tcattctatg atgcccaggc 80040tggagtgtgg tggtgcgatc tcggctcact gcagtctctg cctcccaggt tcaagtgatt 80100cccacgtcag cctcccagga agctggaatt acaggcgagg gccaccactg ccagctaatt 80160tttgtatttt ttggtagaga tggggtttca ccatgttggc cagactggtc tcaagctcct 80220gacctcaagt gatccacctg ccttggcctt ccaaagtgct gggattacag gcgtgagcta 80280ccacacccgg ctgatatttc tttttaaaat aacttacctt cttttgaaag taatacatgt 80340ttaatgaaca gaatttaagg aaaatataaa aaaacgaaat aatctttgta atcaaactac 80400tgaaaagaaa accaaagtta cattttggtg catattcttt ttcattttca tcattgtaat 80460ttgcatttct ttgattactt gtgagacact cctttcattt acttaatagg tttatatgac 80520ttgcctattc agagattttg cagctttacc attttctgca aatgatagca acttcttttt 80580gtttgtttgt ttgtggagac agagtctcgc tctgtcactc aggcaggaat gcagtggtgg 80640aatcttggct cattgcaact attgcctcct gggttcaagc gattttcctg cctcagcctc 80700ccaagtagct gggattacag gagtgtgcca ccatgcccgg ctaatttttg tatctttagt 80760agagatgggg ttttgccatg ttggccgggc tgatcttgaa ctcctggcct caagcggtcc 80820ccctgtctcg gcctcccaaa gtgctgggat tacaggcgtg agccaccgta cccagccagt 80880agttacttct tatattctag aaaaaattct actcatgatc aagtctccat gaggaaagag 80940actttaattg aagatcatgg ggcttgcaga ccaatatgat aaaatagttc attgtttcta 81000aaagtattac tgagtgttga tggcagatat gaaccctttt gtttttgtag gaaaatgtta 81060cccgtattct ccatttgaat tcagtttaga tttgttagga atcgcagctt aagctttgcc 81120atctgggagt gtttgggaca gttttgcaga caaaattgca aaagtgccta aggaatgcag 81180ctggcattca gacctgctct gtgctcagta ctctgtggac agacactgtt cagcacttgt 81240tgatcagaag gtttagaaag agaactttca aagttggttt ttaattaaag catttaatag 81300tgtaaataga aagggattaa attttatgac agacaaaaga aagtacagca cccagctggg 81360cgtgggggct cacgcctgta atccagcact atggggggct gaggtgggtg gatcacgagg 81420tcaggagttc aagagttcaa gaacagcctg gccaaggtga tgaaaccctg tctctactaa 81480aactacaaaa attagccggg cgcggtggca ggcgcctgta atcccagcta ctcaggaggc 81540tgaggcagga gaatcacttg aacctggacg gcagaggttg cagtgagcca agattgcacc 81600attgtactcc ggcctgggcc acagagtgac attctgtctc aaaaaaaaaa aaaaaagaaa 81660aaaagaaagt acagcaccca gttatgtccg agtgggtgca tgagagtgac cctgagattg 81720gagacaacgc tgtcacgtgc ttgaagaacg ccacctgaga aagggggcga gaagtggtgt 81780ccgctggtaa ccagaggtgt tggcttagcc atctgcaggg aggagggtgg tctatcacag 81840gtgagtttca tctactttct taagcaaatt aaccttactt ttgtgttagg cttgtcccaa 81900agctgtttta taaatgtgac caaggacaag ctgatccagt agtggccgtg gcaagagatc 81960aaagcagtgt ttacctgaaa cttctcatgc atgagacgca gcctccatct catttctccg 82020tcagcacaat aaccaggtat gctgacccag tggcatcttc acattgtcgg gaaaatgccc 82080tttcctgatg cctttcttta ggctttaatt gaaaacattt tattttctag aaaaaagctt 82140cagctcagga tgtttgagtg taggtcagtc ctttgatagg atattatcat tttgaggatt 82200gaccacacca cctctgtatt taagctctgc cacaatcact cagctgtgac actgtaaatc 82260tcttaatagt ttattacatt ccatgtgctg acagttgtat ttttgtttgt gacacttacg 82320tattatctgt taaaacattt tcactttagt tgtgttacct ttaaagagga ttgtattcta 82380tcatgcctgt tgattttttg gtgagcgggc tattaaagtc agtgttattt agggttatcc 82440actagttcag tgatttgcga gattatcatt cacatttatt gtggagcttt tgaatatcgt 82500gtcaaatggc cacatatatc ccattcttat ctgcttctta ggtgagtggg acacagtgct 82560ttaatgaagc tataatcttc agaattctag cttgcagaga agattgcaga agtgataaga 82620cttgtgcttt ttaattttgt cttttaaatg ttattttaaa aattggcttt atatgatact 82680ctttttttct gctgagtaac agtgttttac aaaacttgga ctaaatgact tctaagctta 82740aatgatcact tgatgctttt tttctgaatt aggaactcag cttatcaaat atcaaagtca 82800taattcctga ataaataacg tcttttttca tgtaaagact gctttaaaaa acacatggaa 82860ggctgggtgc ggtggctcac gcctgtaatc ctaacacttt gggaggccca ggtgggcagg 82920tcgcttgagc tcaggggttc aagaccaccc agggcaacat ggcaaaaccc acctctactc 82980aaatacaaaa aattagccag gcgtggtggc gggcccctgt aatcccagct actcgggagg 83040ctgagggatg agaatcactt gagccccgga ggcagaggtt gcagtgagcc aagattgtgc 83100cattgcactc ccagcttggg ctacagagtg agactctgtc tcaaaaaaag acacacacac 83160aaacaaaaaa aacatggaga catttttttg gccaccttaa tatttcccct cagataattt 83220cctttgttta aactcagaac tggcattttc tctcttggag aagattcagg acaaatactc 83280ctttaagata agtagaagca gtgaaagagg atttgattat caggaatttg ataagcttag 83340aataaattgt tgcttcttaa tgtcatttca gaagatgaat atttattaat agatgccaac 83400tgagatatca ttaaaattga ttactaacta ctacttggaa aagtctccca gttccaaact 83460tcagcaggcc tcttgacaat tcagctgtgg tcaattgggt cttgcgtgat agatacaatg 83520accaattgtg cagcagagtg tgctgcttag ctgcctattc tgttagcatt catgtgttaa 83580cttaaaatca taatctcctt agttttgttg agtgtctccg tggacaagac actgtgaggg 83640atacaaaatc agattggctt tattcaaacc actggggtat tataattcat ttataattta 83700ttttattttt tgcctttttt ccatgtgttc taaaggaatt agagtttgta tataactata 83760atgggggata gaaattgaca tgtgccatga agggaatgca aaaaagtgcc gtgggagatg 83820agaagtggag aaaggaattt cttttttctt ggaagcagga ataacttcat gaagcatgta 83880tttcaactta aacagatagt aggcaacgct gtaaggggag tatggctgca gcaaaagtgt 83940tcggggcaga ctgggaggaa gggagggaat aaattcagcc attgttatgg aataatgatc 84000aaaatttatt ttcagcccgt ttcacttaaa agttgagact gcttaacttt ttttaatctt 84060taatcttaaa cttttaaatg ccatttgatc tttaaaaata tatgttttaa tagtgtattt 84120taagtctcta tatttttgtt attagaatat atagaggcta taacctacta ccaagcataa 84180cagacgtcac tatggaaaat aacctttcaa gagttattgc agcagtttct catgaactaa 84240tcacatcaac caccagagca ctcacagtaa gtctctttct tgatcggtct tactgacatt 84300gtaatagttt ttggtagctt gtatggccag ttagttgtat ggtcatctta cggtgaggtg 84360cttgtcttac agctcttact tatccatgag gcttgctaag aaattgtgct tctgtgaaaa 84420gaatctcagc ttactccagg aatgtaaatg actatgtttt ttctgattat taaagtaata 84480cacgcccaaa ataaaaaaat tcagccaatt taggaagaca caacaattaa aataagccag 84540gcatggtggc tcatgcctgt aatcccagca ctttgggagg ccaaggttgg gggctcactt 84600gaggtcagga gtcggatacc agcctggcca acgtggtgaa accccatctc tactaaaaat 84660acaaaaatta gctgggcgtg gtggcgggcg cctgtaatcc cagctactca ggaggctgag 84720gcaggagaat cgcttgaacc tgggaggtag aggttgcagt gagctgaggt caagccactg 84780cactccagcc tgtgcaatag agcgagactc tgtctcaaaa aaaaaaaaaa aaaaagaaaa 84840gaaaaaagta aactactgtc acctgcattg gtaatgtatc agaagtttaa aatgtctaga 84900ttataattaa ctcagtgacc tggtaatata tactaaggga aaaatattta taatttacat 84960ttttacattt ttattttttt aattttatta tttttttttt gagacagagt tttgctcttg 85020ttgcccaggc tggagtgcaa tggcatgatc tcagctcacc acaacctcca cctcccgggt 85080tcaagcaatt ctcctgcctc agcctcctga gtagctggga ttacaggcat gcaccaccat 85140gcccggctaa ttttgtattt ttagtagaga cagggtttct ccatgttggt caggctggtc 85200tcaaactccc aacctcaggt gatccgccct cctcgacccc ccaaagtgct gggattacag 85260gtgtgagcca ccatgcctgg ccttacattt ttataataag aatttatgtt gctgacatta 85320gaaaagaacc ataatatcca agaatccaag aataattaaa ttatgtacat atgctagtat 85380atagtgtgat gctttggaga atttttaaca atatggagat gtataatctg gattgtaata 85440ttgagtgaaa aaaggcagaa tacaaacctg gtgggggtat agtcggattt cagttaagaa 85500aaataatatt tacatatata catttctcac actggcagat aatcaccaag ataaattttg 85560ggattgtgga tgattttttt cttctttata tttttcagat attctcaaat tttctaaaat 85620gagcaagtat aacttttgtt atcagaaaaa aataatatac aaaagtaatg ttaatttgct 85680ggtgaccagg ttaaaccttt ttatttttat tttttgagat ggaatctcac tctgttgccc 85740aggctagagc acagtggcat gatcttggct cactgcagcc tccgcttcct gggttcaaat 85800gattctctgg ccccagcctc ctgagtggct ggaattacag gcgtgtggca ccacacctgg 85860ctaatttttg tatttttagt agaggtaggg tttcaccagg ttggtcaggc tggtctcgaa 85920ctcctgacct cgtgatccac ccacctcggc ctcccaaagt gctgggatta caggcgtgag 85980ctactgcgcc cagccagacc tttttatttt atttgacaaa agaaatactt ccatgttata 86040gaagactaaa tattgtttgg gctgtctgca gtatggtctt cccttgattt gttcaaaata 86100tcgtaaactt tgcttattta tttttattgt ggccgactgt gtcgggcact gttgtaggct 86160tgggatggaa aaacaggatt cctgccctta gggtttctgc aggctggtca gggagacgat 86220gtggtaagct ggagctcagc tcctaaggat gtgcaggggc agttgagagg cggaagggtg 86280ggagatcatt ccagggtgtg ggcagcacag gaacctctct tcattgggat ataattgcca 86340ttctgataac acgtgtttga ggtgtctaaa gtaggaagtt gtaccatggt gggacagata 86400tcctgtggtt atcatacaca gatctcagtt ttcttctcat tgtttgtact ttttataaag 86460ggtaacagga gatataattc aataaacctt tgtggtgttt gggtgtgatt ttattgtttc 86520tttcttctca gtttggatgc tgtgaagctt tgtgtcttct ttccactgcc ttcccagttt 86580gcatttggag tttaggttgg cactgtgggt atgtattttc ctcagtatat attaatagtt 86640gtctacaaca gtatgacata aacatagtta ttaggatgcc ctttttcttt ctttttaagt 86700cttttatcaa tttggctttt tggaaaaata tctgatggaa tacttgtttc tgctatatta 86760gctgtgtgag actagtgaca ggagctgtgg gaaatgaatg ccaaatgttc ttaggcattg 86820atgggaattt cagggtgtgg tcttcaagtt catttaaggg aattttcata tgctggcaaa 86880aggcttttct cattagcttg actctttcca aaattatttg ctgtgaatta gaagtttagg 86940aacctttttt cacttaattg tgacctagca tacgaaatgg tgatgattta ggaactactg 87000ttcttgtatt aacagctttt atttaaaaat gattttcctc cagtagatgg ccctactagc 87060atctgggaaa taatttcaag tcttctccag cattcaggaa taggctttca ttttgtgtat 87120caattactga gaatgatttt ggtgactcac atcacatttg agaagtaaac ctgcagattt 87180cttgtgtgtg tcagcaaatg accaactgat atttgcttga agtggattac attatctgct 87240ctagaatgat tgctttccca ccttcctcac atacagactg agcagctacg gtttctaatc 87300ataggtctgg cactagactt cacttctggg caactttggc attggagtaa aatgtattaa 87360tttaaagaaa gttaaaaatc cgttcaagta aacatacagt tctaatactt tttacaattt 87420aaaatataga tttaaatgat aaaataaaaa agaaaatatg ggtagacacc ataatcctcg 87480tttctgcatc tgttcacaag gggttgatat ttatgagttc tattctccat atccattcta 87540tgttctctta atgctcagtc agcacctcag gtggttggag ttcaatgctt ggtagtttga 87600cttacactgt cttttctagg ggattgagcc ctgggtagtc ctgcttattt gaggttgcaa 87660tttgtctttc aataactttt actacaagat atggcgtgtt aaaggatacc attggggaac 87720caacataata atatcaggaa aactaaccac gtcagacctg ccccattgtg tatcaagtac 87780actatttttc catagtaata aagagttcac cccagccaat tctcttttat tttgtgcctg 87840tttactcaat ggcattaaca tgcccaaatg tctgggtagc tgtctcatct ccagttcagc 87900agaaccattg tcatatgccc tagtaaaagc attccttcat tggacactta ggccccaata 87960ctttcattca gatctactac ctgatttcat ttctcaaatg atttttatgg agctctgatt 88020tataggaaag atgttagttg attaaaaata aaacaatttc tgagctggta taaaatgtat 88080tgtgacatgc cttcctcttg gaattgcaag agaaaggaag actgttgttt gcttaaaaat 88140tgtctataat ttgactttgc aaatgtctgc ttccagagtg cctccactga gtgcctcaga 88200tgagtctagg aagagctgta ccgttgggat ggccacaatg attctgaccc tgctctcgtc 88260agcttggttc ccattggatc tctcagccca tcaagatgct ttgattttgg ccggaaactt 88320gcttgcaggt actggtactg agttgaaaca gggactccag gacttggatt ttgatttcct 88380tagggggaat gggggtggtg agcatatgag gggaaaatac tataaggtca ttgccagtga 88440tggcttgtcc ctttagtcaa atttcagatg ttacctatat gcataaacac atgcagttgg 88500cagctgttct gtgctgagta ttttaaagta gcctcttccc aatatagccc ctcagttaac 88560tacaagtaaa ctcattttga atttcatttt aatgggcacc atatgccagt actccctcgg 88620gcactgggat gttaagaaag tataatgtat ggacttcatt ctcaagttag ttttagatta 88680gagggggata cacgtaaaca aaagtgcagt ggtcacacag agtggcccta atcactctcc 88740ttgggcagat ttatgggctg gtaggaaaga gcacaacacg gagagggtgt agcaccttgg 88800cgatgataat ggaggatgtg gccagcaagg aagacggagt ccattgaaat tgattttggg 88860agaagttgcc aatctccatg aaagaattgg ggcctgtgct atttgcttca gggggctata 88920ggagagtttc gtgaaaggga ctaaaagatg agtattttaa taagatcatt catccaactt 88980gaacatgggc tggaggagaa ggtagggaga ctcaggagat taatgttgat gctaaggcaa 89040gataatggct ttgggactgt agggaagaca ctgattgtaa gagaatgaag gaggcagaat 89100tgccaggcct ggttcaccaa ctgaacttcg gttgtgaaga caaagaaacc tgggatgact 89160tcacatcctg ggcaggtgtg tggtggtgac agtcatggaa attgggaaca cagatttgtg 89220cgggaaacat cagtttcagt ttgagtttgg cttatcagtt gaatatcagg cacagatgtc 89280tggccaactc tcaacatagg gtcttaaatg acttcagttc cccaagcaat ttgtccttcc 89340catgctattg gggtggagag gtaatgtctg tgcccatatc acagccagtg ctcccaaatc 89400tctgagaagt tcatgggcct ctgaagaaga agccaaccca gcagccacca agcaagagga 89460ggtctggcca gccctggggg accgggccct ggtgcccatg gtggagcagc tcttctctca 89520cctgctgaag gtgattaaca tttgtgccca cgtcctggat gacgtggctc ctggacccgc 89580aataaaggta atgtcccact tgggtgctgg attcatacag ccttaatgac tatgggtttc 89640cagactacct ttgtttagta atctgtccct tctttattct ctttttgctt taaatgaaca 89700aaattgctca gattgtgaca ctaaatttaa catcaaaatg tgaccatgtg gatgggtgca 89760gtggctcgtg cctgttattc cagcactttg ggagactgag gcaagtggat cacttgaggc 89820caagagttcg agaccagcct gggcaacatc acgaaacccc ctctctacta aaaatacaaa 89880aaattagatg ggttgggccg ggcgtggtgg ctcaagcctg taatcccagc actttgggag 89940gccgaggtgg gcggatcacg aggtcaagag atcaagacca tcctggctaa cacagtgaaa 90000ccccgtctct actaaaaata caaaaaaatt atctgagcat ggtggcgggc gcctgtagtc 90060ccagctgctc gggaggctga ggcaggagaa tggcgtgaat ccgggaggcg gagcttgcag 90120tgagccgaga tcgtgccact gcactccagc ctgggtgaca gagcgagact ccgtctcaaa 90180aaaaaaatta gatgggcatg gtggtgcgtg cctgtaatcc cagctacttg ggaggctgag 90240gcaagagagt tgcttgaacc tgggaggcgg agtttgcagt aagccttgat tgtgccgctg 90300cactccagcc tgggtgacag agtcagactc tttccaaaag

aagaaaaaaa tgtgaccatg 90360tgttttatag ctcttttagt atcatcagtc actgttatcc ctaagaggga aatacctagc 90420tttagtttta ggtttccagc attagccaag aaagctcaga attgatgttc ctggccaagt 90480acctcattgc tgtctcctta aatcttggtt aatggctact gtcctggcta gcatagttat 90540ggagcatttc catggttgta gaatgttctg ccaatctcag ggacagtttt gcttttctgt 90600gaagcaataa aatcaacttc aaaacaaatg ttaactattt gtacaatgga tttaagatag 90660accagttcac atactttttt tttttttttt ttttgagatg gagtttcatt cttgttgcct 90720gggctggagt gcaatggtgt gatctcagct cactgcaact tctgcctcct gggttcaaac 90780gattcttctg cctcagcctc tcgaggcaga ttacagctgg gattacaggc atgcaccacc 90840acacccagct aatttttttg tagttttagt agagacgggg tttcaccatg ttggtcaggt 90900tggtctcaaa ctcctgacct gaagtgatct atccgcttcg gcctcccaaa gtgttgggat 90960tacgggcatg agccaccacg cccagcctaa gatagaccag ttcacttact gtttatatct 91020gattactctc tctttgcctt gtcttctacc tttaaaaatc tccctactaa cttcccattc 91080tcctttagct gccatcagtc ttctcccttc tctgcaaaca tctctggaga gtcccagcct 91140cagcccacag agcttcccac tgctctgagg tggaccttgt ttgcaaggct tctttggctc 91200tcttggcctg gaccctgtct actacttcag ccatccttcc ttaacccctg ctggtggttt 91260ctgttgccac actccatagc agcgtttccc gcccagatca tgtctttaca tctctgggca 91320ctgctctggt cctgcctgcc tttccctctt tgtatcctgc aggctgctac ccccatcttg 91380agtgtcctct tcagttggct ttcagagggc ctcctgggtg ttcccttacc cacttgccac 91440tccccagtca ctgggttcag tccttcctgc ccaccagcac atgctttcta ggctctgtcc 91500taggccgtct tctctctttg tagtctctgg gccagtgctg ttctagagag tggcagaatt 91560ttctataacc atggcagtgc tccatagcta tgccaggcaa gacagtagcc actaaacaca 91620tatagctgtt gagcccttga aatgcagcta gtgtgactga agaactgaac cccgattcgg 91680tttaattttc attaaattta aatttaaata accttatgtg ggtagtggct ccagtattgg 91740gcagggcagc ctgagagtcg gggctgttct cctgtcttca gtgtctagat gagggacctc 91800agaggacctg tctctggagc tgcagttcaa tgtagccagc tgccccgtga cacttacata 91860tagctgattt gtggatatgt cagacacggt gtgatgagct cagctttctg tcctcctccc 91920cacatctgcc cctgccccat ttaccccact ttgtgtctta tcaagctaga aacaggtcac 91980cacaagtctt catttccact caccaagtct tttgtttccc ctactaaata ttttgcgaga 92040agaaagtgtg tacctttgta ttcacataca tgtacatgca catatacatg cacatatgca 92100ggggtcccca acctctgtta aaaaccggac tgcaggccgt gcgtggtggc tcacgcctgt 92160aattccagaa ctttgggagg ccgagaccag tgcatcacaa ggtcaggaga tcgagaccat 92220tccggctcac acggtgaaac cccgtctcta ctaaaaatac aaaaaaaaat tagccgggtg 92280tggtggcggg cgcccatagt cccagctacc tgggaggctg atgcaggaga acggcgtgaa 92340cctgggaggc ggagcttgca gtgagccgag attgtgccat tgcactccag cctgggcgac 92400agagcgagac tctgtctcaa aaacaaaaca aaacaaaaaa aaaaaaaacc aggctgcaca 92460ggaagaagtg agcaagcatt accatctgag ctctatctcc tctcaggcca gtggtggcat 92520tagattctca taggagcgtg tatgagttcg ttctcacact tctgtaaaga catacctgag 92580acatataaag aaaagaggtt taattggctc acagttctgc aggctgtaca ggcttctgtt 92640tctgggaagg cctcaggaaa cttgcagtca tggcagaagg tgaaggggaa gtaggcacat 92700cttcacatgg cccacaggaa aaagagagaa ggagagagag agagagacag agagagagag 92760agaaaaagaa agattgagag ggagagagga gggagaaagg agagtgcctg tagggggagt 92820tgctacacaa aggagcacca gggggatggt gctcaaccat tagaaactac ccccatgatc 92880caatcacctc ccaccaggcc ccacctccga cactggagat tacaattcag catgagattt 92940gggtggggac acagagccaa accatatcag agcatgaacc ctattgtgaa ctgcacattt 93000gagggatcta ggttgcatgc tccttatgag aatctaatgc ctgatgatga tttgaggtgg 93060aacagtttca tcccgaaacc atcccccgcc aaccctggtt tgtggaaaaa ttgtcttcca 93120cagaaccggt ccctggtgcc aaaaagtttg gggacctctg cacatatgca tgcacctgta 93180catggacaca taatacatgt acatatgcat actttatatt ctctgccact tctggtccag 93240actgatatac tatctcattt ggattactgc actagccttt tgttttggaa acagcatttt 93300ttaaaaaatt taatttaatt tttttgagat agggtgtcat tctgttgccc agcttggagt 93360gcagtgtcat gatcatagct cactgcggcc tcgatctccc aggctcaagt gatccttctg 93420cctcagcctt ctcagtagtt gggactacag gcatacccac catgcccagc taattttttg 93480attttttttt ttttttgaga cagagtctca gcctgtcgcc caggctggag tgggttggcg 93540cgatctcagc tcactgcaac ttctgcctcc caggttcaag tgattctcct gcctcagcct 93600cccgagtagt tgggattaca ggcgcctgcc accacaccca gctaactttt tgtattttta 93660gtagagacgg ggtttcacca tgttggccag gctggtctcg aacttgtgac ctcgtgatta 93720gcccgcctcg gcctcccaaa gtgctgggat tacaggcgtg agctaccgct cccagccagg 93780aaacagcatt cttgagataa ttcatataat tcacccattt aaagtatata attcattctc 93840tttagtatgc ccacagagtt gtacagccat caccagaatc agttttagaa cccataaagg 93900aactctgtac tctttaccca aaacctccat gcctccagct gcaggcagcc actaacctgc 93960cttctgtctc tgtgactcta cgtcttctgg acattactgt ggatgggctc atacagtcag 94020tgagcttgtg actggtgcct tctaccaagc agggttttca gtgtagcagc ctctctgttt 94080ttcttttttt tttaaattgt gacggaactt ctgcctcccg ggttcaagcg attctcctgc 94140ctcagcctcc cgagtggctg ggactacagg cccatgtcac catgcctggc taattttttt 94200tttttttttt tttagtagag atgggtttca acatgttagc cagggtggtc tcgatctcct 94260gacttcatga tccgcctgcc tcggcctccc aaagtgctgg gattacaggc gtgagccacc 94320atgcccggct aacctttcat ttactgtctg catttcttcc ctgatgcctt ccagtccatg 94380cacccgattg tagccattca tcctattatg gtttaaggtg actgtcttag tcagcatggg 94440ttgccataac aaaataccat agcctgggtg gcttcaacaa cagaatttac ttctcacact 94500tctggaggtt gggaagtcca agatccagga ctttcgcctt gccctcatgt ggtgaggggg 94560tgaggaagct ctgtggggcc tcttatatat ggatgctaat ctcattcatg aggggtctgc 94620cctcatgacc cagtcacctc ccaaaggccc cacctcctaa taccatcacc ctggtaatta 94680agtttcagtg tataaatttg ggggactata gacattgaaa ccataacaag cacttttcta 94740agatcaggga gtgagtaagt agcagagcta ggacctcaat tccacatgtc agtcatcttg 94800ccttcactct gctccatgat ggctgcctcc tagagcattg ggagtctcga tgttctatat 94860gctctcatgt gttgtgtatt ggagatagtt gaggctttat gaatacatct ggatttgttg 94920acttctagct ttgctggtaa ccagctgtga ccttgaataa gttacttcat ctctgagcct 94980gtttcctctt ttagaaacag gagtttaaaa tgctgctttg ggttgggcac ggtggctcat 95040gcctgtaatt ccagcacttt gggaggctga gatgggagga tcactggagc ttggagttcg 95100agaccagcct gggcatcata gtgtgagatc ctgtctcctc aagaaattaa aaaattagct 95160gggtgatgtg gcgtgtgcct gtggtcccat ctactctgga ggctgaggtg ggaggattgc 95220ttgagcccag gaggttgagg ctacaatgaa atatgattgc accccatcct gggtgacgag 95280tgagaccctg tctcaaaaaa gaaaaaaaaa atgctgcttt gtaccccttt catgtcatgg 95340cgtcatggcc aacatagaat gccctggttg tttgctgttg gagggcatgg gcctgggggc 95400tccctgaggg ctccttccat cttcaactca ttctctgtgc acctgttagg aagttgtggg 95460ccagtcccta ccatgtatca ttgtgtgggt aaaagtaaat aaaatgtgta cagtgtctga 95520actgtacata tcagggtcca agaacaaaat gagtgacatg ggttagctct ttttaataaa 95580tggtaaaacc aaatattcta attttcagtt ttgttatact tccatcacat gtttttgttt 95640ttttgttttt tgtttttgtt tttctatttt aggcagcctt gccttctcta acaaaccccc 95700cttctctaag tcccatccga cgaaagggga aggagaaaga accaggagaa caagcatctg 95760taccgttgag tcccaagaaa ggcagtgagg ccagtgcagg taggaaacag cgtggggaag 95820ggagggacat gagtgcagca tctgtcatgt agaaacatag gatttaagta acttggtgtt 95880ttagagaaat aaatataata cacatcagta aagtgagaga aagtttctcc aggtgcggtt 95940caagatatta gaaactaatg actgatgtac acagaccacc ttttggtctg aagcatttct 96000aagtgccact ggctgacatg cagcccctac agcctccagg cttccagccc tagcatggag 96060catcactctc ctatgcttcc ctggttgcag gtgatggctg gagaggcctc ctgattttca 96120gtaagggaag tggtgtagat gcttaggaat agatgtagtg agtgaaaaaa ctgattctga 96180tatgtcaaaa attctgattg gaaatggaat atttacattt ggaagagcta aaggcgagag 96240aaagtgggga taaagtcatc tgagttggag gagcttaaac cattcacaag tttggaggac 96300ctttttttac ccatgaaaag gtcagaacag aaggggctag gatttaggtg tgactgcagt 96360ttattgaatt cccatccata ctgctctcgg tgggcagtgg caggggcagg agaggagcct 96420ggcaaagcat gaagtgactg ctgctgcctc tgctatctgg gacgcctggc cacctgtctg 96480tacagtctcc ctccagaccc attctcacgc tgtctcttgg cacccagggg ccagtgatgg 96540ttctcccatt tgttttgtgt atatagcatt tatatcaagg ctatttattt atttatttat 96600tttatttatt tatttttttg agacagagtc tcactctgtc acccaggctg gagtgcagtg 96660gtgcaatctc ggctcagtgc aagctctgcc tcctgggttc aagcaattct cctgcctcag 96720cctcctgagt agctgggact acaggtgtgc accaccacac ctggctaatt ttttgtattt 96780tttattagtg gagacggggt ttcaccttgt tggccaggat ggtcttgatc tcctgacctc 96840gtgatccgtc cacctcagcc tctcaaagtg ctgggattac aggcatgagt cactgtaccc 96900ggcctattta tttattttta attgacaaaa ttgtatatat ctgtaatata caacatgatg 96960tttgaaatat gtgtacattg gccaggcgtg gtggctcaca cctgtaatcc cagcactttg 97020ggaggctgag gtgggcggat cacgaggtcg ggagttcaag accaaactgg ccagcatggt 97080gaaatcctgt ctctactaaa aataccacaa aaaaaaaaaa aaaaaaaaaa agccgggcat 97140ggtggctcgc gccagtcgtc ccagctactt gggaggctga ggcaggagaa ttgcttgaat 97200ctggcaggtg gaggttgcag tgagctgagt tcatgccact gcactctagc ctgggcgata 97260gagcgagact ccgtctcaaa aaaaaaaaaa aaagaagaaa tacatatgca ttgtggaatg 97320gctaattaac ctgtgcatca cctcacgtat cattgttttg tggtgagaac acttaaaatc 97380tactctttca gtgattttct tgcatatggt acattgctat taactgcagt caccatgcta 97440tacagtagat ctcttgaact cattcctcct gtctataaat gaaattttgt atccttgacc 97500aacacattca aggttttttt tgagatggag tcttcttcac ccaggctgga gtaccatggc 97560acgatctcat ctcactgcaa cctccgcctc ccaggttcaa gcaattctcc tgcctcagcc 97620tcctgagtag ctgggattac aggcacatgc tactgcacct ggctaatttt tgtattttta 97680gtagaagtgg agtttcacca tgttggccag gctggtctcg aactcctgac ctcaagtgat 97740ccgcctgcct tggcctgcca aagtgctggg attacaggtg tgagccactg cacccggcct 97800caagcgtttt aaaagatgct cttttctaag gattgactgt agtacaggag gaagattgac 97860ctgttgaaaa gcctcagcct ttacaagtgt aaaattatca gtatattact atcatctttc 97920tgatgaatta aataaactaa ggactccaag tcaaaagtct tcaaactgaa gtagaatagt 97980tgtatatagt gcttggcact ttaatattta gtatcggttt aatgataatg tttgtgcctt 98040tgccgtcttt aaaacatttt tacatcatcc ctgtttgatt acttggtgtg ctcatgaagt 98100tgttggccac taaggaatct taggctcaga gaggttctgg aattggccag tggtccttga 98160atcagctgct cctatgattc tctaactgat ttctcacaaa gcaaacaagc aatcataaca 98220aaacaactgt gcacactgct cttcttattt tgttatttaa aaagtactta ggctctactt 98280atgtttgtta gtcaatttct cattacttct agttaatcaa aaggtcagag gaaatacttg 98340aatattttca tactagaata ctttaaaaaa tcatgatttc cagtaatctc tttaaaactt 98400ggcaagttat tttgatctaa aagtttatct tttgtgtgca tatttttaaa gcttctagac 98460aatctgatac ctcaggtcct gttacaacaa gtaaatcctc atcactgggg agtttctatc 98520atcttccttc atacctcaaa ctgcatgatg tcctgaaagc tacacacgct aactacaagg 98580tatgggcctc tgcatctttt aaaaatatat atgcacacat acttacgtct aatggatagt 98640tgatgttttt cttatgattt gtaggatgta taagcccttt gagatatgag ttacatttag 98700ttttttcaag tttgtttgtc tttcagcttt gtttatgata gcttctatca tacaggtgtt 98760ttggattttc atattgtttg tactcacagc taagattgat tacagtgaca gagctaggat 98820gtgcagccag gttatagggg gaagtggccc tggtggagtc tggagggatc cgtgtacagg 98880cttccttccc tcccgtgagg ctcacacaaa aatacagcaa catgctggtc ctgcaggtac 98940cctctgccta acatgagcca caattccaga ctcacagaag aaaagcaggt gttcggcata 99000aaccatgtgt ttcaaatagt ctgggcatgg tgagccactt gttatcagct agggaaagtt 99060tatgtcagcg taagaaactg ttcaccagat acccccaaga gccagccttt ctgtctaggg 99120atgttttagt tttttagttc attttttttt ttaactttaa aattttctgt tcatctgcaa 99180tttgttagat atgaagtatg tgtctaattt aatttttgtt tttggttgtc cccaataatg 99240tttacagaag aatttttctg cactaattgg cttgagttac ttacattctc atagttctct 99300agtttcagta gtttcattta ttattttgtt atatcaatct atctgtctgc tcatctatta 99360gaagcatcct tgtttttttt ttttcttttt tagacagagt cttgctctgt ccccaggttg 99420gagtgcagtg gtgcaaccat gcctccctgc agtctcaggg ctcaagtgat cctcccacct 99480cagctcctga gtacctggga ctaccggcat gtgccaccac acccagctaa tttttacatt 99540ttttgtagag acagggtctc cctaagttgc ctgggctggt ctcaagctcc tggcttaagt 99600aatcctccct ccttggcctc ccaaagtgct gggattacag gtgtgagcaa ctgcacccgg 99660ctacaagtat acttcttaat tattgtagct taatggtatt tatgagggga tcagttcccc 99720tgttgttctt tagaattttc tggatattct tctttattga ttttgggatg tgaacaatag 99780aatcaacttc tacttgtaga ttgatttagg gagaacttat acctcagatg ttaagtcacc 99840ctgtccagaa tgtgggatgc tttcctattt gttcagaact ttttaaatta cctcagaagc 99900acatgaaatt taaaggattt taaaaaaaac ttaaagatta tttcacatag ctcttgcaca 99960tttcttgata aatgaatcct caggtattcc tctgtttttg ttactaatag ttacttctta 100020tgggtttttt ttcccctgaa aatcatttat caaacgtatg tggcttattt tctgaaggat 100080gtttgataat tttggaagat atgaaagtct tcatatttta caaggtttga ggtctcttta 100140agctgcatgg ttctcatgtc agctcccaaa gcagaagacg gcatgttgaa aaatgccgta 100200gagaagatac ttcttttcca cctgttttca actcatatca tcttgaattt cagggcacct 100260ttccatgctc ctagtgcttg ctatctgttt attattttcc ttcctgaata ccctgaactc 100320cagcatgttc tgctgtaatt ctggcctccc tggcatcttg gactcctgtt tcctttgctc 100380tgtcatcccc gcggtcagct cctgctgcgc agcttctcag ctgaagtgcg tttggagtgc 100440ctggcgtgtc ttgctggatc tttgagtatt gcctctggtt tccttggttc cttctgctga 100500gttgctcagc gtctccactc cccatttctt gtgtggccct tcctgcactc ctctgattcc 100560ttttgtcttc cctggtttct tgctttggtt tcgagtctcc acagaacttt tgcagctctt 100620ctgaagacct ggaagctttt tcatcttaat tctcatctca tgacctcttt tcccttcttt 100680gagagctaga acttcccatg gtgaacttct ctttccagaa ttccatgcct tcttttccct 100740cccacttacc tgttgtccag gagaggtcag attgctgtgc atattggagg agaacccttt 100800cttccctggg ctcttcatct cacatgacat caccacatca cctcgttcct tggaccctca 100860gtggtgtcac tgctggattt ttctttcctt tggctggcct tagggcacac ccaggttgac 100920tagcgtagtc atggtattta gatccactca cattttcagt ttctgtgtct gtctcttgcc 100980tgcttctgac ttcgcccaga gaaagcttct ctttcacaag ggttcttaga tttatgttca 101040ctgagcacct tcttttctga ggcagtgttt taccaatatt tattttccta gtcagtctcg 101100ccttaccttt cttgttatgc atgtctttgg tcctgaccca ttctctgagt ctgtaaaata 101160gaattgctgt ataatttaat tacatgaaat cctttagaat cttaacacat cttacacctg 101220atttaatatt ttattgtatc caaattgaac caaccctatg tgaatttgac agtgatttct 101280cccagggatc ctagtgtata aggaatagga cttagtattt tctatttttt gatataccac 101340ataccagata ctgattatga tggacattta accctttttt ctcattatga aagaaagtta 101400ggaattattt cttccagtag cgccagtgta acctgaaagc ctttgaaaga gtagtttttg 101460tatagctatc tgaaaggaat ttctttccaa aatatttttc cagtgctgac aacaaacacg 101520cagacacacc ctgcaaggtg agtgtacggc gccgcacagt ggaggcatct gctgcagccg 101580tcgatgtttg tgtctttggt tgtacattat gagatcgtga cagggccagt aaccgtgtgt 101640tctctccttc accttcccaa ggtcacgctg gatcttcaga acagcacgga aaagtttgga 101700gggtttctcc gctcagcctt ggatgttctt tctcagatac tagagctggc cacactgcag 101760gacattggga aggtttgtgt cttgtttttt ctccttgggt tgtggctggc acacttgatg 101820tgcgtcttct gggctgagtt catctaggat ggagcctggt tctccagggt gcctccggga 101880gactcctccc tgccccacgt gcttgcgtca caggacccaa gtctgactct gccttagcca 101940tgaagtttag ggggaagttt ctatttgtat tctatttttg tctgttatca tgtattagct 102000tagacccagt ttagtttgga aaatcagtgg gtttcaaaat gtgtttgtag agtcctttat 102060ttcttaactt gaccttttca agtggaaagg ggcaaaacag acgggtaagg gggcggggcg 102120ggaggtgtga cttgctcttt tgtgcctgag gaagtaacag agctggggtt gacagtcata 102180ttctctgaca cagatagtct ctgacttatc tcacagaaag tcagcggcag agcctgagtt 102240aaaagtctcg tagattttct ttttcttttt tttggtggct aatttcagtt ttatttatat 102300ttgtttattt atttattata ctttaagttc tgggttacat gtgcagaatg tgcagttttg 102360ttacataggt atacacgtgc catgatggtt tgctgcaccc atcaacccat cacctacatt 102420aggtatttct cctaatgtta tccctccccc agtcccctca ctccccatgg gccccggtgt 102480gtgatgttct cctccctgtg cccatgtgtt ctcattgttc aatttccact tgtgagtgag 102540aacatgcggt gtttggtttt ctgatcttgt gatagtttgc tgagaatgat ggtttccagc 102600atcatccatg tgcctgcaaa ggacatgaac tcatcctttt ttatggctgt atagtattcc 102660atggtgtata tgtgccacat tttcttaatc cagtctatca ttgatggaca ttcgggttgg 102720ttccaagtct ttgctattgt gactagtgcc acaataaaca tacatgtgca tgtgtcttta 102780tcgtagaatg atttataatc ctttgggtat atgcccagta atgggattgc tgggtcaaat 102840ggtatttcta gttctagacc tttgaggaat cgccagactg tcttccacaa tagttgaact 102900aatttacact cccaccaaca gtgtaaaagt gttcctattt ttccacaacc tctccagcat 102960ctgttgtttc gtgacttttt aacgatcgcc atcctaactg gcgtgagatg gtatctcatt 103020gtgattttga tctgcatttc tctaatgacc agtggtgatg agcatttttt cgtatgtctg 103080ttggctgcat aaatgtcttc ttttgcgaag tgtctgttca tatcctttgt ccattttttg 103140atggggttgt ttgctttttt ttcgtaaatt tgtttaagtt ctttgtagat tctggatgtt 103200aatcttttgt cagatgggta gattgcaaaa attttatccc attctgtagg ttgcctgttc 103260actctgatga tagtttcttt tgctatgcag aagctcttta gtttaattag atcccgtttg 103320tcaattttgg cttttgttgc cattgctttt ggtgttttag acatgaagtc tttgcctatg 103380cctatgtcct gaatgttatg gcccaggttt tcttctagga tttttatggt cctaggtctt 103440atgtttaagt ctttgatcca tcttgagttg atttttgtgt aaggtataag gaaggggtcc 103500agtttcagtt ttctgcatgt ggctagccag ttttcccaac accatttatt aaatagggaa 103560tcttttcccc attgcttatg tgtgtcaggt ttgtcaaaga tcagatgatt gtagatgtgt 103620ggtggtattt ctgaggcctc tgttctgttc cattggtcta tatatctgtt ttggtaccag 103680taccatgcag ttttggttac tgtagtgttg tagtatagtt tgaagtcagg tagtgtgatg 103740cctccagctt tgttcttcta gcccaggatt gtcttggcta tgcaggctct tttttggttc 103800catatgaagt ttaaaatagt tttttccaat tctgtgaaga aagtcagtga tagcttgatg 103860gggggatagc attgaatcta taaattactt tgggcagcaa ggccattttc acgatattga 103920ttcgtcctat ccatgaacat ggaatgtttt tctatttgtt tgtgtcctct cttatttcct 103980tgagcagtgg tttgtagttc tccttgaaga ggtccttcac atcccttgta agttgtcttc 104040ctaggtgttt cattccctta gtagcatttg tgaatgggag ttcactcatg atttggctct 104100ctgtttgtct gttattggtg tataggaatg cttgtgattt ttgcacattg attttgtatc 104160ctgagacttt gctgaagttg ctaatcagct taaggagatt ttgagctgaa ccaatagggt 104220tttctaaata tacaatcatg tcatctgcaa acagggacag ttttacttcc tctcttccta 104280tttgaatacc ctttattgct ttctcttgcc tgattgcgct ggccagaact tccaatacta 104340tgttgaatag gagtggtgag agagggcatc cttgtcttgt gccggttttc gaagggaatg 104400cttccagttt ttgcccattc agtatgatat tagctgtggg tttgtcataa atagctctta 104460ctatgttgag atacgttcca tcgataccta gtttattgag agtttttagc atgaaaggct 104520gttgaatttt gtcaaaggcc ttttctgcat ctgttgagat aatcatatgg tttttgttgt 104580tggttctgtt tatgtgatgg attacgttta ttgatttgcg tatgttgaac cagccttgca 104640ttccagggat gaagctgact tgattgtggt ggataagctt tttgatgtgc tgctggattc 104700agtttgccag tattttattg aggattttca catcgatgtt catcagggat attggcctaa 104760aattctcttt ttttgttgtg tctctgccag gctttggtat caggatgatg ctggcctcat 104820aaaatgagtt agggaggatt ctctcttttt ctattgattg gaatagtttc agaaggaatg 104880gtaccatctc ctctttgtac ctctggtaga attcggctgt gaatccatcc tggacttttt 104940ttggttagta ggctattaac tattgcctca agtttagaac ctgttatcag tctattcaga 105000gattcagctt ttttctggtt tagtcttggg agggtgtatg tgtccaggaa tttatccatt 105060tcttctagat tttctagttt atttgggtag agatgtttat agtattctct gatggtagtt 105120tgtatttctg tgggatcggt ggtgatatcc cctttatcgt ttttattgag tctatttgat 105180tcttctctct tttcttcttt attagtcttg ctagcggtct acctatttta ttgatctttt 105240caaaaaacca gcacctggat tcattgattt tttttggagg gttttttttc gtgtctctat 105300ctccttcagt tctgctctga tcttagttat tttttgtctt ctgctagctt ttgaatttgt 105360ttgctcttgc ttttctagtt cttttaattg tgatgttagg

gtgttaattt tagatctttt 105420ctgctttctc ttgtgggcat ttagtgctat aaatttccct ctacacactg ctttaaatgt 105480gtcccagaga ttctggtatg ttgtgtcttc gttctcattg gtttccaaga aaatttttat 105540ttctgccttc atttcgttat ttacccagta gtcattcaag agcaggttgt tcagtttcca 105600tgtagttgtg tggttttgag tgagattctc aatcctgagt tctaatttga ttgcactgtg 105660gtctgacaga cagtttgttg tgatttctgt tcttttacat ttgctgagga gtgttttact 105720tccaactatg tggtcagttt tagaataagt gcaatgtggt gctgagaaga atgtatgttc 105780tgttgatttg gggtgcagag ttctgtagat gtctattagg tccgcttggt ccagtgctga 105840gttcaagtcc tggatatcct tgttaatttt ctggctcatt gatctgccta atattgacag 105900tggggtgtta aagtctccca ctattaccgg gtgggagtct ctttgtaggt ctctaagaac 105960ttgcttcatg aatctgggtg ctcctgtatt gggggcgtgt atatttagga tagttagctc 106020ttcttgttga attgatccct ttaccattat gtaatggcct tctttgtctc ctttgaactt 106080tgttgattta aagtctgttt tatcagagac taggattgca atccctgctt tttttttgct 106140ttccatttgc ttgttagatc ttcctccatc cctttatttt gagccaatga gtgtctttgc 106200atgtgagatg ggtctcctga atacagcaca ccaatgggtc ttgactcttt atccaatttg 106260ccagtctgtg tcttttaatt ggggcattta gcccatttac atttaaggtt aatattgcta 106320tgtgtgaatt tgatcctgtc attatgatcc tagttggtta ttttgcccgt taactgatgc 106380agtttcttca tagcgtcagt agtctttaca atttggcatg tttttgcagt ggctggtact 106440ggttgttcct ttccatgttt agtgcttcct tcaggagctc ttgtaaggca ggcctggtgg 106500tgacaaaatc tctgcatttg cttgtctgta aaggatttta tttctcgttc acttatgaag 106560cttagtttgg ctggatatga aattctgggt tgaaaatact ttttttaaag aatgttgaat 106620attggctccc actcttttct ggcttgtagg atttctgcag agagatctgc tgttagtctg 106680atgggcttcc ctttgtgggt aacccgacct ttctctctgg ctgccctttc cttcatttca 106740atcttggtgg atctgatgat tatgtgtctt ggggttgctc ttctcgagga gtatctttgt 106800ggtgttctct gtatttcctg aatttgaatg ttggtctgcc ttgctaggtt ggggaagttc 106860tcctggataa tatcctgaag agtgttttct aacttggttc tattctcccc atcactttca 106920ggtacaccaa tcaaacgtag atttggtctt ttcacatagt cccatatttc ttggaggctt 106980ggttcatttc ttttcactct tttttctcta atcttgtctt ctcgctttat ttcattaatt 107040tgatcttcaa tcactgatat cctttcttct gcttgattga atcggctgtc gaagcttgtg 107100tatacttcac aaaattctcg ttctgtggtt tttagctcca tcaggtcatt taagctcttc 107160tctacactgg ttattctagc cattagtcta acattttttt caaggttttt agcttccttg 107220tgatgggtta gaacatgctc ctttagctcg gagaagtttg ttattaccga ccttctgaag 107280cctacttctg tcaattcatc aaactcattc tccatccagt tttgttccct tgctggtgag 107340gagttgtgat cctttggagg agaagaggtg ttctggtttt tggaattttc agcctttctg 107400ctatggtttc tccccatcat tgtggtttta tctacctttg gtctttgatg ttggtgacct 107460acggatgggg ttttggtgtg ggtgtccttt ttgttgatgt tgatgctatt cctttctgtt 107520tgttagtttt ccttctaaca gacaggcccc tcagctgcag gtctgttgga gtttgctgga 107580ggtccactcc aggccctgtt tgcctgggca tcaccagcag aggctgcaga acagcaaata 107640ttgctgcctg atccttcctc tggaaacatc gtcccagagc acgaaggtgt ctgcctgtat 107700gaggtgtttg ttggccccta ctgggaggtg tctcccagtc aggctacatg ggggtcaggg 107760acccacttga ggcagtctgt tcattatcgg agcttgaatg ccgtaccggg agaaccactg 107820ctctcttcag agctgtcagg cacgtatgtt taaatctgga gaagctgtct gctgcctttt 107880gttcagatgt gcccttcccc cagaggtgga atctagagag gcagtaggcc ttgctgagct 107940gcagtgggct ctgcccagtt cgagcttccc tgctgctttg tttacactgt gagcatagaa 108000ccacctactc tagcctcagc agtggtggac acccctcccc cagccaagct cctgcatccc 108060aggtcgattt cagagtgctg cgctagcagt gagcaaggcc ccatgggcgt gggacccgct 108120gagccaggca caggagagaa tctcctggtc tgctggttgt gaagactgtg ggaaaagtgc 108180agtatttggg caggagtgta ctgctccttc aggtacagtc actcatggct tcctttggct 108240tggaaaggga agtcccccga ccccttgtgc ttcccaggtg aggcaacacc ccgccctgct 108300tcggcttgcc ctccgtgggc tgcacccact gtccagcaag tcccagtgag atgaactagg 108360tacctcagtt ggaaatgcag aaatcacctg tcttctgtgt cgatctcact gggagctgta 108420gactggagct gttcctattc ggccattttg gaagcatccc ttgttttttg aggtggagtc 108480ttgctctgtc gcccaggctg acgtgcatcg gcacaatctc ggcccactgc aacctttgcc 108540tcctggtttc aagcgattct cctacctcag cctccggagt agctgggatt acaggcacct 108600gccaccatgc ctggctaatt ttttgtattt ttagtggaga tggggtttca ccacattggc 108660caggctagtc tcgaactcct gaccttgtga tccacccacc tcagcctcct agagtgctgg 108720gatcacaggt gtcagccacc acgcccagcc atattttcag atctccctct ctttgcccta 108780aaccactgtg cttaataagt agtttttagt ggccagcagt ctccatgtat aacacatttt 108840agcaaaatgg aaaatactat atgttttaaa tttgaacgtg agattatact gaaataaaaa 108900tcatctaact gggattcttt aaatagtaag attttctttt ttgtatgtgg gttttttttt 108960aaccttatta ttatgactgt catatataga aatggctgtt tttcagttac agtcagtgaa 109020tgtatcaaat gctgccttat ccaaataata aaagtaaatt attaataagt cacaatttaa 109080tgaagattga tgttagttga tctttatatt cttgaaatca gccatatggt tgtgtgtgta 109140tgtatatatt tttaaaggta cataaagata ataagctcat ctctgaaaat ttttacattt 109200ggcataagaa taactggata attaagcatc ttattctctg gcctgtgtct ttacagttaa 109260aggtagattt actcacctct ccttttttgt ttttctaagt tcatcttttt tgctgtttca 109320agacagaggc ccattttagc tttctcgcat atccttttgt ttgtactttg gaagcctcac 109380ctgcttaatt gttgagtttt tatccgtggt cttttagagg gggatatgta gggtagaagc 109440tttcacaggt tcttgtttgc acttggcccc tgactgtttt gaggaatctc cctcactgac 109500tcacagcatg gcaaggtttc agatctcttt ctgccacaca gcagttctga ggcagctgga 109560aagatatcca gatgcttaga ttgtcaggcc aggcttgaga tatacaaact attgagcctt 109620atctgtgacc ttgcttaggt gaaggcatca gagcccctgc accaacatgc ataggcctct 109680gcatgtgtgc ggggctgggt gttgaggtct gagcacaagt gtagctggag aggtgagctt 109740gatgtggcga cgggtatgag caggttttct tcagacttct gtgagtttac ctagttccag 109800gatttaaagg cacagagact ttagaattaa aatagaatca ttttcttttt ctaaatagca 109860acactaggaa taaaaaataa taattccaca ttcttgacag gtaatgtttt ttcttgtctt 109920ctaatcctta tttattccat actcattttt atacataatt gaaatgtatt atgcattgga 109980tttttctttt gcattatatt atagacgatt tttcatgtaa ctccttactg ttccatttta 110040tatgttttgt ctggtttaag actttatctg caaaccggga aactgtctct acaaaaagaa 110100aaacaaaaat agttggccgc agtggcatgc gtctgtggtc ccagctactc ggggctgagg 110160tgggaggatt gcttgagcct tgggaggttg aggctgcaaa gagccatgat catgccattg 110220cactccagca tgggtgacag actttatact gtctgttttg ggtgatttga taatgatatg 110280ccctgatgta gtttttttat atcttgtgtt tcttgtgcct gggtttattg aggttgggtc 110340tgtggcttca tagtattttt aaagtttgga aaattttagg ccattctttc tttctttctt 110400tctttttttt ttttttgaga cagtgtctcg ctctgtcgcc tgcgttggag tgcagtgaca 110460ctatcttggc tcactgcaag ctctgcctcc tgggttcacg ccattctcct gcctcagcct 110520cctgagtagc tgggactaca ggcgcctgcc accacgcctg gctaattttt tgtattttta 110580gtagagacga ggtttcactg tgttagccag gatggtctca atctcctgac ctcgtgatct 110640gcccgcctgg gcctcccaaa gtgctgggat tacaggcgtg agccactgca cccagctagg 110700ccattatttc ttcaaagatt ttttttctgc cctgcctccc tccttttttc cctctcttaa 110760aggggctgtg atttcctgaa tgattgctta gtgttgtccc atagcttact gatgctcttt 110820tcagtgtttg attgttttat gtgttttctg ttttgtatag tttctattat tgtgttttca 110880agttctctga tcttttcttc tacagtgtct actctgttgt taatctgtta atctgttgtt 110940aatcctgtcc agcgtatttt tttttttgtt tttgaaacag tctcactctg ttgcccaggc 111000tggagtttag tggtgcgata tcagctcact gcaacctcca cctcccaggc tcaagcaatt 111060cttctgcctc agcctcccga gtagctggga ctataggcac gtgccaccac acctggctaa 111120tttgtgtatt tttattagag atggggtttc accatgttgg ccaaactggc cttgaactcc 111180tgacctcagg tgattcatcc gcctcggtct cccaaagtgt tgggattata ggcatgagcc 111240accgtgtctg gcccctgttc agtgtatatc actaattttg tttttatctc tagaagtttg 111300atttaggtct tttaaaaatg tctccctgtg tttctgttta gctttgtgaa cacaattgta 111360ataactgttt taatatcctt ctctgctagt tctaagatct tctaataact tcccagttct 111420tggtgtttct cattggttga ttgatactcc tcgttttggg ttgtattttc ctgcctcttt 111480gtatggctgc caatttttta ttggatgccc aaccttgtga attttacttt gttggatgct 111540atatattttt gtgttcccat agatcttctt gagctttgtt ctgaggttag ttgagttaca 111600tatagatggt ttactctttt gggtcttgct ttataatttg tcagatgggt tggagcagtg 111660cttagtttag gactaatttt ttttttggac taattattcc tctttaggaa taattaggta 111720ccatgcttag gaggcaagac catcctgagt actctaccta atgaaccaga aagtttgggt 111780tttccagtcc gcctgctgag aacagtgact ttctagccct gtgtgagcgc tgagctctgc 111840tccttctaat cctttccaat gcttctttcc ctggcctcag ggagttttct cacacacata 111900tctctgctga gtactcgaga gggaccttcc ccagatctcc agagctctct ctgtcttgtt 111960ttctcttctc tggtgctctg tcttatgaac tgtggctgtc ttggtctcct tagattctca 112020gcacctcttc aattcagagg gttgcctgtc cctcctcctt gtgccacagc ctaggaactc 112080tctcaaagca gcgagttggg gcagccatag ggctgactta gtctctcgtc tcccagggat 112140cactgtcctt cattgctcat gtccagtgtc ttgaggactc tgggttttgt ctgttttgtt 112200ttttggtttg ctttggttgt ctcaggcagg agggtaaacc cagtccctca ccctcattgt 112260gctcagtagt ggaagtctca ctctattaca ttagatatta gtatttgtag cagagccctg 112320gttccctggt acttggggag ctcttgaaag gccagaaaca gcatgctttc tcaccttttc 112380cagggcttca gtttctggtg cacatcaagc attccataca catttgttaa agtcctttgt 112440tagacaagta gtgattcaca ggttctattt gtaatttttt cagttaacat gtattgggta 112500tctgctggga gctagtaaaa acaaaaagtg gtgtgtgaca aattcaattc tgacaagaac 112560aaccttaaac acttagaata tactttgagc atatcagaat tttaaaaatg tgtggccctt 112620gagtatttga aaccaacaag aatctattgc ttattagtag aggatatttt gttaaacaag 112680tggagagaga ggcattttca gtctaattgg tgttggcttt tagcagctga tggaaaccag 112740ttcgtgatta gccaggcagt ggtgaaacag gctgtgcatt ctgaatgcct aggtatctag 112800gcattcagaa tggtggcgct ctttgagtta gcatcttctt ctttcttgat tctttttttt 112860ttttttttga gatggacttt cgctcttgtt gcccaggtaa caactccagt gcaatggcgc 112920catctcggct cactgtaacc tctgcctccc tggttcaagc gattctcctg cctcagcctc 112980tcaagtagct gggattacag gtgtgcgcca ccacgcctgg ctaattttgt atttttggta 113040gagatggggt ttcactatat tggtcaggct ggtcttgaac tcctgacctc aagtgatgca 113100cctgcctcga tctcccaaaa tgctgggatt acaggcgtga gccaccactc ccagcccctt 113160cttgattctt gaaaaggaca ttgggtgctg tacatctcgt tatagatgtt gataaaaatg 113220cttgtgagaa gagtaacatt aaggtagtta tttggtcatt tttgcagatt attttaagac 113280aattctagga ctgatttgtg gtaaatcaca cattgctgta tcatagttgt gttcactgaa 113340catattcagg ggctctacag atgcagggct cttagctgct ttgcacactt ctgaattcct 113400gccctgcgaa caggactgga tacctaatag acaacaggta cttgataaca gtttattgaa 113460ttaatgagtg aatgaacaga tacataaatg catgaaagaa tggttgtaat gtatataact 113520tggatttcaa gactttttac tgactgttca aaataagaaa ttgaaaactt tcctctgatt 113580ttcctctact atttacacaa tttaaatgga agttatcttg taccttcaat ttctgtctag 113640gattcgtaca ataacgggtc atctctgagt cgcttaatgt ctcacttgtc tttctacagt 113700gtgttgaaga gatcctagga tacctgaaat cctgctttag tcgagaacca atgatggcaa 113760ctgtttgtgt tcaacaagta agagcttcat tcttttcctc ttctgttaag acgttcgggt 113820atgacagcaa aacgctgcta ctccttaaga ggcaggcgct gttggcataa tcagctggga 113880ggattgtggg gtccagcgca gcactttttg gctcagtcca tgattgagcc aagaggccat 113940ccttcccttc actccccagg aggacgaggt ctgtcactgt ggagggcaga ggacaccaga 114000agctcctctg caacctcgct agttaacttc cagtccctcg gagtttctgt ttagaatgct 114060caatctcatt tagaattgca aggaaaccca aaacgcctat ttaaggtaca aacagcactt 114120catacaatat ctcatgaggt attaatagtg attcacagga agaatttcac gctgtgagtc 114180tttgctaaca tatccagtta tttacagatg gatttgatat ttgtgtggga gattcttaaa 114240agtgttgttc acgccacatt gttgatgcct catttttttc actgtagttg ttgaagactc 114300tctttggcac aaacttggcc tcccagtttg atggcttatc ttccaacccc agcaagtcac 114360aaggccgagc acagcgcctt ggctcctcca gtgtgaggcc aggcttgtac cactactgct 114420tcatggcccc gtacacccac ttcacccagg ccctcgctga cgccagcctg aggaacatgg 114480tgcaggcgga gcaggagaac gacacctcgg ggtaacagtt gtggcaagaa tgctgtcgtt 114540ggtggaagca cgaaagagca agcaggaaat actttgtaaa agaataaaaa cgaaaaatgt 114600tagcgaacat cttctaatag tctgctgtat tcagagaact ctaggagata tatatggttg 114660atgcaaagat gatttaaggc atagcccggc cttccaagaa gtgtgtggcc agtgagtgag 114720atgggcttgg gacttacaca tctcagaggt gggggtagag gaggaggaac actgagtggg 114780ctgagaagca gccagctctc attgccaaag tgtgtcagca aaccagaatg cagttcataa 114840tgtccccacc cattcaaagc acaggacctg tagagtggtg tggcatgtgt tggtggcact 114900tttcaggcct gtaacaagga tgaaagaaca gcttcatagc agcacagtag tgctggtgtt 114960cagaggtgtg tgaaggccat agaagcatct tggatatatt accttgtgtt ttgtcagctt 115020tatgactaga agtctctttt cacttaaatt tgtttttttt ttttttgaga cggagtcttg 115080ctctgtcgcc caggctggag tgcagtggtg caatctcagc tcactgcaag ctctgcatcc 115140tgggttcatg ccattctcct gcctcagcct cccgagtagc tgggactaca ggcgcctgcc 115200atcacgcctg gctaactttt ttttgtattt ttagtagaga cggggtttca ccatgttagc 115260caggatggtc tcgatctcct gacctcgtga tctgcccgtc ccggcctccc aaagtgctgg 115320gattacaggc gtgagccacc gcgcccggcc tcttttcact taaatttatg tttgtgtttt 115380taatgcctag tatacaggac ttcttaaatt gccttaagta tgaacaggta tttgagttgc 115440taatctgtat agtagcaata atagaatccc ttgtttttcc ttttataaat ttagcgatta 115500aatagctaca attaaaacac tagagtcagg agtcaaggaa aatacccatg ttccaggctg 115560tatgttagtg atgtacttac tatatattgg agtttcagga gtaagtctgt ttcaatgctt 115620tctgtaacca tttggggtat taataagcat gtgagtgtgt gcatgtttgg gttaatttca 115680tatatgtttc ttagaaggga tatcattgat gtaaatattt taaaggcttg tcctccaaaa 115740aaatcatgta atttcttcta aattactgat cttttaaatg accttcacct ttctctcaaa 115800tctcacttaa gactgggctg agtagtcagt ttcctgtagc agaaaaaagc tcagacttga 115860gtagccttct gcgagtgagg agacttgatg gctgtcaggc agctgtaaac tctaaataga 115920gtgtcattat ctgaagaggg cgatgctgcc acactgagtg gcctttcaag ttgtttctca 115980atctgacacg ttctgatcgt gtgaatgtga aattggtttg agcaggagta tatctgagtg 116040cagaggagat tatttaaaga tattctcatt ctctgcttcc cttttattcc catttggcag 116100atggtttgat gtcctccaga aagtgtctac ccagttgaag acaaacctca cgagtgtcac 116160aaagaaccgt gcagataagg taaatggtgc cgtttgtggc atgtgaactc aggcgtgtca 116220gtgctagaga ggaaactgga gctgagactt tccaggtatt ttgcttgaag cttttagttg 116280aaggcttact tatggattct ttctttcttt ttttcttttt tatagaatgc tattcataat 116340cacattcgtt tgtttgaacc tcttgttata aaagctttaa aacagtacac gactacaaca 116400tgtgtgcagt tacagaagca ggttttagat ttgctggcgc agctggttca gttacgggtt 116460aattactgtc ttctggattc agatcaggtt tgtcactttt atctttcatc catcatacct 116520gttcctaatt tagtacaaat taccctaaaa gacactgaaa tctactttaa agaaatgtgg 116580tctgcatgtt tccctcatca gttgctgctg cttatctttt tcatgcacct agctggtgca 116640gaaggcctgg ggcatagcca gcctcagcaa gtcagcatcc ttgccccagc tccctggact 116700caaggctaac ctggggttgg ctgttaggga tttccaaagg tttgtcccat ccacttgcct 116760cccctccaaa ataagtttga atttaaattg tgagatacaa ttaagattta ttgtttgggg 116820aacatttttg caaaatctag agttagttta aacagattat caattattac cataattgat 116880catctgcagt ttcaagctat ctaacaggtt cacttacctc tttaaaaagg aatggaattt 116940agcaggacag taactgagac ccgtgctcct ggagtccatg tgggagctgt gtggctctgc 117000acaagcattt gcacgcttcc cctcttgact gcattacctt cctcctatag ttgctgtggg 117060caccagattc tggctagtcc tgtcccttca tgatgcacat tttcctcaag attcgtccca 117120gttaaatcac tgcagatgaa actgcctttt catcgtcaaa atttaactgt catttttgag 117180ccgtgatctt gggctacttt cttatgtggg gtaggaatat ttgtgagtta gaaatattac 117240acttctctat ttccttctag acgtaaatct gttaatcctg tcagcactgt tactcacctg 117300aaagggtctg tttccctagg agaactgagg gcactcggtc aacactgatt ttccacagtg 117360ggtattgggg tggtatctgc ttgttttttt tgttgttgtt gtttgttttt ttttgttttt 117420tttttgagat ggagtctcgc tctgtcaccc aggctggagt gcaggggtgc gatctcggct 117480cactgccagc tccgcctcag aggttcacgc cattctcctg cctcagcctc ccgagtagct 117540gggactacag gcacccacca ctacgccagg ctaatttttt gtatttttag tagagacgag 117600gtttcactgt gttagccagg atggtctcca tctcctgacc tcgtgatctg cccgcctcgg 117660cctcccaaag tgctgggatg acaggcgtga gccaccgcgc ccggcctggg gtctgctttt 117720aatgaaggag gcatcaaggg gtgggctttg cgttggcctg atgctttcat ctttctttca 117780caaaacctgt ccgaagaaaa tccgtctaaa tgggccattg ctctcctcag gaaatagtca 117840ttgggaactt cttttccttt cctttgacac taggaggctg actggggaga agccctggtc 117900tatggctgtg ggcagcaggg gctgagagga gcaggctctc aggggggcac gggtacccca 117960agggaagcca gagccctgat ttgttccatt ctagtaagaa caaagactgc tctggtttca 118020tgtttgttct gattgccttt catcaaccgg tcccctttct cccagttctt aagattcagt 118080acagtgacag ttttatgaac aagaatagaa cactagaaca gacaaaccat tgaactctat 118140gctgataaag atttattgag ctcctgctgt atgtttgcat tctgcccaga ggctctgaga 118200aaaccaggcc atatgctcca tgctttatcc atggaagctc cccgtcaggt tgggaaagct 118260gacagctgca gggaatacag tgtgacacaa aactggctcc catgcagccc ttacgtgtcg 118320cctctcagat ggttggggga cgaaggtcga ctcctttggg tatcttatta ctaaaccagt 118380ttcagggaat ctgtgccacc ctatctgcca ttaacgtgaa cagatgagtc cccaaggtgt 118440aattttgggt attgtctgat gtctcttgga atttattatt tgtttttcca atgagatttc 118500acctcagggt atagtaaagt tgttgagggg attcctggat gtgttctgca attatctagg 118560ctgatttcag aatagagtta tgcttatagt caaatttatc agctgtcaag aattttattt 118620aaaatttatg cagataagca ggaggaaaag aagcctggtt tttacatttt aatcctatta 118680ttgatgtgaa attttatttt ccttcctgta ggtgtttatt ggctttgtat tgaaacagtt 118740tgaatacatt gaagtgggcc agttcaggta atagcatttt attattttag atttttttct 118800tcttcttgtg tacttacatg taatttaggt tattaagtga atgtttaaac tactgttagg 118860catttttgct gttttcttta aatggaaatc tgactaacat actgtgcatt tttgcttctc 118920ttaaaaatta atgtatatct caagacttgt ttggaagtag ttatgtatct gaaaattcca 118980tatgttgtca gtattcattg cacatttcaa agcatttaat tgtgttgaca gatggtggaa 119040tgaaatcttg tggtggagca ctagttttta aatcttctta gagaaagcag ttttatataa 119100tgttgtcttt agtaattatt atgcatttgt attctctgca gctttttctt gctagatgtt 119160gaggttttaa tacttcttgc tagtccatta caggtttata attattaaaa gttaaaattc 119220ttttagtacc taaaatgctt aataaacatt gtaattagga aaatttagtg cagaaggaaa 119280gtgttcccag attccctggg gtctggaaac atagtgttta ttctaattac atgacacctc 119340cactgtgttt tggggcaagt tactgtttct cttttgagtt tcaatttctt caagagcaaa 119400gaggcagagg agagctagga agatcgtagc tgctgtgccc ctgtgccgtc gggtgccttc 119460tacctgctgc ctccgaacct ttacacatgt ccctgctctg cgcgagggca cagatgggat 119520gcactgtggc aggggtgggg ttagagtaga tcacggacac ctgttagctt gatgtgtgct 119580tgctgtcaag gttgaatcat gaattatttt atgttgctta tattgatatg tatcttaatt 119640ttaaaagaaa ggtctaaatg gatgtttttg tttttaggga atcagaggca atcattccaa 119700acatcttttt cttcttggta ttactatctt atgaacgcta tcattcaaaa cagatcattg 119760gaattcctaa aatcattcag ctctgtgatg gcatcatggc cagtggaagg aaggctgtga 119820cacatggtaa cgggacacac ctttcactgt cgtcttcggt gtcgtgatgt gcttggcagt 119880gttcgttttc atatacccac tttgaacgtt gtcagtggca gccatgtgct tctcaggctc 119940tgcatgtgtg tctgtgtatg tgaaggtact ggttagagac gtttcaaaag agaagagagc 120000atattcttta ctctcagcaa tttgtaatct tctcagggaa aaaaattcaa gaaacagtaa 120060gataacctaa ggtacagata gattctgaat ataaagttcc tgttcattca catgaaacgc 120120taaaagttct tcacttgatc ttagccaaaa ggccaagaag cgatgcaaca ctaaaaattc 120180ttaaatcgaa cttgccgtga attaaatttt gatctctcat ccagtggtat tggagatata 120240gtttgacttg ggttcagggc tttctgtttt gcctgatgat tttgctggag cttaaataag 120300gaacccagga gatggccagc tgtgcaagcc cccagcctgt ggaaggagct agtgtggttt 120360tatgaatgag ttgcaaatct ttctttgagc tttttgaact gatcttccag cattgcccta 120420ttgacccctc cctgactcct ttgctggaat ctgtaggctt

ttgaactttg acagggacac 120480atcctaagac ccttgcaaac tcccagatgt gagaatggca ctactactta gagtcttttc 120540gactcagcgt gtgtgcagaa gagcatcaac cgggctgtgt tgcgaggcag ggccttggct 120600gacctctcag tgtttacata gctaagccag ttagtgtttg ccacggcctc acaagggctt 120660cagattcaca cagccaaagt atagattatt aaaggcatag gtgtttggtt tcctggactt 120720ggagggtctt tggacagaaa atcagtaggc aaccacaccc agtactttgt gctgggaagc 120780ttggtcatct gtgagagggt cagagagtat acccatgcgt gcatgccacc gaagggtcag 120840tgagtattcc tgtgtgtgca tgtctcaggg ccggagagag tatgtgtcac tgagaggtca 120900gagtgtttgt gtgtgtgtca aagagggttg cattgtgccc ttcactgagg ggtcagaggg 120960tgcctcgcgt gtgtgtgtgt gtacgtgtgt gtgtgtcact gaggggtcag agtgtgcctg 121020tgtgtgtgct tgtgtgtgcg tacatgtcac tgaggggtca gagtgtgcct ctgtgtgtgt 121080gctcatgtgt gtgcatacgt gtcactgagg ggtcagagtg tgcctctgtg tgtgctcatt 121140tgtgagcgta tgtgtcactg agggggtcag agtgtgcctc tgtgtgtgtg ctcatgtgtg 121200agcgtatgtg tcactgaggg ggtcagagtg tgcctctgtg tgtgtgctca tgtgtgagcg 121260tatgtgtcac tgaggggtca gtgttcctat gtgctcatga cattgagggt cagagtgtgc 121320ctgtgtgcca atgaaaggca tttcttatat ttttttatat gtggtcatag tagaccagtt 121380aatttatttt gactcctgtg ttagaccaaa ataagacttg ggggaaagtc ccttatctat 121440ctaatgacag agtgagttta cttaaaaaag cataataatc cagtggcttt gactaaatgt 121500attatgtgga agtctttatt gtcttttcag atgaatcaag tagattattc ttgagaccag 121560gaatgttgct gttttggtta tttggaaagt tttatcattt tcaaattgac ttttgaattt 121620gagtcacctt ttttcagaag tggtgttaaa ttataggagc cctaggtttt ttttcttttt 121680ttagaagtca tcacaaaatg atcagtgttc agaggaagag ctttgacctt ccacatggta 121740taatgattga taaccttaat tcatctctta ccataaacca agtatgtgta agggttttct 121800ttatttcttg aaagcatttt gtagatgttg agagcagttt tccaaatgta atttccatga 121860aatgcctgat aagggtaccc ttttgtcccc acagccatac cggctctgca gcccatagtc 121920cacgacctct ttgtattaag aggaacaaat aaagctgatg caggaaaaga gcttgaaacc 121980caaaaagagg tggtggtgtc aatgttactg agactcatcc agtaccatca ggtaagagga 122040atgtatgttg gaactgtcgt ggatacttta ttgacccgtg cagatggaag gaagtgccat 122100gtggtaacgc tcactgttaa ctgtgttact ttgaaccagg tttgggcttt ctggggcctg 122160ggtagatgcc ggtgcagggg gatggggagg gaggcggggg gtgggggggt gtggtggagt 122220tggggaggtg cagtggcagg aggtgttgtt ggtgtgtatc cttttttttt ttttgagatg 122280gagtctctct ccgtcgccca ggctggagtg tggtggcacg atcttggctc attgcaagct 122340ccacctcccg ggtttaagca attctcctgc ctccacctcc cgagtagctg ggattacagg 122400catgcaccac catgcccagc aaattttttt ttttgtattt ttagtagaga tggggtttca 122460ccatgatggc caagctgttt cgaactcctg acctcaagtg atcctcctgc cttggcctcc 122520caaagtgcta ggattacagg cgtgagccac catgcccagc ctggtgttta tctttaaagt 122580gggcacagcc acaggagttc acctgactcc tggtctgaga gtcacgagat cgttcaagat 122640agtgaggccc tcttttccaa aacgaggacc aaaaatcaat tgacagtgtt ggtcaagatg 122700gtagaaacct taaaatgata gaaatctcaa ctctgaaata aaaactttat ttgtatattt 122760atttaccact attttgacat agggctaagg tctttttctt tgagctgatt tctggttttg 122820ttttcttaaa gtggcataag aattcaaaga cattttgagg aaggctgagt gcagaaatct 122880ctctttttaa atgacttctc ctttctttta acttgcactg ttgtctagcc ctcacttatt 122940ttgtcaattc tttttagctg tttgtctttg aatcttcata aagccatagc ttttctcata 123000agaagcagca ctttctttgt tcattcatat tttaatgaac ccctgtagta tttaattaaa 123060tacttaatgc ctaattaaat cacataattg caatgcaaaa gtacatgtat cataaagagg 123120tctgaaaatg agcaactggc aagcaggtgg tggcaggcag agctgcttgg gtgggtgggt 123180gtcatggaga ggagttcatc agccacatgt tcagtgagct ctggatatgt ctgtttagaa 123240atgatcacta ataaacttgt gctcaaccat gtatacctct gggaagcagg tgctcttcag 123300tagattgcct ctgcagagaa cacagaattg aagtgaatgt ccacaaaggc aatgagccac 123360ctgcagaata gtttagtcaa ggctgtgttt gaagtttgcc aaagattaat atacatttga 123420ttttcatgtt gtgccttttc tctgattgtg aaatattaca aattctatac aaataacaat 123480gatggcaaat cctcctgagc aaagtgtgca ccttgtatgt gccctagagg aacttgtgtt 123540tcgttctgat tcccctacat ttctcatgtc atagagtggg ggttgcatta gtgtccccct 123600gtcctcgctg ggatcacatc tgtttggatc ctagagtctt ccagctgaac tgggacaagt 123660ataacagacg gacacgtagg ggtggaaagg cgtctcttgg cagcagactt tctaattgtg 123720cacgctctta taggtgttgg agatgttcat tcttgtcctg cagcagtgcc acaaggagaa 123780tgaagacaag tggaagcgac tgtctcgaca gatagctgac atcatcctcc caatgttagc 123840caaacagcag gtttgtcccc gcagccttgg cttgttgttg catagtgatg gtagcttaag 123900gtccttgtga aaggtgggtg gctggaatca gctcttcctt cagtcctaat ctgtgccttg 123960atagcagttc tccgtgctag tcatgggaca gctgacttca tttcttctca caatgccatc 124020tcaggttggt attgcccacc tactttacag gggggatccc acagctccga gaggttatgg 124080aggtgatcag gcagcacaca gctttagagt gctggggtga gggcgggcca aggctaactc 124140taaagcccga acccttacct cctacactgc ctcctgcatt ctggtcaacc cagtgtttta 124200tttggtggtt agatttttgt ttttgttacc ttactgcttg taatttagca gttttccttt 124260cctttccctt cctttccttt ccgacagggt ctcactctgt cacccaggct agagtgcagt 124320cgtgtaatct cactgcaaca acctctgcct cccaggttca accaattctc ccacctcagc 124380ctcctgagta gcaaggacca caggtgtgca ccactacgcc tggctagttt tttgtatttt 124440tagtagagat gaggtctcgc tgtgttgccc aggctggttt taaactcctg ggcgcaagtg 124500atccaccaac cttggcctgc caaagtgctg gcattacagg tgtgagccac ctcgcctggc 124560ctattcatca ctaatcagaa tttctatgat caaatgacat gaatcattgt ttccacaact 124620gcagtggaag gaaatggcct ggcagtgcca gtttcagaag cagcctgccc ccagtcaggc 124680acaggccact gtgcccccag tgtagcagca cctctgtagc tcacagagaa gggtggtggg 124740gacctccttg aggcagctct gccagaaaat ctcatgagct gcctggcaca gcttgaggtt 124800gccttttaag tggactcagc aaatacatgt ttgttcatct tgattataca caataaacaa 124860ctactctgta tagtacgagt agtccgtggt ttttggcatt tgatttaaac ttagaggcat 124920gtgatattga tgttactgcc ttcatgactg cacccccatt ctgatttcat aatggaatgt 124980tatcttgaga ccagttagac aacaggacag ggatcttggc ttctggtgag attgacagca 125040gttttagtgt ggtcagggtc tccctgccta cagatggttt tagaatggtg ccctggaagc 125100tttatcccat tcttttctgt gcgtaatctg agtagagtgg agatcgaagg cctgaataca 125160tagtaaatac ctgacttaat atctgccgca atggaaattg tgtgatacaa catttatgaa 125220acgcttagtg cagcacctgc caggtagctc accacaggtg catgttgcat tcagaagtag 125280tgctagatac tatcctgtta ctggcagtgc atacatcagt gatcaaagca gattaaagaa 125340agaccccctg ccttcttgga gtgaagattt tgttgggatg cgggtaaggg gacagacaat 125400agaaaagcaa gtgagtgaag tctataccat ggcggctgat caggaacacc gtacagaaga 125460atccaggagg gaagagagtt aggtggtgtc tgcggtggga gtggcattgt tcagctggtg 125520atgagaagaa gctttggtga tctggtgaca tttgagtgaa tttgcagaaa ggaaagatac 125580aagcctagga gatacctggg gaaggaacat tccaggcaga gcaaatagca gtgcaaaggc 125640cctggcgggg ggcggacatg ctgttagggt acaagcaatg agggtggagg agtggggcag 125700ccatggggag ggaagggagt gaggcctggt ggggtgaggc cagtgtggag gagccttgag 125760agggtttgcg ctgatgtggt gtaggtttta gcaggatcat tcttattcct gagttgagaa 125820tagccttgag ggggaggtga gggcagagca gggccaccca tgtgagaccc ggcactggag 125880tggaatggcc caagtcagca tcccttggca gcatgaaagc aaaaccagca aggtttgctg 125940gtggcttaga tgtggcatgt gagagagagc agggctttgg gggtgatttc agggtgagga 126000cagggtggct gtggacaagg tagggcagac attgggggca gcaggaggtc agagcctgtc 126060tggatgtagc agttgagacc ccataggtgc ctaatgaggt gaggccagca tcaggtgtat 126120gagcctggag ttgtcgagag actgtggggc agggggtcag catctgagat gtccactcac 126180agtggaccca gactggctgg agaggaggag gagcttgaat accgagcctg ctgagtccca 126240gctccaaggt caggtaggtg aggggagcca gtgctggggc agggggagta ggcaggtgtg 126300gggttcctaa agccaagatt ttttttaagg cattttgtgc aggagggcga catctgctgt 126360cagcaccttg ggaacttggc ccaggtttgg cagcaccgag ggcactgatg agtgcttttg 126420gaggagcaaa gggagccaaa ccctaatggg aatgtgttcc tgaaaggaca ggagagagac 126480ttgggaaaag gttttacttg aagagggaac ggagaaatag ggcagtagcc agaggaggag 126540aggagtcggc aatgggttaa gttggcagaa atgaaggcct gtttacgcac tgagggcaga 126600agcaacaggg aggatcagtt catgacacag gagacacaaa tcgccgttgt ggtgttcaca 126660gacatgggtt aggattggct gcatggatga cagagcactg tgggttctcc cagagttgct 126720ggggaggagg cagagttggt gagcacaggc gagggtccag gatgcaggaa tcctggagct 126780caagtcagtt gttcccttgt tgtaagatgt ggccagtgtt gtgagcttca catctgtgcc 126840ttgaaaaaca ccacatctgt ttgcagagtt gtttactatg tatacacact cagtagaaac 126900aaaaattgga aacagtcagt gcccaccatc aataagtaat ggttgaacac actgtggtat 126960aagcttagac tattttagct tgggctattt tgcatgatta aaaatgttct ggccaggtgt 127020ggtggctcat gcctgtaatc ccagcacttt gggaggccaa ggcaggcaga ttgcttgagc 127080tcaggagttt gagaccagcc tgggcaacat ggtgaaaccc tgtctctact agaaatacaa 127140aaagtagctg ggtgtggtgg tgtgcgcctg tagtcctggc taactcagga ggctgaggtg 127200ggaggatcac ttgagcccat tcgtgcgcca ctgcactcct ggggcacaga gtgagactct 127260gttagaaaga gagagagaga aagaagagag agggagggag gaaggaagga aggaaataaa 127320tggaagaaat ggaagggagg aaggggaggg aggaaggaag aaaggaagtt cagccagttg 127380ccttgggagt tctccattgc actgggttaa gtgagaagag cagagacgtt tatgattttt 127440caaaacaact aaaacaaaac ctctgtgggt gagggggcaa ggatatggct ataggaacat 127500ggggcagatt aagaaaggga tatacacaca ccacttagca tttgttacaa ctgttgtggg 127560agggatggag tgcagaaaaa gaaaaaaaaa agtgcacacc atcccatgta tgtgtataca 127620aagggacgct tggaagactg gtccccaaaa tgttggtaat gattgtgtca gggtgctgca 127680gtgctagttg attttttttc acacttttgt atatttgagt cttttacaga aagcatttat 127740tatttatgta ataaaaatct aaatgacaag atttctgtta tgggaaaaat gtagctatac 127800agtgttgttg taaaaatgtt tgcttggttc accactgaac ttaaaatgct tttaaatgag 127860ggaaggtgac gatgagatga ttatgatgat ttgcccttga gttacatagc tggtgtacag 127920gaagctgtcg tttcttttgg cttacgtaga aatgtttgtg gtgtctaatt ccacagatgc 127980acattgactc tcatgaagcc cttggagtgt taaatacatt atttgagatt ttggcccctt 128040cctccctccg tccggtagac atgcttttac ggagtatgtt cgtcactcca aacacaatgg 128100tgagtctctc gcctggctca gcagatgaat ctggacggct tgttcaggct ctgattactg 128160ggaccacccc cagaatgtct gagtcagtca gtttgggtag ggcttcttga gagtttgctt 128220tttttttttt tttttttttt ggtgtggggg tggtgcggaa cagagtctca ctctgtcgcc 128280caggctggag tacagtgtca tgatctcggc tcactgcaag ctctgccttc cagcttcaca 128340ccattctcct gcctcagcct cccgagttgc tgggactaca agcgcccacc accacgcccg 128400gctaattttt ttgtattttt agtagagatg gggtttcacc gtgttagcca ggatggtctt 128460gatctcctga cctcgtgacc cgcccatctc agcctcccaa agtgctggga ttacaggcgt 128520gagccaccgc acccggcctt tttatttttt ttggagatgg agccttgctc tgtcacccag 128580gctggagtac agtggcgcta cctcgactca ctgcaacctc cgcctcccgg gttcaagcaa 128640ttttcctgcc tcagcctccc gagtagctgg gactacaggt gcgtgccact gtgcccggct 128700aattttttgt atttttagta gagacggggt ttcactgtgt tagccaggat ggtcgcgatc 128760tcctgacctt gtgatccgcc cgcctcggcc tcccaaagtg ttgggattac aggtggctct 128820cgcaccaagc caagagtttg catttttagc aaattcccag gtgaaactaa tgcctgcttt 128880tctgggagca cactttggga ctcagtgata gagaggttta ttggtaggat agtaaaatag 128940gagttatttt ctttcacaaa attggcaatt gggggaaatt taatcttcct tttttcttca 129000gctgtgactt atgtattatg tttattttag gcgtccgtga gcactgttca actgtggata 129060tcgggaattc tggccatttt gagggttctg atttcccagt caactgaaga tattgttctt 129120tctcgtattc aggagctctc cttctctccg tatttaatct cctgtacagt aattaatagg 129180ttaagagatg gggacagtac ttcaacgcta gaagaacaca gtgaagggaa acaaataaag 129240aatttgccag aagaaacatt ttcaaggtat gctttctatc tgagcctata actaacccat 129300gccttttggg aagtcacgtg atgtttcaca gtcagtaagt ctggaataat acctggtctt 129360gcttcacttc tgagttgggt aaagaagtct gtatcagtgt aattttctaa tccgtcctgc 129420attatctatg gctcttggtt catacctgtc ttgaagttct gtcatgttct gtctcttgtc 129480ctcagtagag atgctacagc agtggctcgc ctcaggcagg gcagggcagt ggggtggctg 129540tcctgggggc aggcagtagg ggcacgctga cgtcagggaa gttgaaaccc aagagaagcc 129600agtaaaagtg agtctcagat tgtcaccatg tgctggcagt tttacacgct gtcagtaata 129660aaagtcttct ccctgcaggg cagcctgcct ccaataaata cgtgtagtat caaatcctgt 129720cttccctcat aaattgtttg gaagctcccc aaggacagtg atgaggcact cgtaagtgct 129780tgctgcctag atgggtccct ctccaccttt gctagattct gagcattcac tgagttagag 129840ctgcttctgc aaatgtgctg cttctgctaa gtggctgtga cttcatgcag ccttcacttg 129900gtttgtcatc agtggagatg ccctgtgttg tcgaaggaga taagcccagt aagcctgctg 129960ggcacctttt ggtttgcagg ttcagcaggc agcccatggc tttccctgtg tcgcattgaa 130020gcagctggct aaaattgatg atacattaaa ttcctgtgac agatgatcag cttgtatttg 130080tgtaatggtg tacagttcac aaagcttaaa aaaatgctac ctgccatttc atcctcagtg 130140aggaaggtga tacacagaga gaccaagtga ctgtgtccac ggcgacggcg ctctgcattt 130200cactttagcg gttaatgtac tctacctata tttttacttt atatttacca tatatctttt 130260catgtatact tggcgtaagt gctttatagt agtcacctaa ttcactgtca tcttttttgt 130320ttcttggaag gtttctatta caactggttg gtattctttt agaagacatt gttacaaaac 130380agctgaaggt ggaaatgagt gagcagcaac atactttcta ttgccaggaa ctaggcacac 130440tgctaatgtg tctgatccac atcttcaagt ctggtaggtg aatcacatta gtcttcctgg 130500agtgtctcgt tccccattct gcactataca ctctcagagt gtaggagctg tgctgcccgg 130560tagaaactct gccttgccca gtgtgccagt tgaaaatatt tgttgctgta agagtacacc 130620tgataccatg tgacccagca gttccactct tgggtatata cccaaaagaa tggaaagcag 130680ggtggtgaaa agatatttgc atgccagcat tcatagcagc attattcacg atagctaaaa 130740tgtggaacca actgaagtgt ccctcgatgg atgaatggat aagcaaaatc tggtgtatat 130800ttacagtgga atattattca gccttaaaaa aaggacattc tgacacatgc tacaacatgg 130860gtgaccctta aggacattat gctaaatgaa ataagccagt cacaaaagga caaatactat 130920gtgattccac ttacatgagg gacctggagt agttaattca tagatataga aagtagaatg 130980gtggttgcca ggggctgcag gggaggggag ttatttttac aagatgaaga gagttattct 131040agaaatgaat ggtggtgatg gttgtataac attatgaatg tacttaatgc tactgaactg 131100tacagttaaa aatagttaag aggaccaggt gtcatggctc atgcctgaaa tccaagcact 131160ttgagaggcc aaggcaggag gattgcttga gccaaggagt ttgagaccag cctcagcaac 131220atggtaggac cccatctgta caaacaaact agccggggat agtggtgtgc atgtggtccc 131280agctactcag gagactgagg ctggaggatc gcttgagccc aggaggttaa gtctctagtg 131340agatgtgttc atgccactgc actccagcct cggctataga gtaagaccct gcctcaaaaa 131400aacaaaacaa aacaagacaa gagccaaaaa tggttaagat gggccaatca cagtggctta 131460tgcctgtaat cccaacactt tgggaggtca aggtaaaagg atcacttgaa gccaggagct 131520tgggaccagc ctgagcaaca tatcgagacc cctatctcta caaagaaaat caaaaactag 131580ctagatatgg tgggcacatg cctgtagtcc cagctacttg ggaggctgag gtgggaggat 131640ctcttgagct caggagttcg aggctgcagg gagctattat tgcactccag cctgggctac 131700agaatgatac cctgcctctt attaaaaaaa aatccaaaaa aaaaaaaaag taaacctgag 131760agcttcctcc tcctgtgtta aatttggagg ccaagatgtt tttgttactt ttacaaatga 131820tcaaggacgg tgaaggttgg gcatggtagc tcacacctga aatcccagca ctttgggagg 131880ctgaggcggg gtgatcgctt gagcttgaga ccagcctgga caacatagca agagacccca 131940tctccacaaa aataaaaaaa taaaaaaaaa tagccaggag tagtggcatg agcctgagcc 132000caggaggtca agctgtagtg agccatgatc atgccactgc actccagcct gggcgagatc 132060gagaccatgt ctctagagaa agaaaatgac aaggacagtg aacccaagaa agtcataaga 132120tgccagctgt gcagcaagca tggaaagcag ccagtccaaa ttaggacagt gtgttttcca 132180agaagaacga tcgtttgtaa tgagaatgct ttgctttaaa taaatgacta aatagctaga 132240agcctagttc taggggatag gcacgtcttt cttctctcaa gaaaatagaa aggcaattct 132300aatttctagt aacagcaaac agcattaagt catggtccaa atatgaggca aaccaaaatg 132360tggcttgatt gttcagcagt tgatctgttg gaagcccttg atattaaaaa ggttctcctt 132420taagcggctt aggagtcacg atcaaagacc tatagaaaga gatgccatcc ttctaggatc 132480cttggctctc ttgggaacta gattcagata gtcataatgt aaatactgct tgagctttct 132540ttctttcttt ctttctttct tttttttttt gagacagagt ttcactcttg ttgcccatcc 132600tggagtgcaa tggtgccatc tcggctcacc gcaacctctg cctcccaggt tcaagcaatt 132660ctcctgcctc agcctcccga gtagctggga ttacgggcat gcaccaccac gcctggctaa 132720ttttttgtat ttttagtaga gacagggttt ctccatgttg aggctggtct cgaactcctg 132780acctcaggtg atccacccgc ctcggcctcc caaagtgctg ggattacagg tgtgagccac 132840cgcacccggc ccgagctttc atttttgaaa tcaatgtatg actgaaacac tgaagactta 132900ctgacttaat tatggtttca gaacagaatg aaaatgtctt cggttctgat gaatataaaa 132960ggaaaactaa ccaagttaat ttggcaagta gatggtagag atagaggtgg ggagtggaag 133020gggaactaaa atcttcacct agcattgttg ggattatatg gttacatcat ctgaagttga 133080cagaccaaaa tatagaggct tcagaggtct ccaaatagaa ctaaacatgt aattcagatt 133140gttaggaggt agtataaatg agctaaatct catctttatt acggtagagt taatgggtga 133200tgtctaaagt tgtctgaagt ctataaatca tgacaaatta tgatgtggtg attgtattca 133260acagtctttc agttgcaggg ataaaacccc agtttaaact agagtaagag aaagaatgtg 133320ttggtttaag ctcctggaaa gtgcaggcaa gggtagttgg taggactgca tctagtgttg 133380taattctgtg gtctgcattg tatatttatg catctcagct ctgctttctt cttttcattt 133440atataatttt taaattttat tttaaagata gggtctcact ttgtcgccta ggctgaagtg 133500cagtggcatg aagtgcagtg cgaggctcac tctagcctcg aactcctggg ctctagagtt 133560cttcctgcct cagccttcta agtagctgag acaataggca tgtaccaaca tgcctggata 133620ggttttaaaa tttttttgta gaaatggaag tcttgctgtg ttgcccaggc gggtctttaa 133680ctcttagctt caggcgatcc tcctgcctct gcctcccaaa atgctgaggt tataggtgtc 133740acccaccacg cccagtctca tctctgcttc ctgtgttagt tttgttctct ggtgggctgt 133800tttcacatga ccgaagatga cctctagcag gctgtgttct cagcccctca agtaggccta 133860tgtgattggc cttgcatgag taatatgggt gaccataaac ccctgaatgc tctggtccac 133920atgggccaaa tgggagactg gacagcattc cattgatgag gaggtggggc tggtctccgg 133980gagtaaggga gaggagcaca tgcagtaact gatggtctgc tgcaagggat agcagcacag 134040cagttagaat tttggaggta actaccagaa ctgaaaacag aaatgataac aagtagttgc 134100cttaaaaagg gatgggagca gggtgctttt gtgatcaaag ctcctttctc ttactggatt 134160tttgtacaca ttttgcatac atatcttaga gtaaaagata gcattttcag ccttggtcca 134220tttgaggata ctcttggcgt ggcccgcctc catgctagca ggctctggtt gtgccaagtt 134280cagttgagca tcctggctct tgcctgcacg gaacttccag tcagtgcgtc agtatcacaa 134340gtcttgatat ttcctatgaa gaagaacagt agtgcagtga cagacgaaat gggtgggcag 134400gcagaggcag gatttctgag ggagagaagt agctagcttt ttgcagagaa gagttccggc 134460acccaagaga gcagctgaga gtacaggcag gcaggcagga tgccggtagg gcccggccgc 134520acggcgccac agaatcctgg agaaaggggc ctcttcatgg cctctgcatt cagctgctgt 134580caccctccgc acaggccatg gccaaaattt aattttcata gtggactcta gtttttgagc 134640cttacttgct attattgaaa taattttctt gtttcttttt aaagatcttc ggattatgct 134700tcactgacca ctgtaataag tttaaagttg agaaaatatg gcttgttaat gaatgatagg 134760tcaattttag tatgttggtc attttaatat tttgccacca gttggtttgg atttgatgcc 134820aggaggagac agcctcattt ctaaggacta gtcttgcctt tgtgggataa gggtggtgtg 134880ttctgtgtcc ttctacatgt ccgagcgatc tctgtgcagc tcaaatgtgg tcactgtctt 134940attgcgctga tttcctctcc ttccatctca caattgaggc aaaatattgt tactgttgaa 135000gtgttgtcca ataggacttc cagcagagac aggatgtctg cactgtctaa tttagttgcc 135060tttagccaca tgtggtgttc tgtacctgaa atgtggctgg tctgattgga tagcttaatt 135120tataatttta tttaatttta attaacttaa atttaaacag ctctgtgtgg atagtggctc 135180ctgtatgaga cagtgcaggt ctgttgagaa gcagctttac tggtgggagt ggagggcttg 135240gagagggcac gtgggtttcc tgctggtatc ttttgacctt atttaatctg cccaacattt 135300gcaagtaagt tgtgtgtgtg tgtatatata aatgtgtgtt tctgtcttct tgtttccttt 135360gactgcattt atttgaaaga cactaggtgg cagaattact gtatttgatt ggtttcaaga 135420taagagttga aataattcat ctcgtgtttt tatataagta aggtgtgttt agcatgtaaa 135480attggtaata tgtattcacg tactgcttaa acaaaggcta

tgaattccac ccataaaccg 135540aaaatgaaga cctttaaatt tgtccatttc aggcgtgggt acttcttaaa taatacctgg 135600ttcaggaact agtcagaatg gcacccttga ctttttgttt cctgcttttc ctcttgttgg 135660gagaggaggg tattcatccc aaagtggttt gcctatttca cattccatct aggataagca 135720gaatagccaa gaaagatagc tgtcctcctg tttacaacat ttggggtaac cagcatccct 135780ctcttttggt ccaagataga ctggtttaga aacagatgat ggcaccagag gcccaggagg 135840tggaaacatc agctttgttt gttgtccatg tggctgaatt agagctgtct ggccttgtag 135900cctcaacacg gccttccagc tttgctcacc gtgattttca aggacacatc ttgtgctctt 135960ccctgcctgc catccagact atacccagtc agggtggcag gagctgctgc cccttcctcc 136020ctgagtcctg gtcgtgggtg gtggagatgt gccatgacgc tcacggaggc atgctcaccc 136080cttcctctgt ggcagagggg atggctgcac gacagctctt ccctgtcctt tccaaagcgt 136140ctgtggttcc actttttggg gcaaagcagg aatactggaa gagagagaaa gtggtccttt 136200ctatagtaat aaagttgaca ttgattcaag ttcatgcttg gggaaaggac agggctacta 136260acaattataa tgctgggagc aatggaattt tctcatgggt atgtggtagg tttaatttta 136320attatcccag ttaattctta gaactgctct gtgaagtatt tcccgctttg tgcttaagtt 136380ctaaaagatc ctgtgccaaa accaagaatg aaaacccaag cattctttct tgcccatcga 136440tctttctctc atcaggccac ttcttgggtt gatagtggtg agtgtagccg ctgccacttt 136500cagaataccc accatgggcc ccagtcactg tgtggcgtgg agaagagatg gttctctctg 136560tgtcatagct gaacaagccc agcccagaga ggtttctgcc ctaggagctc tcgatggtgg 136620aattgggatg cgatcccaca tcctgcctgt tttgaaaaca gcattcttta tttccaattc 136680ctgcttccat tgttcctttt aatatttctt tgtttagctc acaaaaacac ggcttgcgga 136740gctgctgcgt gcagctgtag ctgtttctct gggtgcagcc tgcatccgcc ttcctgcccg 136800cctcctttcc tgcactgcca tcgtggtctc cgggcacttg gtccctttct cttcccctga 136860gtccctttgg ctcccctgtg ccacccttgt gatccacagg ctctgccttc tttctgtctc 136920agactgctgc tcatcactac tcgggaccct aggaagggag gttccaccga gaagcatctt 136980ctcatctcag ccacgttctc agtgccactg ttgtctttgt taggtaatgg tagctactgt 137040aacaaataaa ccaacatttc catggcttca caccagagaa ggttgtttct tggttttatg 137100acaatgtatt gagggtgttc ttggttcacg gatggttttc ctccatgtgg gaattcgggg 137160acccaggctc ctttccttct tttggttctg ttctccaggc cttcacatcc tctgtgtctg 137220gttggggaca aggagaggga aggtaaagaa ggctttgtgg ccttggataa gtgacaggca 137280tgcctttgct ggtgttctct cgtggtgaca ggtcacagcc ccaccctgta aaaggggact 137340gagagacgtc gtcctgctgc ttcccagcag cagcactgtg gtctctgatg tgttttctgt 137400gaggataaaa acaggtgatt ccaggatgag gaaagtcagg gaaacccttg gaaggagggg 137460accaggcggg tgtcaccatg ggattagtgg tggcttcaga atgagctgca gcgagtgcca 137520tgccttctaa agcttttgct attctgatat gcccacacca tgcccagcag gtgtctgcct 137580tgctctccgc agagagagtg atgaatcctt ctcatgagcc tctgtccagt tgttcctccc 137640tccacctgga agggaccctg ggttcctcat aacatcccag cggaacaggg gaccttctat 137700cctgtcccca agttcatcct catcctcctg ccggcttcct ggcccctctt atgtctgctt 137760cctgacgcca catccttctg gattctctgg aattgaattt tgcctttgat gcttatttaa 137820aaatatccat tgcaggccag gtgtggtggc tcacacctgt aatcctgtgc actttgggaa 137880gccaaggtgg gcagattgct tgagcccagg agtttgagat tagcctgagc aacatgttga 137940aatcctgttt ctatagaaaa tacaaaaatt agctgggcat ggtggcgcac acctatactc 138000ccagctactc aggaacctga gacaggagga tcaattgagc cccggaggcc aaagctacag 138060tgggctgtga tcgtgccact gtactccagt ctggtcaaac agagtgagac cctgtctgaa 138120aaaaaaaaaa aaatccattg catacttcac cgtagcgaaa catgtatgtc ttacctttcc 138180tttcctgcct gtagctgctc ttttacactt aacagccaca ctaagccagc cttaaatgaa 138240aaacaaacca gcacttcctg tgccctcctg cttccttcat gaggggtccc tccctctgtg 138300tacactccat tctcattgcc catggtggtt tgtttccctc ttgtttctca agccatggca 138360gcctgcctct tgccctcttt actaaaaagg cctttgcaga ggctgcctgt gttctttctt 138420tctaggtctc tctcatccta ggccctccag cttgattctg tggagctgcc ctcttgtcac 138480tcagtagctt gtggggtctt ctctgtctag ccacttaatt gattgtgttc ctcgagttgc 138540tgtccatggt ctctcgttac tgttttctct gtgtttctgc ctctctcctt ggccttggta 138600ggtccatccc ctttgtgacc ttggctgttg ctctcatgga caactttctc ttgctggtcc 138660ttgtagtcct ggcatccagc ttctcgacac gggacttgtc ctgccagtac ctcagacttg 138720cacttaaaat tgaactagca ccactgtcac tctccagggc ctcttcttgt taattagatc 138780attagggatg ttcagaatcc cagcatcata gtatgttcct cctcccgcta ccccaggaac 138840cctaacctta cctcctcctc tctatctact aggaggtggc cctcagagtc cgtctcatct 138900tccacctgaa cttccctaat aggctccagc agctgccacc ccgggggctg agtacttcct 138960ccatgccttg tgcagtgctg agccctttac ctgggttctc ctgtttgctc cttattacag 139020ccctgcgaac agatactgct cttaattcca tcttacacct aaggaagctg aggccccagg 139080taaggtgcat ccaaggtcac ccaggtagta gacagtagag ccacgatctg aaccaggcag 139140tctgattcag agcctgtgtt gacactcagc cacctagaac acagcttgga ttgtgggttt 139200ctattacctg ttcaaaaccc ctacatcccg ggtctgtccc tgcacgtgct ctgtggcctg 139260gctgcatctt ccttgaaggc agtgcatgcc tcttcactca gggggcccat gcaggaacag 139320agggccccac agaaggatga ggccagtgca gaatgggctg gaggggacaa tgctgaccag 139380gaagcaagtg tagagaaatc ccaggaaacc tggaggagcc agagacaagg cattagaact 139440cctcgtcgtg acctggtctg cattctctga gtgtgctgct tctgttagct cgcttccttg 139500gtctcaggtt atagtttaag gcattgtgga gccctaaaaa gcctgtactc tgtttttacc 139560tgttttagga ccctttcact ttggggatgt gttgattttt tttttttttt tttttttttt 139620tttgagatag agtctcgctc cattgcccag gctagagtgc agtggcacga tcttggccac 139680tgctgcccct gcctcctggg ttcaagcaat tcttgtgctc ccgcctccca aatacctggg 139740attacaggca cccgccacca cactcggcca atttttgtat ttttagtgga gacagggttt 139800taccatgttg gtcaggctgg tctcgaactc ctgacctcaa gtgatctgcc caccttggcc 139860tcccaaagtg ctgtgattat aggcgtgagc caccacaccc ggcctgaaat ttaaatcaga 139920aataaaattt tgatcccaac agtgatgcca ggcagcccag atctggggga gagggtggcc 139980ttggccagct gggcctttct ctgtttccca agtcttgctg cctctccctg ctgggctttg 140040cagcctgtgc atgtctctgt gcctttgacc ttgtttatcc aaaggagagg atagaatgaa 140100gtcatgattc ctggagccct gagaaggatg ctgtggagaa atttgccggt agaatctagc 140160tgagtgtgtt gctgaggtgc cagcattgtg tgtggggagg ctgaccgctt ggcctgccta 140220ggcccaggat gctccatggc cgggcacaga ggccacttgg ctgtcaggtg tcaggagcct 140280gcagagggca cacagagcct ggaccgcagg ggggtcctgc tttctcacct ggcctccttc 140340agcatttctg tccctcagtc cttagcaagc ccaggagctg ttgagtttgg caggtgccga 140400gtgctgttcc tgcctgtgta gctgtggctc agtcctgtgg gggccccgct gtggcccgag 140460tgcagtgatt cgaggcgctg agtgttccct gactccttct ccaggagctg tgttcagact 140520ttcgcagctc ttggcttgga gctcctggag ggcttggcat tgccgaccaa tgtggaggtc 140580gacagtgaga gaggaggaat gctagctttc ttgaccagtc cattaaataa gtgggatatt 140640ggccaggcac ggcggctcac gccttaatcc cagcactttg ggaggctgag gcgggtggat 140700cacgagctca ggagttcaag accagcctgg ccaacatggt gaaaccccct ctatactaaa 140760aatacaaata ttagctgggc gtggtggcag gcgcctgtaa tcctagctac ttgggaggct 140820gaggcaggag aacagcttga aaccggaagg tggagtttgc agtgagccaa gattgcgcca 140880ctgcactcca acctgggcaa caagagcaaa actctatctc aaaaaaaaaa aaaaaagtag 140940gatatctgtt tctgcttaga aaaatcagaa ttttctaaat gccaggtgtt ctgaatacgt 141000aagtatggga gacgactcag cctgtttcat ttttatgtaa aatcttcgcg tagccatgtg 141060gcactggacc gagatgaaag caaagacatt tctccttaac tttgtttcta ggaatgttcc 141120ggagaatcac agcagctgcc actaggctgt tccgcagtga tggctgtggc ggcagtttct 141180acaccctgga cagcttgaac ttgcgggctc gttccatgat caccacccac ccggccctgg 141240tgctgctctg gtgtcagata ctgctgcttg tcaaccacac cgactaccgc tggtgggcag 141300aagtgcagca gaccccgaag taggttcata atgccccaca gcccagggcg ccagcccagc 141360accctgtcct gagactccca gtaacctgag ctttggccac cgttaaagca ttttcatttt 141420ccattttttg tgagggcttg tgaaatttct gctgcatatt aatattcctt tcatggacag 141480catattattg ggacaaacat gcggtccagc taaaggcatt caaaatagca gttgctttct 141540aaatgcgatt ttctttggca ggttctttga caccattgca tcttgtggga tatgcttgtc 141600atgctctgtg gctcctacta agttctagtc cttaaattgg ttccatagcc agacatgttg 141660caatgtctta acctcattat aaagtaaatg tggttctggt tatccttaga taatgaagta 141720acagtgtagc aaatttcaaa acctcttgga aatgttattt taccattcaa aaaggcttac 141780taaggttctc gttatgggtg gccctctttt tgcaaaaggt tttcaggctt aagctccatt 141840tctaggtgct ccaacactcc attatttgta tatgtatgga aataaaagct gtgaccaccc 141900ccaaccctgg cccccgccca gctgaatcct cagcacagta tttctggaag gctcaagatc 141960ccacgctggg gaaaagaagt tctggagaca aaagagggca ggtgctgccg tgcctctctg 142020ctcagtatgg atactggacc ttgtgctgcc agggctccca gtagggccag ttcatggcac 142080tcagctggaa agtccactgt tgggaggcat tcttaaccat ccactctgtg ccgtatgtag 142140tggggtctgg tcattctgtt ggaggagaca gaccagtgac gacatttgaa atgcttggtg 142200gatgtcttag gcctgttacg atgactgagc actgtggggg caggagacag aaagtcagtg 142260tctcctagtt ctgtgctgct ttaacgtgca tagaaatcag ctgcggattc agcagatcac 142320tccttttctg acagatgggc ctgcttactc tgatgttata tcagaaagct ctgaatctgg 142380gaattgtgtc ccctgaattg gagtaacaga aatgcttaga tgatgagtgt ttaaaagaaa 142440taaaccaaag gtaaatttag tttggaattc agcaagcgtc ttcattcagc cctctgaggg 142500caaactacag ctttttgtaa atgtaggtaa attctgtgac tgtttcgtga ccccctctga 142560tccagttttc ctttataacc ttctgtattg ttccttctat tatcctgaaa taacattaat 142620agattaggct gggcgtggtg gctcatgcct ataatcccag caccttggga agccaaggcg 142680ggcagatcac ctgaggccag gacttcgaga ccagcctggc caacatgatg aaatgctgtc 142740tctactgaaa ataacaaaaa ttagccgagc atggtgacag gtgcctgtag tccctgctac 142800tcagaaggct gaggcgggag aatcgcttga acctaggagg aaaaggttgc agtgagctga 142860gatcgcgcca ctgcactcta gcctgggtga cagagtgaga ctccatctca aaaaaaaaaa 142920aaaaaaaaaa aaattaatgg atcaatggat ttttaaccta ataattaaat ttcaaaaaat 142980atcgttcttt aatggtaatg taaaggtaaa attaagataa tatgtaacaa gcatgtgagt 143040gtctaaggtg tccccgtggt ggaaggaaaa aataaatccc cataagtgtc caagatgccc 143100atagagagca gagctgttct ggtttaaacc cctgctctta gcactgtgtt tttccagctg 143160tgggtggtgg gggatgagta tctttttatt tccatgagat gagaaaaatg aattactaga 143220agtgtgaaat acaaaacaca gctgctcttt ttttagccat agactcagca gccataaaat 143280tgctgtatcc agttgcagaa attcctgctg cttactcttg accctctctc ggtttgtgtg 143340catctcctct caggctggct cccagatggg agctggctcc aggcgacact gggtgctctg 143400ctccaggagg tccttatgtg ggtcctgccc tagcctagcc cctctcttat ggactctgtc 143460actgtgggtt tatgattcac tctcaatctg tcttacctct tggtgaactg ttagagtcct 143520gcctatactt tggcgcttgt gggtgtgttg tggtacacat gatgtgttgg tcacttccca 143580gctcatcttg ttctgagtca ccctagattt gggacattca ttcgccacca gtaccgggcg 143640gtgtatggcc tgagatttgg gggggcttgt gctgctacaa attggggctg aatttgagtt 143700gacagtggac cttctttatg tctactgctc atatttgaat tgcaaatact gcctcttctc 143760tttcagaggc tcattaccct atagctgtat tattgcaaag tgcacaatta cagcttgagt 143820gtaagtcaca ctgcgctggc aggacggccc actgagaaag ggcacgtttc ctgttcgtta 143880gttttcacat tgacacataa tttacaatac agtaaaatgt acttttctat caactgtagt 143940cagtaacagc ccccctcccc caaccacatc aagatataga ggagtgctgt cacttcaaac 144000agttccctct tcctctgcca catcctgccc ctccccaggt ctaaccacca atccgtgctc 144060tgtccctctg ttcagcccat tgcagaaggc catagaaata gaatctatag gctaggtgtg 144120gtggctcatg cctgtaatcc cagtattttg agaggctgaa gtgggaggat gacttgaggc 144180tgggagttca agactagcct gggctgccta gcaagacccc atctccagaa aaaaaaaatt 144240taaaaattac aatcacgtcc ctgtagttca gctgcttggg aggctgaggc aggaggatca 144300cttgagctca ggagttagag gttacagtga gctatgatcg tgccactgtg ctccagccta 144360ggtgacacag caagacgttg tctctgggga aaaaagaaag aaacggaacc acgcggtgtg 144420cagccttctg agtctggccc ctttcggtga gcagtgtcta aagttctgtc gcgtgttgcc 144480cacgcgtcgg tggctcgctc cttgcaactg ctgagcattg tatggctagg ctgtagtttg 144540ttttcacttc accagttggg aaacagagaa aaggcacttt ttaaaaagtt taaatctgta 144600gaattttggt ttttaccagt tctcttctaa atcctgaggg attacaggaa aagttgttgt 144660atttcagaat attcttagct tgatgtgacc tctgtccccg ttaaggccct ttgccgcaat 144720gggaaggacg tcgctcggtc agaccctgaa ggtcagaggg gcagtttggg agtgtgtcaa 144780cattttaact gtatggacta gagccaagag tctcaaggtt tataattccc acgtattcaa 144840aaagaaaaaa acaataaagt gagaagtcag tgtagagtga aataacctgt gttagtgggg 144900aagaagtgtt tttaaacagg atttccataa cgtataacat caacatgttt agagtggtga 144960tgtttcattg ggaaacgaac agtaaaacat gaaagcaggg aggttttcat tctggcagtt 145020ggcaactttc acggcagatg gagaatttca aaagcaattg ctcaattatc aaacatagcc 145080agtgtgagtt ctgaaataaa ggtgctgatt gaatgtgcag ctttatggtg gattttgcta 145140ttcaggcaag cattttaatt ttctgcctgt taaattctgt tttctttagt ttttcatatg 145200tggtttattg tagcttagga atagataact gagagtatat attacacata caacattctg 145260atatggcaat atttaaaaca acttgtctgt tttagaacta gaattaaaca taatcatctt 145320cagtattttg caaataagct cactgccatc cagaaacatt gtcaatgcat ctgttgctcc 145380ttctagaaga cacagtctgt ccagcacaaa gttacttagt ccccagatgt ctggagaaga 145440ggaggattct gacttggcag ccaaacttgg aatgtgcaat agagaaatag tacgaagagg 145500ggctctcatt ctcttctgtg attatgtcgt aagtttgaaa tgcctgtaaa cggggttgag 145560ggaggtgggg accaggagaa catcctgtgt agatgacact tgcatggacc ctctggaacc 145620cagaccgccc ggtgtcctgc caagctccat cgaaactaaa tctagaatga atgtttactt 145680ctgctgtgac atataattgg agaccaggcc tggccttcca gtcactggat tctaagttgg 145740actgtgagag tttttgcagc tgactcattt atcaaatgcc cggctattgg ctcacgccta 145800catgatgctg ggtatgtttg ttaatttgag ggaagcaatg gaataataat aactaatgat 145860ttaaaaaaca aagtaagtgc attgactgta gtggggttct gattttaaat ttttttaaaa 145920attaatacca ggagcagtgg cttatgccta aattccagca actcgagagg ctgaggtagg 145980aagatcactt gagcccagga gtttgagaca agcctgggct atggtgtgag acacccatct 146040ctaaaaaaat aaaaaataaa aaattatcca agtgtggtgg ctcgtgcctg taatcacagc 146100tctttgagaa gctgagggcg gaggatggct tgagcctggg agttcgagac cagcctggca 146160acacagagaa accctgcctc taccaaaaaa agaaagagag gaagaaagaa aaattagcct 146220ggcgtggtgg tgcatgcctg tggtcccagc cacctgagag actgagaagg gaggattgct 146280tgagcccaga agtttgaggc tgcagtgagc tgtgactgtg tcactgcact ccggcctggg 146340tgacaaggcg agacccctgc tctaaaataa tttttttaag ttaatttgta gaaaaggtgt 146400tagatgttct ttgtcacatt ttatgatgga ttcctgttta aatgccgttc tctttaaaga 146460aaaaaaaata acttgtggga gtttttaacc ataaaactag catcacatat ttaccatgga 146520gaatttacaa aaaaacaaat aaacggagga aaataaaacc tcctgtaatc atactactca 146580gagataactt gctgttagat tttggtctag atttaatact ttttctatat ttatattaaa 146640aatatttaaa acatatgcat ttctttgtca caaacatggt atcttataga tactactgtc 146700acatagcaaa acagtgttaa atattctgaa tcagaaaagg aagccgactc tccaactgaa 146760agaggtgtta tcctagagac tttttctggt gatgacaatt tattaatagt cactttttgc 146820tttactttct ctattgaagt agtttttcta ttttgttcta cttttaagga taatataatt 146880tataatgctg tttttcacag aaatataaga aaaaagatac taattttata agttaataaa 146940gtttgatcat cccaaatcca aaaatctgaa atccaaaatg ctccaaattc tgaagctttt 147000tgagtgctga cattatgttc aaaggaaatg ttcattggaa ggtttcagat tttcggattt 147060agggagctca acaaataagt ataatgcaca tatttcaaaa cctgaaaaaa atcctaaatt 147120cagaatactt ctgatcccaa acatttcaga taagggttat tcaacctgta ctgtcagatg 147180atcccaaatg aaaaatatta atcgttaacc aaatatcaag gaattgatca cattttacag 147240tttctgccta ggattatgaa tcaagatgaa aaggctctgc atgtttaaaa atatatattt 147300ttattttctt ataaatctta aatatctaca cttaagattt atttgatatg tgggatccat 147360tcatattttg gattcaacag ttctgtcaaa actgtggcag tgatagggga ttcttttttt 147420cccactgaac tatcacaaaa ttggaaaaag agtaattgga gaaccccact ggcttagccg 147480gcccgaagcc cgggagaggg caggcagtgc tgtggatggg gtcatcccag cgcaacgctg 147540cccctgctac ctgcggatct cgctgaggcc tgcctttgtc ctttgaccct tggccatttg 147600ttagtgtctc tgagagctgg actgctgtac cctacttccc cagggggcct aacttcacac 147660agcctctgcc gcagtgcgtg gttggaggtg acggccttgg taaatcgagt ttcctacctc 147720ctcaattatt tgtgctcata cactgtatat ttttagtgag gtttatattt gggatgtgtt 147780ttctccttct taccctttct ggcctttcta tggcattaat acctggtctc ttcttgtgta 147840cttgaaaatg aatctctcat catatttttc cttagtgtca gaacctccat gactccgagc 147900acttaacgtg gctcattgta aatcacattc aagatctgat cagcctttcc cacgagcctc 147960cagtacagga cttcatcagt gccgttcatc ggaactctgc tgccagcggc ctgttcatcc 148020aggcaattca gtctcgttgt gaaaaccttt caactgtacg tcttcatcct gccgactatt 148080gccagttgca gttttccctg ccttaaaaat ggagtattga aatttttaac tttaatttct 148140gatttgcaaa atagtcatct tttgttcttt tccttcttgc tgttagccaa ccatgctgaa 148200gaaaactctt cagtgcttgg aggggatcca tctcagccag tcgggagctg tgctcacgct 148260gtatgtggac aggcttctgt gcaccccttt ccgtgtgctg gctcgcatgg tcgacatcct 148320tgcttgtcgc cgggtagaaa tgcttctggc tgcaaattta caggtattgg gaagagaaac 148380cctgatattg atttatattg aaaatttagc aggccaagca aaacaggtgg ctggcttttt 148440cctccgtaag tatggtcttg acatggtcac cgatagaaac atggaaacat ctgcaaactt 148500gccgttactc gtgtgtccga tctgactgtt tcttgtattt ttttctagtc tgcccttact 148560aggatgaact gtacacatca gttcatcctt tttaaatgag catgaggtta ttttgggttg 148620ttaggtgtta caaacacact aatgtgtttt tgtctattag agcagcatgg cccagttgcc 148680aatggaagaa ctcaacagaa tccaggaata ccttcagagc agcgggctcg ctcagaggta 148740atgctggaaa cacaggtcgt ccttgtgtta ggacaaccca ggatataaag gatatagatt 148800tgtacgggaa taaattcaca ggacaagaaa tcgatgtgcc ttataggtgg gtttactgca 148860gaagtgccat aatagaacct tcctactttt aaaacaacca gatctcactt tctaaagagt 148920aaaggatgac cggcaggatc acgtctgtga cgtgagtgga ggcagtttgc actcctggtg 148980gctgtttgag aggtagcatt tagaatgcct gtattcactg tcctgtgatg agtgggaaaa 149040taggttatca ggtttatctt agcaaaatca aagcatgtca tctaattgct aaacaagagt 149100tggcaaatct gagagacatt actcaatcct tggcatgcag gacttacatc tgcatcctgt 149160tgccatttta tgtcttcaaa gcatttaatc atttagttgt gtttgcaaag tctttgagaa 149220gcctttgtca gaaatcccta catctcctat gtgagtgtat ttccatgact gcagaataag 149280ttaaactttt acctttttcc ttcccttgcg gggcggggtg gggggcaggg attgtgtgtg 149340tgagagggag agagagacag cagagaagga gaatataatt atcatgctgt gtactttgag 149400ctgaaactgc aaaaaaggaa aaacacacaa aaattattat gcttttcagt ctttagagta 149460ccttgtctat tatgcttttc agtctttaga gtaccttgtt gatggtgttt ttaaatggga 149520ttgggcacaa ttaggtggac agtttgggat gatttttcag tctgtagggc caagctcttt 149580tgtaatttgc attatgaagt tgtcactctc atagcagatg gcgggagata aactattatt 149640actttttgac cctagactta gtcttcagtc cagatgaggg agattaaaag attataaata 149700tcttgtgcca gatgaggtga ttttattttg aaatgaccat gaattcctat cagttgtctt 149760actgggatat ttgatagtgg aatttgtgca tttgagtctt agatgatctg ttttacattt 149820attaagaaag cctttattag cttttatact gtgtattgcc tgttgcagtg tttgagtata 149880aatgaaattt ctggaaaata ttaatggagt acaaactgtg atacttaaaa gtaaactagg 149940gcctgcattt gtatcatgac ctgtttgagt attgatgaga agatagctgt gaagaaaaag 150000gtttaaacaa gtgtattttc ctttaagaag ccactaatag tgcatctcct tagagtgtat 150060atttctagaa tcctagtgtg cagagtttag actaagacta aaaaaaaaaa aaaacaaatt 150120atactgtaat ttcattttta tttgtatttt agacaccaaa ggctctattc cctgctggac 150180aggtttcgtc tctccaccat gcaagactca cttagtccct ctcctccagt ctcttcccac 150240ccgctggacg gggatgggca cgtgtcactg gaaacagtga gtccggacaa agtaagtgtc 150300cagcgtgtct gcatgggagg cacagggcgc tgagtgcctc tgtcacctgt ggcagataca 150360gagagtgcag aggaggtgcc gtggacccaa ggagttctgg cgctcggctc ggctcagtga 150420agctgtggtt agagacgtgg ggggccatca aggtctgagg gagccaagca gtgctgatgt 150480gggacccttt tggtaggagt gtggggtgag tagttagtgg gtgaatcaag gaatagtcgg 150540ccgtggcctg caggcccctg actgcacagg ccttcaagca

catgtcaatg ccgttagcct 150600ccctccatct cctcatacct tctggccacc tgtgagttgc actgccactg ccagccattc 150660tggtatgttg tcagcacctc cactgctcat acctcatggt tagggaccac ctggagcctt 150720ggtagagcct tggtagagcc ttggtactct actttcctgg acaaagttca gcttatgaat 150780atgaatttag atttcaaaaa ccagcagccc aagtataaga aagcgaaggt tcagtcctgc 150840cttcttaggc tctattcgct aagcacctgc cctgccctgg ttgctgggga gagatgagta 150900aagcagacaa cccaggagag gatggcaaag gggccgctaa cccttagtgg tttagctata 150960tttggaaggc ctattggaag ttcaccaggt gaagggggag gctgtgaggg tgcccaggca 151020ggtaacagaa gtccaaaggg gaaaacctgt ggtgtggtga gccgtatagc cacagcctgc 151080cggccggcag ccctctcagc ctagtgcggt gttcccaagc actggcctag gcctgtagct 151140ccagggatgt gaagtcccct tgaacgccgc ccatcatgtt ccccttatcc atttttttct 151200tcccaggact ggtacgttca tcttgtcaaa tcccagtgtt ggaccaggtc agattctgca 151260ctgctggaag gtgcagagct ggtgaatcgg attcctgctg aagatatgaa tgccttcatg 151320atgaactcgg tacgggggga gcagtggagg caaggaatcc tcagcttttc ttgtgacttc 151380caagtgggat ttgtctcatc atcatgtgac ccacttgttg acaacacatg ttggggactc 151440cagtctgggc agggacggga tgtcggagag actccactct gaatggggcc gggaagtggg 151500gaggactcca tttcagatgg ggtcgggaca tgggggttat gctgatcgag acagaaaagc 151560acattgtttc agccacatta gaatccacgg aggtgttgtt ttgaaatcca gctggcccca 151620aggctgggtg tatggtttgg gatgagaact atctggcctc cactggagga acaaacacag 151680gatgttatca tctaagctcc atggccaaga cagaatggaa gtcaaggttg cgtatttgcc 151740gtagacttca acacagtgtc gtaatgcgtg acgtcaataa cttgtttcta gtgtcttgga 151800agttgatctt tagtcgtaaa agagaccctt ggatgcagcg agatttcctc tactcacacc 151860tctgttagat gtagtgaggt tcttcacccc ccaaccccag atgtcagagg gcaccctgcg 151920cagagctagg aggccatgca aagccttggt gtccctgtcc ctcacccgtg ggcaggtcct 151980gtgagcagtg ggggggccac ctcttgggta tggtgcagcc atggcccaag cagggcttct 152040tctcagacct actaggacgg gagaaacctc ctggtgcttt agccctgcgt tgatatgcag 152100caaatgggag ggaagtgggc acctgggagg acaaatgcct gtagaggccg ggagtgacgg 152160caggtgttca tgaaaagaga ccttgtgggg agggcaacac aacagtgtgt tctgatgtac 152220tgaagagctc aactgaaaac aacaggagaa ttagcccaaa atccatttac taaaattgtt 152280tatctttttt tttttttttg agacaaagtc tcgctgttgt cccccaggct ggagtgcaat 152340ggcgctatct tggctcactg caacctccgc ctcctgggtt catacgattc tcctgcctca 152400gcctcccaaa tagctggtat taacaggcat gcaccaccac gcccggctaa tttttgtatt 152460tttagtagag acgggatttc accatgttgg ccaggctggt ctcaaactcc tgacctcagg 152520tgatccgccc acctcggcct cccaaagtgc tgggattata ggcctgagcc accacgcccg 152580gcctaaaatt gtttatctta agattcatgc agtgaaagct aacttactga gtgataaatt 152640tgcttagtga tctgtttatt aggttttcca aatttgctaa ttgggctttg aacagctgta 152700aaagttctga ctgtaaaaga aagcttcaac ttttggcatt catgatgctt ttctgagtat 152760taaactaaga tagatgtttt acctgaagga tcggccacca atctttaaat ggctaaacaa 152820aagggttgct aaaacataat ccaaattgac ataagaaata ccatttttcc aaccaaaatt 152880ttggcattca tatggctact tttacgtatt tcagctgcat ttgaacatct ttttcaaact 152940ttagggtggt tggtgtatca ctgaggtctt ggatgacact ttagctttga ttttgttttt 153000atgaattaaa attgtcatac caaaattttt atttcaagca aatccaagag cataaaaaat 153060taaaatatta cttaaaatac taagagagaa cagatatata ttttactaag catatgttga 153120atgaaattgt tcaaatattt ataacaggca tagagtagaa ttttcttaaa aatatttttg 153180atggtatacc aatttgtatt ttctcagaaa catttgcctt attctttttt ctgttgtgtt 153240tttcttacct gattgaaagc tcataatctg ttgttattgt ttgttaacct ttaatgctct 153300gatttcagga gttcaaccta agcctgctag ctccatgctt aagcctaggg atgagtgaaa 153360tttctggtgg ccagaagagt gccctttttg aagcagcccg tgaggtgact ctggcccgtg 153420tgagcggcac cgtgcagcag ctccctgctg tccatcatgt cttccagccc gagctgcctg 153480cagagccggc ggcctactgg agcaagttga atgatctgtt tggtaattaa aattaaaatt 153540tatcttattt ttaaaaagca ttccagggcc agtatagtac tttgcaccaa gtaaatgtac 153600aataaaggca gtggatctaa tacattgaaa gcgtttacag aggtagctaa agagcagcac 153660gggtgtcctc ggctcagaat ttcttcctgt gtgtttgcca ctttgccatt cattgacatg 153720gtcatggaca tagggctcta agcccttgag gaaggctggg ccagacctca ggggagatgc 153780agccccaaac cacgtgcagt cctgtggacg gatgtgtaga tgtgccactg aggaacaatg 153840tcttgagctt tcatcagatt ctcagagaat tgcttgactg cctttcgaag ttgatgcatc 153900tgtgctcacg tttgcaccca cccacgaggt ccttctgttt caggggatgc tgcactgtat 153960cagtccctgc ccactctggc ccgggccctg gcacagtacc tggtggtggt ctccaaactg 154020cccagtcatt tgcaccttcc tcctgagaaa gagaaggaca ttgtgaaatt cgtggtggca 154080acccttgagg taagaggcag ctcgggagct cagtgttgct gtggggaggg ggcatggggc 154140tgacactgaa gagggtaaag cagttttatt tgaaaagcaa gatctctgac cagtccagtc 154200acttttccat ctcagcctgg cagtaagtct tgtcaccgtc aagttattgt agccatcctt 154260caccctcacc tcgccactcc tcatggtggc ctgtgaggtc agccaggtcc ccttctcatc 154320tgcacctacc atgttaggtg gatcctaatt ttagagacat gaaaaataat catctggaag 154380tactttatgt cttaagttgg cctggacatg tcagccaagg aatacttact tggtttgtgt 154440tagtgcttgt aattcgcccc cagaatgtgt acacgttctg gatgcattaa agtctggcct 154500gtatccttaa agggccatcg ctgtgctgcc tgccctcagc aaggacacac tttgcagacc 154560cacagaggct ccgcctccac ctcacaccaa agaaagggag gagtccaaag ggcatcagtg 154620ccattactca caaaatgata aatacaccct tattctgaac cacgtggagt catatggttt 154680gtgatccctg tccttcaggt ttcagcttag tggggaagtg ggaaagtcag cgtgtgatca 154740cagcacaggg tgattgctgc tgattatatt atgtgcctgc tgtatgcagg atgaaatact 154800ttatatgcgt catcttattt gactctcaca accccctgtg agataggctc tgttactccc 154860atttgacagg tgaggaaagc aaggcttaga gaatttcagt gacttgccca ggtcctctga 154920gctaggaagt agccattctg gcatttgaac ccaaggcctg ctatccctag aacccacgct 154980ctcaaattca acctatgaca gaggcaagcc ctggtgctgt gggagcccca aggaagagcc 155040tctggcctgg tggccacgta gcccaggaga gatttctaca ggagcccaca gcgctgaagg 155100agagagaggc agcagagtaa gggggctttg tggcagagag gggactggca ctttggggaa 155160taggtgggtc aggactgaat gtaatggagc catgtcagag ctgtccttct ggaagggcaa 155220gggcacctgg acgcgctgcc cctcagtgct ttggacggtt ccacaactgt gattcacacg 155280gcttccccaa acgaaggtac acgagtgggc attctgtgac tcggtacttc cctttaggcc 155340ctgtcctggc atttgatcca tgagcagatc ccgctgagtc tggatctcca ggcagggctg 155400gactgctgct gcctggccct gcagctgcct ggcctctgga gcgtggtctc ctccacagag 155460tttgtgaccc acgcctgctc cctcatctac tgtgtgcact tcatcctgga ggccggtgag 155520tccccgtcca tgaacggtgg gttcctatca tagttcctgt ctgcttcacc atgtttttat 155580tttgtgctgc ctgtttgcca ggtactaagc taggaattgg ggatggagag gtagataaaa 155640tatgcatcag gaagggctgg gccccatctc ttactctcca atatattgga gtctacactg 155700gaatttaact ggaatttgct tttttagtca ttttatttag attttgaagt ttcagctttc 155760atcaaaaata cctctaaact ttatgtctct gtgatctttg gtcttagctg ttttatgtat 155820ttagtcttat atgatcataa gattaataac attacattca gaagattatt tgttttctgt 155880cagagttaaa atgtttgttt ttatactgca ttgtaatatt aacgtactgt aaaataaaag 155940tggcttgttc ttttcaagga acagtatcct caacaagggt cattagccac aatttttaaa 156000aaattggacg tcatagttta catgttagag ggcgttttga agctttgtat ttttaaatta 156060aatgttatag agtgatgttt tcatgtttca taattgtttt catctgtgca tttgtagcca 156120acttgaaaac aaagatccag ggattactac ttaaaagcca gacttcttgg aggttatagt 156180gatgattttg atagtatctt gagccgtctc ataataacct cagggtgaga gatggccaac 156240aggagacagt cgagggactt agaaatctga atgaaatctg aagttcaaat cttcagacat 156300ataccactaa ccaagagatt ggtacctcag tctagtattg tctgtttgtc taaaattggt 156360tctaaggaat ctaggctagt ctgtctatcc ctttcaactt ttgtgaggct gcacaaatgt 156420aaaatgttga ataaaaagca ctgatggaag tgtgtagaaa ttcttctctt tgttctgttg 156480taattttagt tgcagtgcag cctggagagc agcttcttag tccagaaaga aggacaaata 156540ccccaaaagc catcagcgag gaggaggagg aagtagatcc aaacacacag agtaagtctc 156600aggacccatt tttttcttac atgttgttcc tccaggactt aaaaatcatt cacagagacg 156660tgcaccgcgg tgagtgtgga ctcctggaag cgcaccgtag ctccgctgtg tcctgctgct 156720cctccctagc tgtcagggag gctgtagtcc attgctttgc cagctctttt gtttccgagt 156780gaacacctta tccgtacaca tgcggctgtc tctgacccta cagaccagct gggatgccac 156840tgggggagcg ctcccttccc cccgcacttc ccacactctg cagttattct gagatccttg 156900agggcaggga acaggtttgt cttctttgtg ttctcagaaa ttaatgctcg gcctctggtc 156960agcaagcaac aaccttttgt tgagtgataa tgaataaata aatgtttccc acatgagtat 157020tcagtaacct cagtgtcagg ttcagccatc tgttttggtg gatatttaaa agaaaattcc 157080gcttttccta cagaaaaaaa aaaaaatcca aatcccagtg atttaagcca gttatagact 157140tagacatata ctacggcttt tcatgcactt tcctcccaat tctagagtag gtattttact 157200aggaaaatgg tggcagtgcc tgttgggagg aagattcttt ggccaagtgt cttttgttct 157260tgccagggcc cctaggctgc tggggtgctt cagcttcttt agcccagtgt ctggtgggga 157320atggcccctg ttgcctgtcc cacagaggtg ggggtgcctc acctggagcc tgtccacaca 157380ttttacacag cacgcttacc tggagcatca ggcatctttt ccatgctctg tggctcagga 157440aacacgcctt ttcaatcatg agtgcaccag tgcttttggg ctttttctcc ccgcttttgt 157500gcaatcctgg ttgtggatgg agttttcctg tctttagtct tctgcatagt acttttctct 157560tctggttccc ggttcaaggt tttgtaatta gagaatgacc cagaagcaat ggcattttaa 157620tgcacagcca aggacttctc tgaatttgta tctcaaacct ctgtgggtcc ttcaggcttc 157680agtttgtgat ttcatgattt cttgttgcta cctaaggaat atgaaaacac ccacctccct 157740actctgcatc ttccagccga gtggcacctc aggctgtgga tcctgtgctt ctgtggtgag 157800gataagaata gtgccaaccg tgtggattga aatcaatcag ttaatccctc catgtaaagc 157860acctggaacg gatgacagtc ttgttatgaa tactcaacaa atgctatcat gatttttagt 157920tagatttcca ttgctttaaa acagttgaga catcttggcg gtttgagtta gagcaacggg 157980ccctgaagtg ggttctgttt gggtgaagat gattatgctt attccccatg gccctcttta 158040ggcaagagtg ggaagctttc tttgtttttt taatcacctc gataggacgt tacttcttaa 158100aggtcatcca ataaatatta ataggccggg cgcggtggct cacgcctgta atcccagcac 158160tttgggaggc cgaggcgggc ggatcacgag gtcaggagat cgagaccatc ccagctaaaa 158220cggtgaaacc ccgtctctac taaaaataca aaaaattagc cgggcgtagt ggcgggcgcc 158280tgtagtccca gctacttggg aggctgaggc aggagaatgg cgtgaacccg ggaggcggag 158340cttgcagtga gccgagatcc cgccactgca ctccagcctg ggcgacagag caagactccg 158400tctcaaaaaa aaaaaaaaat attaataaag ccaactcgtt agcgtggggc ttaattgctt 158460aagtccaatg agaagtcctt ctctatccta ggaagttgcc caaactgtag aatctcgtgg 158520cctgtgggta atagccacgt aatacacact cactgcctca acaaatcata ttttagtagg 158580tatgatattc tagactcaag acaccattct gtggatcttc ccaagggtgt gaagtgtcca 158640cagcgtctgc cttgggagtt tccatgccca ccagaaccat gccccaagcc cctcaagcac 158700tctgacctag gaaagccagt gaagcaagga tgacaacatg gccctttgat actagctgag 158760ggacagacac aggtcctggg agaccagaga aagacgaggg gcagaggagg tgtcctaaag 158820gaagtctgag gctgaggagc cacaggatgg cttccagctg tcacaggctg ctgctggcct 158880tatcacagag agtgggccag agggctggga accaaggcca gagctcaggt tcaggaccat 158940tccagcaatc ccagcagaaa atggggagaa ttgtatggta taggcggata tgaaggtaga 159000atctgcaggc cttcagtggc caactcagag tctaagtgga ttccacagtt acagcttgag 159060cagctggttg taggtcatgc tttctacact gggcatatag gatgtgtttt ttaaaaagtc 159120ctctcttaac cgttgcttgt ttagatccta agtatatcac tgcagcctgt gagatggtgg 159180cagaaatggt ggagtctctg cagtcggtgt tggccttggg tcataaaagg aatagcggcg 159240tgccggcgtt tctcacgcca ttgctaagga acatcatcat cagcctggcc cgcctgcccc 159300ttgtcaacag ctacacacgt gtgcccccac tggtgagtct gctcgttcct tgcagaagac 159360caagtacggt gaaaggcacc ggtaggccct gggctgggca cacgtgagag ggcgggacag 159420aatccccgca gcccagaggc tgcctgctgt ggttctggtg cccactgtgg ttctggtgcc 159480aggctgcttt cctcaggcac cacgtgtgga ggtcgctagt agaaatactg ggttttctaa 159540aatgaactga ggccctacat ccctaagaga ttagtgttag acctgattct agagcaacta 159600gaccactttg cttaatagca gaccagaaac cacaccccct cgagtgagtg agattttcct 159660ttggagataa ttcatgtttt tctacacagt tttgcagttg tcttcagaat tggtttaaag 159720taggtgttat tgccaggcgc agtagctcat gcctgtaatc ccagcacttt gggaagccaa 159780ggtgggcgga tcacttgagg tcaggatttc gagaccagcc tggccaacat ggtgaaaccc 159840catctctact aaaaatataa aaattagcca ggtgtggtgg tgtacgcctg taatcccagc 159900tactcaggag actgagacag gagaatcgct tgaacccagg aggcgaaggt tgcagtaagc 159960cgagatcgcg ccactgcact ctagcctggg caacagagca agactccgtc tcaaaaaaaa 160020aaaaggtagg tgttattgat cagaaccctt gtttcagata acatgaggag cttagcttga 160080ggagagtgag ggttgatgga gggggactga cttctgccca gtgaaatggc atcatctccc 160140accagcccgc tgaaataaga tgatggggcc tgttccttag ggcctgcagc atcctcaggc 160200aggaaagaaa ggccgacctg gcagggtgtg agccagcagg tgtaggtcag ggagaatgga 160260gccaggtccc agggaagagg cttgtggctg cctgagaagg gtgcgtgcct gcctgtgtgt 160320gtgtgtgcac gtgtgtgtat gtatgctgga gagtctaggg aggcttgctc caaggacgca 160380gtattgtttg atcctgagag ataaggattc tgccgcaggg aatgaaggta ttccagatgg 160440cgggcttatt ccgaagaaga ggccagtgcc tggcggtgct ggaagcagtt gcagaacagg 160500gagttgtagg ctttcctggg aagagagcag caggggtgct ggagaagcag gccacacttg 160560ctgcatgggg ttgctctcgg ccccactctt ggtgcacagc gagtcactgt gggttcatta 160620gcatctggtt atgagacagt aactgctcct ttggaggggc tcgtggagac catgcaggag 160680ggcacggtct tgaggtcatg ccgtccagag cacacctgag gataggccag gacgggctgc 160740acgctgtagg taaaattcct ccagcaagct cttcactggc attgaggagt tccctgagtg 160800cggtcatctg gaaggcagct gtaacaggca ctgcagtctc tccctgggtg ggtaccagag 160860aggagcatag gggagcataa ccgatttaaa gagagggctt tcctgtggtg aggtaagaga 160920ttagctggtc attatcatag agccccctct gcctttgtgc agatgggctg tgggaatcct 160980ggggttccgt tgggtccttt gtcacctcac tgaaggcatg taagctgagc tggccagacc 161040gtgagctgat cctgccactt gaacagcatc aagcctgcct ctggattctt ctgtgcatgg 161100cacttgtctg agcacctcac gcacagagaa ctggacttca gagtttacag aaataagctg 161160tatggttcat tttcatgcct gcttgccaat aaacatatct gagctgaacc tcattgaacg 161220cctgccttta ttctagcaca gcacctgctg tttgtgggcg aggggtgctg tctctaactc 161280ctgcctgctt ctcccagcac tccctgagtg gggtgtgcca gcagcctcag gatgaggaca 161340ggaagtggga gggcagagca gatttgggag ggccacttga tggggaagga agtcccagga 161400agcagttgga gctgttttct gggggagaag gtgccagctc tgggacagtg ttggggtagt 161460gaggagggag cccagtggag agaagtcggg cttcctgctt cctcacagta tgtctgtcct 161520gactcaactc ggatgatgtc acttcctttt catcttctca ggtgtggaag cttggatggt 161580cacccaaacc gggaggggat tttggcacag cattccctga gatccccgtg gagttcctcc 161640aggaaaagga agtctttaag gagttcatct accgcatcaa cacactaggt actcttgggg 161700cctctccttc aggtcaccat tgtcggacat ctaccgggag gaaatccaga gcccccagta 161760ctgggatctt ctcatttgac tccagaaaag atttaagcat gataataata caaacctatg 161820tgaatacatt ttgcagtgtt ggcaaaactc cttttatact gagaaaatag atcccagttc 161880ctgtgttttg tggcttgaat cccagctttg tgtattccgg gcttgtttga agtcaggaaa 161940ggttcatgtg tagtggacaa cgtgagacca aattctgcct tagattttgc atttaggcta 162000aacagtggca gcacttgtct cagaatgttt tcttgtgttc accagtctga tcctgttgtg 162060tctcagtggt ccattttctc atatgggaac aagcagacgg gagcagatgg agtcaggttt 162120cttggcactc gccttcccca gagcctagag gcagcatggg gagaaagcag gcttggggct 162180cagacagtcc tggtctgctt ccagccctcc tacctgagca gcgcagggca agtccgtcta 162240acctctagag accctcagtt ttgtcatatg taaaatgggg gtcgtgtcta tttcatagaa 162300ttgttgcaga tttagaaatt acatttctaa acaaatgtta ccccttattt ctaaataagt 162360gtctaaatga ataagtcacc acttttgccc ctatttgatg gcaagaggtg tgatcttgtg 162420gtgggactgt aatcagtcag ttctcagtga ctgtgccctg ctgtggtgtt tcctggaatg 162480ttcctgtctt gtcctagaaa gtctggcagg ggcaccctga ctccactgtc cagtcctctc 162540cccagtccct cgggcttctg cagatttgag gcttgtttgg atcccagaag gttgtggcag 162600gagacacctt gcctctactt tcccctttat aattcaatgt ccaaagagag ccctgagcag 162660gtacctcacg ccagctgcct cacggagctc ctcctcttcc tggctgtgag gatcggtatc 162720agtggcctcc tgctctctcc cccttgccta acacgagcac ctttgcttac ttgggtgccc 162780ttgctcttga actgcccatc ggacgtgcgt gacccaagac tgtgccgcag tccttgcctt 162840gtctgtgctc attttctttg ttcatttttt tccctgtaac gtaaattgtt atatttgtct 162900gtatctgtgt ctgaatcagt cctgcacgct ctccttctct ctgtctcttg ttctttcttt 162960accccgttta tcacggggac cccgatgtcc attgctctag ttctcctgtc ctaagcaccc 163020catcccgtct ctctggcctt accacaagtg gcgtggctgc ctcagacatc atgatgggga 163080catgaagcac agctgtcaga aacaactgtt cgttagatac actcgaatgc agctcatcaa 163140tagggatgga gggtctgtcg gatgtatttt cactgaatcc ccgttcctac cttgatacac 163200tctttttaat ctattcttct agacaggtca gaggaaccat tactttgact tttaaatttt 163260tagcagcttt attgaggtag aattcacata ctacagattt cacccactct aagcggacag 163320cttggtggcc attagtttta tccacagagt tgtgcagcca gctgcacagt ctcagggctg 163380gactccaggg aagattttag cccatttagt gagtggggca gaagtggccc tggccctgca 163440cgaggttgcc tgcatgggcg tccctgccct gtccctgtgt ctgctccact gggggttgac 163500caggctgcca gggccgactt gggcctgtgc cacctgcctc tcatgtgtct cggacagtgc 163560agccgatgtc tatacttcgg tttcctcaat gatgaaatgg aggggatagt gttccccgca 163620tcatagaact gtgtgaggtt taagggactc actgcccttg gcgtggagcc ttctccaggg 163680gccgtgctgt gtcggcgtag ctgtcagctc tccgttacag gcttgagaag ggttgacact 163740ctctcatgta acatttatat ttctaggctg gaccagtcgt actcagtttg aagaaacttg 163800ggccaccctc cttggtgtcc tggtgacgca gcccctcgtg atggagcagg aggagagccc 163860accagaagta aggccacacc ctgtgctggt tggcacatgg gcagttatgg ccgcttgcag 163920gcctttggtg gggaataaaa taaggcagca agctggtgtt ctttttttct cttaccttat 163980ttttgaaaga gtagctgaat ggtgtcttga ctgatattcc agagcaggga caaagcctgc 164040tgaggtctgg gggctgcgat taccaatggc tggaatgcat tttattacgg tgcattccat 164100gttaaggatc aatacgattg tgccctttct ggaaaatatc ttttagttta tcaatattca 164160gaggagtgta ggttgaatta aaatgaaaag gcactttata aaggccatga gtagtacctg 164220gtttcatttt tctaatgtct tgcagagatt ttatcaggct tcttgaagtg ttcacgtaca 164280ttacgctaac acgatattaa taataactgt gctctggtac agcggagcca gcagaatggg 164340aagttgtgga atgcaggccc ttgattctga tagaaggtgt ggtttgaact cacagaaatg 164400acagtttgga gggtagacat atgtcacaag tcatcaagat tgtctttaaa ttcatgcata 164460gaagctaaca gggtgtcata agcaaggcct gtaaaatgta tgagggaatt caaagataat 164520ttattaaaaa gtaattcatg tttggagttt tgtgcccaaa ggagtccttg atttgaaaaa 164580tgggcttttg cccatcagat tgtttcaggg cccgtgtgtg cggaggccct gccttgtgcc 164640ccgtgagctc agcctgacag aaatcctttg gtagcactta aggctcctct tcctcccatt 164700gaggcaggga agactctggg ttctgcaggc agaggtggtt gtgggtgtct tgctgctctt 164760gttgacatgt gggctctcct tccaggaaga cacagagagg acccagatca acgtcctggc 164820cgtgcaggcc atcacctcac tggtgctcag tgcaatgact gtgcctgtgg ccggcaaccc 164880agctgtaagc tgcttggagc agcagccccg gaacaagcct ctgaaagctc tcgacaccag 164940gtttgcttga gttcccacgt gtctctggga catagcaggt gctggggaca gtgggttccc 165000cgctgaagcg tccagcagct tcaaccaggc cgttttcctt cattgctaga attgaaaaca 165060ccgtccgtgt ggcctgtgca ggagatgcag acccaaaggt ggcctcctgg tcagtgagaa 165120gctggaaacg tgacaggaac tgacgtgggg ttattgagca tttaggggaa gacgttagca 165180gagcaggaat gagcaggcaa ctagtagaac acccacttaa gggctcacgg acaggtgctc 165240acttaggaag tgagtttcat ttggtattac accaggttcc tttaggcaaa gcggagggaa 165300agttctggtg tttttcactt gtaagatttt gaaggaaaca aaacactctt tacctttttt 165360ctaaaatgta ggtttgggag gaagctgagc attatcagag ggattgtgga gcaagagatt 165420caagcaatgg tttcaaagag agagaatatt gccacccatc atttatatca ggcatgggat 165480cctgtccctt ctctgtctcc ggctactaca ggtacctgag ggaaagggtg cgggggagcg 165540gttgtacttg ggctagaatg agagaagact ggcatgctca ccacaccagt gatgcgggaa 165600gacctgagtg tggtctgagt tggaggctgt ggtgctaaat

acgctgcccc tttcataagc 165660aggagtctta gtcaggccca gggaggaagt aaaatctgga aatgaatgag aagcattctc 165720tcctgccagt caagaaatga gaagcgaaag aattctcacg ggctgtaaga ccagcaggat 165780ttaaaagttg aattagttgc ttatgttaag aactcaacca agttcatcta cacaagctga 165840atctccagct tttcctaaga aaccatgtgt ggcagtggct gcagggcagg gcacagctgg 165900gcctgagcac cccgctccct gcacctctcc cctccctggg ccctgcctgt cactgcccac 165960tctcccacca agccttccgg ttgtgtgcct gccctatcac aggcatcgga gcttgtcacc 166020tggtttaaaa gaagagagtt gtgtggggat ttgggatgca cgtttttcac tcaaaagtat 166080tttagcgtag agctctgtga ttccgtagct atttaggagt ttaagcacct tgaaggcttt 166140aattgcagaa agttctatgt ggacgtgcaa tgtgttatac gcagtgtcta tgagactcaa 166200atgtttatta gggcgttgaa gtaaactgag cacttggagg gccatggatc cagccttcaa 166260ggagctcata agtcaggagg acccaggagc aatgacctgt catagaaggc agaaaagagg 166320ggcacagagg tgggtgggag gcatacacag gcagctcctg gagctccaag gggagcaagt 166380gcttccaggg aagggggcgt ggaggcccct ttggaggagg caagttgatc tggggtctgg 166440cagagggtta gctggggaca tttagcggga ggctggtgcc cgggaattgg ggggatgccc 166500agcagaaaga catgaggagg ctggcctggg gcgtgggggg gtgtgaaagg ttaagtgggg 166560gcattatcct gctcccgctc ctgccggctg tatctggtca gcctgggcac cgaggtgggg 166620ttctggaagg cactgttcac caaaatgctt atctgggtcc cccagagagc ttgcctgcct 166680ggactgtcgg ctcgcctgca actgctgact cctaagcttt tgcagctcag cccacaacca 166740gttcctattc acagaggtgg gagctgaggg gtgacaagtg actgctgcag tcttatttgt 166800catagagaaa aagtgacaga gtccagcttg cccactggcc ctgccagctt aactggttat 166860aaagtgacaa atccccaaga cccacagggc tctgcacaac ctgggccctc ctgccagtgg 166920cggcgagggc aggtggctca cggctgggtg cctgtctggg caggagctgg gctggtatgg 166980ggtgggcctg cggccctgcc cccctgtgca gatcaagact cagggtgctg gtgttcacag 167040gtgccctcat cagccacgag aagctgctgc tacagatcaa ccccgagcgg gagctgggga 167100gcatgagcta caaactcggc caggtcagtc tcgcgccccc gccgcctggc ctctgtccgt 167160ttctgtcctc agactttggc gcttgacaca cccaggagaa aagctcagtg cactttttaa 167220atgaaaggaa gttttccttt tttttaaaaa aaaatttaat gttcattgtt tttatctgtt 167280ttattcctag gtcccgcaag cagaggaagc attagttttg tttttattta tgttctgtat 167340tccagaaagt agttaagaga cctcacatgt agcgatagag atgtgtgtaa gagacagtga 167400gagggcgtga cttggactta agcaaggacc gtgagacaca aaaagggggg tgaggacaga 167460gtggagtcag ctgaaatgct caggaggaag tagacgccat gaagggccat ggtatggggg 167520gccgcaggcg tggccgtgag tgtccctggg gccagctctt ggggggctcc ctgagtgtcc 167580ctgtccctgt ggccagttct gggtgggagc cccgtgtgca ggcagacagc tcggccactt 167640cctagcaggt cacattggtc tgtgcttctg tttcctcctc agataagtga agggattcaa 167700gggtctgggt gtggtggcta acacctgtaa tctataacat tttaggaggc tgaggcagga 167760ggcttacctg agctcaggag gttgaggctg cagtgagcca tgattgcacc actgcactcc 167820agcctgggca acagaccagt actctgtccc ttaaaaaaaa atgtaaacag aaacgtaggg 167880ccatttgcat atgatggcac atggcgtgga gccctacagg tgtatgctgg gcggggcccg 167940gctgtgctgg ccgacttgca cctttccctc caccccggtg ctgtgtcttt cgctcaccgg 168000gttcctgatt tagtgaaagc agttgtgcag gacagttctc tttgtagctt ttgtttctgt 168060ggaaatgggt cagaatatgg tgtttagaaa cacttatgag ctctgagagt ttcctcttct 168120gagttcctgg cctgcagcct tcacagcaga aaccctgtga tgtcacaagc ctgtttctgt 168180tccctgctct ctgcctgtac tgtcctgttt tgtgcctgcc ggtttcagtg acaggaagca 168240gggagctact ggaccagcct gtatttttct agacatagtt ggaaaaagaa gtcccactct 168300tctgtccttt cacctttgac agatgtttcc accccaagat aagtgaaaat gaccaatagg 168360atgcactgta tttttcatga aagtgtttct gaagggcagg ctgagagtga gaggcctggg 168420gctcactggg tgcctctggc cttgtcctgg gcccagggac actggtctgt gcccgaggta 168480ttccctatcc ccccaacccc gctgcatttg gccacatcct tcaatgtttg cgttgtgtcc 168540agcgtccgca aaccaactgt catgggatca tactggggct gaagtacggt cccacccctg 168600ccctgtctgg ggctgaagta cagtgccacc cctgccctgt ctggggctga aggacagtgc 168660cacccctgcc ctgtctgggg ctgaagtaca gtgccacccc tgccctgtct ggggctgaag 168720gacagtgcca ccccttccct gtctggggct gaaggacagt gccacccctg ccctgtctgg 168780ggctgaagga cagtgccacc cctgccctgt ctggggctga aggacagtgc cacccctgcc 168840ctgtctgggg ctgaaggaca gtgccacccc tgccctgtct ggggctgaag gacagtgcca 168900cccctgccct gtctggggct gaaggacagt gccacccctg ccctgtctgg ggctgaagga 168960cagtgccacc cctgccctgt ctggggctga aggacagtgc cacccctgcc ctgtctgggg 169020ctgaaggaca gtgccacccc tgccctgtct ggggctgaag gacagtgcca cccctgccct 169080gtctggggct gaaggacagt gccacccctg ccctgtctgg ggctgaagga cagtgccacc 169140cctgccctgt ctggggctga aggacagtgc cacccctgcc ctgtctgggg ctgaaggaca 169200gtgccacccc tgccctgtct gggatgttta gcccctagat gccactggac tgagccgcta 169260cttgcttttg ggaaagaggg gtgggggtta ggggtctggg cgaggggagt gcaggggctc 169320ctccttggcc tgagagctgt tcatacagac tcctcgccca ctccctgcag ggtgctgggt 169380cccagggggg aaatggccct tggtgccaag aacgtgagtt ggggctagtg ccagtgatga 169440tggagaacag ctttttatgg gcacacagcc cacagcactg tgccaagtgc tcgaggcttc 169500ccgagaacca ggcagaaagg aggacagtcg aggtgtgctg actgcgtggt ggctgcgtga 169560tctagagcgc gggtcacaaa ggcgcgaggg agctctggcc ttgggtttac cgcaatgact 169620gccagtgcgg gagactggaa aaggaatctc acgtattggt tccgtgtttt ggggactcca 169680ttcagatgtc acttaggagt gaaagcatcc cttcgtagag cctctttctg tgtcaccctc 169740ctcagctgct cctggggttg actggcccct gattcatgcc tttagcatgt gctggagctt 169800cccagcagct gtccagcccc tgccccaccc tctctgtggg ctcccttgcc cgtaacctgg 169860ggtgtctgaa cgacccttgc taaggggcag actgttagac ggtaggcatg tgctgagtcc 169920cagtggccac acccacccac caggagcctg gcactgtggc cgcagcactg agcagtgccc 169980cgtttctgtg gcaggtgtcc atacactccg tgtggctggg gaacagcatc acacccctga 170040gggaggagga atgggacgag gaagaggagg aggaggccga cgcccctgca ccttcgtcac 170100cacccacgtc tccagtcaac tccaggtttt ccaatggcct ttttcttttt aacagaaatt 170160tgaaatttct tatcagtcat ttgatttgtt tgaggtgctt cttgaaatga gcctctcatc 170220tcatgtactt ggaaaatacc catctcgcat attccacagg aaacaccggg ctggagttga 170280catccactcc tgttcgcagt ttttgcttga gttgtacagc cgctggatcc tgccgtccag 170340ctcagccagg aggaccccgg ccatcctgat cagtgaggtg gtcagatccg taagtgagcc 170400ttcccattcc cctcacacct gcacgtgcca cacgcaccac acacgccaca caccccacac 170460acacacaccg cccacacaca tgccacttgc acacacaccc ctcatgcatg caacacacac 170520acaggccaca cgcaccatag acaccacaca cacatgccac atgcacacac atacacggca 170580tgcaccatac acacaacaca cacagcacac atgccacaca cacacgccac accacatgca 170640ccacacacat gccacatgca cacacactcc acatgcatgc accacacaca cacacacaca 170700ccacacacac cacatgcacc acaccacaca ggttacatgc acacaacaca cacatgccac 170760gtgcacacac cccacacacc acatgtatgt gccacacaca gcacacaacc acacacatgc 170820accacacaca tgccacatgt gcatgcacca gacacatggc acacactaca cacacgccac 170880gtgcacacac cccacacaca tgtacgcacc acacacatgc cacacacaca tgcaccacac 170940acatgccaca tgtacacaca tgtatataca caccccacac cacacacaca ccacttgcac 171000accacgcaca cacaccacat gcgcacacac acaccacata cgccacatgt acacaccata 171060cacacaccat acatgcacca cgtgtaccac gcacccacac agacacagca cacgcataca 171120ccacacacac acgcacacat gcgtcccgca cagtaatgtc tcttgggtgt aagaacacga 171180cttgccagta gtagcgttct ggatgcgttg cctggattct aacagcgcga ttctcccctt 171240gccctcctgg ttttccacat ctccagcttc tagtggtctc agacttgttc accgagcgca 171300accagtttga gctgatgtat gtgacgctga cagaactgcg aagggtgcac ccttcagaag 171360acgagatcct cgctcagtac ctggtgcctg ccacctgcaa ggcagctgcc gtccttggga 171420tggtaagtga caggtggcac agaggtttct gtgctgaagc cacgggggcc catctgcctt 171480gggacctggt gttggccaga ggtgccgggt gcggctgcct ccttccaaga gttgacccga 171540accggactcc acggcccacg tgagctgcag tgcttctcag atggaggggg ttcagcgacg 171600gtcagtgcca ttcacaggtc actgtgatgt gggttgtggc ggccaagcca tggtttgggg 171660tcccgtatcc ctgggcttat gacatcattg tagtagccca tccccacaga accacggtgt 171720gtggtggcgc tgaggcatcg tagatggtgg aaatgctact ggcttcccca tgctctgccc 171780tgaggcctga ctgcctcact ccccttctca gttatgttcc aggccccccg agcttcctgg 171840ctggacagct tctctcctgg gggccgtttt gtcacagtga ccctgtgttt ctagtcccaa 171900atctgggtgc tatagtctct ttttagcgtg gtggttgtct tagtcttttt tggctgctac 171960cacaagttac cttagactgg gtaatttata aacagtggaa atttacttct caccgttctg 172020ggggctggaa gttttcatgg tcaaggtgcc agcagatttg gtgtgtgatg agggctgctc 172080tctgcttcat agatggcatc ttctggctgg gtcctcacgg tggaaggagt gaacaagctc 172140cctcaggcct tttagaaggg ccccaatcca caagggctct cccatcatga cctcatcacc 172200tcccaaggcc ccaccttctt gtactgtggc actgcaaatt aggtgtcagt gtaggagttt 172260caggagggat agaaacattc agaccatccc agcggtcaag tgttcatcct cttgagttcc 172320tccttattct gcttctggtt tatcaggatt cagccagtgc agcatggtac ctgtattctg 172380tggcacatca ccacatggta tttgccaagt atccatcacc tgcacacgtg aaatcattgc 172440ccgtgggtcc cgacatctgg cgaagcatat tcaaggatgg cagaactgtc agagctggca 172500cctctggttc cttgtcatgt ggcattacct agtaatccat tttatgatag caatggaaac 172560tcatttcttc aacaaacacc tgagtggctg ccgtgtgcca gccgtctggg gcccttggtg 172620agaatggcat ggtggtgccc atcagggcct gcctagcccg tgctctggac gggctcctgt 172680gtgtcaggaa cgacaatgct gtcatgacgg tgaatgattt ttttttttgc catcactcca 172740gccgctaaca tttgcggagc tcttcctccc gcacccccac ctgacaaggc caagggtgac 172800cttggcccca ccctaggcgg ccaaggtcag aggttagctg gcttgtctgg gtcacacaaa 172860atgcagcaga ggttgaggtg agcacatgtc cgtgacctgg agcctgactc cctctctgcg 172920agtcttgact gctcttgcct agactctgtc ctccccgagc ccaaacgcca gtcatcttcc 172980cttgtgggtg tccttcagcc tggtgccatg ctggtgactc agcagccgtc cagggagtgg 173040aaacaattga gtgtgtgggt tccctgtgtg ggcatctctc ttcacggcga acaccctctg 173100ggtgttgccc acacgatgtc aaagcggctc ttggaagggg tccttctcct ttgtgggaag 173160tttcagctgc tgggctaact tgaattgtaa ctgtggtttt gtgctcaggc ccagatcccc 173220ctaggcaagt gttgtgccat cagtaatcaa atgagaaata atcattttga aaagcagatc 173280ctaaggcagg atggtcatgg acactcactc ccagctcttt gtgcactcat gctttctgga 173340agatggccat cctctgtgaa ggttttcagc gcgtcatgct tggtacccac gtatccagag 173400catgtcgttt tgaggtattt gcccaccgtt gtgaaatccg tgccacccga gagcaggtcc 173460tgatgtgggg ctttcagaag tgggacctgg ggccgtacgc agtccttagg gaggggccgt 173520gtggcgttgt gcgtgtgagg ggatagcaca gggtgaggtg ggggcccaag aaggaagtga 173580cccacaaaga acagcctcct cttttggtcc ttgttcctgg gatggctggg agtggcttct 173640gtgtcgtccg gccatttccc ctgcggagag gctcctacca ctgccgagaa cctcatcatt 173700ccacaaaaac aagaggccgc ctggccatcc agcgctccat gggaattctg tgtccccata 173760gtcttgggct gaaggagggt gacattcctt gctgacttct gcaggggtct cctcactgtt 173820aaagagcaga ttgaaagtga agaacgtggg ctaagtgttt aggtcgatat ttaaccctgc 173880taggttttgg atactaagtg aaattgaggc cattttggtt gaagttgaca gaaaccacta 173940tcagggatcc ccaagactac cccaggcttt tctagaaaga ctctcagcta agatgtgtta 174000tggtaaaagc acacaaaaca aaatcagcaa agaaaattag caagggcaga ggcccatggg 174060gcgatgtccc gaggacacca ggcttgagct tccagaatcc tctcccagcg gggtcgtgca 174120ggacgcactt aactccccgc acagtgagcc gtgacagcgc gtgtgcagtg tcgtcgccag 174180gaaagcacac tagagactcg gtgccagggt ttttactggg ggctgggcac atgggcaccc 174240tctgcctgcc tcgtgcccag actctggact cccggaggga aggcaagttc tcagcaccaa 174300ccctggtgcc cacacaagca gctgagcaca gggagcccct cctcagtgag gatggtgggc 174360accgtcccaa caccagccag gggccagcct tgcacacagg cctctcagga tggtctccgg 174420cctgctgtgt agtctcttct gcacacaagc gtgagggcag cgcccccgcc tcggctgtgg 174480ggaggagcca ctgggacgtg agctctggtg gcatgcagca gcttttgtct gtgtgtgcct 174540aggacaaggc cgtggcggag cctgtcagcc gcctgctgga gagcacgctc aggagcagcc 174600acctgcccag cagggttgga gccctgcacg gcgtcctcta tgtgctggag tgcgacctgc 174660tggacgacac tgccaagcag ctcatcccgg tcatcagcga ctatctcctc tccaacctga 174720aagggatcgc ccagtgagtg ggagcctggc tggggctggg gcgggggtct cagaatgagc 174780tgtgaaggaa gcagcatcac cctctccaag tgcccaggct cctggccaga tggcaggcca 174840ggtatcagtg ggaacccagg tgggtgccat ggctgaggtc agtgagacgc aagagcacag 174900gtgcgtccta gaggcttcct cgggcacctc cagcgagctg gagctctcgc ctctgctgct 174960gtctcatgtg gcgcttagca cactctccca cgtgcccatt cctgactctg ctctcgaggc 175020catcggctct cattctctgc tcccagaacc ctgttattac ccaggctagc ctcctctctg 175080caccttcccc gccctggccc agtacctccc tcttgtttcc actgtgattc cgacctcacc 175140ttatcttaaa gctgctggac ggcaggttct gtacacacgt gtccttgaca aagcacggct 175200ggtgccgcaa cccctcagcg agcaagtcaa gctcttcaca gcgatgtctt acaagcgcag 175260agggctctgt gacaccctgg tctcaccgcc actcttccaa agtcgcagag gctttagcag 175320agatgggccc agcctctctg agtcataggc ttctgcacac gggagctgtc tttagaggga 175380gggtggaatt tcatcagcca cccacatggg ggagttgagg gcaagaatta ggagcaaaga 175440tgggaagggg tctgggagga atggccagtg atcccctttg acaagtgggc aggaaacggg 175500ggctaggtca aagttgagtg gaagacctgg agggagacgg gaaggtctct gtaggcacag 175560ttcagacagg agggaggtgt gagccagggc acatgccggt ggccgtctgg caggatttgg 175620gacatgctgg agcagggaca gcggctcatc aggggccatt gccctcatcc aggccagagt 175680gtcacaagcc cgtggggagg cccttctcgc ctgtcatcct tgctgggcag tgggtgctgt 175740gctagcagga caggcggacg gctggcaact gtctctgcat ccctggagcc tggcataggg 175800ccaagtcaca cggggcacag gcctgcaaat caggcacata tgttggtgca gtgacgtgat 175860tttggggggc agccccagaa caggccccag acacaggcca aagccctgcc tgtgctggtg 175920tgttgggctg ttctatggct cttgctgtgg gcatggagga ctcagggaag gagagttgag 175980gtggtccagg agttgcgttt gggatgcaga gagcttgtgg catccaggta gaaatggtgc 176040gtggggctga cctcagcacc atgggcagag gggccgtgtc acgtgcctcc gaggtggagg 176100tgggaccacg tggtgacaga tatacgcatc actgggcacg tttttgtggg tgttgggggg 176160catcgtattg gctcctctgt tcacagtggc cactcattca gtccctggct accaggtcct 176220cactgtgcca tggggaaggc cggcgctgtc gggggatcac agaaggcagc acgtcatgat 176280ggcatgtgcc atgaaggaaa agcacagggc actcaggaag tagaggggac tggcctgggg 176340tgtgggaatc tagggcctcg ttgagggaca gagagaggaa gtgtgtggtg gccagcatgg 176400aggtggccac aggggaggct gagttaggcc gagagggcag ggcgttgggg aggtagacgg 176460gctcagccac tcagggagtg gtcaagcaga ggctgaaggg tcaggccagg ttgcaggggc 176520ctgggggagc cactcagggt aggcgctccc gggagcccgc ctggcccata gctctacact 176580cccgcgtggg gccggacatg ctgtgaagcc ctctccacgt tggatggggg tggctgagcc 176640tggatgctgt ctcccgtttt cagctgcgtg aacattcaca gccagcagca cgtactggtc 176700atgtgtgcca ctgcgtttta cctcattgag aactatcctc tggacgtagg gccggaattt 176760tcagcatcaa taatacaggt gagtgggccc tggctgtctt cctctgcaca cggggagtgg 176820gcttcccttc tcttttcctt gcaggatcat accagtgggc cagttttgac ttggtcggga 176880ggaggcatga acacctgaga ctgtgcagcg attctttgac acagaggcct ttctccctgt 176940gcagatgtgt ggggtgatgc tgtctggaag tgaggagtcc accccctcca tcatttacca 177000ctgtgccctc agaggcctgg agcgcctcct gctctctgag cagctctccc gcctggatgc 177060agaatcgctg gtcaagctga gtgtggacag agtgaacgtg cacagcccgc accgggccat 177120ggcggctctg ggcctgatgc tcacctgcat gtacacaggt gagcatgtac acggtgccca 177180taaggccagc ccaagtcctg ttcaagggag gcaggagcat gctcactcaa gggacctcga 177240ctaggtgccc tctgatttca cacttctggt gttgccccaa gccggcccca tcaccttgca 177300agaaaggctc tggagccccc agggctggag tacctggtca gggttgaccg tccctgtggt 177360cactcatccc atgtggctga gctgggctgg gtcctgggca agcaaggggc tgatatcacc 177420tgctttcaga tctccaggga ctcactggac ccctgtgtac aaagcactgt ctacagagcc 177480tattgggttg tatagaggta accttcgtac tgaacacttt tgttacagga aaggagaaag 177540tcagtccggg tagaacttca gaccctaatc ctgcagcccc cgacagcgag tcagtgattg 177600ttgctatgga gcgggtatct gttctttttg ataggtaaga agcgaagccc catccctcag 177660ccgttagctt ccctagaact ttggcctgaa gctgtgcttt tgtgtgtgtc tgctgatccc 177720ctggcgctgt tgctggagtc ctgccagtga ttccccacca cagcctgacc atgggctgcc 177780ttggctcagg gttccactgg cgagctggtg gtccttggac cccagcactc aggtgtagcg 177840ttgaccagtt ccaaggttgt cccagtgcct gcccatctct cctgagggct cagggacagt 177900acctggcagt tgggggtgtg gcagggggca ggaatgacca gcctctggga gggtggggca 177960gaagcctgta cagtgaggag gagctggctc agcctggctg cctatcgtga gaggggagcc 178020cacggggctg tgggaggggg gccgtggtgc ctgtgagcag ggtgaggagc agcggcagga 178080ggatgaaggt ggaacccaca catgcatctt tgagacccgt gtggtcagtg gcttctgccc 178140cccaccaccc cccactgctg tgcgtgcata gaattggctt ccctcacctg ctctggaagt 178200gggttaggag cttggtaggg ctttttctca aggacaaggg cccctgattt gctctcaggc 178260ctcagtcctg gcgacatggt ggatctggag ccttgttgca ctgccttgcc tgtgctctcc 178320aatcagggtg gccagtgggg agccatttgg cttttctcaa gagcatactc aggtggacct 178380tgctccactg tttgaccaga tgaggcattc tgaacagcca agcctgtgct ggtctgtttt 178440catgttgatt tttttttttc ttttcttttt gagatggagt ttttcccttg tcacccaggc 178500tggagtgcaa tggtgtgatc tcggctcact gcaacctccg cctcccgggt tcaagtgatt 178560ctcctgcctc agcctcccta gtagctggga ttacaggcac acaccaccat gcccagctaa 178620tttttgtgtt tttagtagag acggggtttc accgtgttgg ctgggctggt ctcgaactcc 178680tgaactcaag tgatccaccc tccttggcct cccaaagtgc tgggattgca ggcgtgagcc 178740actgcgcccg gcccccatgt cgatttttaa atgcacctct gcatcgttct tcagtcccca 178800tatgctcact gagcaccact gcgactggca gacgggcaca gggaggcgcc acgaccagtc 178860ctggccttca aggggcttgt ggtctagtgg gcccaatgct aggtggcgag tgctccaaag 178920agtgtggtgc acgccttccg cttgaccgct ctccagacgc cacagggagg cacctcgcag 178980ctgaccacag atttctctct gtggagcagt gtcttcagag cggctgccat gccactgctg 179040ggcgagggtc tgcgggcggg tagagccagg agcacctgtg aggaagtgca ctgccatttt 179100cgtagctgct tcccgtgtgt ctcagttaca cacggctggc atgtgtgcac tgatgagacg 179160ggaacgtgat ggttgctttt cagcactgaa agggatactg ctcagggggc gtgtttcagg 179220atctggttag ggaagaagca gcgagagcac agatggggcc ctgtgtggta acaagaaaaa 179280agtcctggtt gacaacagtg ccacgaagcg ttagaacaca tagggatgtt tgtggagcat 179340ttgcatgtgg aaagcagcaa aaacataatg ggaacgggtt cttttgttat gatttttaaa 179400aatctctttt gtaacatcct tcccgctgcg ccgtttctgc atattccttt atgtagcttt 179460caaactcctc ttaggagttc tggtccctac agggcgtggg agcccaggct ttacgtagct 179520ttcaaactcc tcttaggagt tctggtccct acagggtgtg ggagcccagg gcctgtgccg 179580agcagcctgc ctccacgagc tagacagagg aagggctggg gttttgcctt tttagtctca 179640aaattcgtac tccagttgct taggctctga ctttccccac ttggaaagtc cctcacggcc 179700gagggtccct cccagccctg atttcacatc ggcattttcc ccagtattag agccaaggcc 179760ctccgcgggc aggtggggca gctgtgggag ctggtgccag tctctgacct gcgtccctcc 179820tcccaggatc aggaaaggct ttccttgtga agccagagtg gtggccagga tcctgcccca 179880gtttctagac gacttcttcc caccccagga catcatgaac aaagtcatcg gagagtttct 179940gtccaaccag cagccatacc cccagttcat ggccaccgtg gtgtataagg tgaggttgca 180000tgtgggatgg ggatggagtg ggaaagcctg gaggtggagt tgcctccgac ttcccagcag 180060attcgccagc agagcccagc tcctccgctt taaagcagca atgcctctgg cccccacccc 180120acccccgcca cccaggcgca gcaggtgctt cccgtccccc cagccctgac actcaggcac 180180ctgcttgctc cttgcaggtg tttcagactc tgcacagcac cgggcagtcg tccatggtcc 180240gggactgggt catgctgtcc ctctccaact tcacgcagag ggccccggtc gccatggcca 180300cgtggagcct ctcctgcttc tttgtcagcg cgtccaccag cccgtgggtc gcggcgatgt 180360atcctctctg ggtccctggt gctggccccg tttcccttgt caacaccgag gctcatgttt 180420catgataagg ttttgaaacc taacctttgc aaaaacccca cagatgccag ggtgacaggc 180480cctcagcccc agggaagtaa aatgctgaca ggggtacaga aaggagcacg tccagacatt 180540tgctgaccag ggcctctcag aggggccggt gtatggcagg agggtcgcag ctgaggggcc 180600tttctgtgga gggcctgggt gaggggagcg agggtgggcg gtggtctctg cagacgtccc 180660gcccactcgc gggctctgtg tggctgggct tctcctgaca

ctgcttctca ttagctttgg 180720tcattgtgcc tcgatcgccc tctcggggaa aggcttaagt aaagatccag ttcccacccc 180780cagatgctgg ctgccaggag tttccctttc cacagccctt ccccaagaca gaccacaaga 180840gcctccaagc agcacagttg tcctggtgct gacagcacag ccttgcccgg cgtgcctggc 180900acggctctgc cctcactgca ttggagcagg gctagtggag gccagcggaa gcaccggcca 180960ccagcgctgc acaggagcca ggccaggtga gtgctgccga gtgggtgccc tgcctgcagg 181020gcatccagcc agccaagggt tgcaggaatg gaggtggagg cgctgatgca gctggaggca 181080tccaggtggc ccttccgggg ctctgctcgc tctccaggct ccctggaccc ctttgtagac 181140tgtttcagga gaggaactcc caggtgagga cagggaggca gcattcccct catttgccgg 181200cctttttcct taactcctgc accagcctcc cacatgtcat cagcaggatg ggcaagctgg 181260agcaggtgga cgtgaacctt ttctgcctgg tcgccacaga cttctacaga caccagatag 181320aggaggagct cgaccgcagg gccttccagt ctgtgcttga ggtggttgca gccccaggaa 181380gcccatatca ccggctgctg acttgtttac gaaatgtcca caaggtcacc acctgctgag 181440cgccatggtg ggagagactg tgaggcggca gctggggccg gagcctttgg aagtctgcgc 181500ccttgtgccc tgcctccacc gagccagctt ggtccctatg ggcttccgca catgccgcgg 181560gcggccaggc aacgtgcgtg tctctgccat gtggcagaag tgctctttgt ggcagtggcc 181620aggcagggag tgtctgcagt cctggtgggg ctgagcctga ggccttccag aaagcaggag 181680cagctgtgct gcaccccatg tgggtgacca ggtcctttct cctgatagtc acctgctggt 181740tgttgccagg ttgcagctgc tcttgcatct gggccagaag tcctccctcc tgcaggctgg 181800ctgttggccc ctctgctgtc ctgcagtaga aggtgccgtg agcaggcttt gggaacactg 181860gcctgggtct ccctggtggg gtgtgcatgc cacgccccgt gtctggatgc acagatgcca 181920tggcctgtgc tgggccagtg gctgggggtg ctagacaccc ggcaccattc tcccttctct 181980cttttcttct caggatttaa aatttaatta tatcagtaaa gagattaatt ttaacgtaac 182040tctttctatg cccgtgtaaa gtatgtgaat cgcaaggcct gtgctgcatg cgacagcgtc 182100cggggtggtg gacagggccc ccggccacgc tccctctcct gtagccactg gcatagccct 182160cctgagcacc cgctgacatt tccgttgtac atgttcctgt ttatgcattc acaaggtgac 182220tgggatgtag agaggcgtta gtgggcaggt ggccacagca ggactgagga caggccccca 182280ttatcctagg ggtgcgctca cctgcagccc ctcctcctcg ggcacagacg actgtcgttc 182340tccacccacc agtcagggac agcagcctcc ctgtcactca gctgagaagg ccagccctcc 182400ctggctgtga gcagcctcca ctgtgtccag agacatgggc ctcccactcc tgttccttgc 182460tagccctggg gtggcgtctg cctaggagct ggctggcagg tgttgggacc tgctgctcca 182520tggatgcatg ccctaagagt gtcactgagc tgtgttttgt ctgagcctct ctcggtcaac 182580agcaaagctt ggtgtcttgg cactgttagt gacagagccc agcatccctt ctgcccccgt 182640tccagctgac atcttgcacg gtgacccctt ttagtcagga gagtgcagat ctgtgctcat 182700cggagactgc cccacggccc tgtcagagcc gccactccta tccccaggcc aggtccctgg 182760accagcctcc tgtttgcagg cccagaggag ccaagtcatt aaaatggaag tggattctgg 182820atggccgggc tgctgctgat gtaggagctg gatttgggag ctctgcttgc cgactggctg 182880tgagacgagg caggggctct gcttcctcag ccctagaggc gagccaggca aggttggcga 182940ctgtcatgtg gcttggtttg gtcatgcccg tcgatgtttt gggtattgaa tgtggtaagt 183000ggaggaaatg ttggaactct gtgcaggtgc tgccttgaga cccccaagct tccacctgtc 183060cctctcctat gtggcagctg gggagcagct gagatgtgga cttgtatgct gcccacatac 183120gtgaggggga gctgaaaggg agcccctcct ctgagcagcc tctgccaggc ctgtatgagg 183180cttttcccac cagctcccaa cagaggcctc ccccagccag gaccacctcg tcctcgtggc 183240ggggcagcag gagcggtaga aaggggtccg atgtttgagg aggcccttaa gggaagctac 183300tgaattataa cacgtaagaa aatcaccatt ccgtattggt tgggggctcc tgtttctcat 183360cctagctttt tcctggaaag cccgctagaa ggtttgggaa cgaggggaaa gttctcagaa 183420ctgttggctg ctccccaccc gcctcccgcc tcccccgcag gttatgtcag cagctctgag 183480acagcagtat cacaggccag atgttgttcc tggctagatg tttacatttg taagaaataa 183540cactgtgaat gtaaaacaga gccattccct tggaatgcat atcgctgggc tcaacataga 183600gtttgtcttc ctcttgttta cgacgtgatc taaaccagtc cttagcaagg ggctcagaac 183660accccgctct ggcagtaggt gtcccccacc cccaaagacc tgcctgtgtg ctccggagat 183720gaatatgagc tcattagtaa aaatgacttc acccacgcat atacataaag tatccatgca 183780tgtgcatata gacacatcta taattttaca cacacacctc tcaagacgga gatgcatggc 183840ctctaagagt gcccgtgtcg gttcttcctg gaagttgact ttccttagac ccgccaggtc 183900aagttagccg cgtgacggac atccaggcgt gggacgtggt cagggcaggg ctcattcatt 183960gcccactagg atcccactgg cgaagatggt ctccatatca gctctctgca gaagggagga 184020agactttatc atgttcctaa aaatctgtgg caagcaccca tcgtattatc caaattttgt 184080tgcaaatgtg attaatttgg ttgtcaagtt ttgggggtgg gctgtgggga gattgctttt 184140gttttcctgc tggtaatatc gggaaagatt ttaatgaaac cagggtagaa ttgtttggca 184200atgcactgaa gcgtgtttct ttcccaaaat gtgcctccct tccgctgcgg gcccagctga 184260gtctatgtag gtgatgtttc cagctgccaa gtgctctttg ttactgtcca ccctcatttc 184320tgccagcgca tgtgtccttt caaggggaaa atgtgaagct gaaccccctc cagacaccca 184380gaatgtagca tctgagaagg ccctgtgccc taaaggacac ccctcgcccc catcttcatg 184440gagggggtca tttcagagcc ctcggagcca atgaacagct cctcctcttg gagctgagat 184500gagccccacg tggagctcgg gacggatagt agacagcaat aactcggtgt gtggccgcct 184560ggcaggtgga acttcctccc gttgcggggt ggagtgaggt tagttctgtg tgtctggtgg 184620gtggagtcag gcttctcttg ctacctgtga gcatccttcc cagcagacat cctcatcggg 184680ctttgtccct cccccgcttc ctccctctgc ggggaggacc cgggaccaca gctgctggcc 184740agggtagact tggagctgtc ctccagaggg gtcacgtgta ggagtgagaa gaaggaagat 184800cttgagagct gctgagggac cttggagagc tcaggatggc tcagacgagg acactcgctt 184860gccgggcctg ggcctcctgg gaaggaggga gctgctcaga atgccgcatg acaactgaag 184920gcaacctgga aggttcaggg gccgctcttc ccccatgtgc ctgtcacgct ctggtgcagt 184980caaaggaacg ccttcccctc agttgtttct aagagcagag tctcccgctg caatctgggt 185040ggtaactgcc agccttggag gatcgtggcc aacgtggacc tgcctacgga gggtgggctc 185100tgacccaagt ggggcctcct tgtccaggtc tcactgcttt gcaccgtggt cagagggact 185160gtcagctgag cttgagctcc cctggagcca gcagggctgt gatgggcgag tcccggagcc 185220ccacccagac ctgaatgctt ctgagagcaa agggaaggac tgacgagaga tgtatattta 185280attttttaac tgctgcaaac attgtacatc caaattaaag gaaaaaaatg gaaaccatca 185340gttgttgctg tgtgaggctt gctttgcttc atgagaacct agaccttgct gagctggagt 185400cttaggaagc agtctcctaa gtgcttctcc agcaggggca gaaactgtcc caccagctaa 185460catctggcat tatggagggt cccccaggca gctgccagca gggacaggcc ccgtgttttc 185520tgtagccagg gatgaggaag tggccccagg gcatgggcct ggctgggtgc ttctgcaagg 185580gccttcccaa accacagtac aggtggtctt cctgccctgc agatgggagc tgtgggagct 185640gctggagctg ctggagcctt catggtcaag tgacatcata agcttatatg acatacacaa 185700gcctcaggac ttggcccatg gcactgaagc aggtcatcag gcccagcaca gagactagag 185760ctgtgttctc acagggccca ccacccttcc acctccttgg ccattgacac ctgcgtccct 185820ggcccagctg ctcccaggta acccccaaag cagctggcac atcccacctc tggtgtggcc 185880ggggctgctg tgtgtccgca gggcctgccc cgtctattct agcttgtttg tcctgtctga 185940accagcgcct actccaagaa gcctctgctc agcccagcgg ggatgcttct aagctccgga 186000cgagcctctc ggaagccttg gtgattggtg gtgtagtcat cttgggatgc agatgtctta 186060ccaacctgca agaacaaaaa ccctgtggct tcctctggtg cagggtattt agtcaatgtt 186120tgctgaggtc ccgtctggtt ctggctaatt ggcaggggtc gtccacccat tctttccctg 186180ctctgctgtc tgtgccagga gagacggggg ccagtcggcc aaggggccag ctcctgctgc 186240ctgctcctct tgggcacgtg cgggggcccc ctttctctga gcagggatag ggatcagtct 186300gccggaggga tgtggtggac aggcctaaag catttggggc ggggcatgcc acttgagctc 186360cctaaatctg tctcctcata ggtgacaccg ctccagggcc ccccagtggc ctctcctttc 186420agagctacct aaattctggt cacttcagag aaatggagca cccccttctc cctggtccag 186480gtgtggacag cctggcacac tgagcacacc tggcatggct ggtaatttca gaaagaagag 186540gggccggggt ccagtgggaa gcagcggtga acccctcgtg agtgggcttt gcagtccctc 186600cccatgccac ggcagagctg ccctcaacac agccttcctc ttcctcatcg gagagcacac 186660cctgtcccct tgccgagctg tgccctgtgc cttcggtggt atttgatttt ggctgctact 186720ggctttgttg ggatctggaa gtcgcttccc ctgcgtggtg cgtggagcac tgtaagtcag 186780atgagggaag tagccagggt gaggtgagta ccgggtggag ccgccactga agggactggg 186840taggggggcc ttgcctctac atgatgtgac acagccaacc gaggacagag gaagccccgt 186900tcctgggggt gtggggtgca cccctcaggg aagcctgcag tggggcctga ggaaaggcat 186960cctccgcgag cccacgagtc tggtccatga gcaccgtgac agtgtctgtg ggtagaggtg 187020gacccggcct tgtgtcatca ccaggacctc ttttgggaaa ccatgtggac atcgcttgcg 187080ggtcccccag gctctgcagc cccagcagcc tggctgcctt ttgggcaagt ggcttgagcc 187140acagaggacc cagtcctgtt gcagccacat cctctggggg ggcccgccag tgtggccggc 187200tttctccacc ctacaccagg cctccaggtg tcctggtcgg gggtgtctgg gccctgggtg 187260ggccctgtgg acctgtgagg tcagggtcag ggcatcactg gaggcagagg gctgaagttg 187320tgggtctggg ttccccttgt gtgcacaggc ccctgccctc catgcttggt caggcagcta 187380cccccaaaac tgctaggaca ggctggtcct gaggtggatc ctggcccctg taccctctgg 187440acagcccacc cgcccaacct tctaccctgc cccagcggcg gcagtgttgg ccacatcctt 187500cccctcctgg ccccaattgc tctggggaag tccaggctcc ggagcctgcc caggggcccc 187560ccgtgatttg ggcccaggac tccacgtggt tctctgcctt cacccaagcc ctgaactcct 187620cagctgccaa atccccaccc atctgcacag gctgtgctca ccactgctgc tcctggaagg 187680tgcccctcag tgggacgccc acctcctctc tgggcttctg tgtttgggag ccctgctgcc 187740cccacccttg gtcagtcccc atgtcctgct ggcctgtcag gcagggcaga aaatccaccc 187800agaaatgctg agcaggatga gagtctagtt gggcccagcc tcattattta gaagggatgg 187860aggcctaggg agcatgcttc tagcctgagc ccagcagggc cccgcccatg tcccaggtct 187920gcaccaggga cagctcctgc cgaggcctga cctgcccctt ctccctcagg tgctgctggt 187980tgaccagcct ctggccctag gagaccccgt agcgactgag ggtcccagca ggccatgcag 188040ctttgccaag gtacgagccc ctccccagca ggggacagat gtggggaccc tcccaggcag 188100gagcagctgg gtgcctggtg ctgccatctg ctgcctgcct ggttcttgtc ctcacattgg 188160aggtcagtgt gagggctctg cctcgggaaa ggccatggag cttgccctgt ccagggcctc 188220ccatgtgcac tgagcctggg aagagagggt tggagttgag ccttttaccc tgggaatgct 188280gcctggagga tggtgcgggt gtggggtggc accctgccag gcagggccct gcctccctgc 188340gcccactgga actcgggcag gcaggggtgt aggtgcctcc tctagagccg tccggtgggg 188400gcccccggca gtggtggtgg tgtccactgg ccagcagctg ccccttcagc caggacagta 188460ggcctgacgc tgtccccagc agctccaagg tggatttgtg gaagggggta gagggcacgt 188520agaggcccca tgacctcccc agggttctgg gagggctgtg cccccttagc cagcaccatg 188580ctgggtgata tagtcagatc ctgttacccc tgttgtggag gtgaggaaac aggttagtgg 188640ggaggacatg actaaggtcc atgctgagtc gctagagctg cacccagaac cactgctggg 188700accccatgcc tttctgctta ccccttgtgc cgggagatgc caagagatgc tgggagccag 188760ccccacctct gcccttggag tcatggctac ggaaagggca ttcggaccgg tccctgacct 188820caccggggag ggccgaaccc tgttcctgag gagccagggc ttcctagagg aggtaggcct 188880tctagtcact ccttcatctg caggcactcc acagagctct ctgtgccagc ccccagcacg 188940gagggctgac cttagtcgag tggagatgcc ccagtgccag gcagtaggga tgatgtctcc 189000tgaggcccag atggaaggga ctggactagt ctcatggggc tgatggtggg gccaggcctt 189060gaccagggac ccagtgtagg gggtgcagag acccctctga gttcctcaca catccctggg 189120gccctcccca tacacttcct atcctgactg cgggcaagag ggagccccag ttcgccttcc 189180ctatgctggg cacccacagt ggggctgggc acccccgcca tgcccctgcc ctgtccttcc 189240cctgagagcc tcggtcccac ctccaaggtg cctcagagga cagcaggggc agcgggcaga 189300ggccgagatg cctcctcatt ccaggctcag ctgcccttct tggggcagcc cacacctgag 189360agtctcctgc agttggtcag gcctgaggag ggcagggggg tgcctgctgt ccctctgctg 189420accacagtgg catttagcct gggcaccgcg cccagcacag tccatgctgc acaggtgccg 189480tgggctccac agagccctgc ctgacatgca tgtgttacgt ttcgggtgcc gatgcccttg 189540ggcggcactt ctccgggcag aacccccagg ccaccgctcc ggttccggtt ccgctgcatc 189600tggggctctc ggcaggctgt ggtcctccgg ccagcctggg ggcatctcag tccctcagcc 189660ccacaggggc ctgccccgca gcctgggcct cgagccccgt ctccgcacgc tgtgccgaat 189720ctggctgccc atcagctccc tgcgtaccca gactgtgccc tgccatgccc gtggctcttc 189780ccaggagtgc cctgtggcct ccccctggct tgctgggctg attccctcct gtgtctcaaa 189840cagagctcac ctttgccatc actgctgtcc tcaccggccg gtgccagagg cccgtgtctg 189900tgtaccctgt gtctgcacct ctgggcaggg cctggctctg accaacccgg gcttccagtg 189960tccacagacc taaggcccag ggcgcctggg ggctggagca agagaagcaa aaggagccaa 190020gggtgggggt ttggggttct tgtgagggcc cagccccagg accccaggac caggacaccc 190080aggagcccca gggcccagcc ccagttcaga aggcaggggc cttctgaggg agcttaaggg 190140tcccacagcc caggaccccc accagggcca gtggccagcg ttgggggact cagcctcctc 190200gtcgctcgtc ctctctgttt ctcccacctt ttgccccctt tctccttgcc tgttcccacc 190260cgaggccccc tcttggcctg cgtgagccgg ggcggcactg aactgggggc cgatccgcct 190320gggcggcggt gagaggcagg gccgggagcc gggccgctgg gtttgggcct ggcccgctcg 190380ccgcaatatt gatggcccgt cagtgcagcc ctgattcctg tgctttcagt taaaaggttt 190440ctgttgttgt agcttatgca gttgctctgt tgctatggaa acgtgacatc aaaatgacgt 190500ttcccgttta aaagctttta actaaattcc tgcctgtcag atgtaggccc cattttgagc 190560gtggagctgc cttcgagcga gcgtgagcgg cgcctcccgc ccatggtgcg tggggccggg 190620ccggggccct cgctgagcgc gctctctcac cccacaggcg cctccggcat ggcggcggcc 190680gaggggcccg gctacctcgt gtctccccag gcggagaagc accggcgggc ccgcaactgg 190740acggacgccg agatgcgcgg cctcatgctg gtctgggagg agttcttcga cgagctcaag 190800cagaccaagc gcaacgccaa ggtgtacgag aagatggcca gcaagctctt cgagatgacc 190860ggcgagcgca ggctgggcga ggagatcaag atcaagatca ccaacatgac cttccagtac 190920aggtgggcga gcgggcagtg tgggccccac caggacgggc gggcccgggc gtggcgggcc 190980gctcctgact ttcttggagc tctgagtcgg gacgatgtgt gggtcgtggc ctgcctgtcg 191040gtctcctctg gccgggtatg ggcagaaccc cacggggtga gacggggccc acggaaaccg 191100tgtgtgcagc cttccattgg ggaagtgggg aaactgaggc ccagcaaggg caggaaacca 191160gtctaagagc tgaggggtag caggggtggg gctggtgctg ggcagaggcc aggatggctc 191220ccaggacgta tgggcggtct gggcactgtc cctcggaggc agcaacactc atggtggtgc 191280ccactgacct cacaccctgc tcccccatag ggaggcggcg gctgccagtg ccctccccac 191340caccaagctc ccaagctcag caggggtttc aggggcctac tgcgtcattg gggaaattga 191400gactgcaagt gagaaggagg ctcagtgctc tgcgacttgg agcatccact gagcctctgc 191460catgagccgg tgagccccac tggggctggc cctagggtca cggtggggta tttccagaaa 191520tcaccaggtg aggtgcagga ccagccagcg catgggtggg gcttacggtg cgaagaagaa 191580agaggtggag gcctgccctg gcccaggact cccagcgtgg gggctcccgg cctggcccca 191640cctctgctcc tgctacatgg caggtgggcc cttcctgccc tggcaacctg cagggaaggc 191700cggaggggac cacccagcca gggagatgtt ggcgtctagg aggggacagg tgtggtccca 191760cacacccagc atcttaaagt gcgtgggtcc ccagcccatt aggacagggt cccgggtggg 191820caggggtcat ggtggggtga aggtctcagg cacaggcaag gtcacaggtg cggtgagggt 191880cttgcagggt gtgaaggtca taggtgtgcg gtgaaggtca caggtgtggg gtgatggttt 191940tgggtgtggg gagggtcttg cacggagcga gggtggcagc aagagctgga agctgcaggg 192000ggagaatggc agcagagagc acccggccct gtgggcggcc tggacagggc tgggcctggg 192060gctgccggag agcctgtcag cttccaggat gggagtggcc tcactcagct gctccacctc 192120cgggtcaggc aggtgagcct ggggcagaga ggctgagagc acctgagcca cttgtgggag 192180aggccacccc cactgccccc ctcaggcgag gagccggcct ccagcacagc agaagggaac 192240ccccagtccc cagccctagt gggagtgggg aagaggccca gcaaggcccc ggacagaccg 192300ccagcctgtg aggtctccgc tttcagttgc gttgatttga ttttttctga gccttgaagg 192360aggggtccgg ggcctggccc tgcccaaagg cccctaggca ggccccaaag ccgggaccta 192420gggtgctgag catgacggat gttgggtttg agcggctggc ttgcgacgtg agggctgagg 192480tgtgagcctg ggtatcttca gaggttcggt ggacacaggc agctgcccgc ggccccactg 192540ttcccgtggc ctcctagtcc tgctcaggca cctggtgagg aagggacgca gagggcagtg 192600ggaggtggcc acgactgttc cagcaggctc ccctctgact caggaattca cgggcaccac 192660ctccctggct ggctctggtt ggtgtctggc caggttattc attatttatg ctgaaagcct 192720cttcagagtc ccaggggagg gtttctgtct ccattcctgg aggctgagag atgagggtgc 192780agcagagtgg gggcctccac tccagaccct gcagtctggg ctggccaagg gctgcaccgg 192840tgcactgcac gtcatggctg atgaagcact tccacaccgc agcccctcag agctgccaca 192900gtcagcctta gttcaccgag ggggaagctg aggcccagag catgagaggg acttgcccag 192960ggccacatag tccttagcag aggaagctgt ggctgggtga ctcgatcttt gtcctttttc 193020tttatacccg cagtctcccc atagcagagg cttttctttt ttttttcttt ttcttttttt 193080tttttttaca agaactcttt atatattaag gctgttgggc tgaagaagcc tgagagggtg 193140gctggttctg tggagcatgg tttgttgaag tacagtttgg gggcctccta cactgagaat 193200aggccttttc tcgtttctcc aaagagtggg ctggctcaag tagggcagag agagaagcct 193260ggggcagagg ttagggatgg gcacccagcg cctgccctca cacgctctgt gctggtgtct 193320tcacagccac gtgccaccct gggcagcatc ccctgctcac catctggctg tgcctgtttg 193380ctgggggcac ctcattcaga atccagctta ttgtttccaa cggccaatgg ccacaccctg 193440gcaggtagca agagtaggag agaggagaca cccactccga gcacaggttg ggtttggagc 193500ccggccttgg ggcactctgt cactcaaagg cagagtgggg agtgggcact gggccttagg 193560aggtactggg tccagtgagg cagagatgcc cctgccccac ccccaccttg tggcttcttc 193620cctggcctgg ccagagctgt ctggccgcca tggggccctg tgtctcctgc cttgacctcc 193680cagagggcag ccgaggccca ggggaggcct ggggacttag cctctcaggg caggacctgt 193740ctgcaggagt aggtgggtgc tgggggtccc agtggtaatg aggcatcagg cagtgtggga 193800aggggcccat ccggcccacc ccagggcctc tgggcaggtt gcaggttgta gcgctggatc 193860taggctcctg cccagactgt aggttcaacc aagaatggca tgggagccca gcctgctgtt 193920tgctttatta aatctgccct gtagctgggg gaggggctta ctttgatcat cactatgtca 193980ttgatataaa aatagaggct cagagaggtg aatgaacctg cccaaagtca cacagcaaag 194040tgtggagatg agatactgac tcagggctgt ggacactgaa gcctgtgctc taacgccagt 194100ggctgtcgct ccctgaggca ttctctcccg aacaacacag ttattatatt acaaaatatt 194160atcactatat ttatatatct tataatacct tattattaca ataaaacctt attactctac 194220ctttcaaaat gaattattta aaaagcagta tttgctcatt gcagagagtc tagaaactat 194280agaaaagcaa gggaaaagca ataggaccag ccccaaggtc ccagcatgca cagataacct 194340tagtaatact gggacgtgtg cttccttttt aacatctgag cccgtgtagg tcctgaagcc 194400cagcttcttt ctaagtccat tgtcatcttg accctggagc ctggccgatt ttgctgggga 194460ggcccttgcc agccgagagc ggctcctgcc tgtgccggcg tggcgcgccc ctctgctgag 194520gctgggcagg acaggggctg ggccagctct gtttctcacc cttggctctt gtgtctctcg 194580tttcaggaaa ttaaaatgca tgacagatag cgagtccgcc ccgcccgact ggccctatta 194640cctagccatt gatgggattc tggccaaggt ccccgagtcc tgtgatggca aactgccgga 194700cagccagccg ccggggccct ccacgtccca gaccgaggcg tccctgtcgc cgcccgctaa 194760gtccacccct ctgtacttcc cgtataacca gtgctcctac gaaggccgct tcgaggatga 194820tcgctccgac agctcctcca gcttactgtc ccttaagttc aggtagtgtg tctgcttgtc 194880cttcccctgc cctggggtat ctcagccccc accatttaga gaaagggact gggagtggca 194940aggccggcgg cggcggccac agtggttgca gaggccgtgg ctgcgggcag cgcctccagg 195000gacaggcggc ctcagaccag ggagggcttt agtgtccaca ggcagaccga gtttgtctcc 195060cagctccatc acttttgagc tgcacggaaa gttccttgac ttctctggcc tcagtctccc 195120tcctataaaa tgggggtaaa tcagtacctt tctcagaggg tggctgggag catcacagga 195180gagaagacgc agcatggggc ccggcacacg gagggagacc aagccccaga ccccagaatg 195240cgccccctgg cctcccttag cccacacaga ccccaccctc acaggctagc tgccctctca 195300gcactgggga gggtgtcggg ctgcacctca tcacgtgttg ccgtgggcat gacccgtccc 195360ctctgccatc catcccacac ctcagacccg tcccgtgctg gccacgtgac tgtgcctgca 195420agatgctcac agggcagccg ggagccaggc agcatgcagg acagacacct gcggggtggg 195480cctggggagc ccagagaagg tgcttttgag gaggggacat ttggggtggg ctttcaaggt 195540aaaatagaag ttggccattt ggaggcaaga acaggaagat tgtggatttg agtcacagct 195600tctcccctgc cctggtcttc aagtctttct gacaggaggt gtcagaaaag tatctttagt 195660agagaaggcg tctccgagga gggtccctct catgccgggg gccgctgctt gactcaggat 195720ttctcattga agacctgaga caaaaacgct tttgctggca

gctagaagga accagcagga 195780ggcctgagat ttgtggctgt tgttcccgtg gactgagccc agttctcaga ctcagctgcc 195840tggggccttg cacaggactg gggcgtgggg gctgccctcc ctgatcaggc ccaaagcgcg 195900gatctcacgc ccctgaggtt ggctgtaccc tctcagctca gagcagagtg tgggccaggg 195960atgagcaggc actggagcag ggccctgggg tctgtgggtt ttggcagctc cctgcccttc 196020agggaggtct gctgagacca cgggtggccc ctaccccagc agcagagctc tcaggaggcg 196080cccacagggc tggactgcct ttactcacca cctctaccag agctctgagg tcctggggag 196140agagcccagg cctcttgtgg gccccacacc ctctaggtgc ctgtccttct gcctctctac 196200caaggtgtgc cggccccatt tctaggccgc cgggagataa gggggctcac atctcaggcc 196260cttccttctg ggacctcagt ttccccatct gcctaaggcc gggtggggct ggtggtcttg 196320gcttccctac aggggtcctg agtactctgc actacccagc accccccacc cctgccttca 196380tctctccctg ggggtggtct ctccacccct ggcccccaac tggggctgag cccccacctg 196440cccagtttgg tgggtgaagg gtgctccctg gcaggatatg cccctctgca gcccagaaca 196500tcccaccctt tccagaccga aggggtgtgg attgtcctgg gaccctggtc attggggtca 196560tccgctagtc gcaaaggacg gcaatgcctg tggcctctct ttctttcttt ttcttttttt 196620ttttttttga gacggagtct cgctcttgtg cagagagcag tggcgcgatc ttggctcact 196680gcaacctccg cctcgtgggt tcaagcgatt ctcctgcctc agcctcccga gtagctggga 196740ttacaggcac ccgccacaac gcctggctaa tttttgtatt tttagtagag atggggtttc 196800accatgttgg ccaggctggt cttgaactcc tgacctcagg tgatccacct gcctctgcct 196860cccaaagtgc tgggattaca ggcataagcc tccacacccg gccacccctg ttactttctg 196920tcaaaggcgg tgggttctgg cccctccttt gcacatggaa tatgagaccc tgagtaagtg 196980acctgactcc ctggggcctc agtttcccca tttgcccagt aggattgtcg ggagggtccg 197040gtgaggcccc tggtgtgccc aggctctgtg gccagcacgt ccacagccgg cactgtcctt 197100ccaggtcgga ggagcggccg gtgaagaagc gcaaggtgca gagctgccac ctgcagaaga 197160agcagctgcg gctgctggag gccatggtgg aggagcagcg ccggctgagc cgcgccgtgg 197220aggagacctg ccgcgaggtg cgccgcgtgc tggaccagca gcacatcctg caggtgcaga 197280gcctgcagct gcaggagcgc atgatgagtc tgctggagag gatcatcacc aagtccagcg 197340tctaggccag caggcggcgg cggcggcggg gccgggcggc tggtggtact gctcaggcca 197400cccagggcag gccactcagg ccaggcgggc aagggggccg ccccgcgagc ggagaccgcc 197460ttccacctgg cctctggcag gatgtccctt ctgaggggta ttttgaggaa cccccaggcc 197520ctggggaccg tgaggctcca gtctccagca tgaatgccct tcctcggaca caggccaggg 197580cctctggggt tcactccgag taagaacgtc ctagagccac tctccagtgt cgttactatc 197640aatgatactt gacgtggctt tgatattaaa cgtatacttt ttcattcttg cctggaacgc 197700acagtttgct gttgctggct tggtgaggat gccctgattg atggatcccg aaaatgaaag 197760cagatggaaa cgggttgggg caggctggag ctgggggagc tctctcctga agggaaccct 197820gtgtcctccc tcaccaggac ctctgcgtct ctccttaaat ggcctctgac gcctgatgaa 197880aaccccagcg accttccagg aggcttttat tcagctctgt ttggagcatc aggtgtttcc 197940actgcctcct tagcaatgac actaataaaa gtcgtaacac ctgttcacat gcacagccct 198000gttgagtgtt ctgggtgctg gagatatcat ggtggatgac acaaaggccc tggcctcttg 198060gagcttatgc tcccatgcgg ggaagacaca tgggtcagta gagaaatggt tgcaggttgt 198120gataagtgct ggaagggagg ggttggcctg aggacacgga ggcagacata cgtggagctg 198180ggaacagtgg ccacacaggg aacggccagt gcgaaggccc agaggcagag gacactggag 198240caagcccagg agcagctagg aggctggtgg ccagcagcca ggccacggaa gcccgtgcag 198300cccgtgggga ggagtgttca tgcttttcaa gcttagtggg agtcttttgg ccagtgcagc 198360tctgggtctg acatcggtgg gggacagagg ggtggtggag cggccacagc tgcaagctca 198420cctcactgcc ggcccttcca ccagtttcaa actctttcta gaagctccag ctttcccaaa 198480gctgaattct ctatgagcct ccttggccgg gactcgggcg tctggttgcc ctggctgcaa 198540aggaggctgg ggccaggtgt gtttgagtca cctcctggaa ttaggcaagt tgctgcccaa 198600atagaaggtt gttggcaggt gggtcagcag gtgaacagca tggtttgact cagggttcag 198660aaaaatctcc ctctggctgc caagcgagca ggccgtggag acaggtgcag aggcaggtgt 198720ggcagcaggc atcctgccag gcagtgctgc agtcatcctg cgacaagcag cagcagctca 198780tcctaccctc tagggggtct tgaggtcagc caggcaagag agcagcttgg actccactgg 198840gtgtgggacc agcctgtgga ccatggtggt gtggagggtg ccctcggcct gcctgtgtga 198900aggagaggcc ggcgtgttct gtggagccca aaggggagct gggcaagcag gattcacttc 198960actctgaggg tcctggagct cccaccctcc tcagccatct ccccagagcc tgtgtgccga 199020ggactcggcc catgttgctg tgggatgaga ggcagagtgt cgtgagggtg taaggagcgg 199080cggcagtggt gggaggaggg agcagcagcc agcgctacgg tgccagtttc cagctgccag 199140atgacgccgc tgaccctgtg gttgagaaga gatgcacaga gccagctctt gcaagccagt 199200gtggctgcca tagcacctgc cgagaagcag aaggaagggt ggccccagga ggacagagga 199260tgcgggcaca tctgatgcgg gcctgagttt tgggagcttt tgctctagcc agtttccagc 199320tccgggaccc acccgcctcg taggcaagac accacccaag aaatcatttg cttaacaaac 199380acactgggct ccaactggac acctgtgcca ccctagatgc tgggaaccca gccatgacac 199440aggcacctgc ccccagctgc tgaccactga ggctggctag cagctcccat ggggccagtg 199500tggggttccc cagcctccta acagggagcc agtcacaagc cctcgagagg gaagggtgcc 199560cgcggccctg gcaggaaggt taggctggac gctcccacaa gacataacag atggaggttc 199620taaatgatgt agcaacttct tcaccctgaa actgctgtag agtcagccat gacgcaccgg 199680tacttcagta actgccaggc atccgggaca gcacaccgcg agtcgctgct gtgcttgggt 199740tagaagtggt ttggtctgtt ttcttctcgc cctctctaat cagagtcagt gattcatgcc 199800cttccatcac cttagagaag gggcaggcgc tgcccgacct tctccaggct ggagcagcat 199860cgcctcatgt cagcagaact cagctgtaga atatcgtggg gttggtgcct ttcatcagca 199920gcatgtcctt aacaactttc tgatttcttc cttagttgtt ggtccattaa ggagaaaaaa 199980aatgatctca gccattgcta aaatatttga taagattcag caaagcagca tgttaacatt 200040gaaaactaga atcaggagcc aggcagatgt gcttgctttt cacctgtagt atttcatgtt 200100gttttgacgt ttttagctaa tgcattaaga taaataaaca aaagccgggc acggtggttc 200160acgcctgtaa tcccagcact ttgggaggct gaggcgggag gatcctctga ggtcaggagt 200220tcaagaccag cctgaccaac atggagaaac ctcgtcatta ctaaaaatac aaaattagct 200280gggcgtggtg gtgcatgcct gtaatcccag ctacttggga ggctgaggca ggagaatcgc 200340ttgaacccgg gaggcggagg ttgcagtgag ctgagattgc accactgcac tccagcctgg 200400gtgacagtga aactcggtct caaaaaaaaa aaaaaattaa aaaaagataa ataaaataag 200460caggataaga aatgaagaaa gtagagttac ctttgttttc agatttcatt tttgtatacc 200520cagaaagcca aatgtacaaa agactgggag ctctttaaac cagcttaaac ttgttgaaaa 200580tgaggatgaa gaaatatccc attcagagtt ggaatgaatt taacccagaa ggaacaggac 200640ctctactgaa gagaactatg cagtcttact gaaaaatcta aataatacct gagcgctgga 200700gaaacttcgc acactcctga aagctccaaa gtcaatgtca tcattttatt aatgtcattc 200760caaacatagt ctcaataata tcacttcttg gttttgacat ggacgcgatg atgtttaaat 200820tcatatgaaa aaagaacggg gccaaaagtc caaggccagt cagcgtgaga agaccgctcg 200880gcctccctcg gagtcgggga gttggaaccg cagactgaga tcatgtggct gctggaggcc 200940aggacgaacg tcgggaaatg gagactcctg cgttgctggt gggatgtggt gcagccgctt 201000ccaggagcaa tttggtgtcc cgtcctaaag ctgaagaaac gcatttcctc tggtcagtgc 201060cactcctaga caggccaccc tgcggcagcc gtcctcaaac tggtctgagg acccctcaac 201120gctcttaaaa atcattaaaa gtgggccagg tgcggtggct cacacctgta atcccagcac 201180tttgggaggc caagacaggc ggatcacgag gtcaggacat tgagatcatc ctggctaaca 201240cggtgaaacc ccgtctctac taaaaataca aaaaattagc cgggcgtggt ggcgggcgcc 201300tgtagtccca gctacttggg aggctgagcc aggagaatgg cgtgaaccca ggaggtggag 201360cttgcagtga gctgagatca ctccactgca ctccagcctg ggcagcagag cgagactctg 201420tctcaaaaaa aaataataaa taaataaata aaaataaaat aaaataaaat tcattaaaag 201480tgccaaagaa cttttgctta tgtgagttct aatgaccaat attaatacac attagaatat 201540cttattagaa attaaacctg agacctttag aaaacatgta ttcatttcaa aatagcaata 201600aacccatgac atattaacat aaataacaat tgtatgaaaa atatattttc caaaacaaaa 201660agttttcggg agaagtgtgg catagtttta catggtcgta aatctctggc ttaagagaag 201720cccactggcc tctcagcagg ctctgggtcc gtccactttg ggggtgtttt ggttgtgaag 201780tataggagtg aatggagaag ctcattctta cccagatgtg tatttgaaaa gaaaaggaac 201840attttaataa cctttgcaaa taatcggtat attcttccgt gatcctattc caacactgga 201900caggtggtgg tttgtttttt ttttttggag acggagtccc gctctgtcac tcaggctgga 201960gtgcagtggc gcgatttcag ctcactgcaa gctccgcctc c 202001213481DNAHomo sapiens 2gctgccggga cgggtccaag atggacggcc gctcaggttc tgcttttacc tgcggcccag 60agccccattc attgccccgg tgctgagcgg cgccgcgagt cggcccgagg cctccgggga 120ctgccgtgcc gggcgggaga ccgccatggc gaccctggaa aagctgatga aggccttcga 180gtccctcaag tccttccagc agcagcagca gcagcagcag cagcagcagc agcagcagca 240gcagcagcag cagcagcagc aacagccgcc accgccgccg ccgccgccgc cgcctcctca 300gcttcctcag ccgccgccgc aggcacagcc gctgctgcct cagccgcagc cgcccccgcc 360gccgcccccg ccgccacccg gcccggctgt ggctgaggag ccgctgcacc gaccaaagaa 420agaactttca gctaccaaga aagaccgtgt gaatcattgt ctgacaatat gtgaaaacat 480agtggcacag tctgtcagaa attctccaga atttcagaaa cttctgggca tcgctatgga 540actttttctg ctgtgcagtg atgacgcaga gtcagatgtc aggatggtgg ctgacgaatg 600cctcaacaaa gttatcaaag ctttgatgga ttctaatctt ccaaggttac agctcgagct 660ctataaggaa attaaaaaga atggtgcccc tcggagtttg cgtgctgccc tgtggaggtt 720tgctgagctg gctcacctgg ttcggcctca gaaatgcagg ccttacctgg tgaaccttct 780gccgtgcctg actcgaacaa gcaagagacc cgaagaatca gtccaggaga ccttggctgc 840agctgttccc aaaattatgg cttcttttgg caattttgca aatgacaatg aaattaaggt 900tttgttaaag gccttcatag cgaacctgaa gtcaagctcc cccaccattc ggcggacagc 960ggctggatca gcagtgagca tctgccagca ctcaagaagg acacaatatt tctatagttg 1020gctactaaat gtgctcttag gcttactcgt tcctgtcgag gatgaacact ccactctgct 1080gattcttggc gtgctgctca ccctgaggta tttggtgccc ttgctgcagc agcaggtcaa 1140ggacacaagc ctgaaaggca gcttcggagt gacaaggaaa gaaatggaag tctctccttc 1200tgcagagcag cttgtccagg tttatgaact gacgttacat catacacagc accaagacca 1260caatgttgtg accggagccc tggagctgtt gcagcagctc ttcagaacgc ctccacccga 1320gcttctgcaa accctgaccg cagtcggggg cattgggcag ctcaccgctg ctaaggagga 1380gtctggtggc cgaagccgta gtgggagtat tgtggaactt atagctggag ggggttcctc 1440atgcagccct gtcctttcaa gaaaacaaaa aggcaaagtg ctcttaggag aagaagaagc 1500cttggaggat gactctgaat cgagatcgga tgtcagcagc tctgccttaa cagcctcagt 1560gaaggatgag atcagtggag agctggctgc ttcttcaggg gtttccactc cagggtcagc 1620aggtcatgac atcatcacag aacagccacg gtcacagcac acactgcagg cggactcagt 1680ggatctggcc agctgtgact tgacaagctc tgccactgat ggggatgagg aggatatctt 1740gagccacagc tccagccagg tcagcgccgt cccatctgac cctgccatgg acctgaatga 1800tgggacccag gcctcgtcgc ccatcagcga cagctcccag accaccaccg aagggcctga 1860ttcagctgtt accccttcag acagttctga aattgtgtta gacggtaccg acaaccagta 1920tttgggcctg cagattggac agccccagga tgaagatgag gaagccacag gtattcttcc 1980tgatgaagcc tcggaggcct tcaggaactc ttccatggcc cttcaacagg cacatttatt 2040gaaaaacatg agtcactgca ggcagccttc tgacagcagt gttgataaat ttgtgttgag 2100agatgaagct actgaaccgg gtgatcaaga aaacaagcct tgccgcatca aaggtgacat 2160tggacagtcc actgatgatg actctgcacc tcttgtccat tgtgtccgcc ttttatctgc 2220ttcgtttttg ctaacagggg gaaaaaatgt gctggttccg gacagggatg tgagggtcag 2280cgtgaaggcc ctggccctca gctgtgtggg agcagctgtg gccctccacc cggaatcttt 2340cttcagcaaa ctctataaag ttcctcttga caccacggaa taccctgagg aacagtatgt 2400ctcagacatc ttgaactaca tcgatcatgg agacccacag gttcgaggag ccactgccat 2460tctctgtggg accctcatct gctccatcct cagcaggtcc cgcttccacg tgggagattg 2520gatgggcacc attagaaccc tcacaggaaa tacattttct ttggcggatt gcattccttt 2580gctgcggaaa acactgaagg atgagtcttc tgttacttgc aagttagctt gtacagctgt 2640gaggaactgt gtcatgagtc tctgcagcag cagctacagt gagttaggac tgcagctgat 2700catcgatgtg ctgactctga ggaacagttc ctattggctg gtgaggacag agcttctgga 2760aacccttgca gagattgact tcaggctggt gagctttttg gaggcaaaag cagaaaactt 2820acacagaggg gctcatcatt atacagggct tttaaaactg caagaacgag tgctcaataa 2880tgttgtcatc catttgcttg gagatgaaga ccccagggtg cgacatgttg ccgcagcatc 2940actaattagg cttgtcccaa agctgtttta taaatgtgac caaggacaag ctgatccagt 3000agtggccgtg gcaagagatc aaagcagtgt ttacctgaaa cttctcatgc atgagacgca 3060gcctccatct catttctccg tcagcacaat aaccagaata tatagaggct ataacctact 3120accaagcata acagacgtca ctatggaaaa taacctttca agagttattg cagcagtttc 3180tcatgaacta atcacatcaa ccaccagagc actcacattt ggatgctgtg aagctttgtg 3240tcttctttcc actgccttcc cagtttgcat ttggagttta ggttggcact gtggagtgcc 3300tccactgagt gcctcagatg agtctaggaa gagctgtacc gttgggatgg ccacaatgat 3360tctgaccctg ctctcgtcag cttggttccc attggatctc tcagcccatc aagatgcttt 3420gattttggcc ggaaacttgc ttgcagccag tgctcccaaa tctctgagaa gttcatgggc 3480ctctgaagaa gaagccaacc cagcagccac caagcaagag gaggtctggc cagccctggg 3540ggaccgggcc ctggtgccca tggtggagca gctcttctct cacctgctga aggtgattaa 3600catttgtgcc cacgtcctgg atgacgtggc tcctggaccc gcaataaagg cagccttgcc 3660ttctctaaca aacccccctt ctctaagtcc catccgacga aaggggaagg agaaagaacc 3720aggagaacaa gcatctgtac cgttgagtcc caagaaaggc agtgaggcca gtgcagcttc 3780tagacaatct gatacctcag gtcctgttac aacaagtaaa tcctcatcac tggggagttt 3840ctatcatctt ccttcatacc tcaaactgca tgatgtcctg aaagctacac acgctaacta 3900caaggtcacg ctggatcttc agaacagcac ggaaaagttt ggagggtttc tccgctcagc 3960cttggatgtt ctttctcaga tactagagct ggccacactg caggacattg ggaagtgtgt 4020tgaagagatc ctaggatacc tgaaatcctg ctttagtcga gaaccaatga tggcaactgt 4080ttgtgttcaa caattgttga agactctctt tggcacaaac ttggcctccc agtttgatgg 4140cttatcttcc aaccccagca agtcacaagg ccgagcacag cgccttggct cctccagtgt 4200gaggccaggc ttgtaccact actgcttcat ggccccgtac acccacttca cccaggccct 4260cgctgacgcc agcctgagga acatggtgca ggcggagcag gagaacgaca cctcgggatg 4320gtttgatgtc ctccagaaag tgtctaccca gttgaagaca aacctcacga gtgtcacaaa 4380gaaccgtgca gataagaatg ctattcataa tcacattcgt ttgtttgaac ctcttgttat 4440aaaagcttta aaacagtaca cgactacaac atgtgtgcag ttacagaagc aggttttaga 4500tttgctggcg cagctggttc agttacgggt taattactgt cttctggatt cagatcaggt 4560gtttattggc tttgtattga aacagtttga atacattgaa gtgggccagt tcagggaatc 4620agaggcaatc attccaaaca tctttttctt cttggtatta ctatcttatg aacgctatca 4680ttcaaaacag atcattggaa ttcctaaaat cattcagctc tgtgatggca tcatggccag 4740tggaaggaag gctgtgacac atgccatacc ggctctgcag cccatagtcc acgacctctt 4800tgtattaaga ggaacaaata aagctgatgc aggaaaagag cttgaaaccc aaaaagaggt 4860ggtggtgtca atgttactga gactcatcca gtaccatcag gtgttggaga tgttcattct 4920tgtcctgcag cagtgccaca aggagaatga agacaagtgg aagcgactgt ctcgacagat 4980agctgacatc atcctcccaa tgttagccaa acagcagatg cacattgact ctcatgaagc 5040ccttggagtg ttaaatacat tatttgagat tttggcccct tcctccctcc gtccggtaga 5100catgctttta cggagtatgt tcgtcactcc aaacacaatg gcgtccgtga gcactgttca 5160actgtggata tcgggaattc tggccatttt gagggttctg atttcccagt caactgaaga 5220tattgttctt tctcgtattc aggagctctc cttctctccg tatttaatct cctgtacagt 5280aattaatagg ttaagagatg gggacagtac ttcaacgcta gaagaacaca gtgaagggaa 5340acaaataaag aatttgccag aagaaacatt ttcaaggttt ctattacaac tggttggtat 5400tcttttagaa gacattgtta caaaacagct gaaggtggaa atgagtgagc agcaacatac 5460tttctattgc caggaactag gcacactgct aatgtgtctg atccacatct tcaagtctgg 5520aatgttccgg agaatcacag cagctgccac taggctgttc cgcagtgatg gctgtggcgg 5580cagtttctac accctggaca gcttgaactt gcgggctcgt tccatgatca ccacccaccc 5640ggccctggtg ctgctctggt gtcagatact gctgcttgtc aaccacaccg actaccgctg 5700gtgggcagaa gtgcagcaga ccccgaaaag acacagtctg tccagcacaa agttacttag 5760tccccagatg tctggagaag aggaggattc tgacttggca gccaaacttg gaatgtgcaa 5820tagagaaata gtacgaagag gggctctcat tctcttctgt gattatgtct gtcagaacct 5880ccatgactcc gagcacttaa cgtggctcat tgtaaatcac attcaagatc tgatcagcct 5940ttcccacgag cctccagtac aggacttcat cagtgccgtt catcggaact ctgctgccag 6000cggcctgttc atccaggcaa ttcagtctcg ttgtgaaaac ctttcaactc caaccatgct 6060gaagaaaact cttcagtgct tggaggggat ccatctcagc cagtcgggag ctgtgctcac 6120gctgtatgtg gacaggcttc tgtgcacccc tttccgtgtg ctggctcgca tggtcgacat 6180ccttgcttgt cgccgggtag aaatgcttct ggctgcaaat ttacagagca gcatggccca 6240gttgccaatg gaagaactca acagaatcca ggaatacctt cagagcagcg ggctcgctca 6300gagacaccaa aggctctatt ccctgctgga caggtttcgt ctctccacca tgcaagactc 6360acttagtccc tctcctccag tctcttccca cccgctggac ggggatgggc acgtgtcact 6420ggaaacagtg agtccggaca aagactggta cgttcatctt gtcaaatccc agtgttggac 6480caggtcagat tctgcactgc tggaaggtgc agagctggtg aatcggattc ctgctgaaga 6540tatgaatgcc ttcatgatga actcggagtt caacctaagc ctgctagctc catgcttaag 6600cctagggatg agtgaaattt ctggtggcca gaagagtgcc ctttttgaag cagcccgtga 6660ggtgactctg gcccgtgtga gcggcaccgt gcagcagctc cctgctgtcc atcatgtctt 6720ccagcccgag ctgcctgcag agccggcggc ctactggagc aagttgaatg atctgtttgg 6780ggatgctgca ctgtatcagt ccctgcccac tctggcccgg gccctggcac agtacctggt 6840ggtggtctcc aaactgccca gtcatttgca ccttcctcct gagaaagaga aggacattgt 6900gaaattcgtg gtggcaaccc ttgaggccct gtcctggcat ttgatccatg agcagatccc 6960gctgagtctg gatctccagg cagggctgga ctgctgctgc ctggccctgc agctgcctgg 7020cctctggagc gtggtctcct ccacagagtt tgtgacccac gcctgctccc tcatctactg 7080tgtgcacttc atcctggagg ccgttgcagt gcagcctgga gagcagcttc ttagtccaga 7140aagaaggaca aataccccaa aagccatcag cgaggaggag gaggaagtag atccaaacac 7200acagaatcct aagtatatca ctgcagcctg tgagatggtg gcagaaatgg tggagtctct 7260gcagtcggtg ttggccttgg gtcataaaag gaatagcggc gtgccggcgt ttctcacgcc 7320attgctaagg aacatcatca tcagcctggc ccgcctgccc cttgtcaaca gctacacacg 7380tgtgccccca ctggtgtgga agcttggatg gtcacccaaa ccgggagggg attttggcac 7440agcattccct gagatccccg tggagttcct ccaggaaaag gaagtcttta aggagttcat 7500ctaccgcatc aacacactag gctggaccag tcgtactcag tttgaagaaa cttgggccac 7560cctccttggt gtcctggtga cgcagcccct cgtgatggag caggaggaga gcccaccaga 7620agaagacaca gagaggaccc agatcaacgt cctggccgtg caggccatca cctcactggt 7680gctcagtgca atgactgtgc ctgtggccgg caacccagct gtaagctgct tggagcagca 7740gccccggaac aagcctctga aagctctcga caccaggttt gggaggaagc tgagcattat 7800cagagggatt gtggagcaag agattcaagc aatggtttca aagagagaga atattgccac 7860ccatcattta tatcaggcat gggatcctgt cccttctctg tctccggcta ctacaggtgc 7920cctcatcagc cacgagaagc tgctgctaca gatcaacccc gagcgggagc tggggagcat 7980gagctacaaa ctcggccagg tgtccataca ctccgtgtgg ctggggaaca gcatcacacc 8040cctgagggag gaggaatggg acgaggaaga ggaggaggag gccgacgccc ctgcaccttc 8100gtcaccaccc acgtctccag tcaactccag gaaacaccgg gctggagttg acatccactc 8160ctgttcgcag tttttgcttg agttgtacag ccgctggatc ctgccgtcca gctcagccag 8220gaggaccccg gccatcctga tcagtgaggt ggtcagatcc cttctagtgg tctcagactt 8280gttcaccgag cgcaaccagt ttgagctgat gtatgtgacg ctgacagaac tgcgaagggt 8340gcacccttca gaagacgaga tcctcgctca gtacctggtg cctgccacct gcaaggcagc 8400tgccgtcctt gggatggaca aggccgtggc ggagcctgtc agccgcctgc tggagagcac 8460gctcaggagc agccacctgc ccagcagggt tggagccctg cacggcgtcc tctatgtgct 8520ggagtgcgac ctgctggacg acactgccaa gcagctcatc ccggtcatca gcgactatct 8580cctctccaac ctgaaaggga tcgcccactg cgtgaacatt cacagccagc agcacgtact 8640ggtcatgtgt gccactgcgt tttacctcat tgagaactat cctctggacg tagggccgga 8700attttcagca tcaataatac agatgtgtgg ggtgatgctg tctggaagtg aggagtccac 8760cccctccatc atttaccact

gtgccctcag aggcctggag cgcctcctgc tctctgagca 8820gctctcccgc ctggatgcag aatcgctggt caagctgagt gtggacagag tgaacgtgca 8880cagcccgcac cgggccatgg cggctctggg cctgatgctc acctgcatgt acacaggaaa 8940ggagaaagtc agtccgggta gaacttcaga ccctaatcct gcagcccccg acagcgagtc 9000agtgattgtt gctatggagc gggtatctgt tctttttgat aggatcagga aaggctttcc 9060ttgtgaagcc agagtggtgg ccaggatcct gccccagttt ctagacgact tcttcccacc 9120ccaggacatc atgaacaaag tcatcggaga gtttctgtcc aaccagcagc cataccccca 9180gttcatggcc accgtggtgt ataaggtgtt tcagactctg cacagcaccg ggcagtcgtc 9240catggtccgg gactgggtca tgctgtccct ctccaacttc acgcagaggg ccccggtcgc 9300catggccacg tggagcctct cctgcttctt tgtcagcgcg tccaccagcc cgtgggtcgc 9360ggcgatcctc ccacatgtca tcagcaggat gggcaagctg gagcaggtgg acgtgaacct 9420tttctgcctg gtcgccacag acttctacag acaccagata gaggaggagc tcgaccgcag 9480ggccttccag tctgtgcttg aggtggttgc agccccagga agcccatatc accggctgct 9540gacttgttta cgaaatgtcc acaaggtcac cacctgctga gcgccatggt gggagagact 9600gtgaggcggc agctggggcc ggagcctttg gaagtctgcg cccttgtgcc ctgcctccac 9660cgagccagct tggtccctat gggcttccgc acatgccgcg ggcggccagg caacgtgcgt 9720gtctctgcca tgtggcagaa gtgctctttg tggcagtggc caggcaggga gtgtctgcag 9780tcctggtggg gctgagcctg aggccttcca gaaagcagga gcagctgtgc tgcaccccat 9840gtgggtgacc aggtcctttc tcctgatagt cacctgctgg ttgttgccag gttgcagctg 9900ctcttgcatc tgggccagaa gtcctccctc ctgcaggctg gctgttggcc cctctgctgt 9960cctgcagtag aaggtgccgt gagcaggctt tgggaacact ggcctgggtc tccctggtgg 10020ggtgtgcatg ccacgccccg tgtctggatg cacagatgcc atggcctgtg ctgggccagt 10080ggctgggggt gctagacacc cggcaccatt ctcccttctc tcttttcttc tcaggattta 10140aaatttaatt atatcagtaa agagattaat tttaacgtaa ctctttctat gcccgtgtaa 10200agtatgtgaa tcgcaaggcc tgtgctgcat gcgacagcgt ccggggtggt ggacagggcc 10260cccggccacg ctccctctcc tgtagccact ggcatagccc tcctgagcac ccgctgacat 10320ttccgttgta catgttcctg tttatgcatt cacaaggtga ctgggatgta gagaggcgtt 10380agtgggcagg tggccacagc aggactgagg acaggccccc attatcctag gggtgcgctc 10440acctgcagcc cctcctcctc gggcacagac gactgtcgtt ctccacccac cagtcaggga 10500cagcagcctc cctgtcactc agctgagaag gccagccctc cctggctgtg agcagcctcc 10560actgtgtcca gagacatggg cctcccactc ctgttccttg ctagccctgg ggtggcgtct 10620gcctaggagc tggctggcag gtgttgggac ctgctgctcc atggatgcat gccctaagag 10680tgtcactgag ctgtgttttg tctgagcctc tctcggtcaa cagcaaagct tggtgtcttg 10740gcactgttag tgacagagcc cagcatccct tctgcccccg ttccagctga catcttgcac 10800ggtgacccct tttagtcagg agagtgcaga tctgtgctca tcggagactg ccccacggcc 10860ctgtcagagc cgccactcct atccccaggc caggtccctg gaccagcctc ctgtttgcag 10920gcccagagga gccaagtcat taaaatggaa gtggattctg gatggccggg ctgctgctga 10980tgtaggagct ggatttggga gctctgcttg ccgactggct gtgagacgag gcaggggctc 11040tgcttcctca gccctagagg cgagccaggc aaggttggcg actgtcatgt ggcttggttt 11100ggtcatgccc gtcgatgttt tgggtattga atgtggtaag tggaggaaat gttggaactc 11160tgtgcaggtg ctgccttgag acccccaagc ttccacctgt ccctctccta tgtggcagct 11220ggggagcagc tgagatgtgg acttgtatgc tgcccacata cgtgaggggg agctgaaagg 11280gagcccctcc tctgagcagc ctctgccagg cctgtatgag gcttttccca ccagctccca 11340acagaggcct cccccagcca ggaccacctc gtcctcgtgg cggggcagca ggagcggtag 11400aaaggggtcc gatgtttgag gaggccctta agggaagcta ctgaattata acacgtaaga 11460aaatcaccat tccgtattgg ttgggggctc ctgtttctca tcctagcttt ttcctggaaa 11520gcccgctaga aggtttggga acgaggggaa agttctcaga actgttggct gctccccacc 11580cgcctcccgc ctcccccgca ggttatgtca gcagctctga gacagcagta tcacaggcca 11640gatgttgttc ctggctagat gtttacattt gtaagaaata acactgtgaa tgtaaaacag 11700agccattccc ttggaatgca tatcgctggg ctcaacatag agtttgtctt cctcttgttt 11760acgacgtgat ctaaaccagt ccttagcaag gggctcagaa caccccgctc tggcagtagg 11820tgtcccccac ccccaaagac ctgcctgtgt gctccggaga tgaatatgag ctcattagta 11880aaaatgactt cacccacgca tatacataaa gtatccatgc atgtgcatat agacacatct 11940ataattttac acacacacct ctcaagacgg agatgcatgg cctctaagag tgcccgtgtc 12000ggttcttcct ggaagttgac tttccttaga cccgccaggt caagttagcc gcgtgacgga 12060catccaggcg tgggacgtgg tcagggcagg gctcattcat tgcccactag gatcccactg 12120gcgaagatgg tctccatatc agctctctgc agaagggagg aagactttat catgttccta 12180aaaatctgtg gcaagcaccc atcgtattat ccaaattttg ttgcaaatgt gattaatttg 12240gttgtcaagt tttgggggtg ggctgtgggg agattgcttt tgttttcctg ctggtaatat 12300cgggaaagat tttaatgaaa ccagggtaga attgtttggc aatgcactga agcgtgtttc 12360tttcccaaaa tgtgcctccc ttccgctgcg ggcccagctg agtctatgta ggtgatgttt 12420ccagctgcca agtgctcttt gttactgtcc accctcattt ctgccagcgc atgtgtcctt 12480tcaaggggaa aatgtgaagc tgaaccccct ccagacaccc agaatgtagc atctgagaag 12540gccctgtgcc ctaaaggaca cccctcgccc ccatcttcat ggagggggtc atttcagagc 12600cctcggagcc aatgaacagc tcctcctctt ggagctgaga tgagccccac gtggagctcg 12660ggacggatag tagacagcaa taactcggtg tgtggccgcc tggcaggtgg aacttcctcc 12720cgttgcgggg tggagtgagg ttagttctgt gtgtctggtg ggtggagtca ggcttctctt 12780gctacctgtg agcatccttc ccagcagaca tcctcatcgg gctttgtccc tcccccgctt 12840cctccctctg cggggaggac ccgggaccac agctgctggc cagggtagac ttggagctgt 12900cctccagagg ggtcacgtgt aggagtgaga agaaggaaga tcttgagagc tgctgaggga 12960ccttggagag ctcaggatgg ctcagacgag gacactcgct tgccgggcct gggcctcctg 13020ggaaggaggg agctgctcag aatgccgcat gacaactgaa ggcaacctgg aaggttcagg 13080ggccgctctt cccccatgtg cctgtcacgc tctggtgcag tcaaaggaac gccttcccct 13140cagttgtttc taagagcaga gtctcccgct gcaatctggg tggtaactgc cagccttgga 13200ggatcgtggc caacgtggac ctgcctacgg agggtgggct ctgacccaag tggggcctcc 13260ttgtccaggt ctcactgctt tgcaccgtgg tcagagggac tgtcagctga gcttgagctc 13320ccctggagcc agcagggctg tgatgggcga gtcccggagc cccacccaga cctgaatgct 13380tctgagagca aagggaagga ctgacgagag atgtatattt aattttttaa ctgctgcaaa 13440cattgtacat ccaaattaaa ggaaaaaaat ggaaaccatc a 13481320DNAArtificial SequencePrimer 3ctccgtccgg tagacatgct 20423DNAArtificial SequencePrimer 4ggaaatcaga accctcaaaa tgg 23529DNAArtificial SequenceProbe 5tgagcactgt tcaactgtgg atatcggga 29620DNAArtificial SequenceSynthetic Oligonucleotide 6tctctattgc acattccaag 20720DNAArtificial SequenceSynthetic Oligonucleotide 7gccgtagcct gggacccgcc 20819DNAArtificial SequenceSynthetic Oligonucleotide 8taacactcga ttaaccctg 19919DNAArtificial SequenceSynthetic Oligonucleotide 9taacacttga ttaaccctg 191019DNAArtificial SequenceSynthetic Oligonucleotide 10gttaacactc gattaaccc 191119DNAArtificial SequenceSynthetic Oligonucleotide 11gttaacactt gattaaccc 191219DNAArtificial SequenceSynthetic Oligonucleotide 12gctagttcat cccagtgag 191319DNAArtificial SequenceSynthetic Oligonucleotide 13gctagttcac cccagtgag 191419DNAArtificial SequenceSynthetic Oligonucleotide 14tggaaatggg tttttccac 191519DNAArtificial SequenceSynthetic Oligonucleotide 15tggaaatggc tttttccac 191619DNAArtificial SequenceSynthetic Oligonucleotide 16tttaaccgtg gcatgggca 191719DNAArtificial SequenceSynthetic Oligonucleotide 17tttaaccgta gcatgggca 191819DNAArtificial SequenceSynthetic Oligonucleotide 18ttcaagctag taacgatgc 191919DNAArtificial SequenceSynthetic Oligonucleotide 19ttcaagccag taacgatgc 192019DNAArtificial SequenceSynthetic Oligonucleotide 20acttcaagct agtaacgat 192119DNAArtificial SequenceSynthetic Oligonucleotide 21acttcaagcc agtaacgat 192219DNAArtificial SequenceSynthetic Oligonucleotide 22gcagctaggt taaagagtc 192319DNAArtificial SequenceSynthetic Oligonucleotide 23gcagctaggc taaagagtc 192419DNAArtificial SequenceSynthetic Oligonucleotide 24aataagaaac acaatcaaa 192519DNAArtificial SequenceSynthetic Oligonucleotide 25aataagaaat acaatcaaa 192619DNAArtificial SequenceSynthetic Oligonucleotide 26cagaggaggc atactgtat 192719DNAArtificial SequenceSynthetic Oligonucleotide 27cagaggaggt atactgtat 192819DNAArtificial SequenceSynthetic Oligonucleotide 28cacagtgcta cccaacctt 192919DNAArtificial SequenceSynthetic Oligonucleotide 29cacagtgctc cccaacctt 193019DNAArtificial SequenceSynthetic Oligonucleotide 30taattttcta gactttatg 193119DNAArtificial SequenceSynthetic Oligonucleotide 31taattttctg gactttatg 193219DNAArtificial SequenceSynthetic Oligonucleotide 32gctacaacgc aggtcaaat 193319DNAArtificial SequenceSynthetic Oligonucleotide 33gctacaatgc aggtcaaat 193419DNAArtificial SequenceSynthetic Oligonucleotide 34gagctacaac gcaggtcaa 193519DNAArtificial SequenceSynthetic Oligonucleotide 35gagctacaat gcaggtcaa 193619DNAArtificial SequenceSynthetic Oligonucleotide 36agagagaacg agaaggctc 193719DNAArtificial SequenceSynthetic Oligonucleotide 37agagagaaca agaaggctc 193819DNAArtificial SequenceSynthetic Oligonucleotide 38agcccctctg tgtaagttt 193919DNAArtificial SequenceSynthetic Oligonucleotide 39agcccttctg tgtaagttt 194019DNAArtificial SequenceSynthetic Oligonucleotide 40gagcccctct gtgtaagtt 194119DNAArtificial SequenceSynthetic Oligonucleotide 41gagcccttct gtgtaagtt 194219DNAArtificial SequenceSynthetic Oligonucleotide 42tgagcccctc tgtgtaagt 194319DNAArtificial SequenceSynthetic Oligonucleotide 43tgagcccttc tgtgtaagt 194419DNAArtificial SequenceSynthetic Oligonucleotide 44atgagcccct ctgtgtaag 194519DNAArtificial SequenceSynthetic Oligonucleotide 45atgagccctt ctgtgtaag 194619DNAArtificial SequenceSynthetic Oligonucleotide 46gatgagcccc tctgtgtaa 194719DNAArtificial SequenceSynthetic Oligonucleotide 47gatgagccct tctgtgtaa 194819DNAArtificial SequenceSynthetic Oligonucleotide 48tgatgagccc ctctgtgta 194919DNAArtificial SequenceSynthetic Oligonucleotide 49tgatgagccc ttctgtgta 195019DNAArtificial SequenceSynthetic Oligonucleotide 50atgatgagcc cctctgtgt 195119DNAArtificial SequenceSynthetic Oligonucleotide 51atgatgagcc cttctgtgt 195219DNAArtificial SequenceSynthetic Oligonucleotide 52taatgatgag cccctctgt 195319DNAArtificial SequenceSynthetic Oligonucleotide 53taatgatgag cccttctgt 195419DNAArtificial SequenceSynthetic Oligonucleotide 54agaatacggg taacatttt 195519DNAArtificial SequenceSynthetic Oligonucleotide 55agaatacagg taacatttt 195619DNAArtificial SequenceSynthetic Oligonucleotide 56ggagaatacg ggtaacatt 195719DNAArtificial SequenceSynthetic Oligonucleotide 57ggagaataca ggtaacatt 195819DNAArtificial SequenceSynthetic Oligonucleotide 58ttagtaatca attttaatg 195919DNAArtificial SequenceSynthetic Oligonucleotide 59ttagtaacca attttaatg 196019DNAArtificial SequenceSynthetic Oligonucleotide 60agttagtaat caattttaa 196119DNAArtificial SequenceSynthetic Oligonucleotide 61agttagtaac caattttaa 196219DNAArtificial SequenceSynthetic Oligonucleotide 62gaaggaatgc ttttactag 196319DNAArtificial SequenceSynthetic Oligonucleotide 63gaaggaatgt ttttactag 196419DNAArtificial SequenceSynthetic Oligonucleotide 64ctaaaactaa cttgagaat 196519DNAArtificial SequenceSynthetic Oligonucleotide 65ctaaaaccaa cttgagaat 196619DNAArtificial SequenceSynthetic Oligonucleotide 66atctaaaact aacttgaga 196719DNAArtificial SequenceSynthetic Oligonucleotide 67atctaaaacc aacttgaga 196819DNAArtificial SequenceSynthetic Oligonucleotide 68ggtgggcagg aaggactga 196919DNAArtificial SequenceSynthetic Oligonucleotide 69ggtgggcaga aaggactga 197019DNAArtificial SequenceSynthetic Oligonucleotide 70cctaaatcaa tctacaagt 197119DNAArtificial SequenceSynthetic Oligonucleotide 71cctaaattaa tctacaagt 197219DNAArtificial SequenceSynthetic Oligonucleotide 72tccctaaatc aatctacaa 197319DNAArtificial SequenceSynthetic Oligonucleotide 73tccctaaatt aatctacaa 197419DNAArtificial SequenceSynthetic Oligonucleotide 74gaaaatgtga gtggatcta 197519DNAArtificial SequenceSynthetic Oligonucleotide 75gaaaatgtgc gtggatcta 197619DNAArtificial SequenceSynthetic Oligonucleotide 76gtaaggcgag actgactag 197719DNAArtificial SequenceSynthetic Oligonucleotide 77gtaaggcaag actgactag 197819DNAArtificial SequenceSynthetic Oligonucleotide 78aggtaaggcg agactgact 197919DNAArtificial SequenceSynthetic Oligonucleotide 79aggtaaggca agactgact 198019DNAArtificial SequenceSynthetic Oligonucleotide 80ctgagcggag aaaccctcc 198119DNAArtificial SequenceSynthetic Oligonucleotide 81ctgagcgaag aaaccctcc 198219DNAArtificial SequenceSynthetic Oligonucleotide 82ggctgagcgg agaaaccct 198319DNAArtificial SequenceSynthetic Oligonucleotide 83ggctgagcga agaaaccct 198419DNAArtificial SequenceSynthetic Oligonucleotide 84aaggctgagc ggagaaacc 198519DNAArtificial SequenceSynthetic Oligonucleotide 85ttccctaaaa acaaaaaca 198619DNAArtificial SequenceSynthetic Oligonucleotide 86ttccctagaa acaaaaaca 198719DNAArtificial SequenceSynthetic Oligonucleotide 87gattccctaa aaacaaaaa 198819DNAArtificial SequenceSynthetic Oligonucleotide 88gattccctag aaacaaaaa 198919DNAArtificial SequenceSynthetic Oligonucleotide 89cttttctatt gtctgtccc

199019DNAArtificial SequenceSynthetic Oligonucleotide 90cttttctgtt gtctgtccc 199119DNAArtificial SequenceSynthetic Oligonucleotide 91tgcttttcta ttgtctgtc 199219DNAArtificial SequenceSynthetic Oligonucleotide 92tgcttttctg ttgtctgtc 199319DNAArtificial SequenceSynthetic Oligonucleotide 93aagggatgcc gacttgggc 199419DNAArtificial SequenceSynthetic Oligonucleotide 94aagggatgct gacttgggc 199519DNAArtificial SequenceSynthetic Oligonucleotide 95accttcctca ctgaggatg 199619DNAArtificial SequenceSynthetic Oligonucleotide 96accttcctcg ctgaggatg 199719DNAArtificial SequenceSynthetic Oligonucleotide 97caaaccactg tgggatgaa 199819DNAArtificial SequenceSynthetic Oligonucleotide 98caaaccactt tgggatgaa 199919DNAArtificial SequenceSynthetic Oligonucleotide 99aataaattgt catcaccag 1910019DNAArtificial SequenceSynthetic Oligonucleotide 100aataaattgc catcaccag 1910119DNAArtificial SequenceSynthetic Oligonucleotide 101tcacagctat cttctcatc 1910219DNAArtificial SequenceSynthetic Oligonucleotide 102tcacagctaa cttctcatc 1910319DNAArtificial SequenceSynthetic Oligonucleotide 103gcacacagta gatgaggga 1910419DNAArtificial SequenceSynthetic Oligonucleotide 104gcacacagtg gatgaggga 1910519DNAArtificial SequenceSynthetic Oligonucleotide 105cagaacaaag agaagaatt 1910619DNAArtificial SequenceSynthetic Oligonucleotide 106cagaacaaac agaagaatt 1910719DNAArtificial SequenceSynthetic Oligonucleotide 107gcttacatgc cttcagtga 1910819DNAArtificial SequenceSynthetic Oligonucleotide 108gcttacacgc cttcagtga 1910919DNAArtificial SequenceSynthetic Oligonucleotide 109cagcttacat gccttcagt 1911019DNAArtificial SequenceSynthetic Oligonucleotide 110cagcttacac gccttcagt 1911119DNAArtificial SequenceSynthetic Oligonucleotide 111aagaagcctg ataaaatct 1911219DNAArtificial SequenceSynthetic Oligonucleotide 112aagaagccta ataaaatct 1911319DNAArtificial SequenceSynthetic Oligonucleotide 113catacatcag ctcaaactg 1911419DNAArtificial SequenceSynthetic Oligonucleotide 114catacattag ctcaaactg 1911519DNAArtificial SequenceSynthetic Oligonucleotide 115cacatacatc agctcaaac 1911619DNAArtificial SequenceSynthetic Oligonucleotide 116cacatacatt agctcaaac 1911719DNAArtificial SequenceSynthetic Oligonucleotide 117gtcacataca tcagctcaa 1911819DNAArtificial SequenceSynthetic Oligonucleotide 118gagactatag cacccagat 1911919DNAArtificial SequenceSynthetic Oligonucleotide 119gagactataa cacccagat 1912019DNAArtificial SequenceSynthetic Oligonucleotide 120tagaggacgc cgtgcaggg 1912119DNAArtificial SequenceSynthetic Oligonucleotide 121tagaggatgc cgtgcaggg 1912219DNAArtificial SequenceSynthetic Oligonucleotide 122catagaggac gccgtgcag 1912319DNAArtificial SequenceSynthetic Oligonucleotide 123catagaggat gccgtgcag 1912419DNAArtificial SequenceSynthetic Oligonucleotide 124cacatagagg acgccgtgc 1912519DNAArtificial SequenceSynthetic Oligonucleotide 125acgtgtgtac agaacctgc 1912619DNAArtificial SequenceSynthetic Oligonucleotide 126acgtgtgtat agaacctgc 1912719DNAArtificial SequenceSynthetic Oligonucleotide 127tgttcagaat gcctcatct 1912819DNAArtificial SequenceSynthetic Oligonucleotide 128tgttcagaac gcctcatct 1912919DNAArtificial SequenceSynthetic Oligonucleotide 129aaacggcgca gcgggaagg 1913019DNAArtificial SequenceSynthetic Oligonucleotide 130aaacggcaca gcgggaagg 1913119DNAArtificial SequenceSynthetic Oligonucleotide 131agaaacggcg cagcgggaa 1913219DNAArtificial SequenceSynthetic Oligonucleotide 132agaaacggca cagcgggaa 1913319DNAArtificial SequenceSynthetic Oligonucleotide 133agggcgcaga cttccaaag 1913419DNAArtificial SequenceSynthetic Oligonucleotide 134agggcacaga cttccaaag 1913519DNAArtificial SequenceSynthetic Oligonucleotide 135aagggcgcag acttccaaa 1913619DNAArtificial SequenceSynthetic Oligonucleotide 136aagggcacag acttccaaa 1913719DNAArtificial SequenceSynthetic Oligonucleotide 137caagggcgca gacttccaa 1913819DNAArtificial SequenceSynthetic Oligonucleotide 138caagggcaca gacttccaa 1913919DNAArtificial SequenceSynthetic Oligonucleotide 139acaagggcgc agacttcca 1914019DNAArtificial SequenceSynthetic Oligonucleotide 140acaagggcac agacttcca 1914119DNAArtificial SequenceSynthetic Oligonucleotide 141cacaagggcg cagacttcc 1914219DNAArtificial SequenceSynthetic Oligonucleotide 142cacaagggca cagacttcc 1914319DNAArtificial SequenceSynthetic Oligonucleotide 143gcacaagggc gcagacttc 1914419DNAArtificial SequenceSynthetic Oligonucleotide 144gcacaagggc acagacttc 1914519DNAArtificial SequenceSynthetic Oligonucleotide 145ggcacaaggg cgcagactt 1914619DNAArtificial SequenceSynthetic Oligonucleotide 146ggcacaaggg cacagactt 1914719DNAArtificial SequenceSynthetic Oligonucleotide 147agggcacaag ggcgcagac 1914819DNAArtificial SequenceSynthetic Oligonucleotide 148agggcacaag ggcacagac 1914919DNAArtificial SequenceSynthetic Oligonucleotide 149gagcagctgc aacctggca 1915019DNAArtificial SequenceSynthetic Oligonucleotide 150gagcagctgt aacctggca 1915119DNAArtificial SequenceSynthetic Oligonucleotide 151tggtgccggg tgtctagca 1915219DNAArtificial SequenceSynthetic Oligonucleotide 152tggtgccagg tgtctagca 1915319DNAArtificial SequenceSynthetic Oligonucleotide 153aatggtgccg ggtgtctag 1915419DNAArtificial SequenceSynthetic Oligonucleotide 154aatggtgcca ggtgtctag 1915519DNAArtificial SequenceSynthetic Oligonucleotide 155ggggacaggg tgtgctctc 1915619DNAArtificial SequenceSynthetic Oligonucleotide 156ggggacaggt tgtgctctc 1915719DNAArtificial SequenceSynthetic Oligonucleotide 157gcttttcatt gaaaagaaa 1915819DNAArtificial SequenceSynthetic Oligonucleotide 158gcttttcgtt gaaaagaaa 1915919DNAArtificial SequenceSynthetic Oligonucleotide 159ctgcttttca ttgaaaaga 1916019DNAArtificial SequenceSynthetic Oligonucleotide 160ctgcttttcg ttgaaaaga 1916119DNAArtificial SequenceSynthetic Oligonucleotide 161actaggccgg gcatgctgg 1916219DNAArtificial SequenceSynthetic Oligonucleotide 162actaggctgg gcatgctgg 1916319DNAArtificial SequenceSynthetic Oligonucleotide 163agactaggcc gggcatgct 1916419DNAArtificial SequenceSynthetic Oligonucleotide 164agactaggct gggcatgct 1916519DNAArtificial SequenceSynthetic Oligonucleotide 165aaacagctgt tagttccca 1916619DNAArtificial SequenceSynthetic Oligonucleotide 166aaacagccgt tagttccca 1916719DNAArtificial SequenceSynthetic Oligonucleotide 167agaaacagct gttagttcc 1916819DNAArtificial SequenceSynthetic Oligonucleotide 168agaaacagcc gttagttcc 1916919DNAArtificial SequenceSynthetic Oligonucleotide 169gtgctaccca acctttctg 1917019DNAArtificial SequenceSynthetic Oligonucleotide 170agtgctaccc aacctttct 1917119DNAArtificial SequenceSynthetic Oligonucleotide 171cagtgctacc caacctttc 1917219DNAArtificial SequenceSynthetic Oligonucleotide 172acagtgctac ccaaccttt 1917318DNAArtificial SequenceSynthetic Oligonucleotide 173acagtgctac ccaacctt 1817417DNAArtificial SequenceSynthetic Oligonucleotide 174acagtgctac ccaacct 1717516DNAArtificial SequenceSynthetic Oligonucleotide 175cagtgctacc caacct 1617619DNAArtificial SequenceSynthetic Oligonucleotide 176tcacagtgct acccaacct 1917715DNAArtificial SequenceSynthetic Oligonucleotide 177cagtgctacc caacc 1517819DNAArtificial SequenceSynthetic Oligonucleotide 178atcacagtgc tacccaacc 1917919DNAArtificial SequenceSynthetic Oligonucleotide 179tatcacagtg ctacccaac 1918019DNAArtificial SequenceSynthetic Oligonucleotide 180atatcacagt gctacccaa 1918119DNAArtificial SequenceSynthetic Oligonucleotide 181atgctgactt gggccattc 1918219DNAArtificial SequenceSynthetic Oligonucleotide 182gatgctgact tgggccatt 1918319DNAArtificial SequenceSynthetic Oligonucleotide 183ggatgctgac ttgggccat 1918419DNAArtificial SequenceSynthetic Oligonucleotide 184gggatgctga cttgggcca 1918519DNAArtificial SequenceSynthetic Oligonucleotide 185agggatgctg acttgggcc 1918618DNAArtificial SequenceSynthetic Oligonucleotide 186agggatgctg acttgggc 1818717DNAArtificial SequenceSynthetic Oligonucleotide 187agggatgctg acttggg 1718819DNAArtificial SequenceSynthetic Oligonucleotide 188caagggatgc tgacttggg 1918915DNAArtificial SequenceSynthetic Oligonucleotide 189gggatgctga cttgg 1519019DNAArtificial SequenceSynthetic Oligonucleotide 190ccaagggatg ctgacttgg 1919119DNAArtificial SequenceSynthetic Oligonucleotide 191gccaagggat gctgacttg 1919219DNAArtificial SequenceSynthetic Oligonucleotide 192tgccaaggga tgctgactt 1919319DNAArtificial SequenceSynthetic Oligonucleotide 193ctgccaaggg atgctgact 1919419DNAArtificial SequenceSynthetic Oligonucleotide 194attgtcatca ccagaaaaa 1919519DNAArtificial SequenceSynthetic Oligonucleotide 195aattgtcatc accagaaaa 1919619DNAArtificial SequenceSynthetic Oligonucleotide 196aaattgtcat caccagaaa 1919719DNAArtificial SequenceSynthetic Oligonucleotide 197taaattgtca tcaccagaa 1919819DNAArtificial SequenceSynthetic Oligonucleotide 198ataaattgtc atcaccaga 1919918DNAArtificial SequenceSynthetic Oligonucleotide 199ataaattgtc atcaccag 1820017DNAArtificial SequenceSynthetic Oligonucleotide 200ataaattgtc atcacca 1720116DNAArtificial SequenceSynthetic Oligonucleotide 201taaattgtca tcacca 1620219DNAArtificial SequenceSynthetic Oligonucleotide 202taataaattg tcatcacca 1920315DNAArtificial SequenceSynthetic Oligonucleotide 203taaattgtca tcacc 1520419DNAArtificial SequenceSynthetic Oligonucleotide 204ttaataaatt gtcatcacc 1920519DNAArtificial SequenceSynthetic Oligonucleotide 205attaataaat tgtcatcac 1920619DNAArtificial SequenceSynthetic Oligonucleotide 206tattaataaa ttgtcatca 1920719DNAArtificial SequenceSynthetic Oligonucleotide 207ctattaataa attgtcatc 1920819DNAArtificial SequenceSynthetic Oligonucleotide 208cagtagatga gggagcagg 1920919DNAArtificial SequenceSynthetic Oligonucleotide 209acagtagatg agggagcag 1921019DNAArtificial SequenceSynthetic Oligonucleotide 210cacagtagat gagggagca 1921119DNAArtificial SequenceSynthetic Oligonucleotide 211acacagtaga tgagggagc 1921219DNAArtificial SequenceSynthetic Oligonucleotide 212cacacagtag atgagggag 1921318DNAArtificial SequenceSynthetic Oligonucleotide 213cacacagtag atgaggga 1821417DNAArtificial SequenceSynthetic Oligonucleotide 214cacacagtag atgaggg 1721516DNAArtificial SequenceSynthetic Oligonucleotide 215acacagtaga

tgaggg 1621619DNAArtificial SequenceSynthetic Oligonucleotide 216tgcacacagt agatgaggg 1921715DNAArtificial SequenceSynthetic Oligonucleotide 217acacagtaga tgagg 1521819DNAArtificial SequenceSynthetic Oligonucleotide 218gtgcacacag tagatgagg 1921919DNAArtificial SequenceSynthetic Oligonucleotide 219agtgcacaca gtagatgag 1922019DNAArtificial SequenceSynthetic Oligonucleotide 220aagtgcacac agtagatga 1922119DNAArtificial SequenceSynthetic Oligonucleotide 221gaagtgcaca cagtagatg 1922219DNAArtificial SequenceSynthetic Oligonucleotide 222gctgcaacct ggcaacaac 1922319DNAArtificial SequenceSynthetic Oligonucleotide 223agctgcaacc tggcaacaa 1922419DNAArtificial SequenceSynthetic Oligonucleotide 224cagctgcaac ctggcaaca 1922519DNAArtificial SequenceSynthetic Oligonucleotide 225gcagctgcaa cctggcaac 1922619DNAArtificial SequenceSynthetic Oligonucleotide 226agcagctgca acctggcaa 1922718DNAArtificial SequenceSynthetic Oligonucleotide 227agcagctgca acctggca 1822817DNAArtificial SequenceSynthetic Oligonucleotide 228agcagctgca acctggc 1722916DNAArtificial SequenceSynthetic Oligonucleotide 229gcagctgcaa cctggc 1623019DNAArtificial SequenceSynthetic Oligonucleotide 230agagcagctg caacctggc 1923115DNAArtificial SequenceSynthetic Oligonucleotide 231gcagctgcaa cctgg 1523219DNAArtificial SequenceSynthetic Oligonucleotide 232aagagcagct gcaacctgg 1923319DNAArtificial SequenceSynthetic Oligonucleotide 233caagagcagc tgcaacctg 1923419DNAArtificial SequenceSynthetic Oligonucleotide 234gcaagagcag ctgcaacct 1923519DNAArtificial SequenceSynthetic Oligonucleotide 235tgcaagagca gctgcaacc 1923619DNAArtificial SequenceSynthetic Oligonucleotide 236accatgatat ctccagcac 1923719DNAArtificial SequenceSynthetic Oligonucleotide 237accatgacat ctccagcac 1923819DNAArtificial SequenceSynthetic Oligonucleotide 238ccaccatgat atctccagc 1923919DNAArtificial SequenceSynthetic Oligonucleotide 239ccaccatgac atctccagc 1924019DNAArtificial SequenceSynthetic Oligonucleotide 240ttaacactcg attaaccct 1924119DNAArtificial SequenceSynthetic Oligonucleotide 241tagttcatcc cagtgagaa 1924219DNAArtificial SequenceSynthetic Oligonucleotide 242ctagttcatc ccagtgaga 1924319DNAArtificial SequenceSynthetic Oligonucleotide 243gaaatgggtt tttccacat 1924419DNAArtificial SequenceSynthetic Oligonucleotide 244ggaaatgggt ttttccaca 1924519DNAArtificial SequenceSynthetic Oligonucleotide 245taaccgtggc atgggcagt 1924619DNAArtificial SequenceSynthetic Oligonucleotide 246ttaaccgtgg catgggcag 1924719DNAArtificial SequenceSynthetic Oligonucleotide 247cttcaagcta gtaacgatg 1924819DNAArtificial SequenceSynthetic Oligonucleotide 248agctaggtta aagagtcac 1924919DNAArtificial SequenceSynthetic Oligonucleotide 249cagctaggtt aaagagtca 1925019DNAArtificial SequenceSynthetic Oligonucleotide 250taagaaacac aatcaaaga 1925119DNAArtificial SequenceSynthetic Oligonucleotide 251ataagaaaca caatcaaag 1925219DNAArtificial SequenceSynthetic Oligonucleotide 252attttctaga ctttatgat 1925319DNAArtificial SequenceSynthetic Oligonucleotide 253aattttctag actttatga 1925419DNAArtificial SequenceSynthetic Oligonucleotide 254gagaatacgg gtaacattt 1925519DNAArtificial SequenceSynthetic Oligonucleotide 255tgggcaggaa ggactgaac 1925619DNAArtificial SequenceSynthetic Oligonucleotide 256gtgggcagga aggactgaa 1925719DNAArtificial SequenceSynthetic Oligonucleotide 257ccctaaatca atctacaag 1925819DNAArtificial SequenceSynthetic Oligonucleotide 258cttttccgtg ctgttctga 1925919DNAArtificial SequenceSynthetic Oligonucleotide 259acttttccgt gctgttctg 1926019DNAArtificial SequenceSynthetic Oligonucleotide 260aacttttccg tgctgttct 1926119DNAArtificial SequenceSynthetic Oligonucleotide 261gctgagcgga gaaaccctc 1926219DNAArtificial SequenceSynthetic Oligonucleotide 262attccctaaa aacaaaaac 1926319DNAArtificial SequenceSynthetic Oligonucleotide 263gcttttctat tgtctgtcc 1926419DNAArtificial SequenceSynthetic Oligonucleotide 264cttcctcact gaggatgaa 1926519DNAArtificial SequenceSynthetic Oligonucleotide 265ccttcctcac tgaggatga 1926619DNAArtificial SequenceSynthetic Oligonucleotide 266aaccactttg ggatgaata 1926719DNAArtificial SequenceSynthetic Oligonucleotide 267aaaccacttt gggatgaat 1926819DNAArtificial SequenceSynthetic Oligonucleotide 268acagctatct tctcatcaa 1926919DNAArtificial SequenceSynthetic Oligonucleotide 269cacagctatc ttctcatca 1927019DNAArtificial SequenceSynthetic Oligonucleotide 270gaacaaagag aagaatttc 1927119DNAArtificial SequenceSynthetic Oligonucleotide 271agaacaaaga gaagaattt 1927219DNAArtificial SequenceSynthetic Oligonucleotide 272gaagcctgat aaaatctct 1927319DNAArtificial SequenceSynthetic Oligonucleotide 273agaagcctga taaaatctc 1927419DNAArtificial SequenceSynthetic Oligonucleotide 274tgatctgtag cagcagctt 1927519DNAArtificial SequenceSynthetic Oligonucleotide 275ttgatctgta gcagcagct 1927619DNAArtificial SequenceSynthetic Oligonucleotide 276gttgatctgt agcagcagc 1927719DNAArtificial SequenceSynthetic Oligonucleotide 277atagaggacg ccgtgcagg 1927819DNAArtificial SequenceSynthetic Oligonucleotide 278gtgtgtacag aacctgccg 1927919DNAArtificial SequenceSynthetic Oligonucleotide 279cgtgtgtaca gaacctgcc 1928019DNAArtificial SequenceSynthetic Oligonucleotide 280ttcagaatgc ctcatctgg 1928119DNAArtificial SequenceSynthetic Oligonucleotide 281gttcagaatg cctcatctg 1928219DNAArtificial SequenceSynthetic Oligonucleotide 282ggacagggtg tgctctccg 1928319DNAArtificial SequenceSynthetic Oligonucleotide 283gggacagggt gtgctctcc 1928416DNAArtificial SequenceSynthetic Oligonucleotide 284gggatgctga cttggg 1628517DNAArtificial SequenceSynthetic Oligonucleotide 285acacagtaga tgaggga 1728610081DNAMus musculus 286gcactcgccg cgagggttgc cgggacgggc ccaagatggc tgagcgcctt ggttccgctt 60ctgcctgccg cgcagagccc cattcattgc cttgctgcta agtggcgccg cgtagtgcca 120gtaggctcca agtcttcagg gtctgtccca tcgggcagga agccgtcatg gcaaccctgg 180aaaagctgat gaaggctttc gagtcgctca agtcgtttca gcagcaacag cagcagcagc 240caccgccgca ggcgccgccg ccaccgccgc cgccgcctcc gcctcaaccc cctcagccgc 300cgcctcaggg gcagccgccg ccgccaccac cgccgctgcc aggtccggca gaggaaccgc 360tgcaccgacc aaagaaggaa ctctcagcca ccaagaaaga ccgtgtgaat cattgtctaa 420caatatgtga aaacattgtg gcacagtctc tcagaaattc tccagaattt cagaaactct 480tgggcatcgc tatggaactg tttctgctgt gcagtgacga tgcggagtca gatgtcagaa 540tggtggctga tgagtgcctc aacaaagtca tcaaagcttt gatggattct aatcttccaa 600ggctacagtt agaactctat aaggaaatta aaaagaatgg tgctcctcga agtttgcgtg 660ctgccctgtg gaggtttgct gagctggctc acctggttcg acctcagaag tgcaggcctt 720acctggtgaa tcttcttcca tgcctgaccc gaacaagcaa aagaccggag gaatcagttc 780aggagacctt ggctgcagct gttcctaaaa ttatggcttc ttttggcaat ttcgcaaatg 840acaatgaaat taaggttctg ttgaaagctt tcatagcaaa tctgaagtca agctctccca 900ccgtgcggcg gacagcagcc ggctcagccg tgagcatctg ccaacattct aggaggacac 960agtacttcta caactggctc cttaatgtcc tcctaggtct gctggttccc atggaagaag 1020agcactccac tctcctgatc ctcggtgtgt tgctcacatt gaggtgtcta gtgcccttgc 1080tccagcagca ggtcaaggac acaagtctaa aaggcagctt tggggtgaca cggaaagaaa 1140tggaagtctc tccttctaca gagcagcttg tccaggttta tgaactgact ttgcatcata 1200ctcagcacca agaccacaat gtggtgacag gggcactgga gctcctgcag cagctcttcc 1260gtacccctcc acctgaactc ctgcaagcac tgaccacacc aggagggctt gggcagctca 1320ctctggttca agaagaggcc cggggccgag gccgcagcgg gagcatcgtg gagcttttag 1380ctggaggggg ttcctcgtgc agccctgtcc tctcaagaaa gcagaaaggc aaagtgctct 1440taggagagga agaagccttg gaagatgact cggagtccag gtcagatgtc agcagctcag 1500cctttgcagc ctctgtgaag agtgagattg gtggagagct cgctgcttct tcaggtgttt 1560ccactcctgg ttctgttggt cacgacatca tcactgagca gcctagatcc cagcacacac 1620ttcaagcaga ctctgtggat ttgtccggct gtgacctgac cagtgctgct actgatgggg 1680atgaggagga catcttgagc cacagctcca gccagttcag tgctgtccca tccgaccctg 1740ccatggacct gaatgatggg acccaggcct cctcacccat cagtgacagt tctcagacca 1800ccactgaagg acctgattca gctgtgactc cttcggacag ttctgaaatt gtgttagatg 1860gtgccgatag ccagtattta ggcatgcaga taggacagcc acaggaggac gatgaggagg 1920gagctgcagg tgttctttct ggtgaagtct cagatgtttt cagaaactct tctctggccc 1980ttcaacaggc acacttgttg gaaagaatgg gccatagcag gcagccttcc gacagcagta 2040tagataagta tgtaacaaga gatgaggttg ctgaagccag tgatccagaa agcaagcctt 2100gccgaatcaa aggtgacata ggacagccta atgatgatga ttctgctcct ctggtacatt 2160gtgtccgtct tttatctgct tcctttttgt taactggtga aaagaaagca ctggttccag 2220acagagacgt gagagtcagt gtgaaggccc tggccctcag ctgcattggt gcggctgtgg 2280cccttcatcc agagtcgttc ttcagcagac tgtacaaagt acctcttaat accacggaaa 2340gtactgagga acagtatgtt tctgacatct tgaactacat cgatcatgga gacccacagg 2400tccgaggagc tactgccatt ctctgtggga cccttgtcta ctccatcctc agtaggtccc 2460gtctccgtgt tggtgactgg ctgggcaaca tcagaaccct gacaggaaat acattttctc 2520tggtggactg cattccttta ctgcagaaaa cgttgaagga tgaatcttct gttacttgca 2580agttggcttg tacagctgtg aggcactgtg tcctgagtct ttgcagcagc agctacagtg 2640acttgggatt acaactgctt attgatatgc tgcctctgaa gaacagctcc tactggctgg 2700tgaggaccga actgctggac actctggcag agattgactt caggctcgtg agttttttgg 2760aggcaaaagc agaaagttta caccgagggg ctcatcatta tacagggttt ctaaaactac 2820aagaacgagt actcaataat gtggtcattt atttgcttgg agatgaagac cccagggttc 2880gacatgttgc tgcaacatca ttaacaaggc ttgtcccaaa gctgttttac aagtgtgacc 2940aaggacaagc tgatccagtt gtggctgtag cgagggatca gagcagtgtc tacctgaagc 3000tcctcatgca tgagacccag ccaccatcac acttttctgt cagcaccatc accagaatct 3060atagaggcta tagcttactg ccaagtataa cagatgtcac catggaaaac aatctctcaa 3120gagttgttgc cgcagtttct catgaactca ttacgtcaac aacacgggca ctcacatttg 3180gatgctgtga agccttgtgt cttctctcag cagcctttcc agtttgcact tggagtttag 3240gatggcactg tggagtgccc ccactgagtg cctctgatga gtccaggaag agctgcactg 3300ttgggatggc ctccatgatt ctcaccttgc tttcatcagc ttggttccca ctggatctct 3360cagcccatca ggatgccttg attttggctg gaaacttgct agcagcgagt gcccccaagt 3420ctctgagaag ttcatggacc tctgaagaag aagccaactc agcagccacc agacaggagg 3480aaatctggcc tgctctgggg gatcggactc tagtgccctt ggtggagcag cttttctccc 3540acctgctgaa ggtgatcaat atctgtgctc atgtcttgga cgatgtgact cctggaccag 3600caatcaaggc agccttgcct tctctaacaa accccccttc tctaagtcct attcgacgga 3660aagggaagga gaaagaacct ggagaacaag cttctactcc aatgagtccc aagaaagttg 3720gtgaggccag tgcagcctct cgacaatcag acacctcagg acctgtcaca gcaagtaaat 3780catcctcact ggggagtttc taccatctcc cctcctacct caaactgcat gatgtcctga 3840aagccactca cgccaactat aaggtcacct tagatcttca gaacagcact gaaaagtttg 3900gggggttcct gcgctctgcc ttggacgtcc tttctcagat tctagagctg gcgacactgc 3960aggacattgg aaagtgtgtt gaagaggtcc ttggatacct gaaatcctgc tttagtcgag 4020aaccaatgat ggcaactgtc tgtgtgcagc agctattgaa gactctcttt gggacaaact 4080tagcctcaca gtttgatggc ttatcttcca accccagcaa gtctcagtgc cgagctcagc 4140gccttggctc ttcaagtgtg aggcccggct tatatcacta ctgcttcatg gcaccataca 4200cgcacttcac acaggccttg gctgacgcaa gcctgaggaa catggtgcag gcggagcagg 4260agcgtgatgc ctcggggtgg tttgatgtac tccagaaagt gtctgcccaa ttgaagacga 4320acctaacaag cgtcacaaag aaccgtgcag ataagaatgc tattcataat cacattaggt 4380tatttgagcc tcttgttata aaagcattga agcagtacac cacgacaaca tctgtacaat 4440tgcagaagca ggttttggat ttgctggcac agctggttca gctacgggtc aattactgtc 4500tactggattc agaccaggtg ttcatcgggt ttgtgctgaa gcagtttgag tacattgaag 4560tgggccagtt cagggaatca gaggcaatta ttccaaatat atttttcttc ctggtattac 4620tgtcttatga gcgctaccat tcaaaacaga tcattggaat tcctaaaatc atccagctgt 4680gtgatggcat catggccagt ggaaggaagg ccgttacaca tgctatacct gctctgcagc 4740ccattgtcca tgacctcttt gtgttacgag gaacaaataa agctgatgca gggaaagagc 4800ttgagacaca gaaggaggtg gtggtctcca tgctgttacg actcatccag taccatcagg 4860tgctggagat gttcatcctt gtcctgcagc agtgccacaa ggagaatgag gacaagtgga 4920aacggctctc tcggcaggtc gcagacatca tcctgcccat gttggccaag cagcagatgc 4980atattgactc tcatgaagcc cttggagtgt taaatacctt gtttgagatt ttggctcctt 5040cctccctacg tcctgtggac atgcttttgc ggagtatgtt catcactcca agcacaatgg 5100catctgtaag cactgtgcag ctgtggatat ctggaatcct cgccattctg agggttctca 5160tttcccagtc aaccgaggac attgttcttt gtcgtattca ggagctctcc ttctctccac 5220acttgctctc ctgtccagtg attaacaggt taaggggtgg aggcggtaat gtaacactag 5280gagaatgcag cgaagggaaa caaaagagtt tgccagaaga tacattctca aggtttcttt 5340tacagctggt tggtattctt ctagaagaca tcgttacaaa acagctcaaa gtggacatga 5400gtgaacagca gcatacgttc tactgccaag agctaggcac actgctcatg tgtctgatcc 5460acatattcaa atctggaatg ttccggagaa tcacagcagc tgccactaga ctcttcacca 5520gtgatggctg tgaaggcagc ttctatactc tagagagcct gaatgcacgg gtccgatcca 5580tggtgcccac gcacccagcc ctggtactgc tctggtgtca gatcctactt ctcatcaacc 5640acactgacca ccggtggtgg gcagaggtgc agcagacacc caagagacac agtctgtcct 5700gcacgaagtc acttaacccc cagaagtctg gcgaagagga ggattctggc tcggcagctc 5760agctgggaat gtgcaataga gaaatagtgc gaagaggggc ccttattctc ttctgtgatt 5820atgtctgtca gaatctccat gactcagaac acttaacatg gctcattgtg aatcacattc 5880aagatctgat cagcttgtct catgagcctc cagtacaaga ctttattagt gccattcatc 5940gtaattctgc agctagtggt ctttttatcc aggcaattca gtctcgctgt gaaaatcttt 6000caacgccaac cactctgaag aaaacacttc agtgcttgga aggcatccat ctcagccagt 6060ctggtgctgt gctcacacta tatgtggaca ggctcctggg cacccccttc cgtgcgctgg 6120ctcgcatggt cgacaccctg gcctgtcgcc gggtagaaat gcttttggct gcaaatttac 6180agagcagcat ggcccagttg ccagaggagg aactaaacag aatccaagaa cacctccaga 6240acagtgggct tgcacaaaga caccaaaggc tctattcact gctggacaga ttccgactct 6300ctactgtgca ggactcactt agccccttgc ccccagtcac ttcccaccca ctggatgggg 6360atgggcacac atctctggaa acagtgagtc cagacaaaga ctggtacctc cagcttgtca 6420gatcccagtg ttggaccaga tcagattctg cactgctgga aggtgcagag ctggtcaacc 6480gtatccctgc tgaagatatg aatgacttca tgatgagctc ggagttcaac ctaagccttt 6540tggctccctg tttaagcctt ggcatgagcg agattgctaa

tggccaaaag agtcccctct 6600ttgaagcagc ccgtggggtg attctgaacc gggtgaccag tgttgttcag cagcttcctg 6660ctgtccatca agtcttccag cccttcctgc ctatagagcc cacggcctac tggaacaagt 6720tgaatgatct gcttggtgat accacatcat accagtctct gaccatactt gcccgtgccc 6780tggcacagta cctggtggtg ctctccaaag tgcctgctca tttgcacctt cctcctgaga 6840aggaggggga cacggtgaag tttgtggtaa tgacagttga ggccctgtca tggcatttga 6900tccatgagca gatcccactg agtctggacc tccaagccgg gctagactgc tgctgcctgg 6960cactacaggt gcctggcctc tggggggtgc tgtcctcccc agagtacgtg actcatgcct 7020gctccctcat ccattgtgtg cgattcatcc tggaagccat tgcagtacaa cctggagacc 7080agcttctcgg tcctgaaagc aggtcacata ctccaagagc tgtcagaaag gaggaagtag 7140actcagatat acaaaacctc agtcatgtca cttcggcctg cgagatggtg gcagacatgg 7200tggaatccct gcagtcagtg ctggccttgg gccacaagag gaacagcacc ctgccttcat 7260ttctcacagc tgtgctgaag aacattgtta tcagtctggc ccgactcccc ctagttaaca 7320gctatactcg tgtgcctcct ctggtatgga aactcgggtg gtcacccaag cctggagggg 7380attttggcac agtgtttcct gagatccctg tagagttcct ccaggagaag gagatcctca 7440aggagttcat ctaccgcatc aacaccctag ggtggaccaa tcgtacccag ttcgaagaaa 7500cttgggccac cctccttggt gtcctggtga ctcagcccct ggtgatggaa caggaagaga 7560gcccaccaga ggaagacaca gaaagaaccc agatccatgt cctggctgtg caggccatca 7620cctctctagt gctcagtgca atgaccgtgc ctgtggctgg caatccagct gtaagctgct 7680tggagcaaca gccccggaac aagccactga aggctctcga taccagattt ggaagaaagc 7740tgagcatgat cagagggatt gtagaacaag aaatccaaga gatggtttcc cagagagaga 7800atactgccac tcaccattct caccaggcgt gggatcctgt cccttctctg ttaccagcta 7860ctacaggtgc tcttatcagc catgacaagc tgctgctgca gatcaaccca gagcgggagc 7920caggcaacat gagctacaag ctgggccagg tgtccataca ctccgtgtgg ctgggaaata 7980acatcacacc cctgagagag gaggaatggg atgaggaaga agaggaagaa agtgatgtcc 8040ctgcaccaac gtcaccacct gtgtctccag tcaattccag aaaacaccgt gccggggttg 8100atattcactc ctgttcgcag tttctgcttg aattgtacag ccgatggatc ctgccatcca 8160gtgcagccag aaggaccccc gtcatcctga tcagtgaagt ggttcgatct cttcttgtag 8220tgtcagactt attcaccgaa cgtacccagt ttgaaatgat gtatctgacg ctgacagaac 8280tacggagagt gcacccttca gaagatgaga tcctcattca gtacctggtg cctgccacct 8340gtaaggcagc tgctgtcctt ggaatggaca aaactgtggc agagccagtc agccgcctac 8400tggagagcac actgaggagc agccacctgc ccagccagat cggagccctg cacggcatcc 8460tctatgtgtt ggagtgtgac ctcttggatg acactgcaaa gcagctcatt ccagttgtta 8520gtgactatct gctgtccaac ctcaaaggaa tagcccactg cgtgaacatt cacagccagc 8580agcatgtgct ggtaatgtgt gccactgctt tctacctgat ggaaaactac cctctggatg 8640tgggaccaga attttcagca tctgtgatac agatgtgtgg agtaatgctg tctggaagtg 8700aggagtccac cccctccatc atttaccact gtgccctccg gggtctggag cggctcctgc 8760tgtctgagca gctatctcgg ctagacacag agtccttggt caagctaagt gtggacagag 8820tgaatgtaca aagcccacac agggccatgg cagccctagg cctgatgctc acctgcatgt 8880acacaggaaa ggaaaaagcc agtccaggca gagcttctga ccccagccct gctacacctg 8940acagcgagtc tgtgattgta gctatggagc gagtgtctgt tctctttgat aggatccgca 9000agggatttcc ctgtgaagcc agggttgtgg caaggatcct gcctcagttc ctagatgact 9060tctttccacc tcaagatgtc atgaacaaag tcattggaga gttcctgtcc aatcagcagc 9120catacccaca gttcatggcc actgtagttt acaaggtttt tcagactctg cacagtgctg 9180ggcagtcatc catggtccgg gactgggtca tgctgtccct gtccaacttc acacaaagaa 9240ctccagttgc catggccatg tggagcctct cctgcttcct tgttagcgca tctaccagcc 9300catgggtttc tgcgatcctt ccacatgtca tcagcaggat gggcaaactg gaacaggtgg 9360atgtgaacct tttctgcctg gttgccacag acttctacag acaccagata gaggaggaat 9420tcgaccgcag ggctttccag tctgtgtttg aggtggtggc tgcaccagga agtccatacc 9480acaggctgct tgcttgtttg caaaatgttc acaaggtcac cacctgctga gtagtgcctg 9540tgggacaaaa ggctgaaaga aggcagctgc tggggcctga gcctccagga gcctgctcca 9600agcttctgct ggggctgcct tggccgtgca ggcttccact tgtgtcaagt ggacagccag 9660gcaatggcag gagtgctttg caatgagggc tatgcaggga acatgcacta tgttggggtt 9720gagcctgagt cctgggtcct ggcctcgctg cagctggtga cagtgctagg ttgaccaggt 9780gtttgtcttt ttcctagtgt tcccctggcc atagtcgcca ggttgcagct gccctggtat 9840gtggatcaga agtcctagct cttgccagat ggttctgagc ccgcctgctc cactgggctg 9900gagagctccc tcccacattt acccagtagg catacctgcc acaccagtgt ctggacacaa 9960aatgaatggt gtgtggggct gggaactggg gctgccaggt gtccagcacc attttccttt 10020ctgtgttttc ttctcaggag ttaaaattta attatatcag taaagagatt aattttaatg 10080t 1008128720DNAArtificial SequencePrimer 287attacagtct caccacgccc 2028820DNAArtificial SequencePrimer 288gacaagggaa gacccaagtg 20

Claims

1.-102. (canceled)

103. A compound comprising: a modified oligonucleotide consisting of 15 to 19 linked nucleosides and complementary to a differentiating polymorphism site, wherein the nucleoside at position 6, 7, 8, 9, 10, 11, 12, 13, or 14 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, aligns with the differentiating polymorphism; and wherein at least one of positions 2, 3, 6, 9, 10, 11, 13, or 14 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, comprises a high-affinity sugar modification.

104. The compound of claim 103, wherein the nucleoside that aligns with the differentiating polymorphism comprises a high-affinity sugar modification.

105. The compound of claim 103, wherein the nucleoside immediately adjacent to and at the 5'-side of the nucleoside that aligns with the differentiating polymorphism comprises a high-affinity sugar modification.

106. The compound of claim 103, wherein the nucleoside immediately adjacent to and at the 3'-side of the nucleoside that aligns with the differentiating polymorphism comprises a high-affinity sugar modification.

107. The compound of claim 106, wherein the modified oligonucleotide is 100% complementary to the single nucleotide polymorphism site.

108. The compound of claim 106, wherein the high-affinity sugar modification is a bicyclic sugar.

109. The compound of claim 108, wherein the bicyclic sugar comprises a 4'-CH(CH.sub.3)--O-2' bridge.

110. The compound of claim 108, wherein the bicyclic sugar comprises a 4'-(CH.sub.2)--O-2' bridge.

111. The compound of claim 106, wherein the high-affinity sugar modification is a 2'-O-methoxyethyl nucleoside.

112. The compound of claim 106, wherein at least one of positions 2, 3, 13, and 14 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, comprises a high-affinity sugar modification.

113. The compound of claim 106, wherein each of nucleosides at positions 2, and 13 of the modified oligonucleotide, as counted from the 5' terminus of the modified oligonucleotide, comprises a high-affinity sugar modification.

114. The compound of claim 112, wherein the high-affinity sugar modification is a bicyclic sugar.

115. The compound of claim 114, wherein the bicyclic sugar comprises a 4'-CH(CH.sub.3)--O-2' bridge.

116. The compound of claim 113, wherein the high-affinity sugar modification is a bicyclic sugar.

117. The compound of claim 116, wherein the bicyclic sugar comprises a 4'-CH(CH.sub.3)--O-2' bridge.

118. The compound of claim 106, wherein at least one internucleoside linkage is a modified internucleoside linkage.

119. The compound of claim 106, wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.

120. The compound of claim 113, wherein at least one internucleoside linkage is a modified internucleoside linkage.

121. The compound of claim 113, wherein each internucleoside linkage is a phosphorothioate internucleoside linkage

122. The compound of claim 106, comprising a pharmaceutically acceptable carrier or diluent.