Detection of susceptibility to autoimmune diseases

Abstract

The present invention provides methods and reagents for determining sequence variants present at the IL4 receptor, IL-4 and IL-13 loci, which facilitate identifying individuals at risk for type 1 diabetes.

Description

[0001] The present patent application is a continuation of U.S. patent application Ser. No. 10/267,844 filed Oct. 8, 2002, which is a continuation-in-part of U.S. patent application Ser. No. 10/264,965 filed Oct. 4, 2002, which is a continuation-in-art of U.S. patent application Ser. No. 10/189,956 filed Jul. 3, 2002, which claims priority to U.S. provisional patent application Ser. No. 60/306,912 filed Jul. 20, 2001, all of which are herein incorporated by reference in their entireties.

1. FIELD OF THE INVENTION

[0002] The present invention relates to the fields of immunology and molecular biology. In particular, it relates to methods and reagents for detecting an individual's risk for autoimmune diseases. More specifically, it relates to methods and reagents for detecting an individual's increased or decreased risk for type 1 diabetes.

2. DESCRIPTION OF RELATED ART

[0003] The immunological response to an antigen is mediated through the selective differentiation of CD4+ T helper precursor cells (Th0) to T helper type 1 (Th1) or T helper type 2 (Th2) effector cells, with functionally distinct patterns of cytokine (also described as lymphokine) secretion. Th1 cells secrete interleukin 2 (IL-2), IL-12, tumor necrosis factor (TNF), lymphotoxin (LT), and interferon gamma (IFN-.gamma.) upon activation, and are primarily responsible for cell-mediated immunity such as delayed-type hypersensitivity. Th2 cells secrete IL-4, IL-5, IL-6, IL-9, and IL-1 3 upon activation, and are primarily responsible for extracellular defense mechanisms. The role of Th1 and Th2 cells is reviewed in Peltz, 1991, Immunological Reviews, 123: 23-35, incorporated herein by reference.

[0004] IL4 and IL13 play a central role in IgE-dependent inflammatory reactions. IL4 induces IgE antibody production by B Cells and further provides a regulatory function in the differentiation of Th0 to Th1 or Th2 effector cells by both promoting differentiation into Th2 cells and inhibiting differentiation into Th1 cells. IL13 also induces IgE antibody production by B Cells.

[0005] IL4 and IL13 operate through the IL4 receptor ("IL4R"), found on both B and T cells, and the IL13R, found on B cells, respectively. The human IL4 receptor (IL4R) is a heterodimer comprising the IL4R .alpha. chain and the IL2 receptor .gamma. chain. The .alpha.-chain of the IL4 receptor also serves as the .alpha.-chain of the IL13 receptor. IL4 binds to both IL4R and IL13R through the IL4R .alpha.-chain and can activate both B and T cells, whereas IL13 binds only to IL13R through the IL13R .alpha.1 chain and activates only T cells.

3. SUMMARY OF INVENTION

[0006] The present invention provides methods for detecting an individual's increased or decreased risk for an autoimmune disease such as type 1 diabetes, also known as insulin-dependent diabetes mellitus ("IDDM"). The present invention also provides kits, reagents and arrays useful for detecting an individual's risk for autoimmune diseases such as type 1 diabetes.

[0007] In one aspect, the present invention provides a method for detecting an individual's increased or decreased risk for an autoimmune disease such as type 1 diabetes by detecting the presence of a type 1 diabetes-associated polymorphism in the IL4R, IL4 or IL13 loci in a nucleic acid sample of the individual, wherein the presence of said polymorphism indicates the individual's increased risk for type 1 diabetes.

[0008] In one aspect, the present invention provides a method for detecting an individual's increased or decreased risk for an autoimmune disease such as type 1 diabetes by detecting the presence of a type 1 diabetes-associated polymorphism in the IL4R, IL4 or IL13 loci in a nucleic acid sample of the individual, wherein the presence of said polymorphism indicates the individual's increased risk for type 1 diabetes.

[0009] In one embodiment, the polymorphism is an IL4R polymorphism. In another embodiment, the polymorphism is an IL4 polymorphism. In another embodiment, the polymorphism is an IL13 polymorphism. In another embodiment, an IL4R polymorphism and an IL4 polymorphism are detected. In another embodiment, an IL4R polymorphism and an IL13 polymorphism are detected. In another embodiment, an IL4 polymorphism and an IL13 polymorphism are detected. In another embodiment, an IL4R polymorphism, an IL4 polymorphism and an IL13 polymorphism are detected.

[0010] In another embodiment, the IL4R polymorphism is selected from the IL4R polymorphisms listed in Table 21. In another embodiment, the IL4 polymorphism is the the IL4(-524) polymorphisms listed in Table 21. In another embodiment, the IL13 polymorphism is selected from the IL4R polymorphisms listed in Table 21. In another embodiment, 2 or more IL4R polymorphisms selected from the IL4R polymorphisms listed in Table 21 are detected. In another embodiment, 6 or more IL4R polymorphisms selected from the IL4R polymorphisms listed in Table 21 are detected. In another embodiment, 7 or more IL4R polymorphisms selected from the IL4R polymorphisms listed in Table 21 are detected. In another embodiment, 8 or more IL4R polymorphisms selected from the IL4R polymorphisms listed in Table 21 are detected. In another embodiment, all 10 IL4R polymorphisms listed in Table 21 are detected.

[0011] The individual can belong to any race or population. In one embodiment, the individual is an Asian, preferably a Filipino, or a Caucasian.

[0012] The nucleic acid sample can be obtained from any part of the individual's body, including, but not limited to hair, skin, nails, tissues or bodily fluids such as saliva, blood, etc. The nucleic acid sample can, but need not, be amplified by any amplification method including, but not limited to, polymerase chain reaction ("TCR").

[0013] The polymorphism can be any predisposing or protective polymorphism in the IL4R, IL4 or IL13 loci. In one embodiment of the invention, the polymorphism can be any polymorphism identified as predisposing or protective by methods taught herein. In one embodiment, the polymorphism can be a single nucleotide polymorphism ("SNP") in the IL-4 receptor ("IL4-R"), IL4 or IL13 loci. In another embodiment, specific haplotypes in the IL4R, IL4 and IL13 loci as well as specific combinations of, and interactions between, SNPs at these loci can be indicative of an increased or a decreased risk to an autoimmune disease such as type 1 diabetes.

[0014] The polymorphism can be detected by any method known in the art for detecting the presence of a specific polymorphism in a nucleic acid sample. These methods include, but are not limited to, contacting the nucleic acid sample with one or more nucleic acid molecules that hybridize under stringent hybridization conditions to at least one type 1 diabetes-associated IL4R, IL4 or IL13 polymorphism and detecting the hybridization, detection by amplification of the nucleic acid sample by, for example, PCR, and by direct sequencing of the nucleic acid sample.

[0015] Another aspect of the invention relates to a kit useful for detecting the presence of a predisposing or a protective polymorphism in the IL4R, IL4 or IL13 loci in a nucleic acid sample of an individual whose risk for type 1 diabetes is being assessed. The kit can comprise one or more oligonucleotides capable of detecting a predisposing or protective polymorphism in the IL4R, IL4 or IL13 loci as well as instructions for using the kit to detect susceptibility for an autoimmune disease such as type 1 diabetes. In preferred embodiments, the oligonucleotide or oligonucleotides each individually comprise a sequence that hybridizes under stringent hybridization conditions to at least one type 1 diabetes-associated IL4R, IL4 or IL13 polymorphism. In some embodiments, the oligonucleotide or oligonucleotides each individually comprise a sequence that is fully complementary to a nucleic acid sequence comprising a type 1 diabetes-associated IL4R, IL4 or IL13 polymorphism.

[0016] In some embodiments, the oligonucleotide can be used to detect the presence of a type 1 diabetes-associated IL4R, IL4 or IL13 polymorphism by hybridizing to the polymorphism under stringent hybridizing conditions. In some embodiments, the oligonucleotide can be used as an extension primer in either an amplification reaction such as PCR or a sequencing reaction, wherein the type 1 diabetes-associated IL4R, IL4 or IL13 polymorphism is detected either by amplification or sequencing.

[0017] In certain embodiments, the kit can further comprise amplification or sequencing primers which can, but need not, be sequence-specific. The kit can also comprise reagents for labeling one or more of the oligonucleotides, or comprise labeled oligonucleotides. Optionally, the kit can comprise reagents to detect the label.

[0018] In some embodiments, the kit can comprise one or more oligonucleotides that can be used to detect the presence of two or more predisposing or protective IL4R, IL4 or IL 13 polymorphisms or combinations of predisposing polymorphisms, protective polymorphisms or both.

[0019] In another aspect, the invention provides an array useful for detecting the presence of a predisposing or a protective IL4R, IL4 or IL13 polymorphism in a nucleic acid sample of an individual whose risk for type 1 diabetes is being assessed. The array can comprise one or more oligonucleotides capable of detecting a predisposing or protective IL4R, IL4 or IL13 polymorphism. The oligonucleotides can be immobilized on a substrate, e.g., a membrane or glass. In preferred embodiments, the oligonucleotide or oligonucleotides each individually comprise a sequence that can hybridize under stringent hybridization conditions to a nucleic acid sequence comprising a type 1 diabetes-associated IL4R, IL4 or IL13 polymorphism. In some embodiments, the oligonucleotide or oligonucleotides each individually comprise a sequence that is fully complementary to a nucleic acid sequence comprising a type 1 diabetes-associated IL4R, IL4 or IL13 polymorphism. The oligonucleotide or oligonucleotides can, but need not, be labeled. In some embodiments, the array can be a microarray.

[0020] In some embodiments, the array can comprise one or more oligonucleotides used to detect the presence of two or more predisposing or protective IL4R, IL4 or IL13 polymorphisms or combinations of predisposing polymorphisms, protective polymorphisms or both.

[0021] In certain embodiments, an individual's risk for particular Th1-mediated diseases is diagnosed from the individual's IL4R, IL4 or IL13 genotype. In a preferred embodiment, the Th1-mediated disease is type 1 diabetes. An individual who has at least one polymorphism statistically associated with type 1 diabetes possesses a factor contributing to either an increased or a decreased risk of a type 1 diabetes as compared to an individual without the polymorphism. The statistical association of IL4R, IL4 or IL13 polymorphisms (sequence variants) is shown in the examples.

[0022] The genotype can be determined using any method capable of identifying nucleotide variation consisting of single nucleotide polymorphic sites. The particular method used is not a critical aspect of the invention. A number of suitable methods are described below.

[0023] In one embodiment of the invention, genotyping is carried out using oligonucleotide probes specific to variant IL4R, IL4 or IL13 sequences. Preferably, a region of the IL4R, IL4 or IL13 genes which encompasses one or several polymorphic sites of interest is amplified prior to, or concurrent with, the hybridization of probes directed to such sites. Probe-based assays for the detection of sequence variants are well known in the art.

[0024] Alternatively, genotyping is carried out using allele-specific amplification or extension reactions, wherein allele-specific primers are used which support primer extension only if the targeted allele is present. Typically, an allele-specific primer hybridizes to the IL4R, IL4 or IL13 genes such that the 3' terminal nucleotide aligns with a polymorphic position. Allele-specific amplification reactions and allele-specific extension reactions are well known in the art.

4. BRIEF DESCRIPTION OF THE FIGURES

[0025] FIG. 1 provides a schematic of a molecular haplotyping method;

[0026] FIG. 2 provides an illustration of epistasis between the IL4R SNPs and IL4 and IL13 SNPs.

5. BRIEF DESCRIPTION OF THE TABLES

[0027] Table 1 provides the nucleotide sequence of the coding region of an IL4R (SEQ ID NO: 2);

[0028] Table 2 provides IL4R, IL4 and IL13 SNPs useful in the methods of the invention;

[0029] Table 3 provides probes used to identify IL4R polymorphisms (SEQ ID NO: 3-19);

[0030] Table 4 provides computationally estimated haplotype frequencies compared between Filipino controls and diabetics (SEQ ID NO: 20-24);

[0031] Table 5 provides genotypes of affected and nonaffected individuals;

[0032] Table 6 provides single nucleotide polymorphisms detected;

[0033] Table 7 provides amplicon primers and lengths (SEQ ID NO: 25-36);

[0034] Table 8 provides hybridization probes and titers (SEQ ID NO: 37-53);

[0035] Table 9 provides allele frequency of wildtype allele in HBDI founders;

[0036] Table 10 provides D' and A values for pairs of IL4R SNPs;

[0037] Table 11A provides results of single locus TDT analysis;

[0038] Table 11B provides results of single locus TDT analysis;

[0039] Table 12 provides allele-specific PCR primers (SEQ ID NO: 54-62);

[0040] Table 13 provides IBD distributions for IL4R haplotypes;

[0041] Table 14A provides haplotype transmissions;

[0042] Table 14B provides haplotype transmissions;

[0043] Table 14C provides haplotype transmissions;

[0044] Table 15A provides SNP by SNP allele transmissions;

[0045] Table 15B provides SNP by SNP allele transmissions;

[0046] Table 16A provides a TDT analysis;

[0047] Table 16B provides a TDT analysis;

[0048] Table 16C provides a TDT analysis;

[0049] Table 17A provides a TDT analysis;

[0050] Table 17B provides a TDT analysis;

[0051] Table 18 provides allele frequencies in Filipino controls and diabetics;

[0052] Table 19 provides estimated haplotype frequencies;

[0053] Table 20 provides observed haplotype frequencies;

[0054] Table 21 provides allele frequencies in diabetics and controls;

[0055] Table 22 provides pairwise linkage disequilibrium values for IL4R SNPs;

[0056] Table 23 provides pairwise linkage disequilibrium values for IL4 and IL13 SNPs;

[0057] Table 24 provides genotype frequencies in patients and controls;

[0058] Table 25A provides IL-4R 7-SNP Haplotypes in Filipino diabetics and controls;

[0059] Table 25B provides estimated IL4R 10-SNP haplotype frequencies in diabetics and controls;

[0060] Table 26 provides estimated IL4 and IL13 5-SNP haplotype frequencies in diabetics and controls;

[0061] Table 27 provides correlation between genotype frequencies at IL4R SNPs and five IL4 and IL13 SNPs;

[0062] Table 28 provides epistatic interaction between IL4R SNPs and five IL4 and IL13 SNPs;

[0063] Table 29 provides probes used to identify IL4 and IL13 polymorphisms (SEQ ID NO: 63-68);

[0064] Table 30 provides amplicon primers and lengths for an IL4 promoter and IL13 SNPs (SEQ ID NO: 69-74);

[0065] Table 31 provides amplicon primers and lengths for IL4R promoter SNPs (SEQ ID NO: 75-80);

[0066] Table 32 provides amplicon primers and lengths for IL13 promoter SNPs (SEQ ID NO: 81-86);

6. DETAILED DESCRIPTION OF THE INVENTION

[0067] The present invention provides methods, reagents and kits for detecting an individual's increased or decreased risk for an autoimmune disease. Examples of autoimmune diseases include, but are not limited to, multiple sclerosis, myasthenia gravis, Crohn's disease, ulcerative colitis, primary biliary cirrhosis, type 1 diabetes mellitus (insulin dependent diabetes mellitus or IDDM), Grave's disease, autoimmune hemolytic anemia, pernicious anemia, autoimmune thrombocytopenia, vasculitides such as Wegener's granulomatosis, Behcet's disease, rheumatoid arthritis, systemic lupus erythematosus (lupus), scleroderma, systemic sclerosis, Guillain-Barre syndromes, Hashimoto's thyroiditis spondyloarthropathies such as ankylosing spondylitis, psoriasis, dermatitis herpetiformis, inflammatory bowel diseases, pemphigus vulgaris and vitiligo. In certain preferred embodiments, the autoimmune disease is type 1 diabetes.

[0068] Abbreviations and Terminology:

[0069] The term "IL4R gene" or "IL4R locus" refers to the genomic nucleic acid sequence that encodes the alpha sub-unit of the interleukin 4 receptor protein. The nucleotide sequence of a gene, as used herein, encompasses coding regions, referred to as exons, intervening, non-coding regions, referred to as introns, and upstream or downstream regions. Upstream or downstream regions can include regions of the gene that are transcribed but not part of an intron or exon, or regions of the gene that comprise, for example, binding sites for factors that modulate gene transcription. The gene sequence of a Human mRNA for IL4R is provided at GenBank accession number X52425.1 (SEQ ID NO: 1). The coding region is provided as SEQ ID NO: 2. The genomic sequence for the IL4R gene is included in GenBank accession number AC004525.1 (SEQ ID NO: 88).

[0070] The term "IL4 gene" or "IL4 locus" refers to the genomic nucleic acid sequence that encodes the interleukin 4 protein. The nucleotide sequence of a gene, as used herein, encompasses coding regions, referred to as exons, intervening, non-coding regions, referred to as introns, and upstream or downstream regions. Upstream or downstream regions can include regions of the gene that are transcribed but not part of an intron or exon, or regions of the gene that comprise, for example, binding sites for factors that modulate gene transcription. The genomic sequence for the IL4 gene is provided at GenBank accession number M23442.1 (SEQ ID NO: 89).

[0071] The term "IL13 gene" or "IL13 locus" refers to the genomic nucleic acid sequence that encodes the interleukin 13 protein. The nucleotide sequence of a gene, as used herein, encompasses coding regions, referred to as exons, intervening, non-coding regions, referred to as introns, and upstream or downstream regions. Upstream or downstream regions can include regions of the gene that are transcribed but not part of an intron or exon, or regions of the gene that comprise, for example, binding sites for factors that modulate gene transcription. The genomic sequence for the IL13 gene is provided at GenBank accession number U10307.1 (SEQ ID NO: 90).

[0072] The term "allele", as used herein, refers to a sequence variant of the gene. Alleles are identified with respect to one or more polymorphic positions, with the rest of the gene sequence unspecified. For example, an IL4R allele may be defined by the nucleotide present at a single SNP, or by the nucleotides present at a plurality of SNPs. In certain embodiments of the invention, an IL4R is defined by the genotypes of 6, 7, 8 or 10 IL4R SNPs. Examples of such IL4R SNPs are provided in Table 2, below.

[0073] For convenience, the allele present at the higher or highest frequency in the population will be referred to as the wild-type allele; less frequent allele(s) will be referred to as mutant-allele(s). This designation of an allele as a mutant is meant solely to distinguish the allele from the wild-type allele and is not meant to indicate a change or loss of function.

[0074] The term "predisposing polymorphism" refers to a polymorphism that is positively associated with an autoimmune disease such as type 1 diabetes. The presence of a predisposing polymorphism in an individual could be indicative that the individual has an increased risk for the disease relative to an individual without the polymorphism.

[0075] The term "protective polymorphism" refers to a polymorphism that is negatively associated with an autoimmune disease such as type 1 diabetes. The presence of a protective polymorphism in an individual could be indicative that the individual has a decreased risk for the disease relative to an individual without the polymorphism.

[0076] The terms "polymorphic" and "polymorphism", as used herein, refer to the condition in which two or more variants of a specific genomic sequence, or the encoded amino acid sequence, can be found in a population. The terms refer either to the nucleic acid sequence or the encoded amino acid sequence; the use will be clear from the context. The polymorphic region or polymorphic site refers to a region of the nucleic acid where the nucleotide difference that distinguishes the variants occurs, or, for amino acid sequences, a region of the amino acid where the amino acid difference that distinguishes the protein variants occurs. As used herein, a "single nucleotide polymorphism", or SNP, refers to a polymorphic site consisting of a single nucleotide position.

[0077] "Odds Ratio" ("OR") refers to the ratio of the odds of the disease for individuals with the marker (allele or polymorphism) relative to the odds of the disease in individuals without the marker (allele or polymorphism).

[0078] "Linkage Disequilibrium" ("LD") refers to alleles at different loci that are not associated at random, i.e., not associated in proportion to their frequencies. If the alleles are in positive linkage disequilibrium, then the alleles occur together more often than expected assuming statistical independence. Conversely, if the alleles are in negative linkage disequilibrium, then the alleles occur together less often than expected assuming statistical independence.

[0079] The term "genotype" refers to a description of the alleles of a gene or genes contained in an individual or a sample. As used herein, no distinction is made between the genotype of an individual and the genotype of a sample originating from the individual. Although, typically, a genotype is determined from samples of diploid cells, a genotype can be determined from a sample of haploid cells, such as a sperm cell.

[0080] The term "haplotype" refers to a description of the variants of a gene or genes contained on a single chromosome, i.e., the genotype of a single chromosome. A haplotype is a set of maternally inherited alleles, or a set of paternally inherited alleles, at any locus.

[0081] The term "target region" refers to a region of a nucleic acid which is to be analyzed and usually includes at least one polymorphic region.

[0082] Individual amino acids in a sequence are represented herein as AN or NA, wherein A is the amino acid in the sequence and N is the position in the sequence. In the case that position N is polymorphic, it is convenient to designate the more frequent variant as A.sub.1N and the less frequent variant as NA.sub.2. Alternatively, the polymorphic site, N, is represented as A.sub.1NA.sub.2, wherein A.sub.1 is the amino acid in the more common variant and A.sub.2 is the amino acid in the less common variant. Either the one-letter or three-letter codes are used for designating amino acids (see Lehninger, BioChemistry 2nd ed., 1975, Worth Publishers, Inc. New York, N.Y.: pages 73-75, incorporated herein by reference). For example, 150V represents a single-amino-acid polymorphism at amino acid position 50, wherein isoleucine is the present in the more frequent protein variant in the population and valine is present in the less frequent variant. The amino acid positions are numbered based on the sequence of the mature IL4R protein, as described below.

[0083] "Stringent" as used herein refers to hybridization and wash conditions at 50.degree. C. or higher. Other stringent hybridization conditions may also be selected. Generally, stringent conditions are selected to be about 5.degree. C. lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength and pH) at which 50% of the target sequence hybridizes to a perfectly matched probe. Typically, stringent conditions will be those in which the salt concentration is at least about 0.02 molar at pH 7 and the temperature is at least about 50.degree. C. As other factors may significantly affect the stringency of hybridization, including, among others, base composition, length of the nucleic acid strands, the presence of organic solvents, the extent of base mismatching, the combination of parameters is more important than the absolute measure of any one.

[0084] Representations of nucleotides and single nucleotide changes in DNA sequences are analogous. For example, A398G represents a single nucleotide polymorphism at nucleotide position 398, wherein adenine is the present in the more frequent (wild-type) allele in the population and guanine is present in the less frequent (mutant) allele. The nucleotide positions are numbered based on the IL4R coding region sequence provided as SEQ ID NO:2, shown below. It will be clear that in a double stranded form, the complementary strand of each allele will contain the complementary base at the polymorphic position.

[0085] Conventional techniques of molecular biology and nucleic acid chemistry, which are within the skill of the art, are fully explained in the literature. See, for example, Sambrook et al., 1989, Molecular Cloning--A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.; Oligonucleotide Synthesis (M. J. Gait, ed., 1984); Nucleic Acid Hybridization (B. D. Hames and S. J. Higgins. eds., 1984); the series, Methods in Enzymology (Academic Press, Inc.); and the series, Current Protocols in Human Genetics (Dracopoli et al., eds., 1984 with quarterly updates, John Wiley & Sons, Inc.); all of which are incorporated herein by reference. All patents, patent applications, and publications mentioned herein, both supra and infra, are incorporated herein by reference.

[0086] Association with Type 1 Diabetes

[0087] As IL4R, IL4 or IL13 are a small component of the complex system of genes involved in an immune response, the effect of the IL4R, IL4 or IL13 loci is expected to be small. Other factors, such as an individual's HLA genotype, may exert dominating effects which, in some cases, may mask the effect of the IL4R, IL4 or IL13 genotypes. For example, particular HLA genotypes are known to have a major effect on the likelihood of type 1 diabetes (see Noble et al., 1996, Am. J. Hum. Genet, 59:1134-1148, incorporated herein by reference). The IL4R, IL4 or IL13 genotypes are likely to be more informative as an indicator of predisposition towards type 1 diabetes among individuals who have HLA genotypes that confer neither increased nor decreased risk. Furthermore, because allele frequencies at other loci relevant to immune system-related diseases differ between populations and, thus, populations exhibit different risks for immune system-related diseases, it is expected that the effect of the IL4R, IL4 or IL13 genotypes may be of different magnitude in some populations. Although the contribution of the IL4R, IL4 or IL13 genotypes may be relatively minor by itself, genotyping at the IL4R, IL4 or IL13 loci will contribute information that is, nevertheless, useful for a characterization of an individual's predisposition towards type 1 diabetes. The IL4R, IL4 or IL13 genotype information may be particularly useful when combined with genotype information from other loci.

[0088] Methods for Detecting Risk for Autoimmune Diseases

[0089] The present invention provides methods of determining an individual's risk for any autoimmune disease or condition or any Th-1 mediated disease. Such diseases or conditions include, but are not limited to, multiple sclerosis, myasthenia gravis, Crohn's disease, ulcerative colitis, primary biliary cirrhosis, type 1 diabetes mellitus (insulin dependent diabetes mellitus or IDDM), Grave's disease, autoimmune hemolytic anemia, pernicious anemia, autoimmune thrombocytopenia, vasculitides such as Wegener's granulomatosis, Behcet's disease, rheumatoid arthritis, systemic lupus erythematosus (lupus), scleroderma, systemic sclerosis, Gullian-Barre syndromes, Hashimoto's thyroiditis spondyloarthropathies such as ankylosing spondylitis, psoriasis, dermatitis herpetiformis, inflammatory bowel diseases, pemphigus vulgaris and vitiligo. In certain embodiments of the invention, the methods are used to determine an individual's risk for type 1 diabetes. Preferably, the individual is a human.

[0090] Nucleic Acids

[0091] Accordingly, one embodiment of the invention is an isolated nucleic acid molecule comprising a portion of the IL4R, IL4 or IL13 genes, their complements, or variants thereof. Preferably said variant comprises at least one of the polymorphisms identified herein. Even more preferably, said variant comprises at least one of the polymorphisms identified herein to be associated with type 1 diabetes. Thus, in one embodiment, the nucleic acid molecule comprises at least one of the IL4R, IL4, and/or IL13 polymorphisms provided in Table 2. In a further embodiment, the nucleic acid molecule comprises or consists of primers and probes specific to polymorphisms identified in the IL4R, IL4, or IL13 gene, including but not limited to SEQ ID NOS: 3-19, 25-36, 37-53, 54-62, 69-74, 75-80, and 81-86.

[0092] The isolated nucleic acid molecules may be RNA, mRNA, DNA, cDNA, and may be double- or single-stranded. They may encode the sense strand, the non-coding regions, or the antisense strand. The nucleic acid molecule can include all or a portion of the coding sequence of the gene and can further comprise additional non-coding regions such as introns and non-coding 3' and 5' sequences (including regulatory sequences for example). Additionally, the nucleic acid molecule can be fused to a marker sequence, for example, a sequence that encodes a polypeptide to assist in isolation or purification of the polypeptide.

[0093] An "isolated" nucleic acid molecule, as used herein, is one that is separated from nucleotide sequences which normally flank the nucleic acid molecule and/or has been completely or partially purified from other biological material (e.g., protein) normally associated with the nucleic acid.

[0094] The nucleic acid molecule can be fused to other coding or regulatory sequences and still be considered isolated. Thus, recombinant DNA contained in a vector is included in the definition of "isolated" as used herein. Also, isolated nucleic acid molecules include recombinant DNA molecules in heterologous host cells, as well as partially or substantially purified DNA molecules in solution. "Isolated" nucleic acid molecules also encompass in vivo and in vitro RNA transcripts of the DNA molecules of the present invention. An isolated nucleic acid molecule or nucleotide sequence can include a nucleic acid molecule or nucleotide sequence that is synthesized chemically or by recombinant means. Also, isolated polynucleotides include recombinant DNA molecules in heterologous organisms, as well as partially or substantially purified DNA molecules in solution. In vivo and in vitro RNA transcripts of the DNA molecules of the present invention are also encompassed by "isolated" nucleotide sequences. Such polynucleotides are useful in the manufacture of the encoded polypeptide, as probes for isolating homologous sequences (e.g., from other mammalian species), for gene mapping (e.g., by in situ hybridization with chromosomes), or for detecting expression of the gene in tissue (e.g., human tissue), such as by Northern blot analysis.

[0095] The nucleic acid molecules of the invention can comprise one or more modified nucleotide residues. The modification may be at the base, sugar and/or phosphate moiety and include, for example, halogenation, hydroxylation, alkylation, an attached linker and/or label. The modifications can further comprise, for example, labeling, methylation, internucleotide modifications such as uncharged linkages (e.g., methyl phosphonates, phosphotriesters, phosphoamidates, carbamates), charged linkages (e.g., phosphorothioates, phosphorodithioates), pendent moieties (e.g., polypeptides), intercalators (e.g., acridine, psoralen), chelators, alkylators, and modified linkages (e.g., alpha anomeric nucleic acids). Also included are synthetic molecules that mimic nucleic acid molecules in the ability to bind to a designated sequence via hydrogen bonding and other chemical interactions. Such molecules include, for example, those in which peptide linkages substitute for phosphate linkages in the backbone of the molecule.

[0096] In certain embodiments, nucleic acid molecules of the invention include, but are not limited to, IL4R, IL4 and/or IL13 mRNA, cDNA and/or genomic DNA molecules. The nucleotide sequence of the coding region of a IL4R mRNA is available from GenBank under accession number X52425.1, nucleotides 176-2653 are provided as SEQ ID NO: 2, shown in a 5' to 3' orientation in Table 1, below. The IL4R mRNA is provided as SEQ ID NO: 1. Although only one strand of the nucleic acid is shown in Table 1, those of skill in the art will recognize that SEQ ID NO: 1 and SEQ ID NO: 2 identify regions of double-stranded genomic nucleic acid, and that the sequences of both strands are fully specified by the sequence information provided. The genomic sequence for the IL4R gene is included in GenBank accession number AC004525.1 (SEQ ID NO: 88). The nucleotide sequence of the coding region of a IL4 mRNA is available from GenBank under accession number M23442.1 (SEQ ID NO: 89) and the nucleotide sequence of the coding region of a IL13 mRNA is available from GenBank under accession number U10307.1 (SEQ ID NO: 90).

[0097] Primers And Probes

[0098] By "oligonucleotide" is meant a single-stranded nucleotide polymer made of more than 2 nucleotide subunits covalently joined together. In one embodiment said oligonucleotides are between about 10 and 1000 nucleotide units, in a further embodiment, said oligonucleotides are between about 12 and 100 nucleotides units. The sugar groups of the nucleotide subunits may be ribose, deoxyribose or modified derivatives thereof such as o-methyl ribose. The nucleotide subunits of an oligonucleotide may be joined by phosphodiester linkages, phosphorothioate linkages, methyl phosphonate linkages or by other linkages, including but not limited to rare or non-naturally-occurring linkages, that do not prevent hybridization of the oligonucleotide. Furthermore, an oligonucleotide may have uncommon nucleotides or non-nucleotide moieties. An oligonucleotide as defined herein is a nucleic acid, preferably DNA, but may be RNA or have a combination of ribo- and deoxyribonucleotides covalently linked. Oligonucleotide probes and amplification oligonucleotides of a defined sequence may be produced by techniques known to those of ordinary skill in the art, such as by chemical or biochemical synthesis, and by in vitro or in vivo expression from recombinant nucleic acid molecules, e.g., bacterial or retroviral vectors. As used herein, an oligonucleotide does not consist of wild-type chromosomal DNA or the in vivo transcription products thereof.

[0099] Primer and probe sequences may comprise DNA, RNA (oligonucleotides--see above) or nucleic acid analogs such as uncharged nucleic acid analogs including but not limited to peptide nucleic acids (PNAs) which are disclosed in International Patent Application WO 92/20702 or morpholino analogs which are described in U.S. Pat. Nos. 5,185,444, 5,034,506, and 5,142,047 all of which are herein incorporated by reference in their entireties. Such sequences can routinely be synthesized using a variety of techniques currently available. For example, a sequence of DNA can be synthesized using conventional nucleotide phosphoramidite chemistry and the instruments available from Applied Biosystems, Inc, (Foster City, Calif.); DuPont, (Wilmington, Del.); or Milligen, (Bedford, Mass.). Similarly, and when desirable, the sequences can be labeled using methodologies well known in the art such as described in U.S. Pat. Nos. 5,464,746; 5,424,414; and 4,948,882 all of which are herein incorporated by reference in their entireties. Primers and Probes may be used in a variety of ways and may be defined by the specific use. For example, a "capture probe" is immobilized or can be immobilized on a solid support by any appropriate means, including, but not limited to: by covalent bonding, by adsorption, by hydrophobic and/or electrostatic interaction, or by direct synthesis on a solid support (see in particular patent application WO 92 10092). A "detection probe" may be labeled by means of a marker chosen, for example, from radioactive isotopes, enzymes, in particular enzymes capable of acting on a chromogenic, fluorigenic or luminescent substrate (in particular a peroxidase or an alkaline phosphatase), chromophoric chemical compounds, chromogenic, fluorigenic or luminescent compounds, analogues of nucleotide bases, and ligands such as biotin. A "primer" is a probe comprising, for example, from 10 to 100 nucleotide units and having a hybridization specificity under determined conditions for the initiation of an enzymatic polymerization, for example in an amplification technique such as PCR (Polymerase Chain Reaction), in a process of sequencing, in a method of reverse transcription and the like. One use of a probe is as a hybridization assay probe; probes may also be used as in vivo or in vitro therapeutic amplification oligomers or antisense agents to block or inhibit gene transcription, or translation in diseased, infected, or pathogenic cells.

[0100] All of the oligonucleotides, primers and probes of the present invention, whether hybridization assay probes, amplification oligonucleotides, or helper oligonucleotides, may be modified with chemical groups to enhance their performance or to facilitate the characterization of amplification products. For example, backbone-modified oligonucleotides such as those having phosphorothioate or methylphosphonate groups which render the oligonucleotides resistant to the nucleolytic activity of certain polymerases or to nuclease enzymes may allow the use of such enzymes in an amplification or other reaction. Another example of modification involves using non-nucleotide linkers (e.g., Arnold, et al., "Non-Nucleotide Linking Reagents for Nucleotide Probes", EP 0 313 219 hereby incorporated by reference herein in its entirety) incorporated between nucleotides in the nucleic acid chain which do not interfere with hybridization or the elongation of the primer. Amplification oligonucleotides may also contain mixtures of the desired modified and natural nucleotides.

[0101] The 3' end of an amplification oligonucleotide may be blocked to prevent initiation of DNA synthesis as described by McDonough, et al., entitled "Nucleic Acid Sequence Amplification", WO94/03472 which enjoys common ownership with the present invention and is hereby incorporated by reference herein in its entirety. A mixture of different 3' blocked amplification oligonucleotides, or of 3' blocked and unblocked oligonucleotides may increase the efficiency of nucleic acid amplification, as described therein.

[0102] The 5' end of the oligonucleotides may be modified to be resistant to the 5'-exonuclease activity present in some nucleic acid polymerases. Such modifications can be carried out by adding a non-nucleotide group to the terminal 5' nucleotide of the primer using techniques such as those described by Arnold, et al., supra, entitled "Non-Nucleotide Linking Reagents for Nucleotide Probes", incorporated by reference herein.

[0103] Once synthesized, selected oligonucleotide probes may be labeled by any of several well-known methods (e.g., J. Sambrook, supra). Useful labels include radioisotopes as well as non-radioactive reporting groups. Isotopic labels include .sup.3H, .sup.35S, .sup.32P, .sup.125I, .sup.57Co and .sup.14C. Isotopic labels can be introduced into the oligonucleotide by techniques known in the art such as nick translation, end labeling, second strand synthesis, the use of reverse transcription, and by chemical methods. When using radiolabeled probes hybridization can be detected by autoradiography, scintillation counting, or gamma counting. The detection method selected will depend upon the particular radioisotope used for labeling.

[0104] Non-isotopic materials can also be used for labeling and may be introduced internally into the nucleic acid sequence or at the end of the nucleic acid sequence. Modified nucleotides may be incorporated enzymatically or chemically. Chemical modifications of the probe may be performed during or after synthesis of the probe, for example, through the use of non-nucleotide linker groups as described by Arnold, et al., supra "Non-Nucleotide Linking Reagents for Nucleotide Probes," incorporated by reference herein. Non-isotopic labels include fluorescent molecules, chemiluminescent molecules, enzymes, cofactors, enzyme substrates, haptens or other ligands.

[0105] In one embodiment, the probes are labeled with an acridinium ester. Acridinium ester labeling may be performed as described by Arnold et al., U.S. Pat. No. 5,185,439, entitled "Acridinium Ester Labeling and Purification of Nucleotide Probes," issued Feb. 9, 1993 and hereby incorporated by reference herein in its entirety. TABLE-US-00001 TABLE 1 SEQ ID NO: 2 1 atggggtggc tttgctctgg gctcctgttc cctgtgagct gcctggtcct gctgcaggtg 61 gcaagctctg ggaacatgaa ggtcttgcag gagcccacct gcgtctccga ctacatgagc 121 atctctactt gcgagtggaa gatgaatggt cccaccaatt gcagcaccga gctccgcctg 181 ttgtaccagc tggtttttct gctctccgaa gcccacacgt gtatccctga gaacaacgga 241 ggcgcggggt gcgtgtgcca cctgctcatg gatgacgtgg tcagtgcgga taactataca 301 ctggacctgt gggctgggca gcagctgctg tggaagggct ccttcaagcc cagcgagcat 361 gtgaaaccca gggccccagg aaacctgaca gttcacacca atgtctccga cactctgctg 421 ctgacctgga gcaacccgta tccccctgac aattacctgt ataatcatct cacCtatgca 481 gtcaacattt ggagtgaaaa cgacccggca gatttcagaa tctataacgt gacctaccta 541 gaaccctccc tccgcatcgc agccagcacc ctgaagtctg ggatttccta cagggcacgg 601 gtgagggcct gggctcagtg ctataacacc acctggagtg agtggagccc cagcaccaag 661 tggcacaact cctacaggga gcccttcgag cagcacctcc tgctgggcgt cagcgtttcc 721 tgcattgtca tcctggccgt ctgcctgttg tgctatgtca gcatcaccaa gattaagaaa 781 gaatggtggg atcagattcc caacccagcc cgcagccgcc tcgtggctat aataatccag 841 gatgctcagg ggtcacagtg ggagaagcgg tcccgaggcc aggaaccagc caagtgccca 901 cactggaaga attgtcttac caagctcttg ccctgttttc tggagcacaa catgaaaagg 961 gatgaagatc ctcacaaggc tgccaaagag atgcctttcc agggctctgg aaaatcagca 1021 tggtgcccag tggagatcag caagacagtc ctctggccag agagcatcag cgtggtgcga 1081 tgtgtggagt tgtttgaggc cccggtggag tgtgaggagg aggaggaggt agaggaagaa 1141 aaagggagct tctgtgcatc gcctgagagc agcagggatg acttccagga gggaagggag 1201 ggcattgtgg cccggctaac agagagcctg ttcctggacc tgctcggaga ggagaatggg 1261 ggcttttgcc agcaggacat gggggagtca tgccttcttc caccttcggg aagtacgagt 1321 gctcacatgc cctgggatga gttcccaagt gcagggccca aggaggcacc tccctggggc 1381 aaggagcagc ctctccacct ggagccaagt cctcctgcca gcccgaccca gagtccagac 1441 aacctgactt gcacagagac gcccctcgtc atcgcaggca accctgctta ccgcagcttc 1501 agcaactccc tgagccagtc accgtgtccc agagagctgg gtccagaccc actgctggcc 1561 agacacctgg aggaagtaga acccgagatg ccctgtgtcc cccagctctc tgagccaacc 1621 actgtgcccc aacctgagcc agaaacctgg gagcagatcc tccgccgaaa tgtcctccag 1681 catggggcag ctgcagcccc cgtctcggcc cccaccagtg gctatcagga gtttgtacat 1741 gcggtggagc agggtggcac ccaggccagt gcggtggtgg gcttgggtcc cccaggagag 1801 gctggttaca aggccttctc aagcctgctt gccagcagtg ctgtgtcccc agagaaatgt 1861 gggtttgggg ctagcagtgg ggaagagggg tataagcctt tccaagacct cattcctggc 1921 tgccctgggg accctgcccc agtccctgtc cccttgttca cctttggact ggacagggag 1981 ccacctcgca gtccgcagag ctcacatctc ccaagcagct ccccagagca cctgggtctg 2041 gagccggggg aaaaggtaga ggacatgcca aagcccccac ttccccagga gcaggccaca 2101 gacccccttg tggacagcct gggcagtggc attgtctact cagcccttac ctgccacctg 2161 tgcggccacc tgaaacagtg tcatggccag gaggatggtg gccagacccc tgtcatggcc 2221 agtccttgct gtggctgctg ctgtggagac aggtcctcgc cccctacaac ccccctgagg 2281 gccccagacc cctctccagg tggggttcca ctggaggcca gtctgtgtcc ggcctccctg 2341 gcaccctcgg gcatctcaga gaagagtaaa tcctcatcat ccttccatcc tgcccctggc 2401 aatgctcaga gctcaagcca gacccccaaa atcgtgaact ttgtctccgt gggacccaca 2461 tacatgaggg tctcttag

[0106] SNPs

[0107] In one aspect, the present invention provides a method for detecting an individual's increased or decreased risk for an autoimmune disease such as type 1 diabetes by detecting the presence of one or more IL4R, IL4 or IL13 SNPs in a nucleic acid sample of the individual, wherein the presence of said SNP(s) indicates the individual's increased or decreased risk for type 1 diabetes. The SNPs can be any SNPs in the IL4R, IL4 or IL13 loci including SNPs in exons, introns or upstream or downstream regions. Examples of such SNPs include, but are not limited to those provided in Table 2, below, and discussed in detail in the Examples. In one embodiment, the SNPs present in the IL4R, IL4 or IL13 loci are identified by genotyping the IL4R, IL4 or IL13 SNPs.

[0108] In certain embodiments, the genotype of one IL4R, IL4 or IL13 SNP can be used to determine an individual's risk for an autoimmune disease. In other embodiments, the genotypes of a plurality of IL4R, IL4 or IL13 SNPs can be used. For example, in certain embodiments, the genotypes of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or 31 of the SNPs in Table 2 can be used to determine a individual's risk for an autoimmune disease. In other embodiments, certain combinations of SNPs at either the same or different loci can be used, as described in the Examples, below. TABLE-US-00002 TABLE 2 IL4R, IL4 and IL13 SNPs IL4R SNPs Acc dbSNP WT Var X52425.1 AC004525.1 Formal ID rs# Exon* Variation allele allele (cDNA) (genomic) SNP name P C(-3223)T G A NA 128387 G128387A P T(-1914)C A G NA 127078 A127078G 3 I50V A G 398 94272 A398G 4 N142N C T 676 92548 C676T 4 C92516T C T NA 92516 C92516T 4 A92417T A T NA 92417 A92417T 2234896 7 P249P C G 997 80189 C997G 2234897 9 F288F T C 1114 76868 T1114C 1805011 9 E375A A C 1374 76608 A1374C 9 E375E G A 1375 76607 G1375A 2234898 9 L389L G T 1417 76565 G1417T 1805012 9 C406R T C 1466 76516 T1466C 2234899 9 C406C C T 1468 76514 C1468T 2234900 9 L408L T C 1474 76508 T1474C 1805013 9 S411L C T 1482 76500 C1482T 1805015 9 S478P T C 1682 76300 T1682C 1801275 9 Q551R A G 1902 76080 A1902G 9 V554I G A 1910 76072 G1910A 9 P650S C T 2198 75784 C2198T 1805016 9 S727A T G 2429 75553 T2429G 9 G759G C T 2567 75455 C2567T 1805014 9 S761P T C 2531 75451 T2531C 9 P774P T C 2572 75410 T2572C 1049631 9 3'UTR G A 3044 74938 G3044A 8832 9 3'UTR A G 3289 74693 A3289G 8674 9 3'UTR C T 3391 74581 C3391T SNP WT Var Genbank Chromosomal Formal Description Exon Variation allele allele Access # Position SNP name IL4 SNPs 5' (-524) P (-524) C T M23442.1 5q31 C582T IL13 SNPs (-1512) P (-1512) A C U10307.1 5q31 A(-1512)C (-1112) P (-1112) C T U10307.1 5q31 C(-1112)T Intron 3 -- Intron 3 C T U10307.1 5q31 C4045T R110Q 4 R110Q G A U10307.1 5q31 G4166A P: Promoter region *The exon numbering is based on the scheme disclosed by Ober etal., 2000, Am J Hum Genet 66: 517-526. Other numbering schemes are available and one of skill in the art will be able to identify the SNP and the scheme is used.

[0109] Genotyping Methods

[0110] In the methods of the present invention, the alleles present in a sample are identified by identifying the nucleotide present at one or more of the polymorphic sites. Any type of tissue containing IL4R, IL4 or IL13 nucleic acid may be used for determining the IL4R, IL4 or IL13 genotypes of an individual. A number of methods are known in the art for identifying the nucleotide present at polymorphic sites. The particular method used to identify the genotype is not a critical aspect of the invention. Although considerations of performance, cost, and convenience will make particular methods more desirable than others, it will be clear that any method that can identify the nucleotide present will provide the information needed to identify the genotype. Preferred genotyping methods involve DNA sequencing, allele-specific amplification, or probe-based detection of amplified nucleic acid.

[0111] IL4R, IL4 or IL13 alleles can be identified by DNA sequencing methods, such as the chain termination method (Sanger et al., 1977, Proc. Natl. Acad. Sci,. 74:5463-5467, incorporated herein by reference), which are well known in the art. In one embodiment, a subsequence of the gene encompassing the polymorphic site is amplified and either cloned into a suitable plasmid and then sequenced, or sequenced directly. PCR-based sequencing is described in U.S. Pat. No. 5,075,216; Brow, in PCR Protocols, 1990, (Innis et al., eds., Academic Press, San Diego), chapter 24; and Gyllensten, in PCR Technology, 1989 (Erlich, ed., Stockton Press, New York), chapter 5; each incorporated herein by reference. Typically, sequencing is carried out using one of the automated DNA sequencers which are commercially available from, for example, PE Biosystems (Foster City, Calif.), Pharmacia (Piscataway, N.J.), Genomyx Corp. (Foster City, Calif.), LI-COR Biotech (Lincloln, Neb.), GeneSys technologies (Sauk City, Wis.), and Visible Genetics, Inc. (Toronto, Canada).

[0112] IL4R, IL4 or IL13 alleles can also be identified using amplification-based genotyping methods. Various nucleic acid amplification methods known in the art can be used in to detect nucleotide changes in a target nucleic acid. A preferred method is the polymerase chain reaction (PCR), which is now well known in the art, and described in U.S. Pat. Nos. 4,683,195; 4,683,202; and 4,965,188; each incorporated herein by reference. Examples of the numerous articles published describing methods and applications of PCR are found in PCR Applications, 1999, (Innis et al., eds., Academic Press, San Diego), PCR Strategies, 1995, (Innis et al., eds., Academic Press, San Diego); PCR Protocols, 1990, (Innis et al., eds., Academic Press, San Diego); and PCR Technology, 1989, (Erlich, ed., Stockton Press, New York); each incorporated herein by reference. Commercial vendors, such as PE Biosystems (Foster City, Calif.) market PCR reagents and publish PCR protocols.

[0113] Other suitable amplification methods include the ligase chain reaction (Wu and Wallace, 1988, Genomics 4:560-569); the strand displacement assay (Walker et al., 1992, Proc. Natl. Acad. Sci. USA 89:392-396, Walker et al. 1992, Nucleic Acids Res. 20:1691-1696, and U.S. Pat. No. 5,455,166); and several transcription-based amplification systems, including the methods described in U.S. Pat. Nos. 5,437,990; 5,409,818; and 5,399,491; the transcription amplification system (TAS) (Kwoh et al., 1989, Proc. Natl. Acad. Sci. USA, 86:1173-1177); and self-sustained sequence replication (3SR) (Guatelli et al., 1990, Proc. Natl. Acad. Sci. USA, 87:1874-1878 and WO 92/08800); each incorporated herein by reference. Alternatively, methods that amplify the probe to detectable levels can be used, such as QB-replicase amplification (Kramer et al., 1989, Nature, 339:401-402, and Lomeli et al., 1989, Clin. Chem., 35:1826-1831, both of which are incorporated herein by reference). A review of known amplification methods is provided in Abramson et al., 1993, Current Opinion in Biotechnology, 4:41-47, incorporated herein by reference.

[0114] Genotyping also can also be carried out by detecting and analyzing IL4R, IL4 or IL13 mRNA under conditions when both, maternal and paternal, chromosomes are transcribed. Amplification of RNA can be carried out by first reverse-transcribing the target RNA using, for example, a viral reverse transcriptase, and then amplifying the resulting cDNA, or using a combined high-temperature reverse-transcription-polymerase chain reaction (RT-PCR), as described in U.S. Pat. Nos. 5,310,652; 5,322,770; 5,561,058; 5,641,864; and 5,693,517; each incorporated herein by reference (see also Myers and Sigua, 1995, in PCR Strategies, supra, chapter 5).

[0115] IL4R, IL4 or IL13 alleles can also be identified using allele-specific amplification or primer extension methods, which are based on the inhibitory effect of a terminal primer mismatch on the ability of a DNA polymerase to extend the primer. To detect an allele sequence using an allele-specific amplification or extension-based method, a primer complementary to the IL4R, IL4 or IL13 genes is chosen such that the 3' terminal nucleotide hybridizes at the polymorphic position. In the presence of the allele to be identified, the primer matches the target sequence at the 3' terminus and primer is extended. In the presence of only the other allele, the primer has a 3' mismatch relative to the target sequence and primer extension is either eliminated or significantly reduced. Allele-specific amplification- or extension-based methods are described in, for example, U.S. Pat. Nos. 5,137,806; 5,595,890; 5,639,611; and U.S. Pat. No. 4,851,331, each incorporated herein by reference.

[0116] Using allele-specific amplification-based genotyping, identification of the alleles requires only detection of the presence or absence of amplified target sequences. Methods for the detection of amplified target sequences are well known in the art. For example, gel electrophoresis (see Sambrook et al., 1989, supra.) and the probe hybridization assays described above have been used widely to detect the presence of nucleic acids.

[0117] Allele-specific amplification-based methods of genotyping can facilitate the identification of haplotypes, as described in the examples. Essentially, the allele-specific amplification is used to amplify a region encompassing multiple polymorphic sites from only one of the two alleles in a heterozygous sample. The SNP variants present within the amplified sequence are then identified, such as by probe hybridization or sequencing.

[0118] An alternative probe-less method, referred to herein as a kinetic-PCR method, in which the generation of amplified nucleic acid is detected by monitoring the increase in the total amount of double-stranded DNA in the reaction mixture, is described in Higuchi et al., 1992, Bio/Technology, 10:413-417; Higuchi et al., 1993, Bio/Technology, 11:1026-1030; Higuchi and Watson, in PCR Applications, supra, Chapter 16; U.S. Pat. Nos. 5,994,056 and 6,171,785; and European Patent Publication Nos. 487,218 and 512,334, each incorporated herein by reference. The detection of double-stranded target DNA relies on the increased fluorescence that DNA-binding dyes, such as ethidium bromide, exhibit when bound to double-stranded DNA. The increase of double-stranded DNA resulting from the synthesis of target sequences results in an increase in the amount of dye bound to double-stranded DNA and a concomitant detectable increase in fluorescence. For genotyping using the kinetic-PCR methods, amplification reactions are carried out using a pair of primers specific for one of the alleles, such that each amplification can indicate the presence of a particular allele. By carrying out two amplifications, one using primers specific for the wild-type allele and one using primers specific for the mutant allele, the genotype of the sample with respect to that SNP can be determined. Similarly, by carrying out four amplifications, each with one of the possible pairs possible using allele specific primers for both the upstream and downstream primers, the genotype of the sample with respect to two SNPs can be determined. This gives haplotype information for a pair of SNPs.

[0119] Alleles can be also identified using probe-based methods, which rely on the difference in stability of hybridization duplexes formed between a probe and its corresponding target sequence comprising an IL4R, IL4 or IL13 allele. Under sufficiently stringent hybridization conditions, stable duplexes are formed only between a probe and its target allele sequence and not other allele sequences. The presence of stable hybridization duplexes can be detected by any of a number of well known methods. In general, it is preferable to amplify a nucleic acid encompassing a polymorphic site of interest prior to hybridization in order to facilitate detection. However, this is not necessary if sufficient nucleic acid can be obtained without amplification.

[0120] A probe suitable for use in the probe-based methods of the present invention, which contains a hybridizing region either substantially complementary or exactly complementary to a target region of SEQ ID NOS: 2, 88, 89 or 90 or the complement of SEQ ID NOS: 2, 88, 89 or 90, wherein the target region encompasses the polymorphic site, and exactly complementary to one of the two allele sequences at the polymorphic site, can be selected using the guidance provided herein and well known in the art. Similarly, suitable hybridization conditions, which depend on the exact size and sequence of the probe, can be selected empirically using the guidance provided herein and well known in the art. The use of oligonucleotide probes to detect nucleotide variations including single base pair differences in sequence is described in, for example, Conner et al., 1983, Proc. Natl. Acad. Sci. USA, 80:278-282, and U.S. Pat. Nos. 5,468,613 and 5,604,099, each incorporated herein by reference.

[0121] In preferred embodiments of the probe-based methods for determining the IL4R, IL4 or IL13 genotypes, multiple nucleic acid sequences from the IL4R, IL4 or IL13 genes which encompass the polymorphic sites are amplified and hybridized to a set of probes under sufficiently stringent hybridization conditions. The alleles present are inferred from the pattern of binding of the probes to the amplified target sequences. In this embodiment, amplification is carried out in order to provide sufficient nucleic acid for analysis by probe hybridization. Thus, primers are designed such that regions of the IL4R, IL4 or IL13 genes encompassing the polymorphic sites are amplified regardless of the allele present in the sample. Allele-independent amplification is achieved using primers which hybridize to conserved regions of the IL4R, IL4 or IL13 genes. The IL4R, IL4 or IL13 genes contain many invariant or monomorphic regions and suitable allele-independent primers can be selected routinely from SEQ ID NOS: 1, 88, 89 or 90. One of skill will recognize that, typically, experimental optimization of an amplification system is helpful.

[0122] Suitable assay formats for detecting hybrids formed between probes and target nucleic acid sequences in a sample are known in the art and include the immobilized target (dot-blot) format and immobilized probe (reverse dot-blot or line-blot) assay formats. Dot blot and reverse dot blot assay formats are described in U.S. Pat. Nos. 5,310,893; 5,451,512; 5,468,613; and 5,604,099; each incorporated herein by reference.

[0123] In a dot-blot format, amplified target DNA is immobilized on a solid support, such as a nylon membrane. The membrane-target complex is incubated with labeled probe under suitable hybridization conditions, unhybridized probe is removed by washing under suitably stringent conditions, and the membrane is monitored for the presence of bound probe. A preferred dot-blot detection assay is described in the examples.

[0124] In the reverse dot-blot (or line-blot) format, the probes are immobilized on a solid support, such as a nylon membrane or a microtiter plate. The target DNA is labeled, typically during amplification by the incorporation of labeled primers. One or both of the primers can be labeled. The membrane-probe complex is incubated with the labeled amplified target DNA under suitable hybridization conditions, unhybridized target DNA is removed by washing under suitably stringent conditions, and the membrane is monitored for the presence of bound target DNA. A preferred reverse line-blot detection assay is described in the examples.

[0125] Probe-based genotyping can be carried out using a "TaqMan" or "5'-nuclease assay," as described in U.S. Pat. Nos. 5,210,015; 5,487,972; and 5,804,375; and Holland et al., 1988, Proc. Natl. Acad. Sci. USA, 88:7276-7280, each incorporated herein by reference. In the TaqMan assay, labeled detection probes that hybridize within the amplified region are added during the amplification reaction mixture. The probes are modified so as to prevent the probes from acting as primers for DNA synthesis. The amplification is carried out using a DNA polymerase that possesses 5' to 3' exonuclease activity, e.g., Tth DNA polymerase. During each synthesis step of the amplification, any probe which hybridizes to the target nucleic acid downstream from the primer being extended is degraded by the 5' to 3' exonuclease activity of the DNA polymerase. Thus, the synthesis of a new target strand also results in the degradation of a probe, and the accumulation of degradation product provides a measure of the synthesis of target sequences.

[0126] Any method suitable for detecting degradation product can be used in the TaqMan assay. In a preferred method, the detection probes are labeled with two fluorescent dyes, one of which is capable of quenching the fluorescence of the other dye. The dyes are attached to the probe, preferably one attached to the 5' terminus and the other is attached to an internal site, such that quenching occurs when the probe is in an unhybridized state and such that cleavage of the probe by the 5' to 3' exonuclease activity of the DNA polymerase occurs in between the two dyes. Amplification results in cleavage of the probe between the dyes with a concomitant elimination of quenching and an increase in the fluorescence observable from the initially quenched dye. The accumulation of degradation product is monitored by measuring the increase in reaction fluorescence. U.S. Pat. Nos. 5,491,063 and 5,571,673, both incorporated herein by reference, describe alternative methods for detecting the degradation of probe which occurs concomitant with amplification.

[0127] The TaqMan assay can be used with allele-specific amplification primers such that the probe is used only to detect the presence of amplified product. Such an assay is carried out as described for the kinetic-PCR-based methods described above. Alternatively, the TaqMan assay can be used with a target-specific probe.

[0128] Examples of other techniques that can be used for probe-based genotyping include, but are not limited to, Amplifluor.TM., Dye Binding-Intercalation, Fluorescence Resonance Energy Transfer (FRET), Hybridization Signal Amplification Method (HSAM), HYB Probes.TM., Invader/Cleavase Technology (Invader/CFLP.TM.), Molecular Beacons.TM., Origen.TM., DNA-Based Ramification Amplification (RAM.TM.), Rolling circle amplification (RCA.TM.), Scorpions.TM., Strand displacement amplification (SDA).

[0129] The assay formats described above typically utilize labeled oligonucleotides to facilitate detection of the hybrid duplexes. Oligonucleotides can be labeled by incorporating a label detectable by spectroscopic, photochemical, biochemical, immunochemical, radiological, radiochemical or chemical means. Useful labels include .sup.32P, fluorescent dyes, electron-dense reagents, enzymes (as commonly used in ELISAs), biotin, or haptens and proteins for which antisera or monoclonal antibodies are available. Labeled oligonucleotides of the invention can be synthesized and labeled using the techniques described above for synthesizing oligonucleotides. For example, a dot-blot assay can be carried out using probes labeled with biotin, as described in Levenson et al., 1989, in PCR Protocols: A Guide to Methods and Applications (Innis et al., eds., Academic Press. San Diego), pages 99-112, incorporated herein by reference. Following hybridization of the immobilized target DNA with the biotinylated probes under sequence-specific conditions, probes which remain bound are detected by first binding the biotin to avidin-horseradish peroxidase (A-HRP) or streptavidin-horseradish peroxidase (SA-HRP), which is then detected by carrying out a reaction in which the HRP catalyzes a color change of a chromogen.

[0130] Whatever the method for determining which oligonucleotides of the invention selectively hybridize to IL4R, IL4 or IL13 allelic sequences in a sample, the central feature of the typing method involves the identification of the IL4R, IL4 or IL13 alleles present in the sample by detecting the variant sequences present.

[0131] The present invention also relates to a kit, a container unit comprising useful components for practicing the present method. A useful kit can contain oligonucleotide probes specific for IL4R, IL.sup.4 or IL13 alleles as well as instructions for their use to determine risk for an autoimmune disease such as type 1 diabetes. In some cases, detection probes may be fixed to an appropriate support membrane. The kit can also contain amplification primers for amplifying regions of the IL4R, IL4 or IL13 loci encompassing the polymorphic sites, as such primers are useful in the preferred embodiment of the invention. Alternatively, useful kits can contain a set of primers comprising an allele-specific primer for the specific amplification of IL4R, IL4 or IL13 alleles. Other optional components of the kits include additional reagents used in the genotyping methods as described herein. For example, a kit additionally can contain an agent to catalyze the synthesis of primer extension products, substrate nucleoside triphosphates, reagents for labeling and/or detecting nucleic acid (for example, an avidin-enzyme conjugate and enzyme substrate and chromogen if the label is biotin) and appropriate buffers for amplification or hybridization reactions.

[0132] The present invention also relates to an array, a support with immobilized oligonucleotides useful for practicing the present method. A useful array can contain oligonucleotide probes specific for IL4R, IL4, IL13 alleles or certain combinations of IL4R, IL4 and/or IL13 alleles. The oligonucleotides can be immobilized on a substrate, e.g., a membrane or glass. The oligonucleotides can, but need not, be labeled. In some embodiments, the array can be a micro-array. In some embodiments, the array can comprise one or more oligonucleotides used to detect the presence of two or more IL4R, IL4, IL13 alleles or certain combinations of IL4R, IL4 and/or IL13 alleles.

[0133] The examples of the present invention presented below are provided only for illustrative purposes and not to limit the scope of the invention. Numerous embodiments of the invention within the scope of the claims that follow the examples will be apparent to those of ordinary skill in the art from reading the foregoing text and following examples.

7 EXAMPLE 1

Genotyping Protocol: Probe-Based Identification of IL4R, IL4 and IL13 Alleles

[0134] This example describes a method of genotyping SNPs in the IL4R, IL4 and IL13 loci that are associated with type 1 diabetes. Two different genotyping methods, line blot assays and kinetic thermocycling, were used, depending on the region and gene genotyped.

[0135] Line Blot Assay for Identifying 8 IL4R SNPs, 1 IL4 SNP and 2 IL13 SNPs

[0136] Eight exemplary SNPs in the human IL4R gene (listed in Table 6), one exemplary SNP in the human IL4 gene (Table 2) and two exemplary SNPs in the human IL4R gene (Table 2) were genotyped using this method. Each SNP is described by its position in the reference GenBank accession sequence. For example, SNP 1 of Table 6 is found at position 398 of X52425.1 (SEQ ID NO: 1), where an "A" nucleotide is present. The variant allele at this position has a "G" nucleotide. The SNPs will be referred to by the SNP # in the subsequent text.

[0137] The regions of the IL4R, IL4 and IL13 genes that encompass the SNPs were amplified and the nucleotide present identified by probe hybridization. The probe detection was carried out using an immobilized probe (line blot) format.

Amplicons and Primers

[0138] The pairs of primers used to amplify the regions encompassing the eight IL4R SNPs are listed in Table 7 (SEQ ID NO: 25-36) and those used to amplify the regions encompassing the IL4 SNP and two IL13 SNPs are listed in Table 30 (SEQ ID NO: 69-74). IL4R SNP numbers 3, 4, and 5 (Table 6) were co-amplified on the same 228 basepair fragment. The primers were modified at the 5' phosphate by conjugation with biotin. Reagents for synthesizing oligonucleotides with a biotin label attached to the 5' phosphate are commercially available from Clontech (Palo Alto, Calif.) and Glenn Research (Sterling, Va.). A preferred reagent is Biotin-ON from Clontech.

Amplification Primers

[0139] Amplification of six regions of the IL4R gene, which encompass eight polymorphic sites, the one region of the IL4 gene, which encompass one polymorphic site, and the two regions of the IL13 gene, which encompass two polymorphic sites, was carried out using the primer pairs shown below. All primers are shown in the 5' to 3' orientation.

[0140] The following primers amplify a 114 base-pair region encompassing nucleotide position 398 the IL4R gene. TABLE-US-00003 RR192B (SEQ ID NO: 25) CAGCCCCTGTGTCTGCAGA RR193B (SEQ ID NO: 31) GTCCAGTGTATAGTTATCCGCACTGA

[0141] The following primers amplify a 163 base-pair region encompassing nucleotide position 676. TABLE-US-00004 DBM0177B (SEQ ID NO: 26) CTGACCTGGAGCAACCCGTA DBM0178B (SEQ ID NO: 32) ACTGGGCCTCTGCTGGTCA

[0142] The following primers amplify a 228 base-pair region encompassing nucleotide positions 1374, 1417, and 1466 of the IL4R gene. TABLE-US-00005 DBM0023B (SEQ ID NO: 27) ATTGTGTGAGGAGGAGGAGGAGGTA DBM0022B (SEQ ID NO: 33) GTTGGGCATGTGAGCACTCGTA

[0143] The following primers amplify a 129 base-pair region encompassing nucleotide position 1682 of the IL4R gene. TABLE-US-00006 DBM0097B (SEQ ID NO: 28) CTCGTCATCGCAGGCAA DBM0098B (SEQ ID NO: 34) AGGGCATCTCGGGTTCTA

[0144] The following primers amplify a 198 base-pair region encompassing nucleotide position 1902 of the IL4R gene. TABLE-US-00007 RR200B (SEQ ID NO: 29) GCCGAAATGTCCTCCAGCA RR178B (SEQ ID NO: 35) CCACATTTCTCTGGGGACACA

[0145] The following primers amplify a 177 base-pair region encompassing nucleotide position 2531 of the IL4R gene. TABLE-US-00008 DBM0112B (SEQ ID NO: 30) CCGGCCTCCCTGGCA DBM0071B (SEQ ID NO: 36) GCAGACTCAGCAACAAGAGG

[0146] The following primers amplify a 107 base-pair region encompassing nucleotide position 582 in the promoter region of the IL4 gene. TABLE-US-00009 RR169B (SEQ ID NO: 69) ACTAGGCCTCACCTGATACGA RR170B (SEQ ID NO: 72) CATAGAGGCAGAATAACAGGCAGA

[0147] The following primers amplify a 118 base-pair region encompassing nucleotide position 4045 in intron 3 of the IL13 gene. TABLE-US-00010 DBM0165B (SEQ ID NO: 70) CTCGGACATGCAAGCTGGAA DBM0166B (SEQ ID NO: 73) ACTGAATGAGACAGTCCCTGGA

[0148] The following primers amplify a 187 base-pair region encompassing codon 4166 in exon 4 of the IL13 gene. TABLE-US-00011 DBM0167B (SEQ ID NO: 71) AATCGAGGTGGCCCAGTTTGTA DBM0168B (SEQ ID NO: 74) CCTAACCCTCCTTCCCGCCTA

Amplification

[0149] The PCR amplification was carried out in a total reaction volume of 25-100 .mu.l containing the following reagents:

[0150] 0.2 ng/.mu.l purified human genomic DNA

[0151] 0.2 mM each primer

[0152] 800 mM total dNTP (200 mM each dATP, dTTP, dCTP, dGTP)

[0153] 70 mM KCl

[0154] 12 mM Tris-HCl, pH 8.3

[0155] 3 mM MgCl.sub.2,

[0156] 0.25 units/.mu.l AmpliTaq Gold.TM. DNA polymerase* * developed and manufactured by Hoffmann-La Roche and commercially available from Applera (Foster City, Calif.).

[0157] Amplification was carried out in a GeneAmp.TM. PCR System 9600 thermal cycler (Applera, Foster City, Calif.), using the specific temperature cycling profile shown below. TABLE-US-00012 Pre-reaction incubation: 94.degree. C. for 12.5 minutes 33 cycles: denature: 95.degree. C. for 45 seconds anneal: 61.degree. C. for 30 seconds extend: 72.degree. C. for 45 seconds Final extension: 72.degree. C. for 7 minutes Hold: 10.degree. C.-15.degree. C.

Detection Probes

[0158] Preferred probes used to identify the nucleotides present at the 8 SNPs present in the amplified IL4R nucleic acids are described in Table 3. Two probes are shown for the detection of T1466; a mixture of the two probes was used. Preferred probes used to identify the nucleotides present at the one SNP present in the amplified IL4 nucleic acids and the two SNPs present in the amplified IL13 nucleic acids are described in Table 29. All probes are shown in the 5' to 3' orientation.

Probe Hybridization Assay, Immobilized Probe Format

[0159] In the immobilized probe format, the probes were immobilized to a solid support prior to being used in the hybridization. The probe-support complex was immersed in a solution containing denatured amplified nucleic acid (biotin labeled) to allow hybridization to occur. Unbound nucleic acid was removed by washing under stringent hybridization conditions, and nucleic acid remaining bound to the immobilized probes was detected using a chromogenic reaction. The details of the assay are described below.

[0160] For use in the immobilized probe detection format, described below, a moiety was attached to the 5' phosphate of the probe to facilitate immobilization on a solid support. See Cheng et al., 1999, Genome Res 9:936-949, incorporated herein by reference. Preferably, Bovine Serum Albumin (BSA) is attached to the 5' phosphate essentially as described by Tung et al., 1991, Bioconjugate Chem., 2:464-465, incorporated herein by reference. Alternatively, a poly-T tail is added to the 5' end as described in U.S. Pat. No 5,451,512, incorporated herein by reference.

[0161] The probes were applied in a linear format to sheets of nylon membrane (e.g., BioDyne.TM. B nylon filters, Pall Corp., Glen Cove, N.Y.) using a Linear Striper and Multispense2000.TM. controller (IVEK, N. Springfield, Vt.). The detection of the wildtype allele of SNP #5 (table 6) was carried out using a mixture of two probes as listed; this mixture enables the detection of SNP #5 indiscriminately of another nearby SNP. Probe titers were chosen to achieve signal balance between the allelic variants; the titers used are provided in the table of probes, above. Each sheet was cut to strips between 0.35 and 0.5 cm in width. To denature the amplification products, 20 .mu.l of amplification product (based on a 50 .mu.l reaction) were added to 20 .mu.l of denaturation solution (1.6% NaOH) and incubated at room temperature to complete denaturation.

[0162] The denatured amplification product (40 .mu.l) was added to the well of a typing tray containing 3 ml of hybridization buffer (4.times.SSPE, 0.5% SDS) and the membrane strip. Hybridizations were allowed to proceed for 15 minutes at 55.degree. C. in a rotating water bath. Following hybridization, the hybridization solution was aspirated, the strip was rinsed in 3 ml warm wash buffer (2.times.SSPE, 0.5% SDS) by gently rocking strips back and forth, and the wash buffer was aspirated. Following rinsing, the strips were incubated in 3 ml enzyme conjugate solution (3.3 ml hybridization buffer and 12 .mu.l of strepavidin-horseradish peroxidase (SA-HRP)) in the rotating water bath for 5 minutes at 55.degree. C. Then the strips were rinsed with wash buffer, as above, incubated in wash buffer at 55.degree. for 12 minutes (stringent wash), and finally rinsed with wash buffer again.

[0163] Target nucleic acid, now HRP-labeled, which remains bound to the immobilized amplification product was visualized as follows. A color development solution was prepared by mixing 100 ml of citrate buffer (0.1 M Sodium Citrate, pH 5.0), 5 ml 3,3',5,5'-tetramethylbenzidine (TMB) solution (2 mg/ml TMB powder from Fluka, Milwaukee, Wis., dissolved in 100% EtOH), and 100 .mu.l of 3% hydrogen peroxide. The strips were first rinsed in 0.1 M sodium citrate (pH 5.0) for 5 minutes, then incubated in the color development solution with gentle agitation for 8 to 10 minutes at room temperature in the dark. The TMB, initially colorless, is converted by the target-bound HRP, in the presence of hydrogen peroxide, into a colored precipitate. The developed strips were rinsed in water for several minutes and immediately photographed.

[0164] Kinetic Thermocycling to Identify 2 IL4R Promoter and 2 IL13 Promoter SNPs

[0165] The two IL4R promoter and the two IL13 promoter SNPs were genotyped using allele-specific PCR on a PE9700 thermal cycler (ABI) measuring SyBr Green (Molecular Probes) fluorescence (Higuchi, Fockler, Dollinger, Watson. Biotechnology 11:1026-30 (1993)). For each DNA, two ampifications were set up in parallel. One contained the common primer and one allele-specific primer; the other contained the common primer and the other allele-specific primer. The primers used to genotype the two IL4R promoter SNPs are provided in Table 31 and the primers used to genotype the two IL13 promoter SNPs are provided in Table 32. The amplification of the DNA with a particular allele-specific primer indicated the presence of the corresponding allele. An increase in the fluorescence of SyBr Green was indicative of the accumulation of amplification product. One of skill in the art will be able to correlate the change in fluorescence with the presence or absence of amplification product, and thus, the presence or absence of the corresponding allele.

[0166] The PCR amplification was carried out in a total reaction volume of 100 .mu.l containing the following reagents: TABLE-US-00013 Volume (.mu.l) Component 1 1 M Tris pH 8.0 12 25 mM MgCl2 1 100X dNTPs (50 mM each dA, dC, dG; 25 mM dT; 75 mM dU) 1 20X SYBR Green 1 200 .mu.M ROX 2 1 U/.mu.l UNG 2.5 80% Glycerol 4 100% DMSO 1 20 .mu.M primer 1 1 20 .mu.M primer 2 1 12 U/.mu.l CEA2 Gold DNA Polymerase* 2 10 ng/.mu.l genomic DNA to 100 .mu.l sterile water *Developed and manufactured by Roche Molecular Systems. Alternatively, 1 .mu.l of AmpliTaq Gold DNA Polymerase, also developed and manufactured by Roche Molecular Systems and sold commercially by Applied Biosystems, Inc, (Foster City, Calif.), can be used.

[0167] Amplification was carried out in a GeneAmp.TM. PCR System 9600 thermal cycler (Applera, Foster City, Calif.), using the specific temperature cycling profile shown below. TABLE-US-00014 2 minutes at 50.degree. C. Pre-reaction incubation: 12 minutes at 94.degree. C. 50 cycles: denature: 20 seconds at 95.degree. C. anneal: 20 seconds at 58.degree. C. Final extension: 5 minutes at 72.degree. C.

8 EXAMPLE 2

Association of IL4R SNPs with Type 1 Diabetes in HBDI Families

[0168] This example demonstrates the association of IL4R SNPs with type 1 diabetes in HBDI families.

[0169] IL4R genotyping was carried out on individuals from 282 Caucasian families ascertained because they contained two offspring affected with type 1 diabetes. The IL4R genotypes of all individuals were determined. IL4R genotyping was carried out using a genotyping method essentially as described in Example 1. In addition to the 564 offspring (2 siblings in each of 282 families) in the affected sibling pairs on which ascertainment was based, there were 26 other affected children. There were 270 unaffected offspring among these families.

[0170] The family-based samples were provided as purified genomic DNA from the Human Biological Data Interchange (HBDI), which is a repository for cell lines from families affected with type 1 diabetes. All of the HBDI families used in this study are nuclear families with unaffected parents (genetically unrelated) and at least two affected siblings. These samples are described further in Noble et al., 1996, Am. J. Hum. Genet. 59:1134-1148, incorporated herein by reference.

[0171] It is known that the HLA genotype can have a significant effect, either increased or decreased depending on the genotype, on the risk for type 1 diabetes. In particular, individuals with the HLA DR genotype DR3-DQB1*0201/DR4-DQB1*0302 (referred to as DR3/DR4 below) appear to be at the highest risk for type 1 diabetes (see Noble et al., 1996, Am. J. Hum. Genet., 59:1134-1148, incorporated herein by reference). These high-risk individuals have about a 1 in 15 chance of being affected with type 1 diabetes. Because of the strong effect of this genotype on the likelihood of type 1 diabetes, the presence of the DR3-DQB1*0201/DR4-DQB1*0302 genotype could mask the contribution from the IL4R allelic variants.

[0172] Individuals within these families also were genotyped at the HLA DRB1 and DQB1 loci. Of the affected sibling pairs, both siblings have the DR3/DR4 genotype in 90 families. Neither affected sibling has the DR 3/4 genotype in 144 families. Exactly one of the affected pair has the DR 3/4 genotype in the remaining 48 families.

Statistical Analysis, Methods and Algorithms

[0173] Since the eight SNPs in IL4R are both physically and genetically very closely linked to each other, the presence of a particular allele at a particular SNP is correlated with the presence of another particular allele at a nearby SNP. This non-random association of two or more SNPs' alleles is known as linkage disequilibrium (LD).

[0174] Linkage disequilibrium among the eight IL4R SNPs was assessed using the genotypes of the 282 pairs of parents. These 564 individuals are not related to each other except by marriage. A summary of the calculated frequency of the WT allele for each SNP in this group of 564 individuals (the "HBDI founders") is shown in Table 9.

[0175] The calculation of LD can be performed in several ways. Two complementary methods to assess LD between all pairs of IL4R SNP loci were used. In the first method, the values of two distinct but related metrics for LD, namely D and .DELTA. (Devlin and Risch 1995, Genomics, 29(2): 311-22), using the Maximum Likelihood Estimation algorithm of Hill (Hill, 1974, Heredity, 33(2): 229-39) were calculated. The values for D and .DELTA. for all pairs of IL4R SNPs are shown in Table 10, in the lower left triangular portion. Both D and .DELTA. can have values that range between -1 and +1. Values near +1 or -1 suggest strong linkage disequilibrium; values near zero indicate the absence of LD.

[0176] A second measure of LD uses a permutation test method implemented in the Arlequin program (Excoffier et al., 1995, Mol Biol Evol, 12:921-7, University of Geneva, CH) (Slatkin et al., 1996, Heredity, 76:377-83). This method maximizes the likelihood ratio statistic (S=-2log (L.sub.H*/L.sub.H)) by permuting alleles and recalculating S over a large number of iterations until S is maximized. These iterations allow the determination of the null distribution of S, and thus the maximum S obtained can be converted into an exact P-value (significance level). These P-values are listed in the upper right triangular portion of Table 10.

[0177] Table 10 of pairwise LD shows that there is significant evidence for LD between SNPs 1 and 2, and among (all combinations of) SNPs 3, 4, 5, 6, 7 and 8. SNPs 3 through 8 are known to exist within 1200 basepairs of each other in a single exon (exon 9) of the IL4R gene, and the LD between these SNPs is evidence for very small genetic distances as well.

[0178] The Transmission Disequilibrium Test (TDT) of Spielman (Spielman and Ewens, 1996, Am J Hum Genet, 59(5): 983-9; Spielman and Ewens, 1998, Am J Hum Genet, 62(2): 450-8) was performed on the IL4R genotype data for the 282 affected sib pairs (namely, a family structure consisting of the two parents and the two affected children). The TDT was used to test for the association of the individual alleles of the eight IL4R SNPs to type 1 diabetes. The TDT assesses whether an allele is transmitted from heterozygous parents to their affected children at a frequency that is significantly different than expected by chance. Under the null hypothesis of no association of an allele with disease, a heterozygous parent will transmit or will not transmit an allele with equal frequency to an affected child. The significance of deviation from the null hypothesis can be assessed using the McNemar chi-squared test statistic (=(T-NT) 2/(T+NT), where T is the observed number of transmissions and NT is the observed number of non-transmissions). The significance (P-value) of the McNemar chi-squared test statistic is equal to the Pearson chi-squared statistic with one degree of freedom (Glantz et al., Primer of biostatistics., New York, McGraw-Hill Health Professions Division, 1997).

[0179] The results of the single SNP locus TDT results are shown in tables 11A and 11B. The TDT/S-TDT program (version 1.1) of Spielman was used to perform the counting of transmitted and non-transmitted alleles (Spielman, McGinnis et al., 1993, Am J Hum Genet, 52(3): 506-16; Spielman and Ewens, 1998, supra). The table lists the observed transmissions of the wildtype allele at each SNP locus. Since these are biallelic polymorphisms, the transmission counts of the variant allele are equal to the non-transmissions of the wildtype allele.

[0180] The counts of transmissions and non-transmissions of alleles to the probands only shown in Table 11A do not quite reach statistical significance, at a=0.05. However, it is valid to count transmission events to all affected children. However, when the TDT is used in this way (or, for that matter, with more than one child per family), then a significant test statistic is evidence of linkage only, not of association and linkage. Table 11B shows the TDT analysis when 26 additional affected children are included. The results presented in Table 11B below show that there is a significant deviation from the expected transmission frequencies for alleles of SNPs 3, 4, 5 and 6. Inspection of the "% transmission" values for these SNPs indicates that the wildtype allele is transmitted to affected children at frequencies greater than the expectation of 50%.

[0181] The evidence for strong LD among the eight IL4R SNPs suggested that the transmission of the ordered set of alleles from each parent to each affected child in the HBDI cohort could be detected. This ordered set of alleles corresponds physically to one of the two parental chromosomes, and is called a haplotype. By inferring the parental haplotypes and their transmission or non-transmission to affected children, more statistical information is expected to be obtained than that from alleles alone.

[0182] Haplotypes were inferred using a combination of two methods. As the first step, the GeneHunter program (Falling Rain Genomics, Palo Alto, Calif.) (Kruglyak, et al., 1996, Am J Hum Genet, 58(6): 1347-63) was used as it very rapidly calculates haplotypes from genotype data from pedigrees. Each HBDI family pedigree was then inspected individually using the Cyrillic program (Cherwell Scientific Publishing, Palo Alto, Calif.), to resolve any ambiguous or unsupported haplotype assignments. Unambiguous and non-recombinant haplotypes could be confidently assigned in all but six of the 282 families. The haplotype data for these 276 families were used in subsequent data analysis.

[0183] The IL4R gene has the property that many of the SNPs reside within the 3'-most exon (exon 9), whose coding region is approximately 1.5 kb long. A method was developed for directly haplotyping up to five of these exon 9 alleles (namely, SNPs #3-7) without needing parental genotypes. As many of these SNPs direct changes to the amino acid sequence of the IL4R protein, different haplotypes encode different proteins with likely different functions.

[0184] Haplotypes, in an individual for which no parental genotypic information is known, can be inferred unambiguously only when at most one of the SNP sites of those is heterozygous. In other cases, the ambiguity must be resolved experimentally.

[0185] Two allele-specific primers with one common primer to perform PCR reactions (using Stoffel Gold.TM. polymerase) to separately amplify the DNA from each chromosome, as shown in FIG. 1 below were used. The alleles on each amplicon were then detected by the same strip hybridization procedure, and the linked alleles called directly. The choice of allele-specific (colored or shaded arrows) and common (black arrows) primers depends on which SNP loci are heterozygous. The primers were modified at the 5' phosphate by conjugation with biotin, and are shown in Table 12 (SEQ ID NO: 54-62).

[0186] For each haplotyping assay, two PCR reactions were set up for each DNA to be tested. One reaction contained the common primer and the wildtype allele-specific primer, the other contained the common primer and the variant allele-specific primer. Each PCR reaction was made in a total reaction volume of 50-100 .mu.l containing the following reagents: [0187] 0.2 ng/ml purified human genomic DNA [0188] 0.2 mM each primer [0189] 800 mM total dNTP (200 mM each dATP, dTTP, dCTP, dGTP) [0190] 10 mM KCl [0191] 10 mM Tris-HCl, pH 8.0 [0192] 2.5 mM MgCl.sub.2 [0193] 0.12 units/ml Stoffel Gold.TM. DNA polymerase* *developed and manufactured by Roche Molecular Systems.

[0194] Amplification was carried out in a GeneAmp.TM. PCR System 9600 thermal cycler (PE Biosystems, Foster City, Calif.), using the specific temperature cycling profile shown below: TABLE-US-00015 Pre-reaction incubation: 94.degree. C. for 12.5 minutes 33 cycles: Denature: 95.degree. C., 45 seconds Anneal: 64.degree. C., 30 seconds Extend: 72.degree. C., 45 seconds Final Extension: 72.degree. C., 7 minutes. Hold: 10.degree. C.-15.degree. C.

[0195] Following amplification, each PCR product reaction was denatured and separately used for hybridization to the membrane-bound probes as described above.

Haplotype Sharing in Affected Sibs

[0196] Evidence for linkage of IL4R to type 1 diabetes (as opposed to association) can be assessed by the haplotype sharing method. This method assesses the distribution over all families of the number of chromosomes that are identical-by-descent (IBD) between the two affected siblings in each family. For example, if in a family, the father transmits the same one of his two IL4R haplotypes to both children, and the mother transmits the same one of her two IL4R haplotypes to both children, then the children are said to share two chromosomes IBD (or, to be IBD=2). If both parents transmit different IL4R haplotypes to their two children, the children are said to be IBD=0.

[0197] Under the null hypothesis of no linkage of IL4R to type 1 diabetes, the proportion of families IBD=0 is 25%, IBD=1 is 50% and IBD=2 is 25%, as expected by random assortment (see Table 13). Evidence for a statistically significant difference from this expectation can be assessed using the chi-square statistic.

[0198] Identity-by-descent (IBD) values of parental IL4R haplotypes in the affected sibs could be determined unambiguously in 256 families. In the rest of the families, one or both parents were homozygous and/or the parental source of the child's chromosomes could not be determined. The distribution of IBD is shown in Table 13.

[0199] It is known that the HLA genotype can have a significant effect, either increased or decreased depending on the genotype, on the risk for type 1 diabetes. In particular, individuals with the HLA DR genotype DR3-DQB1*0201/DR4-DQB1*0302 (referred to as DR3/4 below) appear to be at the highest risk for type 1 diabetes (see Noble, Valdes et al., 1996), incorporated herein by reference). These high-risk individuals have about a 1 in 15 chance of being affected with type 1 diabetes. Because of the strong effect of this genotype on the likelihood of type 1 diabetes, the presence of the DR3/4 genotype could mask the contribution of IL4R alleles or haplotypes.

[0200] The distribution of IBD in families was stratified into two groups based on the DR3/4 genotype of the children. The first group contains the families in which one or both of the sibs are DR3/4 ("Either/both sib DR3/4", n=119). The second group contains the families where neither child is DR3/4 ("Neither sib DR3/4", n=137). The IBD distribution in these subgroups is shown in Table 13. There was no statistically significant departure from the expected distribution of IBD sharing in the "either/both sib DR3/4" subgroup of families. There is a statistically significant departure from the expected distribution of IBD sharing in the "neither sib DR3/4" subgroup of families (Table 13). This indicates that there is evidence for linkage of the IL4R loci to IDDM in the "neither sib DR3/4" families.

Association by AFBAC

[0201] Association of IL4R haplotypes with type 1 diabetes was assessed using the AFBAC (Affected Family Based Control) method (Thomson, G., 1995, Am J Hum Genet 57:487-98). In essence, two groups of haplotypes, and the haplotype frequencies in the groups, are compared with each other as in a case/control scheme of sampling. These two groups are the case (transmitted) and the control (AFBAC) haplotypes.

[0202] The case haplotypes, namely those transmitted to the affected children, were collected and counted as follows. For every pair of siblings, regardless of the status of the parents (homozygote or heterozygote) all four transmitted chromosomes were counted. However, the haplotypes in the two siblings in a pair are not independent of each other. The way to make a statistically conservative and valid enumeration is to divide all counts by two.

[0203] The control (AFBAC) haplotypes are those that are never transmitted to the affected pair of children (Thomson, 1995). The AFBAC haplotypes permit an unbiased estimate of control haplotype frequencies. AFBACs can only be determined from heterozygous parents, and furthermore, only when the parent transmits one haplotype to both children; the other, never-transmitted haplotype is counted in the AFBAC population. The AFBAC population serves as a well-matched set of control haplotypes for the study.

[0204] Table 14A shows the comparison of transmitted and AFBAC frequencies for all HBDI haplotypes that were observed at least five times in the complete sample set. Each row represents data on an individual haplotype. However, in all 16 distinct haplotypes were observed in the HBDI data set, although some very rarely. The seven rarest haplotypes are grouped together in the "others" row. Each haplotype is listed by the ordered set of alleles (namely, from SNPs 1-2-3-4-5-6-7-8) present at each of the eight IL4R SNPs as described in Table 6. A "1" denotes the presence of the reference allele, a "2" the presence of the variant allele for each SNP. The "reference" allele for each SNP is that present in GenBank Accession X52425.1 as described in Table 6.

[0205] Tables 14B and 14C show the comparison of transmitted and AFBAC frequencies for all HBDI haplotypes seen in the "either/both sib DR3/4" and the "neither sib DR3/4" subgroups of families, respectively. These tables show that stratifying the families based on the DR3/4 genotype of the children permits the identification of haplotypes that are associated with IDDM. In particular, in the "neither sib DR3/4" subgroup one haplotype (labeled "2 1 2 2 2 2 2 1") is significantly under represented in the pool of transmitted chromosomes (P<0.005).

[0206] From the transmitted and AFBAC haplotype frequency information in Tables 14B and 14C, one can derive by counting the frequencies of transmitted and AFBAC alleles. The locus-by-locus AFBAC analyses are shown in Tables 15A and 15B.

[0207] The data present in Tables 15A and 15B show that there statistically significant evidence, in the "neither sib DR3/4" subgroup of families, that alleles of SNPs numbers 3, 4 5, 6, and 7 are associated with IDDM. The evidence for association is especially strong for SNP #6. In the "either/both sib DR3/4" subgroup, there is the same trend of allelic association, although the trend does not quite reach statistical significance.

Association by Haplotype-Based TDT

[0208] The TDT analysis can be utilized for determining the transmission (or non-transmission) of 8-locus haplotypes from parents to affected children, once the haplotypes have been inferred or assigned by molecular means. Tables 16A, B, and C summarize the TDT results for the HBDI families. Each haplotype is listed by the ordered set of alleles (namely, from SNPs 1-2-3-4-5-6-7-8) present at each of the eight IL4R SNPs as described in Table 6. A "1" denotes the presence of the reference allele, a "2" the presence of the variant allele for each SNP. The "reference" allele for each SNP is that present in GenBank Accession X52425.1 as described in Table 6. Table 16A counts informative transmission events only to one child (the proband) per family, Table 16B counts informative transmissions to the two primary affected children per family, and Table 16C counts informative transmissions to all affected children. The 8-locus haplotype TDT results reach statistical significance when all affected children (2 or more per family) are included.

[0209] The TDT analyses can be performed on families after stratifying for the DR3/4 genotype of the children. The summary of counts of informative transmissions to the two primary affected children per family, in the "either/both sib DR3/4" and the "neither sib DR3/4" subgroups of families, are shown in tables 17A and 17B respectively. Each haplotype is listed by the ordered set of alleles (namely, from SNPs 1-2-3-4-5-6-7-8) present at each of the eight IL4R SNPs as described in Table 6. A "1 " denotes the presence of the reference allele, a "2" the presence of the variant allele for each SNP. The "reference" allele for each SNP is that present in GenBank Accession X52425.1 as described in Table 6.

[0210] As presented above, there is significant evidence of linkage of IDDM to IL4R in the "neither sib DR3/4" subgroup. The data in Table 17B indicate that there is significant evidence of association of IL4R haplotypes to IDDM, in the presence of this linkage. In particular, in the "neither sib DR3/4" subgroup one haplotype (labeled "2 1 2 2 2 2 2 1") is significantly under-transmitted to affected children.

9 EXAMPLE 3

Association of IL4R with Type 1 Diabetes in Filipino Samples

[0211] This example demonstrates the association of IL4R SNPs with type 1 diabetes in a Filipino population.

[0212] As used in this section, "patients" refers to individuals with the disease, namely individuals with type 1 diabetes and "controls" refers normal individuals, those without the disease.

[0213] Ninety patients (n=90) were selected for this study from amongst the Filipino population. The patients included in the study were affected by type 1 diabetes as defined by the recent ADA classification (the Expert Committee on the Diagnosis and Classification of Diabetes Mellitus 1997). The patients were born in the Philippines and all had two Filipino parents. These patients had been characterized for C-peptide levels below 0.3mmol/l and for autoantibodies to islet cell autoantigens (Medici et al., 1999, Diabetes Care, 9:1458-62). Samples were also collected from ninety-four Filipino normal subjects without a family history for diabetes. This was the control group. All patients and controls were from the southern region of Luzon, Philippines. The study was approved by the local Ethics Committee and informed consent was given by patients. In addition, independent samples from a previous study of HLA class II loci in Filipinos (Bugawan et al., 1994, Genetics, 54:331-340) originating from the same region were used, following a statistical test of heterogeneity, to supplement the control samples. These comprised a total of 194 chromosomes taken from family and individual samples.

[0214] The individuals were genotyped as described above. The genotypes of the affected and unaffected individuals are shown in Table 4 (SEQ ID NO: 20-24). Both the actual numbers and the frequencies are provided for each genotype. The data (Table 5) confirm the presence of an association of IL4R SNP variants with type 1 diabetes.

Statistical Methods & Algorithms

[0215] Allele and haplotype frequencies between groups were compared using the z-test. Haplotype compositions and frequencies were estimated from the genotype data using the EM algorithm in the Arlequin program (L. Excoffier, University of Geneva, CH) (Excoffier et al., 1995, Mol Biol Evol, 12:921-7; Slatkin et al., 1996, Heredity, 76:377-83).

Results

[0216] The wildtype allele frequencies for each of the eight IL4R SNPs in the Filipino control and diabetic groups are shown in Table 18. Table 18 provides evidence that the allele frequencies for SNPs #3 and 4 are significantly different between the two groups, and suggests an association to IDDM.

[0217] It is also possible to infer and construct the multi-locus IL4R haplotypes in the Filipino subjects, either computationally by Maximum-likelihood estimation (MLE), or by using molecular haplotyping methods described previously. Table 19 lists the five most frequent computationally estimated haplotypes and their frequencies in the Filipino diabetics and controls, and presents the significance of the differences in frequencies. Each haplotype is listed by the ordered set of alleles (namely, from SNPs 1-2-3-4-5-6-7-8) present at each of the eight IL4R SNPs as described in Table 6. A "1" denotes the presence of the reference allele, a "2" the presence of the variant allele for each SNP. The "reference" allele for each SNP is that present in GenBank Accession X52425.1 as described in Table 6.

[0218] Table 20 lists the observed haplotypes as derived and inferred by molecular haplotyping; the unambiguous seven-locus haplotypes (SNP#1 allele not shown, as indicated by the "x") are compiled. Each haplotype is listed by the ordered set of alleles (namely, from SNPs 1-2-3-4-5-6-7-8) present at each of the eight IL4R SNPs as described in Table 6. A "1" denotes the presence of the reference allele, a "2" the presence of the variant allele for each SNP. The "reference" allele for each SNP is that present in GenBank Accession X52425.1 as described in Table 6. Tables 18 and 19 both provide evidence of a statistically significant difference in the frequency of one or more haplotypes between the Filipino control and diabetic populations, and support the presence of an association of IL4R to IDDM. In particular, the haplotype (labeled "x 1 2 2 2 2 2 1") is significantly under represented in the Filipino diabetics group.

10. EXAMPLE 4

Association of IL4R, IL4 and IL13 SNPs with Type 1 Diabetes in Filipino Samples

[0219] This example demonstrates the association of IL4R, IL4 and IL13 SNPs with type 1 diabetes in the same Filipino population as described above, in Example 3.

[0220] As used in this section, "patients" refers to individuals with the disease, namely individuals with type 1 diabetes and "controls" refers normal individuals, those without the disease.

[0221] The individuals were genotyped as described above.

[0222] Individual SNPs

[0223] The distributions of alleles at the individual SNPs in the IL4R locus (n=10), the IL4 locus (n=1) and the IL13 locus (n=4) among patients and controls are shown in Table 21. Linkage disequilibrium patterns were estimated using maximum likelihood approaches from individual genotype data from unrelated individuals (Slatkin and Excoffier, 1996 Heredity 76:377:383). The patterns of pairwise linkage disequilibrium (LD) for these SNPs inferred among the control population are shown in Tables 22 and 23. Among the individual IL4R SNPs, three (E375A, L389L, and C406R) showed a nominally significant association with type 1 diabetes while the variant allele at two additional SNPs (I50V, p=0.062 and S478G, p=0.064) was decreased among patients (Table 21).

[0224] The two promoter SNPs were not significantly associated with type 1 diabetes, although the variant allele of the -3223 SNP was slightly increased among patients (OR=1.45, p=0.10). With the exception of this promoter SNP and the I50V SNP, with which it is in strong LD, the variant allele at each SNP was under represented among patients. Some of the polymorphic amino acid residues in this chain appear to be biologically important and affect IL-4 receptor signaling (Kruse et al., 1999, Immunology, 96:365-71).

[0225] Of the 10 IL4R SNPs typed, the L389L SNP showed the strongest association with type 1 diabetes in this population, with significantly lower frequencies among patients than controls (OR=0.34; p=0.001). Without being bound by theory, it is believed that because this is a silent (synonymous) polymorphism, it is unlikely that this SNP is responsible for the observed protective effect for type 1 diabetes. This SNP is in very strong LD (Table 22) with the nonsynonymous flanking SNPs (E375A, C406R, S478P and Q551R) and that these SNPs all show a trend toward protection (negative association). The L389L SNP is also in strong negative LD with the -3223 promoter SNP (Table 22).

[0226] In the comparison of genotypes at the individual IL4R SNPs (Table 24), the protective effect is dominant in that the heterozygote for IL4R 389 has an OR=0.29. Among the individual SNPs on chromosome 5q31, only the variant alleles at the two IL13 promoter SNPs were increased among patients (OR=1.58 and p =0.05 for -1512 and OR=1.49 and p=0.12 for -1112) (Table 21) When genotype frequencies are compared, however, the IL13 R 1000Q showed a nominally significant association in this population (p=0.03; Table 24). These data suggest that the variant homozygote, but not the heterozygote, may be at increased risk for type 1 diabetes. In Table 24, each haplotype is listed by the ordered set of alleles (namely, from SNPs 1-2-3-4-5-6-7-8) present at each of the eight IL4R SNPs as described in Table 6. The letters refer to the actual allele (nucleotide) present, as described in Table 6.

[0227] Haplotypes

[0228] IL4R

[0229] IL4R haplotypes were estimated based on an expectation-maximization (EM) method (Excoffier et al., 1995, Mol. Biol. Evol., 12:921-927.) and were directly determined by molecular haplotyping methods, described in Example 4. The molecular haplotyping method allowed the unambiguous assignment of phase for 7 IL4R SNPs (C676T, A1374C, G1417T, T1466C, T1682C, A1902G and T2531C). Using molecular haplotyping of these 7 SNPs, 7 different IL4R haplotypes were determined in this population and their frequencies among patients and controls compared (Table 25A). In Table 25A each haplotype is listed by the ordered set of alleles (namely, from SNPs 2-3-4-5-6-7-8) present at seven of the eight IL4R SNPs as described in Table 6. The letters refer to the actual allele (nucleotide) present, as described in Table 6.

[0230] One specific haplotype (CCTCCGT)) was significantly under represented among patients (OR=0.4, p=0.013). This same haplotype was also found to be protective (significant negative association), by the TDT methods, in the HBDI families, as described in Example 2. This protective effect has thus been observed in two different populations and in two different study designs, namely case/control (Filipino) and TDT, in addition to the biological plausibility (i.e., functional consequences) of these SNPs. This strongly suggests that variants of the IL4R molecule influence the susceptibility to type 1 diabetes. In particular, this specific haplotype of IL4R appears to confer a dominant protective effect.

[0231] In the HBDI families, stratification based on the highest risk HLA genotype (HLA-DRB1*0301-DQB1*0201/HLA-DRB1 *04-DQB1*0302) was necessary to demonstrate the protective effect of the IL4R haplotype. A significant negative association was found only among those families in which neither affected sib was DR3/4, presumably because the effect of the IL4R polymorphism was relatively modest compared to the risk conferred by this high risk HLA genotype, which confers a disease risk higher than DR3/3 or DR4/4 homozygotes. Among Filipinos, a significant protective effect of a specific IL4R haplotype was observed without stratification (Table 25). Without being bound by theory, this may reflect the absence among Filipinos of a higher risk associated with DR3/4 than with DR3/3 or DR4/4 genotypes. The lack of the "DR3/4 effect," well-established by many studies of Caucasian type 1 diabetes, can be attributed among Filipinos to the differing patterns of linkage disequilibrium of DQB1 alleles with DRB1*04 alleles between Asians and Caucasians.

[0232] The molecular haplotyping approach used in this example did not assign phase for the two promoter SNPs and the I50V SNP in the IL4R locus. Consequently, the EM approach was used to estimate frequencies for 10-SNP haplotypes for these 3 individual SNPs and the 7-SNP haplotype previously determined by molecular methods (Table 25B). In Table 25B, each haplotype is listed by the ordered set of alleles (namely, from SNPs 1-2-3-4-5-6-7-8-9-10) present at each of the ten IL4R SNPs described in Table 21 (SNPs in order: C(-3223)T-C(-1914)T-I50V-N142N-E375A-L389L-C406R-S478P-Q551R-S761P). The letters refer to the actual allele (nucleotide) present, as described in Table 21.

[0233] Of the 17 10-SNP haplotypes with an estimated frequency >1%, in either group, only one 10-SNP haplotype containing the protective 7-SNP haplotype (H5A or CCA-CCTCCGT) appeared strongly negatively associated (OR=0.0; 95%CI [0-0.5]; p=0.001) with disease. Interestingly, the other haplotype which contained the same 7-SNP haplotype (H5B or CTA-CCTCCGT) was not significantly associated with disease (OR=0.66 p=0.33). This suggests that a specific combination of IL4R promoter SNPs with a particular coding sequence variant contributes to the risk for type 1 diabetes.

[0234] IL4 and IL13

[0235] The IL4 and the IL13 SNPs are in strong LD (Table 23). The estimated frequencies for these 5-SNP haplotypes were compared among patients and controls (Table 26). In Table 26, each haplotype is listed by the ordered set of alleles present at the one IL4 SNP and the four IL13 SNPs as described in Table 21. The order is IL4 C(-524)T-IL13 A(-1512)C-IL13 C(-1112)T-IL13 intron3-IL13 R110Q. The letters refer to the actual allele (nucleotide) present, as described in Table 21.

[0236] The overall distributions were different (p=0.005) and one haplotype, TCTTA, was strongly associated with type 1 diabetes (OR=3.47, p=0.004). Two other haplotypes showed a nominally significant association (p=0.02 and 0.03). One surprising observation was that the IL13 haplotype CTTA appeared to be associated with disease only in combination with the T allele at the IL4 -524 promoter SNP because the CCTTA haplotype showed no disease association. These data could reflect LD between the associated 5-SNP haplotype with some nearby causal gene or suggest that a particular combination of a promoter variant at IL4 and promoter and coding variants at IL13 are responsible for an elevated type 1 diabetes risk (gene-gene interaction).

13. EXAMPLE 7

Dependence of Risk for Type 1 Diabetes Conferred by IL4R SNPs on Genotype at IL4 and IL13 in Filipino Samples: Evidence for Epistasis

[0237] This example demonstrates epistasis, the interaction between SNPs on the IL4R locus on chromosome 16 and those on the IL4 and IL13 loci on chromosome 5.

[0238] Because IL4 and IL13 both serve as ligands for a receptor composed, in part, of the IL4R alpha chain, there is a likelihood of gene-gene interactions between polymorphisms in the IL4R locus on chromosome 16p11 and the five SNPs in the IL4 and IL13 loci on chromosome 5q31. In one approach, the statistical independence for genotypes at the 10 IL4R SNPs and the genotypes at each of the IL4 and IL13 SNPs (Table 27) was examined. Gene by gene interactions at SNPs in different genes were evaluated by assessing whether the genotype frequencies at unlinked loci were independent (i.e., the IL13 and IL4 SNPs on chromosome 5 and the IL4R SNPs on chromosome 16) among patients. These analyses were done for each pair of unlinked SNPs carrying out a chi-square test in contingency tables with marginals defined by genotype counts either in patients or controls. The chi-square values and the corresponding degrees of freedom for each IL4R SNP comparisons were summed and p-value of the sum of chi-squares computed.

[0239] No deviation from independence was found for these SNPs among controls but a significant deviation was found for the IL4 -524 promoter SNP (p=0.001) and the IL13 intron 3 SNP (p=0.019) among patients.

[0240] To assess whether the effect on type 1 diabetes susceptibility due to IL4R SNPs was modified by the IL4 or the IL13 SNPs, epistasis was modeled using a logistic regression model (see below). For each of the five IL4 and IL13 SNPs, we tested whether the effect of the combined IL4R SNP genotypes on type 1 diabetes susceptibility differed depending on the IL4 and IL13 SNPs. The results (Table 28) indicate that there is indeed an epistatic interaction between the IL4R genotypes and IL4 and IL13 genotypes. To address the issue of multiple comparisons, we carried out permutation analysis on this test. In 22/200 permuations one or more of the 5-SNP tests showed a p<0.035, in 13/200 one or more of the SNP tests had p<0.035 and another one had p<0.075. In 9/200 one or more of the 5-SNP tests showed a p<0.035, another had a p<0.075 and another had a p<0.135. Thus, the pattern observed in Table 28 has a probability of p<0.045. The conclusion from this is that the epistatic interaction observed between the IL4 and IL13 SNPs and the IL4R genotypes is statistically significant indicating that, in this data set, the genotypes in the IL4, IL13 region affect the genetic susceptibility to type 1 diabetes conferred by IL4R.

[0241] To illustrate this interaction, the odds ratios for individual IL4R SNPs as a function of the IL4 and IL13 SNP genotype were also calculated. The differences among the Odds Ratios were greatest for the IL4R -3223 SNP and the four IL13 SNPs. The odds ratios with the 95% confidence intervals and the p values from the stratified contingency table analyses are shown in FIG. 2. FIG. 2 shows the Odds ratios, patient and control counts of specific IL4R and IL4, IL13 genotypes. FIG. 2A shows the interaction of IL4R Ala375Glu with and IL4 -524 (promoter). FIG. 2B shows the interaction of IL4R Gln551Arg with IL13 A-1512C. FIG. 2C shows the interaction between IL4R C-3223T and IL13 A-1512C and FIG. 2D shows the interaction between IL4R C-3223T and IL13 C-1112T. The p-value for the association of each genotype combination is shown above each odds ratios bar.

[0242] The most striking observation was that the IL4R -3223 SNP CT genotype had an OR of 8.55 (95% CI=1.05, 69.8) when present with the TT IL13 -1112 genotype and an OR=0.53 (95% CI=0.29, 0.98) when present with the CC genotype. Without being bound by theory, the observation of an interaction between polymorphisms in the IL13 and IL4 genes and polymorphism in the gene encoding the receptor for the products of these two genes represents an interesting and biologically plausible hypothesis that, given the multiple comparisons, requires further testing. A recently published study of asthma patients reported a gene-gene interaction between IL4R and IL13 in the determination of serum IgE levels (Howard et al., 2002, Am J Hum Genet, 70:230-6).

[0243] As described above, the IL4R association data obtained in this Filipino case control study indicate that the 7-SNP haplotype, composed primarily of variant alleles at these SNPs, confers dominant protection to type 1 diabetes (Table 25A), consistent with our recent observations, based on TDT and AFBAC analysis in a set of multiplex Caucasian families (the HBDI registry). The replication of this observation in two different populations and in two different study designs strengthens this inference. The analysis of 10-SNP IL4R haplotypes among Filipinos suggests that a specific promoter variant in combination with specific coding sequence variants may be responsible for the observed protection (Table 25B). Several recent studies have shown that the reference or wild-type allele at several of these IL4R SNPs is associated with atopic asthma and increased IgE levels (Howard et al., 2002, Am J Hum Genet, 70:230-6, Sandford et al., 2000, J Allergy Clin Immunol, 106:135-40). Thus, it appears that the same alleles at IL4R SNPs confer an increased risk to a canonical Th1 (type 1 diabetes) and a Th2 disease (atopic asthma). Without being bound by theory, these associations argue against an effect on Th1/Th2 balance mediated by polymorphism in the IL4R gene and suggest instead that this polymorphism may influence some aspect of immune regulation and homeostasis in Th1 and Th2 pathways and possibly, B cell activation. Conceivably, the observed patterns of disease association reflect the effect of IL4R polymorphisms on the balance between the activation of Th1 and Th2 cells and that of T regulatory cells. In conclusion, the extent of risk for type 1 diabetes may be determined by specific combinations of variants at the IL4R locus and at the genes encoding its two ligands, IL4 and IL13.

Calculations Performed to Achieve the Results of Table 28

[0244] For each IL4R SNP, the homozygote genotype with the highest odds ratio was given a value of 2, the heterozygote was given a value of 1, the other homozygote was 0. A logistic regression was carried out on nine IL4R polymorphisms (S761P did not show the variant) in this way and a new numerical variable "il4r" was derived given by: il4r=a.sub.1G.sub.-3223+a.sub.2G.sub.-1914+a.sub.3G.sub.50+a.sub.4G.sub.1- 42+a.sub.5G.sub.375+a.sub.6G.sub.389+a.sub.7G.sub.406+a.sub.8G.sub.478+a.s- ub.9G.sub.551 where Gi denotes the genotype (0, 1 or 2) at the ith position and aj denotes the coefficient fitted by logistic regression. The coefficients fitted by the regression were a1=0.368; a2=0.053; a3=0.37; a4=0.061; a5=0.66; a6=1.08; a7=0.57; a8=0.54 and a9=0.22. Epistasis was then tested independently for each of the five chromosome 5 SNPs by fitting the following logistic regression model: P(T1DM)=exp(X)/(1+exp(X)) Where X=C+.beta..sub.1il4r+.beta..sub.2G.sub.chr5i+.beta..sub.3(il4r-G.sub.chr5- i) and G.sub.chr5i is the genotype of one of the chromosome 5 SNPs (values 0, 1 or 2).

Permutation Analysis

[0245] Because five different SNPs were compared, it was important to correct for multiple tests. However, a Bonferroni or a Dunn-Sidak correction was not appropriate since the IL4 and IL13 SNPs are not independent (see Table 23). Therefore permutation analysis was carried out, keeping constant the patient and control genotype frequencies, but permutating the IL4 and IL13 genotypes and the IL4R genotypes within the patient and within the control groups separately. In this way, only the epistatic interaction between the two genetic regions was tested and not the individual IL4 or IL13 and IL4R genetic associations. 200 permutations were carried testing for epistasis at all five chromosome 5 SNPs each time. Analyses were carried out using S-Plus version 6.0 Professional (Insightful Corporation).

[0246] Various embodiments of the invention have been described. The descriptions and examples are intended to be illustrative of the invention and not limiting. Indeed, it will be apparent to those of skill in the art that modifications may be made to the various embodiments of the invention described without departing from the spirit of the invention or scope of the appended claims set forth below.

[0247] All references cited herein are hereby incorporated by reference in their entireties. TABLE-US-00016 TABLE 3 Variant Probe Probe Sequence Seq ID No: titer (.mu.l) A398 DBM0081P CCACACGTGTATCCCTGAGAA 3 1.0 398G DBM0082P TCTCAGGGACACACGTGTG 4 4.0 C676 DBM0172P TGGAGTGAAAACGACCCGGCAG 5 1.0 676T DBM0073P CTGCCGGGTCATTTTCGCTCC 6 1.0 A1374 DBM0043RC GAGGGAAGGGAGGGCATTGTG 7 1.0 1374C DBM0139P AGGGAAGGGCGGGCATTGT 8 4.0 G1417 DBM0124P CTCTCCGAGCAGGTCCAGG 9 1.0 1417T DBM0046RC TCCTGGACCTTCTCGGAGAGG 10 1.0 T1466 DBM0076P AAGGTGGAAGAAGGCATGACTCC 11 1.0 DBM0132P AAGGTGGGAGAAGACATGACTCC 12 1.0 1466C DBM0171P GGAGTCACGTCTTCTCCTACCTT 13 2.0 T1682 DBM0157P TGGCTCAGAGAGTTGCTGAAGC 14 2.0 1682C DBM0094P TTCAGCAACCCCCTGAGCC 15 2.0 A1902 KW86 AGTGGCTATCAGGAGTTTGT 16 1.6 1902G KW85 AGTGGCTATCGGGAGTTTGT 17 0.8 T2531 DBM0080P CTCTTCTCTGAGATGCCCGAG 18 0.5 2531C DBM0048RC CTCGGGCATCCCAGAGAAGAG 19 0.5

[0248] TABLE-US-00017 TABLE 4 Computationally estimated haplotype frequencies compared between Filipino controls and diabetics. Seq MLE MLE COUNT: COUNT: Total exp exp haplotype ID No: label frequency frequency O.R. control diabetics count control diabetic chi-square P-value ACATTTAT 20 H-1 0.318 0.329 1.1 59.7 58.5 118.2 60.7 57.5 0.033 0.855 ACAGTTGT 21 H-6 0.074 0.074 1.0 14.0 13.2 27.2 14.0 13.2 0.000 0.987 22 H-3 0.145 0.058 0.4 27.3 10.3 37.6 19.3 18.3 6.727 0.009 GCAGTTAT 23 H-2 0.390 0.441 1.2 73.3 78.5 151.8 78.0 73.8 0.582 0.445 GCCGCCGT 24 H-10 0.032 0.051 1.6 6.0 9.0 15.0 7.7 7.3 0.776 0.378 n = 188 178 366 overall 8.118 0.0987

[0249] TABLE-US-00018 TABLE 5 Affected Genotypes Control Genotypes SNP (n = 89) (n = 94) A398G AA = 21 AG = 40 GG = 28 AA = 32 AG = 41 GG = 21 (50 I/V) (0.236) (0.449) (0.315) (0.340) (0.436) (0.223) C676G CC = 89 CC = 92 CG = 2 (142 N/N) (1) (0.979) (0.021) A1374C AA = 70 AC = 17 CC = 2 AA = 55 AC = 30 CC = 3 (375 E/A) (0.787) (0.191) (0.023) (0.630) (0.341) (0.034) G1417T GG = 78 GT = 10 TT = 1 GG = 63 GT = 29 TT = 2 (389 L/L) (0.876) (0.112) (0.011) (0.670) (0.309) (0.022) T1466C TT = 70 TC = 17 CC = 2 TT = 60 TC = 32 CC = 2 (406 C/R) (0.787) (0.191) (0.023) (0.638) (0.340) (0.022) T1682C TT = 70 TC = 17 CC = 2 TT = 61 TC = 31 CC = 2 (478 S/P) (0.787) (0.191) (0.023) (0.649) (0.330) (0.022) A1902G AA = 50 AG = 35 GG = 3 AA = 50 AG = 36 GG = 8 (551 Q/R) (0.562) (0.393) (0.034) (0.532) (0.383) (0.085) T2531C TT = 89 TT = 94 (761 S/P) (1) (1)

[0250] TABLE-US-00019 TABLE 6 SNPs detected Genbank SNP # LOCUS SNP Variation Access # 1 IL4R A398G I50V X52425 2 IL4R C676T N142N X52425 3 IL4R A1374C E375A X52425 4 IL4R G1417T L389L X52425 5 IL4R T1466C C406R X52425 6 IL4R T1682C S478P X52425 7 IL4R A1902G Q551R X52425 8 IL4R T2531C S761P X52425

[0251] TABLE-US-00020 TABLE 7 Amplicon primers and lengths Amplicon Seq Amplicon Seq Amplicon Left Primer ID Right Primer ID SNP # size, bp Name Left primer sequence No: Name Right primer sequence No: 1 114 RR192B CAGCCCCTGTGTCTGCAGA 25 RR193B GTCCAGTGTATAGTTATCCGCACTGA 31 2 163 DBM0177B CTGACCTGGAGCAACCCGTA 26 DBM0178B ACTGGGCCTCTGCTGGTCA 32 3, 4, 5 228 DBM0023B ATTGTGTGAGGAGGAGGAGGAGGTA 27 DBM0022B GTTGGGCATGTGAGCACTCGTA 33 6 129 DBM0097B CTCGTCATCGCAGGCAA 28 DBM0098B AGGGCATCTCGGGTTCTA 34 7 198 RR200B GCCGAAATGTCCTCCAGCA 29 RR178B CCACATTTCTCTGGGGACACA 35 8 177 DBM0112B CCGGCCTCCCTGGCA 30 DBM0071B GCAGACTCAGCAACAAGAGG 36

[0252] TABLE-US-00021 TABLE 8 Hybridization probes and titers Variant allele Seq WT allele Variant Seq probe SNP WT allele WT allele ID probe allele Varient allele probe ID titer # Probe name probe sequence No: titer (.mu.M) probe name sequence No: (.mu.M) 1 DBM0081P CCACACGTGTATCCCTGAGAA 37 1.0 DBM0082P TCTCAGGGACACACGTGTG 46 4.0 2 DBM0172P TGGAGTGAAAACGACCCGGCAG 38 1.0 DBM0073P CTGCCGGGTCATTTTCGCTCC 47 1.0 3 DBM0043RC GAGGGAAGGGAGGGCATTGTG 39 1.0 DBM0139P AGGGAAGGGCGGGCATTGT 48 4.0 4 DBM0124P CTCTCCGAGCAGGTCCAGG 40 1.0 DBM0046RC TCCTGGACCTTCTCGGAGAGG 49 1.0 5 DBM0076P + AAGGTGGAAGAAGGCATGACTCC 41 1.0 DBM0171P GGAGTCACGTCTTCTCCTACCTT 50 2.0 DBM0132P AAGGTGGGAGAAGACATGACTCC 42 1.0 6 DBM0157P TGGCTCAGAGAGTTGCTGAAGC 43 2.0 DBM0094P TTCAGCAACCCCCTGAGCC 51 2.0 7 KW86 AGTGGCTATCAGGAGTTTGT 44 1.6 KW85 AGTGGCTATCGGGAGTTTGT 52 0.8 8 DBM0080P CTCTTCTCTGAGATGCCCGAG 45 0.5 DBM0048RC CTCGGGCATCCCAGAGAAGAG 53 0.5

[0253] TABLE-US-00022 TABLE 9 Allele Frequency Of wildtype allele in HBDI founders WT Allele SNP # Frequency 1 0.53 2 0.90 3 0.89 4 0.89 5 0.89 6 0.83 7 0.79 8 0.99

[0254] TABLE-US-00023 TABLE 10 Pairwise Linkage Disequilibrium Measures ##STR1## ##STR2## ##STR3##

[0255] TABLE-US-00024 TABLE 11A Results of single locus TDT analysis, to the 282 probands only Transmissions of Non-transmissions McNemar chi- WT allele to of WT allele to Total observed squared SNP # affected (T) affected (NT) Transmissions (T + NT) statistic Significance 1 144 138 282 0.128 N.S. 2 55 39 94 2.723 N.S. 3 62 46 108 2.370 N.S. 4 62 46 108 2.370 N.S. 5 61 46 107 2.103 N.S. 6 90 67 157 3.369 N.S. 7 103 81 184 2.630 N.S. 8 5 7 12 0.333 N.S.

[0256] TABLE-US-00025 TABLE 11B Results of single locus TDT analysis, to 564 primary affected and 26 additional children Non- Transmissions of transmissions of Total observed McNemar WT allele to WT allele to Transmissions chi-squared Transmission SNP # affected (T) affected (NT) (T + NT) statistic Significance T/(T + NT) 1 311 280 591 1.626 N.S. 2 107 90 197 1.467 N.S. 3 130 98 228 4.491 P < 0.05 57.0% 4 130 98 228 4.491 P < 0.05 57.0% 5 128 98 226 3.982 P < 0.05 56.6% 6 189 149 338 4.734 P < 0.05 55.9% 7 212 180 392 2.612 N.S. 8 12 13 25 0.040 N.S.

[0257] TABLE-US-00026 TABLE 12 Allele specific PCR primers and amplicon lengths SNP detected Other SNPs Wildtype Allele- allele- Amplicon spanned by Common Specific Primer specifically Size. bp amplicon Primer Name Common Primer Sequences Seq ID No: Sequence 3 678 4, 5, 6, 7 RR178B CCACATTTCTCTGGGGACACA 54 DBM0149B 7 639 3, 4, 5, 6 DBM0023B ATTGTGTGAGGAGGAGGAGGAGGTA 55 DBN9155B 6 413 3, 4, 5 DBM0023B ATTGTGTGAGGAGGAGGAGGAGGTA 56 DBM0194B SNP detected Variant Allele- allele- Wildtype Allele-specific Primr Specific Primer Variant Allele-specific Primer specifically Sequence Seq ID No: Name Sequences Seq ID No: 3 TTCCAGGAGGGAAGGGA 57 DBM0150B TTCCAGGAGGGAAGGGC 60 7 CACCGCATGTACAAACTCCT 58 DBM0156B CACCGCATGTACAAACTCCC 61 6 GGTGACTGGCTCAGGGA 59 DBM0195B GGTGACTGGCTCAGGGG 62

[0258] TABLE-US-00027 TABLE 13 IBD Distributions for IL4R haplotypes Group (n) IBD = 2 IBD = 1 IBD = 0 P-value All families (256) 30.7% 47.9% 21.5% N.S. Either/both sib DR3/4 (119) 26.1% 50.4% 23.5% N.S. Neither sib DR3/4 (137) 34.7% 45.6% 19.7% 0.03 Expectation 25% 50% 25%

[0259] TABLE-US-00028 TABLE 14A Haplotype transmissions by AFBAC method: all families Counts Observed Frequency Chi-squared IL4R 8-locus Haplotype Transmitted AFBAC Total Transmitted AFBAC Statistic p-value 1 1 1 1 1 1 1 1 259.5 109.5 369 46.3% 41.8% 0.82 0.36 2 1 1 1 1 1 1 1 170.5 71.5 242 30.4% 27.3% 0.60 0.44 2 1 2 2 2 2 2 1 29.5 23.5 53 5.3% 9.0% 3.79 0.05 2 2 1 1 1 1 1 1 23.5 13 36.5 4.2% 5.0% 0.24 0.63 1 1 2 2 2 2 2 1 23 12.5 35.5 4.1% 4.8% 0.18 0.67 2 2 1 1 1 2 2 1 20.5 14 34.5 3.7% 5.3% 1.20 0.27 1 1 1 1 1 1 2 1 19 9 28 3.4% 3.4% 0.00 0.98 2 1 1 1 1 1 2 1 3.5 3 6.5 0.6% 1.1% 0.61 0.43 2 2 1 1 1 2 2 2 4.5 2 6.5 0.8% 0.8% 0.00 0.95 Others 6.5 4 10.5 1.2% 1.5% 0.19 0.67 TOTAL 560 262 822 7.64 0.94

[0260] TABLE-US-00029 TABLE 14B Haplotype transmissions by AFBAC method: "Either/both sib DR3/4" subgroup of familes Counts Observed Frequency Chi-squared IL4R 8-locus Haplotype Transmitted AFBAC Total Transmitted AFBAC Statistic p-value 1 1 1 1 1 1 1 1 254 50.5 304.5 46.7% 41.6% 0.57 0.45 2 1 1 1 1 1 1 1 156 30 186 28.7% 24.7% 0.56 0.45 2 1 2 2 2 2 2 1 35 11 46 6.4% 9.1% 0.99 0.32 2 2 1 1 1 1 1 1 24 7 31 4.4% 5.8% 0.39 0.53 1 1 1 1 1 1 2 1 20 6 26 3.7% 4.9% 0.40 0.52 1 1 2 2 2 2 2 1 18 7 25 3.3% 5.8% 1.59 0.21 2 2 1 1 1 2 2 1 18 4 22 3.3% 3.3% 0.00 0.99 2 2 1 1 1 2 2 2 8 2 10 1.5% 1.6% 0.02 0.89 2 2 1 1 1 2 2 2 8 1 9 1.5% 0.8% 0.31 0.58 2 1 1 1 1 1 2 1 1 2 3 0.2% 1.6% 4.71 0.03 2 1 1 1 1 2 2 1 2 1 3 0.4% 0.8% 0.46 0.50 TOTAL 544 121.5 665.5 10.00 0.44

[0261] TABLE-US-00030 TABLE 14C Haplotype transmissions by AFBAC method: "neither sib DR3/4" subgroup of families Counts Observed Frequency Chi-squared IL4R 8-locus Haplotype Transmitted AFBAC Total Transmitted AFBAC Statistic p-value 1 1 1 1 1 1 1 1 267 62.5 329.5 45.7% 40.7% 0.68 0.409 2 1 1 1 1 1 1 1 185 43 228 31.7% 28.0% 0.53 0.467 2 1 2 2 2 2 2 1 24 15.5 39.5 4.1% 10.1% 8.14 0.004 1 1 2 2 2 2 2 1 30 6 36 5.1% 3.9% 0.38 0.540 2 2 1 1 1 2 2 1 23 10 33 3.9% 6.5% 1.80 0.179 2 2 1 1 1 1 1 1 23 7.5 30.5 3.9% 4.9% 0.26 0.607 1 1 1 1 1 1 2 1 17 5 22 2.9% 3.3% 0.05 0.825 2 1 1 1 1 1 2 1 8 1 9 1.4% 0.7% 0.51 0.473 2 2 1 1 1 2 2 2 3 1 4 0.5% 0.7% 0.04 0.837 1 1 1 1 1 2 2 1 2 1 3 0.3% 0.7% 0.29 0.593 2 1 2 2 1 1 2 1 2 1 3 0.3% 0.7% 0.29 0.593 TOTAL 584 153.5 737.5 12.97 0.226

[0262] TABLE-US-00031 TABLE 15A SNP by SNP allele transmissions by AFBAC method: "either/both sib DR3/4" subgroup of familes Transmitted (n = 540) AFBAC (n = 121.5) allele counts variant allele allele counts variant allele Chi-squared p-value SNP # wt variant frequency wt allele variants frequency Statistic (df = 1) 1 296 244 45.2% 64.5 57 46.9% 0.12 0.730 2 490 50 9.3% 107.5 14 11.5% 0.58 0.446 3 485 55 10.2% 103.5 18 14.8% 2.17 0.141 4 485 55 10.2% 103.5 18 14.8% 2.17 0.141 5 485 55 10.2% 103.5 18 14.8% 2.17 0.141 6 457 83 15.4% 96.5 25 20.6% 1.97 0.161 7 436 104 19.35 88.5 33 27.2% 3.77 0.052 8 532 8 1.5% 119.5 2 1.6% 0.02 0.893 TOTAL

[0263] TABLE-US-00032 TABLE 15B SNP by SNP allele transmissions by AFBAC method: "neither sib DR3/4" subgroup of familes Transmitted (n = 588) AFBAC (n = 158.5) allele counts variant allele allele counts variant allele Chi-squared p-value SNP # wt variant frequency wt allele variant frequency Statistic (df = 1) 1 320 268 45.6% 77.5 81 51.1% 1.53 0.216 2 539 49 8.3% 139 19.5 12.3% 2.36 0.124 3 529 59 10.0% 132 26.5 16.7% 5.50 0.019 4 529 59 10.0% 132 26.5 16.7% 5.50 0.019 5 534 54 9.2% 136 22.5 14.2% 3.41 0.065 6 502 86 14.6% 121 37.5 23.7% 7.38 0.007 7 475 113 19.2% 115 43.5 27.4% 5.10 0.024 8 584 4 0.7% 157.5 1 0.6% 0.00 0.946

[0264] TABLE-US-00033 TABLE 16A TDT analysis of 8-loocus haplotypes to probands only. TOTAL transmitted to NOT transmitted TRANSMISSIONS McNemar chi- P-value haplotype affected to affected OBSERVED squared statistic (1 df) % TRANSMISSION 11111111 140 131 271 0.299 0.585 51.7% 21111111 130 105 235 2.660 0.103 55.3% 21222221 27 39 66 2.182 0.140 40.9% 22111111 19 28 47 1.723 0.189 40.4% 11222221 19 24 43 0.581 0.446 44.2% 22111221 17 23 40 0.900 0.343 42.5% 11111121 18 15 33 0.273 0.602 54.5% 22111222 6 5 11 0.091 0.763 54.5% 21111121 1 5 6 2.667 0.102 16.7% 11111221 1 3 4 1.000 0.317 25.0% 11221221 2 0 2 2.000 0.157 100.0% 21221121 1 1 2 0.000 1.000 50.0% 21221221 0 2 2 2.000 0.157 0.0% 11221111 0 1 1 1.000 0.317 0.0% 11111222 1 0 1 1.000 0.317 100.0% total 382 382 764

[0265] TABLE-US-00034 TABLE 16B TDT analysis of 8-loocus haplotypes to two primary affected children per family. TOTAL transmitted to NOT transmitted TRANSMISSIONS McNemar chi- P-value haplotype affected to affected OBSERVED squared statistic (1 df) % TRANSMISSION 21222221 52 80 132 5.939 0.015 39.4% 21111111 245 225 470 0.851 0.356 52.1% 11111111 283 259 542 1.063 0.303 52.2% 22111111 43 51 94 0.681 0.409 45.7% 11221221 3 1 4 1.000 0.317 75.0% 11222221 44 42 86 0.047 0.829 51.2% 22111221 37 43 80 0.450 0.502 46.3% 11111121 36 30 66 0.545 0.460 54.5% 22111222 11 11 22 0.000 1.000 50.0% 21111121 3 9 12 3.000 0.083 25.0% 21221121 2 2 4 0.000 1.000 50.0% 21221221 0 4 4 4.000 0.046 0.0% 11111221 4 4 8 0.000 1.000 50.0% 11221111 0 2 2 2.000 0.157 0.0% 11111222 1 1 2 0.000 1.000 50.0%

[0266] TABLE-US-00035 TABLE 16c TDT analysis of 8-loocus haplotypes to all affected children. TOTAL transmitted to NOT transmitted TRANSMISSIONS McNemar chi- P-value haplotype affected to affected OBSERVED squared statistic (1 df) % TRANSMISSION 11111111 295 272 567 0.933 0.334 52.0% 21111111 259 236 495 1.069 0.301 52.3% 21222221 55 88 143 7.615 0.006 38.5% 22111111 45 53 98 0.653 0.419 45.9% 11222221 44 43 87 0.011 0.915 50.6% 22111221 40 46 86 0.419 0.518 46.5% 11111121 38 30 68 0.941 0.332 55.9% 22111222 12 11 23 0.043 0.835 52.2% 21111121 3 10 13 3.769 0.052 23.1% 11111221 6 4 10 0.400 0.527 60.0% 21221221 1 5 6 2.667 0.102 16.7% 11221221 3 2 5 0.200 0.655 60.0% 21221121 3 2 5 0.200 0.655 60.0% 11221111 0 2 2 2.000 0.157 0.0% 11111222 1 1 2 0.000 1.000 50.0% TOTAL 805 805 1610

[0267] TABLE-US-00036 TABLE 17A analysis of 8-loocus haplotypes to two primary affected children per family: "either/both sib DR3/4" subgroup. TOTAL transmitted to NOT transmitted TRANSMISSIONS McNemar chi- P-value haplotype affected to affected OBSERVED squared statistic (1 df) % TRANSMISSION 11111111 133 121 254 0.567 0.451 52.45 21111111 105 101 206 0.078 0.780 51.0% 21222221 30 36 66 0.545 0.460 45.5% 22111111 22 26 48 0.333 0.564 45.8% 11222221 19 19 38 0.000 1.000 50.0% 22111221 16 22 38 0.947 0.330 42.1% 22111222 14 14 28 0.000 1.000 50.0% 22111222 8 6 14 0.286 0.593 57.1% 21111121 1 5 6 2.667 0.102 16.7% 11111221 2 0 2 2.000 0.157 100.0% TOTAL 350 350 700

[0268] TABLE-US-00037 TABLE 17B TDT analysis of 8-loocus haplotypes to two primary affected children per family: "neither sib DR3/4" subgroup.. TOTAL transmitted to NOT transmitted TRANSMISSIONS McNemar chi- P-value haplotype affected to affected OBSERVED squared statistic (1 df) % TRANSMISSION 11111111 138 126 264 0.545 0.460 52.3% 21111111 128 112 240 1.067 0.302 53.3% 21222221 22 44 66 7.333 0.007 33.3% 22111111 23 29 52 0.692 0.405 44.2% 11222221 28 20 48 1.333 0.248 58.3% 22111111 21 25 46 0.348 0.555 45.7% 11111121 17 11 28 1.286 0.257 60.7% 22111222 3 5 8 0.500 0.480 37.5% 11111221 2 4 6 0.667 0.414 33.3% 21111121 2 4 6 0.667 0.414 33.3% 11221221 3 1 4 1.000 0.317 75.0% 21221121 2 2 4 0.000 1.000 50.0% 21221221 0 4 4 4.000 0.046 0.0% 11221111 0 2 2 2.000 0.157 0.0% 11111222 1 1 2 0.000 1.000 50.0% TOTAL 390 390 780

[0269] TABLE-US-00038 TABLE 18 Allele frequencies compared between Filipino controls and diabetics. WT Allele WT allele Frequency frquency in SNP # in controls diabetics P-value by z-test 1 0.559 0.461 0.077 2 0.989 1.000 0.480 3 0.793 0.882 0.031 4 0.824 0.933 0.003 5 0.809 0.882 0.075 6 0.814 0.882 0.097 7 0.723 0.770 0.362 8 1.000 1.000 1.000 n = 188 178 chromosomes

[0270] TABLE-US-00039 TABLE 19 Computationally estimated haplotype frequencies compared between Filipino controls and diabetics. MLE MLS frequency: frequency: COUNT: COUNT: exp exp chi- controls Diabetics O.R. control diabetics Total count control diabetic square P-value IL4R 11111111 0.318 0.329 1.1 59.7 58.5 118.2 60.7 57.5 0.033 0.855 11111121 0.074 0.074 1.0 14.0 13.2 27.2 14.0 13.2 0.000 0.987 11222221 0.145 0.058 0.4 27.3 10.3 37.6 19.3 18.3 6.727 0.009 21111111 0.390 0.441 1.2 73.3 78.5 151.8 78.0 73.8 0.582 0.445 21212221 0.032 0.051 1.6 6.0 9.0 15.0 7.7 7.3 0.776 0.378 n = 188 178 366

[0271] TABLE-US-00040 TABLE 20 Observed haplotypes and frequencies compared between Filipino controls and diabetics.. calc. calc. frequency: frequency: COUNT: COUNT: exp exp chi- haplotype controls Diabetics O.R. control diabetics Total count control diabetic square P-value IL4R x1111111 0.707 0.770 1.4 133.0 137.0 270.0 138.7 131.3 0.480 0.489 x1111121 0.074 0.112 1.6 14.0 20.0 34.0 17.5 16.5 1.413 0.235 x1222221 0.154 0.067 0.4 29.0 12.0 41.0 21.1 19.9 6.155 0.013 x1212221 0.032 0.051 1.6 6.0 9.0 15.0 7.7 7.3 0.776 0.378 x1222111 0.005 0.000 0.0 1.0 0.0 1.0 0.5 0.5 0.947 0.331 x2111111 0.011 0.000 0.0 2.0 0.0 2.0 1.0 1.0 1.894 0.169 x1221121 0.016 0.000 0.0 3.0 0.0 3.0 1.5 1.5 2.840 0.092 n = 188.0 178.0 366 overall 14.504 0.024

[0272] TABLE-US-00041 TABLE 21 Allele Frequencies in Patients and Controls Freq in SNP Major Minor Freq in T1D Gene Description allele allele Controls patients p-value.sup.a O.R..sup.b 95% CI IL4R 5' (-3223).sup.c C T 41.9% 51.1% 0.10 1.45 0.96, 2.19 5' (-1914).sup.c C T 44.1% 42.7% 0.86 1.06 0.70, 1.60 I50V.sup.d A G 44.1% 53.9% 0.06 1.48 0.98, 2.23 N142N.sup.d C G 1.1% 0.0% 0.17 0.00 E375A.sup.d A C 20.7% 11.7% 0.02 0.50 0.28, 0.90 L389L.sup.d G T 17.6% 6.7% 0.001 0.34 0.17, 0.67 C406R.sup.d T C 19.1% 11.7% 0.05 0.56 0.31, 0.99 S478P.sup.d T C 18.6% 11.7% 0.06 0.58 0.32, 1.04 Q551R.sup.d A G 27.7% 23.3% 0.34 0.80 0.50, 1.27 S761P.sup.d T 100.0% 100.0% n.a. n.a. IL4 5' (-524).sup.e C T 30.9% 34.4% 0.46 1.18 0.76, 1.82 IL13 5' (-1512).sup.f A C 30.6% 41.0% 0.05 1.58 1.03, 2.42 5' (-1112).sup.f C T 23.1% 30.9% 0.117 1.49 0.94, 2.37 Intron 3.sup.f C T 40.4% 45.6% 0.32 1.23 0.82, 1.86 R110Q.sup.f G A 39.9% 46.1% 0.23 1.29 0.85, 1.95 .sup.aDifferences in allele frequencies between cases and controls were tested using a chi-square test. .sup.bOdds ratios refer to the minority allele. .sup.cAccession sequence AC004525.1; .sup.dAccession sequence X52425.1; .sup.eAccession sequence M23442.1; .sup.fAccession sequence U10307.1

[0273] TABLE-US-00042 TABLE 22 Pairwise Linkage Disequilibrium Between IL4R SNPs -3223 (T) -1914 (T) 50 (G) 142 (G) 375 (C) 389 (T) 406 (C) 478 (C) 551 (G) -3223 (T) -- -0.183*** 0.176*** 0.004 -0.063*** -0.068*** -0.061*** -0.059*** -0.068*** -1914 (T) -1.0 -- -0.068*** -0.005 0.105 0.029* 0.014 0.011 0.027 50 (G) 0.75 -0.81 -- 0.006 -0.049** -0.067*** -0.043** -0.039** -0.079*** 142 (G) 1.0 -1.0 1.0 -- -0.002 -0.002 -0.002 -0.002 -0.003 375 (C) -0.70 0.70 -0.53 -1.0 -- 0.139*** 0.152*** 0.147*** 0.145*** 389 (T) -0.93 0.29 -0.86 -1.0 1.0 -- 0.126*** 0.122*** 0.122*** 406 (C) -0.74 0.13 -0.49 -1.0 1.0 0.89 -- 0.150*** 0.133*** 478 (C) -0.74 0.62 -0.48 -1.0 1.0 0.85 1.0 -- 0.135*** 551 (G) -0.56 0.17 -0.65 -1.0 0.96 0.96 0.96 1.0 -- Top diagonal (D = P.sub.ab - P.sub.a P.sub.b). Bottom diagonal D' (normalized linkage disequilibrium, D' = D/Dmax). All values refer to the minority allele indicated in the table. Statistically significant linkage disequilibrium values are indicated as follows: * p < 0.05; **p < 0.01; ***p < 0.0001

[0274] TABLE-US-00043 TABLE 23 Pairwise Linkage Disequilibrium Between IL4 and IL13 SNPs IL13- IL4-524 (C) IL13-1512 (C) 1112 (T) IL13 intr (T) IL13 110 (A) IL4-524 (C) -- 0.062*** 0.069*** 0.024 0.063*** IL13-1512 (C) 0.29 -- 0.163*** 0.057*** 0.058** IL13-1112 (T) 0.41 1.0 -- 0.077*** 0.078*** IL13 intr (T) 0.13 0.31 0.54 -- 0.201*** IL13 110 (A) 0.34 0.31 0.55 0.84 -- Top diagonal (D = P.sub.ab - P.sub.a P.sub.b). Bottom diagonal D' (normalized linkage disequilibrium, D' = D/Dmax). Statistically significant linkage disequilibrium values are indicated as follows: * p < 0.05; **p < 0.01; ***p < 0.0001

[0275] TABLE-US-00044 TABLE 24 Genotype Frequencies in Diabetics and Controls. genotype controls T1DM Fisher's exact test OR 95% CI IL4R_3223 CC 29.0% 24.1% 0.1584 0.78 (0.40, 1.51) CT 58.1% 51.7% 0.77 (0.43, 1.38) TT 12.9% 24.1% 2.15 (0.99, 4.68) IL4R_1914 CC 31.2% 36.0% 0.6636 1.24 (0.67, 2.29) TC 49.5% 42.7% 0.76 (0.43, 1.36) TT 19.4% 21.3% 1.13 (0.55, 2.33) IL4R 50 AA 34.7% 23.6% 0.1846 0.58 (0.30, 1.11) AG 43.2% 44.9% 1.08 (0.60, 1.93) GG 22.1% 31.5% 1.62 (0.84, 3.13) IL4R 142 CC 97.9% 100.0% 0.4978 CG 2.1% 0.0% IL4R 375 AA 62.1% 78.7% 0.038 2.25 (1.17, 4.33) AC 34.7% 19.1% 0.44 (0.23, 0.87) CC 3.2% 2.2% 0.70 (0.12, 4.32) IL4R 389 GG 67.4% 87.6% 0.002 3.43 (1.60, 7.37) GT 30.5% 11.2% 0.29 (0.13, 0.63) TT 2.1% 1.1% 0.53 (0.05, 5.93) IL4R 406 CC 2.1% 2.2% 0.0585 1.07 (0.15, 7.76) TC 33.7% 19.1% 0.46 (0.24, 0.92) TT 64.2% 78.7% 2.05 (1.06, 3.97) IL4R 478 CC 2.1% 2.2% 0.0904 1.07 (0.15, 7.76) TC 32.6% 19.1% 0.49 (0.25, 0.96) TT 65.3% 78.7% 1.96 (1.01, 3.79) IL4R 551 AA 52.6% 57.3% 0.5613 1.21 (0.68, 2.16) AG 38.9% 38.2% 0.97 (0.53, 1.76) GG 8.4% 4.5% 0.51 (0.15, 1.76) IL4-590 CC 10.5% 12.4% 0.8143 1.20 (0.48, 2.98) CT 40.0% 42.7% 1.12 (0.62, 2.01) TT 49.5% 44.9% 0.83 (0.47, 1.49) IL13INT3 C CC 33.7% 31.5% 0.5413 0.90 (0.49, 1.68) CT 50.5% 46.1% 0.84 (0.47, 1.49) TT 15.8% 22.5% 1.55 (0.74, 3.25) IL13 110 AA 12.6% 27.0% 0.0252 2.55 (1.19, 5.49) GA 54.7% 38.2% 0.51 (0.28, 0.92) GG 32.6% 34.8% 1.10 (0.60, 2.03) IL13_1512 AA 48.4% 34.8% 0.1363 0.57 (0.31, 1.03) AC 41.9% 48.3% 1.29 (0.72, 2.32) CC 9.7% 16.9% 1.89 (0.78, 4.57) IL13_1112 CC 60.2% 51.7% 0.2429 0.71 (0.39, 1.27) CT 33.3% 34.8% 1.07 (0.58, 1.97) TT 6.5% 13.5% 2.26 (0.81, 6.31)

[0276] TABLE-US-00045 TABLE 25A Molecular IL-4 R Haplotypes in Filipino Diabetics and Controls 7 SNP Frequency Frequency chi-square Haplotype in Controls in Diabetics p-value O.R. 95% CI CAGTTAT 70.7% 77.0% 0.49 1.38 (0.9, 2.2) CCTCCGT 15.4% 6.7% 0.013 0.40 (0.2, 0.8) GAGTTAT 1.1% 0.0% 0.50 0.0 (0.0, 5.8) CAGTTGT 7.4% 11.2% 0.23 1.57 (0.8, 3.2) CCGCCGT 3.2% 5.1% 0.38 1.62 (0.6, 4.6) CCTTTGT 1.6% 0.0% 0.27 0.0 (0.0, 3.5) CCTGTAT 0.5% 0.0% 0.98 0.0 (0.0, 15.8) total (6 d.f.) 0.076 n = 188 178

[0277] TABLE-US-00046 TABLE 25B Estimated IL4R 10-SNP Haplotype Frequencies in Diabetics and Controls IL4R 10 Locus Controls.sup.a TIDM P- 8 locus H-ID Haplotype N = 188 std dev N = 180 std dev O.R. 95% CI value.sup.b H-5A CCACGTCCGT 5.8% (1.7%) 0.0% 0.00 (0.0, 0.8) 0.001 H-5B CTACCTCCGT 8.5% (2.3%) 5.8% (2.2%) 0.66 (0.3, 1.6) 0.33 H-3A CTGCCTCCGT 0.9% (0.6%) 0.0% 0.00 (0.0, 6.0) 0.21 H-3B TCGCCTCCGT 0.0% 0.9% (1.1%) n. a. 0.19 Total (23 d. f.) 0.04 .sup.a Maximum likelihood haplotype frequencies were computed using an Expectation-Maximization (EM) algorithm (see Excoffier and Slatkin 1995) as implemented by the Arlequin software program L. Excoffier, University of Geneva, CH). The standard deviation was computed carrying out 100 boostrap replicates. 24 of the possible 512 haplotypes were observed. The four shown here all include the disease associated seven SNP haplotype. The two 8 SNP haplotypes containing the seven SNP haplotype were #designated H-3 and H-5 in reference Mirel et al, (in press) The standard deviation was computed carrying out 100 boostrap replicates. .sup.b Differences in allele frequencies between cases and controls were tested using a chi-square test

[0278] TABLE-US-00047 TABLE 26 Estimated IL4 and IL13 5-SNP Haplotype Frequencies Diabetics and Controls 5 locus Controls TIDM Odds haploytpe N = 188 Sd N = 176* Sd Ratio P-value CACCA 4.1% (1.3%) 3.5% (1.7%) 0.84 CACCG 8.9% (2.2%) 10.6% (2.9%) 1.21 CACTA 1.3% (1.1%) 3.0% (1.8%) 2.26 CATTA 0.0% 0.6% (0.5%) n. a. CCCCG 1.5% (1.0%) 0.9% (0.9%) 0.61 CCTCG 1.1% (0.9%) 5.5% (2.3%) 5.19 0.02 CCTTA 12.6% (2.4%) 9.3% (2.0%) 0.71 CCTTG 0.0% 0.4% (0.7%) n. a. TACCG 33.6% (3.8%) 21.9% (3.9%) 0.55 0.03 TACTA 16.4% (3.3%) 16.3% (3.0%) 0.99 TACTG 5.0% (1.7%) 3.2% (1.4%) 0.62 TCCCG 4.6% (1.8%) 9.8% (2.9%) 2.23 0.06 TCCTA 1.3% (1.1%) 0.0% 0.00 TCCTG 0.2% (0.5%) 0.0% 0.00 TCTCG 5.3% (2.4% 1.7% (1.2%) 0.31 0.07 TCTTA 4.1% (2.1%) 12.8% (2.2%) 3.47 0.004 TCTCA 0.0% 0.6% (0.7%) n. a. Total (16 d. f.) 0.005 * due to missing genotypes. (order of the SNPs: IL4-524, IL13-1512, IL13-1112, IL13INT3, IL13R110)

[0279] TABLE-US-00048 TABLE 27 Test For Independence Between Genotype Frequencies at IL4R SNPs and Genotype Frequencies at Five IL4 and IL13 SNPs. Chromsome 5 SNP controls T1D IL4-524 0.15 0.001 IL13-1512 0.77 0.78 IL13-1112 0.93 0.73 IL13 110 0.99 0.91 IL3 intron 3 0.99 0.019

[0280] TABLE-US-00049 TABLE 28 Epistasis Between IL4R and Five IL4 and IL13 SNPs Walds Nominal .beta..sub.3. Std. Error Odds ratio Chi-sq p-value IL4-524:il4r -0.22 0.10 0.81 4.55 0.033 IL13-1512:il4r 0.04 0.15 1.04 0.06 0.811 IL13-1112:il4r 0.10 0.16 1.10 0.37 0.545 IL13 110:il4r 0.14 0.09 1.15 2.24 0.135 IL13INT3:il4r 0.17 0.09 1.18 3.19 0.074 The Overall p-Value For All Five Tests By Permutation Analysis Was p < 0.045 (see text).

[0281] TABLE-US-00050 TABLE 29 Probes For IL4 and IL13 Seq ID Conc. SNP Allele Probe Probe Sequence No: (.mu.M) IL4P C582T C KW66 AACATTGTCCCCCAGTGC 63 1.2 IL4P C582T T KW89 AGCACTGGGGAACAATGTTC 64 0.9 IL13 Int 3 C DBM0161P TTCTACTCACGTGCTGACCT 65 1.0 IL13 Int 3 T DBM0164P GGTCAGCACATGAGTAGAACG 66 0.3 IL13 Ex 4 R110Q DBM0136P TCAGTTGAACCGTCCCTCG 67 1.0 IL13 Ex 4 R110Q DBM0181P GAGGGACAGTTCAACTGAAAC 68 1.0

[0282] TABLE-US-00051 TABLE 30 Amplicon Primers and Lengths Amplicon Forward Seq ID Seq ID Exon Size Primer No Sequence Reverse Primer No Sequence IL4P 107 RR169B 69 ACTAGGCCTCACCTGATACGA RR170B 72 CATAGAGGCAGAATAACAGGCAGA IL 13 in 3 118 DBM0165B 70 CTCGGACATGCAAGCTGGAA DBM0166B 73 ACTGAATGAGACAGTCCCTGGA IL 13 ex 4 187 DBM0167B 71 AATCGAGGTGGCCCAGTTTGTA DBM0168B 74 CCTAACCCTCCTTCCCGCCTA

[0283] TABLE-US-00052 TABLE 31 Amplicon Primers and Lengths for IL4R Amplicon Seq ID Exon Size Allele Primer No Sequence T(-1914)C 49 T DBM0659 75 ACTGACTTATCTTTACTGTCACTTCT DBM0661 76 GCAAGACAGCCACCAACCC T(-1914)C 49 C DBM0660 77 TGACTTATCTTTACTGTCACTTCC DBM0661 76 GCAAGACAGCCACCAACCC C(-3223)T 42 C DBM0672 79 CCTGCTCCCAGGACTGAC DBM0667 80 CCCAGACTTTATCTGTGACTGCTC C(-3223)T 42 T DBMO671 81 CCTGCTCCCAGGACTGAT DBM0667 80 CCCAGACTTTATCTGTGACTGCTC

[0284] TABLE-US-00053 TABLE 32 Amplicon Primers and Lengths for IL13 Amplicon Seq ID Exon Size Allele Primer No Sequence A(-1512)C 44 A DBM0650 82 GGAAACAGGCCCGTAGA DBM0652 83 GAGTGCCGCTACTTGGCC A(-1512)C 43 C DBM0651 84 GAAACAGGCCCGTAGC DBM0652 83 GAGTGCCGCTACTTGGCC C(-1112)T 55 C DBM0656 85 TCTGGAGGACTTCTAGGAAAAC DBM0658 86 TGCAGCCATGTCGCCTTT C(-1112)T 55 T DBM0657 87 TCTGGAGGACTTCTAGGAAAAT DBM0658 86 TGCAGCCATGTCGCCTTT

[0285]

Sequence CWU 1

1

90 1 3597 DNA Homo sapiens 1 ggcgaatgga gcaggggcgc gcagataatt aaagatttac acacagctgg aagaaatcat 60 agagaagccg ggcgtggtgg ctcatgccta taatcccagc acttttggag gctgaggcgg 120 gcagatcact tgagatcagg agttcgagac cagcctggtg ccttggcatc tcccaatggg 180 gtggctttgc tctgggctcc tgttccctgt gagctgcctg gtcctgctgc aggtggcaag 240 ctctgggaac atgaaggtct tgcaggagcc cacctgcgtc tccgactaca tgagcatctc 300 tacttgcgag tggaagatga atggtcccac caattgcagc accgagctcc gcctgttgta 360 ccagctggtt tttctgctct ccgaagccca cacgtgtatc cctgagaaca acggaggcgc 420 ggggtgcgtg tgccacctgc tcatggatga cgtggtcagt gcggataact atacactgga 480 cctgtgggct gggcagcagc tgctgtggaa gggctccttc aagcccagcg agcatgtgaa 540 acccagggcc ccaggaaacc tgacagttca caccaatgtc tccgacactc tgctgctgac 600 ctggagcaac ccgtatcccc ctgacaatta cctgtataat catctcacct atgcagtcaa 660 catttggagt gaaaacgacc cggcagattt cagaatctat aacgtgacct acctagaacc 720 ctccctccgc atcgcagcca gcaccctgaa gtctgggatt tcctacaggg cacgggtgag 780 ggcctgggct cagtgctata acaccacctg gagtgagtgg agccccagca ccaagtggca 840 caactcctac agggagccct tcgagcagca cctcctgctg ggcgtcagcg tttcctgcat 900 tgtcatcctg gccgtctgcc tgttgtgcta tgtcagcatc accaagatta agaaagaatg 960 gtgggatcag attcccaacc cagcccgcag ccgcctcgtg gctataataa tccaggatgc 1020 tcaggggtca cagtgggaga agcggtcccg aggccaggaa ccagccaagt gcccacactg 1080 gaagaattgt cttaccaagc tcttgccctg ttttctggag cacaacatga aaagggatga 1140 agatcctcac aaggctgcca aagagatgcc tttccagggc tctggaaaat cagcatggtg 1200 cccagtggag atcagcaaga cagtcctctg gccagagagc atcagcgtgg tgcgatgtgt 1260 ggagttgttt gaggccccgg tggagtgtga ggaggaggag gaggtagagg aagaaaaagg 1320 gagcttctgt gcatcgcctg agagcagcag ggatgacttc caggagggaa gggagggcat 1380 tgtggcccgg ctaacagaga gcctgttcct ggacctgctc ggagaggaga atgggggctt 1440 ttgccagcag gacatggggg agtcatgcct tcttccacct tcgggaagta cgagtgctca 1500 catgccctgg gatgagttcc caagtgcagg gcccaaggag gcacctccct ggggcaagga 1560 gcagcctctc cacctggagc caagtcctcc tgccagcccg acccagagtc cagacaacct 1620 gacttgcaca gagacgcccc tcgtcatcgc aggcaaccct gcttaccgca gcttcagcaa 1680 ctccctgagc cagtcaccgt gtcccagaga gctgggtcca gacccactgc tggccagaca 1740 cctggaggaa gtagaacccg agatgccctg tgtcccccag ctctctgagc caaccactgt 1800 gccccaacct gagccagaaa cctgggagca gatcctccgc cgaaatgtcc tccagcatgg 1860 ggcagctgca gcccccgtct cggcccccac cagtggctat caggagtttg tacatgcggt 1920 ggagcagggt ggcacccagg ccagtgcggt ggtgggcttg ggtcccccag gagaggctgg 1980 ttacaaggcc ttctcaagcc tgcttgccag cagtgctgtg tccccagaga aatgtgggtt 2040 tggggctagc agtggggaag aggggtataa gcctttccaa gacctcattc ctggctgccc 2100 tggggaccct gccccagtcc ctgtcccctt gttcaccttt ggactggaca gggagccacc 2160 tcgcagtccg cagagctcac atctcccaag cagctcccca gagcacctgg gtctggagcc 2220 gggggaaaag gtagaggaca tgccaaagcc cccacttccc caggagcagg ccacagaccc 2280 ccttgtggac agcctgggca gtggcattgt ctactcagcc cttacctgcc acctgtgcgg 2340 ccacctgaaa cagtgtcatg gccaggagga tggtggccag acccctgtca tggccagtcc 2400 ttgctgtggc tgctgctgtg gagacaggtc ctcgccccct acaacccccc tgagggcccc 2460 agacccctct ccaggtgggg ttccactgga ggccagtctg tgtccggcct ccctggcacc 2520 ctcgggcatc tcagagaaga gtaaatcctc atcatccttc catcctgccc ctggcaatgc 2580 tcagagctca agccagaccc ccaaaatcgt gaactttgtc tccgtgggac ccacatacat 2640 gagggtctct taggtgcatg tcctcttgtt gctgagtctg cagatgagga ctagggctta 2700 tccatgcctg ggaaatgcca cctcctggaa ggcagccagg ctggcagatt tccaaaagac 2760 ttgaagaacc atggtatgaa ggtgattggc cccactgacg ttggcctaac actgggctgc 2820 agagactgga ccccgcccag cattgggctg ggctcgccac atcccatgag agtagagggc 2880 actgggtcgc cgtgccccac ggcaggcccc tgcaggaaaa ctgaggccct tgggcacctc 2940 gacttgtgaa cgagttgttg gctgctccct ccacagcttc tgcagcagac tgtccctgtt 3000 gtaactgccc aaggcatgtt ttgcccacca gatcatggcc cacgtggagg cccacctgcc 3060 tctgtctcac tgaactagaa gccgagccta gaaactaaca cagccatcaa gggaatgact 3120 tgggcggcct tgggaaatcg atgagaaatt gaacttcagg gagggtggtc attgcctaga 3180 ggtgctcatt catttaacag agcttcctta ggttgatgct ggaggcagaa tcccggctgt 3240 caaggggtgt tcagttaagg ggagcaacag aggacatgaa aaattgctat gactaaagca 3300 gggacaattt gctgccaaac acccatgccc agctgtatgg ctgggggctc ctcgtatgca 3360 tggaaccccc agaataaata tgctcagcca ccctgtgggc cgggcaatcc agacagcagg 3420 cataaggcac cagttaccct gcatgttggc ccagacctca ggtgctaggg aaggcgggaa 3480 ccttgggttg agtaatgctc gtctgtgtgt tttagtttca tcacctgtta tctgtgtttg 3540 ctgaggagag tggaacagaa ggggtggagt tttgtataaa taaagtttct ttgtctc 3597 2 2478 DNA Homo sapiens 2 atggggtggc tttgctctgg gctcctgttc cctgtgagct gcctggtcct gctgcaggtg 60 gcaagctctg ggaacatgaa ggtcttgcag gagcccacct gcgtctccga ctacatgagc 120 atctctactt gcgagtggaa gatgaatggt cccaccaatt gcagcaccga gctccgcctg 180 ttgtaccagc tggtttttct gctctccgaa gcccacacgt gtatccctga gaacaacgga 240 ggcgcggggt gcgtgtgcca cctgctcatg gatgacgtgg tcagtgcgga taactataca 300 ctggacctgt gggctgggca gcagctgctg tggaagggct ccttcaagcc cagcgagcat 360 gtgaaaccca gggccccagg aaacctgaca gttcacacca atgtctccga cactctgctg 420 ctgacctgga gcaacccgta tccccctgac aattacctgt ataatcatct cacctatgca 480 gtcaacattt ggagtgaaaa cgacccggca gatttcagaa tctataacgt gacctaccta 540 gaaccctccc tccgcatcgc agccagcacc ctgaagtctg ggatttccta cagggcacgg 600 gtgagggcct gggctcagtg ctataacacc acctggagtg agtggagccc cagcaccaag 660 tggcacaact cctacaggga gcccttcgag cagcacctcc tgctgggcgt cagcgtttcc 720 tgcattgtca tcctggccgt ctgcctgttg tgctatgtca gcatcaccaa gattaagaaa 780 gaatggtggg atcagattcc caacccagcc cgcagccgcc tcgtggctat aataatccag 840 gatgctcagg ggtcacagtg ggagaagcgg tcccgaggcc aggaaccagc caagtgccca 900 cactggaaga attgtcttac caagctcttg ccctgttttc tggagcacaa catgaaaagg 960 gatgaagatc ctcacaaggc tgccaaagag atgcctttcc agggctctgg aaaatcagca 1020 tggtgcccag tggagatcag caagacagtc ctctggccag agagcatcag cgtggtgcga 1080 tgtgtggagt tgtttgaggc cccggtggag tgtgaggagg aggaggaggt agaggaagaa 1140 aaagggagct tctgtgcatc gcctgagagc agcagggatg acttccagga gggaagggag 1200 ggcattgtgg cccggctaac agagagcctg ttcctggacc tgctcggaga ggagaatggg 1260 ggcttttgcc agcaggacat gggggagtca tgccttcttc caccttcggg aagtacgagt 1320 gctcacatgc cctgggatga gttcccaagt gcagggccca aggaggcacc tccctggggc 1380 aaggagcagc ctctccacct ggagccaagt cctcctgcca gcccgaccca gagtccagac 1440 aacctgactt gcacagagac gcccctcgtc atcgcaggca accctgctta ccgcagcttc 1500 agcaactccc tgagccagtc accgtgtccc agagagctgg gtccagaccc actgctggcc 1560 agacacctgg aggaagtaga acccgagatg ccctgtgtcc cccagctctc tgagccaacc 1620 actgtgcccc aacctgagcc agaaacctgg gagcagatcc tccgccgaaa tgtcctccag 1680 catggggcag ctgcagcccc cgtctcggcc cccaccagtg gctatcagga gtttgtacat 1740 gcggtggagc agggtggcac ccaggccagt gcggtggtgg gcttgggtcc cccaggagag 1800 gctggttaca aggccttctc aagcctgctt gccagcagtg ctgtgtcccc agagaaatgt 1860 gggtttgggg ctagcagtgg ggaagagggg tataagcctt tccaagacct cattcctggc 1920 tgccctgggg accctgcccc agtccctgtc cccttgttca cctttggact ggacagggag 1980 ccacctcgca gtccgcagag ctcacatctc ccaagcagct ccccagagca cctgggtctg 2040 gagccggggg aaaaggtaga ggacatgcca aagcccccac ttccccagga gcaggccaca 2100 gacccccttg tggacagcct gggcagtggc attgtctact cagcccttac ctgccacctg 2160 tgcggccacc tgaaacagtg tcatggccag gaggatggtg gccagacccc tgtcatggcc 2220 agtccttgct gtggctgctg ctgtggagac aggtcctcgc cccctacaac ccccctgagg 2280 gccccagacc cctctccagg tggggttcca ctggaggcca gtctgtgtcc ggcctccctg 2340 gcaccctcgg gcatctcaga gaagagtaaa tcctcatcat ccttccatcc tgcccctggc 2400 aatgctcaga gctcaagcca gacccccaaa atcgtgaact ttgtctccgt gggacccaca 2460 tacatgaggg tctcttag 2478 3 21 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 3 ccacacgtgt atccctgaga a 21 4 19 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 4 tctcagggac acacgtgtg 19 5 22 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 5 tggagtgaaa acgacccggc ag 22 6 21 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 6 ctgccgggtc attttcgctc c 21 7 21 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 7 gagggaaggg agggcattgt g 21 8 19 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 8 agggaagggc gggcattgt 19 9 19 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 9 ctctccgagc aggtccagg 19 10 21 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 10 tcctggacct tctcggagag g 21 11 23 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 11 aaggtggaag aaggcatgac tcc 23 12 23 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 12 aaggtgggag aagacatgac tcc 23 13 23 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 13 ggagtcacgt cttctcctac ctt 23 14 22 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 14 tggctcagag agttgctgaa gc 22 15 19 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 15 ttcagcaacc ccctgagcc 19 16 20 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 16 agtggctatc aggagtttgt 20 17 20 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 17 agtggctatc gggagtttgt 20 18 21 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 18 ctcttctctg agatgcccga g 21 19 21 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4R polymorphisms 19 ctcgggcatc ccagagaaga g 21 20 8 DNA Artificial sequence Description of artificial sequence Sequences used to determine computationally estimated haplotype frequencies compared between Filipino controls and diabetics 20 acatttat 8 21 8 DNA Artificial sequence Description of artificial sequence Sequences used to determine computationally estimated haplotype frequencies compared between Filipino controls and diabetics 21 acagttgt 8 22 8 DNA Artificial sequence Description of artificial sequence Sequences used to determine computationally estimated haplotype frequencies compared between Filipino controls and diabetics 22 acctccgt 8 23 8 DNA Artificial sequence Description of artificial sequence Sequences used to determine computationally estimated haplotype frequencies compared between Filipino controls and diabetics 23 gcagttat 8 24 8 DNA Artificial sequence Description of artificial sequence Sequences used to determine computationally estimated haplotype frequencies compared between Filipino controls and diabetics 24 gccgccgt 8 25 19 DNA Artificial sequence Description of artificial sequence Amplicon primer 25 cagcccctgt gtctgcaga 19 26 20 DNA Artificial sequence Description of artificial sequence Amplicon primer 26 ctgacctgga gcaacccgta 20 27 25 DNA Artificial sequence Description of artificial sequence Amplicon primer 27 attgtgtgag gaggaggagg aggta 25 28 17 DNA Artificial sequence Description of artificial sequence Amplicon primer 28 ctcgtcatcg caggcaa 17 29 19 DNA Artificial sequence Description of artificial sequence Amplicon primer 29 gccgaaatgt cctccagca 19 30 15 DNA Artificial sequence Description of artificial sequence Amplicon primer 30 ccggcctccc tggca 15 31 26 DNA Artificial sequence Description of artificial sequence Amplicon primer 31 gtccagtgta tagttatccg cactga 26 32 19 DNA Artificial sequence Description of artificial sequence Amplicon primer 32 actgggcctc tgctggtca 19 33 22 DNA Artificial sequence Description of artificial sequence Amplicon primer 33 gttgggcatg tgagcactcg ta 22 34 18 DNA Artificial sequence Description of artificial sequence Amplicon primer 34 agggcatctc gggttcta 18 35 21 DNA Artificial sequence Description of artificial sequence Amplicon primer 35 ccacatttct ctggggacac a 21 36 20 DNA Artificial sequence Description of artificial sequence Amplicon primer 36 gcagactcag caacaagagg 20 37 21 DNA Artificial sequence Description of artificial sequence Hybridization probe 37 ccacacgtgt atccctgaga a 21 38 22 DNA Artificial sequence Description of artificial sequence Hybridization probe 38 tggagtgaaa acgacccggc ag 22 39 21 DNA Artificial sequence Description of artificial sequence Hybridization probe 39 gagggaaggg agggcattgt g 21 40 19 DNA Artificial sequence Description of artificial sequence Hybridization probe 40 ctctccgagc aggtccagg 19 41 23 DNA Artificial sequence Description of artificial sequence Hybridization probe 41 aaggtggaag aaggcatgac tcc 23 42 23 DNA Artificial sequence Description of artificial sequence Hybridization probe 42 aaggtgggag aagacatgac tcc 23 43 22 DNA Artificial sequence Description of artificial sequence Hybridization probe 43 tggctcagag agttgctgaa gc 22 44 20 DNA Artificial sequence Description of artificial sequence Hybridization probe 44 agtggctatc aggagtttgt 20 45 21 DNA Artificial sequence Description of artificial sequence Hybridization probe 45 ctcttctctg agatgcccga g 21 46 19 DNA Artificial sequence Description of artificial sequence Hybridization probe 46 tctcagggac acacgtgtg 19 47 21 DNA Artificial sequence Description of artificial sequence Hybridization probe 47 ctgccgggtc attttcgctc c 21 48 19 DNA Artificial sequence Description of artificial sequence Hybridization probe 48 agggaagggc gggcattgt 19 49 21 DNA Artificial sequence Description of artificial sequence Hybridization probe 49 tcctggacct tctcggagag g 21 50 23 DNA Artificial sequence Description of artificial sequence Hybridization probe 50 ggagtcacgt cttctcctac ctt 23 51 19 DNA Artificial sequence Description of artificial sequence Hybridization probe 51 ttcagcaacc ccctgagcc 19 52 20 DNA Artificial sequence Description of artificial sequence Hybridization probe 52 agtggctatc gggagtttgt 20 53 21 DNA Artificial sequence Description of artificial sequence Hybridization probe 53 ctcgggcatc ccagagaaga g 21 54 21 DNA Artificial sequence Description of artificial sequence allele- specific PCR primer 54 ccacatttct ctggggacac a 21 55 25 DNA Artificial sequence Description of artificial sequence allele- specific PCR primer 55 attgtgtgag gaggaggagg aggta 25 56 25 DNA Artificial sequence Description of artificial sequence allele- specific PCR primer 56 attgtgtgag gaggaggagg aggta 25 57 17 DNA Artificial sequence Description of artificial sequence allele- specific PCR primer 57 ttccaggagg gaaggga 17 58 20 DNA Artificial sequence Description of artificial sequence allele- specific PCR primer 58 caccgcatgt acaaactcct 20 59 17 DNA Artificial sequence Description of artificial sequence allele- specific PCR primer 59 ggtgactggc tcaggga 17 60 17 DNA Artificial sequence Description of

artificial sequence allele- specific PCR primer 60 ttccaggagg gaagggc 17 61 20 DNA Artificial sequence Description of artificial sequence allele- specific PCR primer 61 caccgcatgt acaaactccc 20 62 17 DNA Artificial sequence Description of artificial sequence allele- specific PCR primer 62 ggtgactggc tcagggg 17 63 18 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4 and IL13 polymorphisms 63 aacattgtcc cccagtgc 18 64 20 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4 and IL13 polymorphisms 64 agcactgggg aacaatgttc 20 65 20 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4 and IL13 polymorphisms 65 ttctactcac gtgctgacct 20 66 21 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4 and IL13 polymorphisms 66 ggtcagcaca tgagtagaac g 21 67 19 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4 and IL13 polymorphisms 67 tcagttgaac cgtccctcg 19 68 21 DNA Artificial sequence Description of artificial sequence Probe used to identify IL4 and IL13 polymorphisms 68 gagggacagt tcaactgaaa c 21 69 21 DNA Artificial sequence Description of artificial sequence Amplicon primer 69 actaggcctc acctgatacg a 21 70 20 DNA Artificial sequence Description of artificial sequence Amplicon primer 70 ctcggacatg caagctggaa 20 71 22 DNA Artificial sequence Description of artificial sequence Amplicon primer 71 aatcgaggtg gcccagtttg ta 22 72 24 DNA Artificial sequence Description of artificial sequence Amplicon primer 72 catagaggca gaataacagg caga 24 73 22 DNA Artificial sequence Description of artificial sequence Amplicon primer 73 actgaatgag acagtccctg ga 22 74 21 DNA Artificial sequence Description of artificial sequence Amplicon primer 74 cctaaccctc cttcccgcct a 21 75 26 DNA Artificial sequence Description of artificial sequence Amplicon primer 75 actgacttat ctttactgtc acttct 26 76 19 DNA Artificial sequence Description of artificial sequence Amplicon primer 76 gcaagacagc caccaaccc 19 77 24 DNA Artificial sequence Description of artificial sequence Amplicon primer 77 tgacttatct ttactgtcac ttcc 24 78 0 PRT Homo sapiens 78 000 79 18 DNA Artificial sequence Description of artificial sequence Amplicon primer 79 cctgctccca ggactgac 18 80 24 DNA Artificial sequence Description of artificial sequence Amplicon primer 80 cccagacttt atctgtgact gctc 24 81 18 DNA Artificial sequence Description of artificial sequence Amplicon primer 81 cctgctccca ggactgat 18 82 17 DNA Artificial sequence Description of artificial sequence Amplicon primer 82 ggaaacaggc ccgtaga 17 83 18 DNA Artificial sequence Description of artificial sequence Amplicon primer 83 gagtgccgct acttggcc 18 84 16 DNA Artificial sequence Description of artificial sequence Amplicon primer 84 gaaacaggcc cgtagc 16 85 22 DNA Artificial sequence Description of artificial sequence Amplicon primer 85 tctggaggac ttctaggaaa ac 22 86 18 DNA Artificial sequence Description of artificial sequence Amplicon primer 86 tgcagccatg tcgccttt 18 87 22 DNA Artificial sequence Description of artificial sequence Amplicon primer 87 tctggaggac ttctaggaaa at 22 88 130020 DNA Homo sapiens 88 aagcttcact ctcacgctct gtggtgtgac atccactcag caaagaggtg caaacctgga 60 cccccagagt gtgaatcaca gacacctctt ataccccagc ccctctgctg ggaatctagc 120 ggcagaaact cccgagctca tgggcggggc cttcctgagg gcaggctccc tttgcagcct 180 gtgtttgcct ctggggccgc tttgggtaca cacatacaca cacacacaca cacacacaca 240 ccatgccaaa gccacctccc aggcccttcc atgccctgcc tagttctgtt ttgttttgtt 300 tgagacggag tctcactctg acgcccaggc tggagtgcgg tggcatgatc ttggctcatt 360 gcaacctctg tctcccaggt tcaaacaatt ctcctgcctc agcctcccga gtagccggga 420 ttacgggcgt gtgccaccac gcccggcaaa tttttttgta tttttagtag agatggggtt 480 tcaccatgtt gcccaggctg gtctcgaaca cctgacctca aacgatccac ctgcctcagc 540 ttcccaaagt gctgggatta gaggcgtgag ccacagtgcc tggcccctgc cctggttctt 600 gttttgtgtt ttgagatgag gtctcgctat attgcccagg ttggtcttaa agccccaggt 660 tcaagcaatc ctcctgcctc agccttccaa agtgttggaa ttacaggcat gagccacctc 720 gccctgccac ctgccctaat tctgaactca cagataaaac tgctcccacc aaccaggccc 780 tcctgcccct gggacccccc tctacctcca gcctggaggg aaattccagg cctccctgga 840 gcttatgaag gggaaaaaac ccaagagggt tttttttttc tgtaactcag tgtcgtccat 900 tcgcctcatt atggggtcag gatcccagaa agtcagggtt aaaaggacat ttgctctaca 960 aatagggaag ctgtcccaga ggtaggtatg cctcatccat ggtcacacag atgggcagag 1020 ctgagccgga acacgaaccc agaagtcccg ggtcccagcc tagctccttc aatacctctc 1080 aaacctatta tcaacctttc acactttagt aaaaaatagc gcaactcaaa acagaactct 1140 aaggtgtagt gaggacatag gccaggtcag atctgatgga aggaggagag ctggcccaga 1200 gggcagggag taggggaccc agcccctacc tataatccta gcactttggg aggccaaggc 1260 aagtggatcg ctggagccca ggagttcgag actggcctgg gcaacatggc aagatccccg 1320 tctctactaa aaatacaaaa attagctggg tgtagtggtg tacacctgtt gtcccagcta 1380 cacgggaggc tgaggtggga ggatcacctg agcccagaga ggtcagggct gcggtgagct 1440 aagattgagt gccaccctaa gcctggggtc ccaggatact cactgctctc agccaggaga 1500 aagctgccac actcctggga gtagttgcca gactggtctg tgatgttgac actgaaaatg 1560 tcgtcggcca tgaagtggaa tacatccatg tggcaggtgt aggtggcatg cgtggcattg 1620 tgggccgacc tgtggaggct gcaggaggtg gcctcgtcct tcagctcttc atactggtct 1680 tgccttcaag gacaaagcag ccggtcatgg ctggaagcca ggggcagggg ggatggtggt 1740 gggctcagtg ctcagggcca gggcagaagc ccagactctc agggctggga gggatttgag 1800 gcagggctgg ggggtggtca cctggtccaa ccaacctttt cccagggaag gtatcacctc 1860 ccctacagca tcctcaatga attgtgtccg ggctcagctt gcacacctcc caggggccag 1920 gaactcacta cctcatttag ggcagcagct ctcaattttg gaaatttggg ccattcttgc 1980 ttggactaag cctaaatctg tccctgttgg tcccaacaac cagatggctg tcctgcagga 2040 agggcgctag gcattttggg tttccttctc tcctttctgt ccaacatcca tttttttgag 2100 acagggtctc actctggccc caggctggag tatagtagtg tgatcatggc ttactgcagc 2160 ctcgaccttc ccaggctcag ctgatcctcc cacctcagcc tcctcagtac ctgggactac 2220 atgcatgcac caccatgccc agctaacttt ttgtattttt tgtagagatg gggtcccacc 2280 atgttgctta ggctggtctc aaactcctgg gctcaagtga tctgccagcc ttggcctccc 2340 aaagtgctgg gattacagga ctgagccacc tcgcccagtc cccaaatcca ctgaatatct 2400 aataatagcc tgtgctgagg gccaggaaaa gagctaaatg aacctcatct cctcctctgg 2460 gtggacttct gggaaagttg gtgggatggg attagggaaa atgctgtggc tttttaactc 2520 ccactggaca aaaatgatgg gttttgaagt atgtatagta gtttaccgga tattaatttc 2580 cctacctaga tgttcaggat ctgaacatga ccactctcta ggcccagtag ggatgtgtgg 2640 actgagttct ctgccaaatt tgctggaata caagttcctg ctgaataaat gctctccaag 2700 cctcccagcc tgaacttgct ccttattccc cctcacctgc ctcactccac tccagccaca 2760 ctgacctcct ccctattcct caaacgcacc agctcttctc ccacagggcc tttgcactgg 2820 cagtgccctc tgcccagctc cctggcttcc tccttctcac tagcagttgt ggtcccttta 2880 cctgaggtcc agtgttctgt ccttgatata gcacccagta ttattagaaa gtatcctgtt 2940 tggggcagaa cacggtggct tatgcctgta atcccaacac tttgggaggc tggccatgcg 3000 gcagaccggc catcccgggg tctcaaaacc atgggcgacc tgtgtgggcg agagagtggt 3060 ctttgaagct ggtggataga ccaaccctgc gggctcctga cttttcaagt cctgtgagcc 3120 ctgagtggtt gtctgaaagg cgggtacagt gggcggatca cttgaggcca ggagttcgag 3180 accaggtggg ctaacatgac aaaaccctgt ctctacttaa aatacaaaaa ttagccaggc 3240 atggtggtgc acacctggat gcaatcccag atacttggga ggttgaggca ggagaattgc 3300 ttgaacccag gaggtggagg ttgcagtgag ccgagatcat gccaccgcac tccagcctgg 3360 gcaacagagt gagactccgt ctccaaaaaa aagaaagaaa gaaagtgtcc tgtttgggcc 3420 tggcatggtg gctcacacct gtaatcccag cactttggga ggctgaggtg ggcagattgc 3480 ttgagcccag gagtttgaat ccagcctggg taacatggca aaaccctgtc tttacaaaaa 3540 ctacaaaaat tatccaggta tggtgatgca tgcctgtagt cccagctact caggaggctt 3600 aggtggaggg attgcttgag cccaggaggt caaggctaca gtgagttctg atcccaccac 3660 tgcactccag cctgggcaac agagcaagac cctgtctcaa aaaaataaaa taaaataaaa 3720 taaaataaaa taaaaaagag aaagcacctt gttggttgtt ttgtgtgtct gtctccccca 3780 gcctccgccc caccgtggta cgtcagttcc aaggcatctg gggcctggtc agtcttgctc 3840 tgctgaatcc ccagtgcctg caacagtgtc tggcaggaac ttaatacatg ttttgttttg 3900 ttttgttttg tttttgagtt gggatcttgc ttttttgccc aggctggagt gcagtggcgt 3960 gatcatagct cactgcagcc tcgacctcct gagcttggtc cccctgcctt agactctcaa 4020 gtagctggga ttacaggatt gtgccaccac gcccgactga tttatttttt gtagagacag 4080 ggttttgtga tgttgcccag gctggtctca aactcctggg ctcaagtgat cctccttcct 4140 tggcctccca aagtgctggg gttagaaaca tgagccactg cacctggcct ataaatggtt 4200 ttgaatgaat gaatgagctg atggataaca gaagctcttc tttatcctga gctgcatagg 4260 gtctaaaagg gacccatgga ctttggagtt gggcggtctg aatttgaatc ccagctccag 4320 cattcactgg ctgtgtgacc ttaggcaagt tgcttaacct ctctgaacct caatgtcctc 4380 atctgtaaaa tgggagtaat cattgtgagg gttgggggtg accattcatg ggaagcgtgt 4440 agttgggggc ctggtaccta agtgcttgaa aaatcatgcg ggaaaccagc catcccgggg 4500 tctcaaaacc atggccgacc tgtgtgggtg agagagtggc ctttgaagct ggtggataga 4560 ccaaccctgc gggctcctga cttttcaagt cctgtgagcc ctgagtggtt gtctgaaagg 4620 cggacacagt ggtcctcagt gcacagtgtc ctcctggggg aatcactgag caggcaccct 4680 gaattgcatc cccggggact ggtgaggccc ggctacttac caggtaaggg tgagcgtgct 4740 ggggtggagg ttccacattt ccaggatgca gatgaccgtc tggaggtaat cggtgtagca 4800 gacgaggtcg gggcagcccc agcctggagg gtcagtgggg agagaggcct tggtgggggg 4860 tggcttggga ctgcagagct tcctctccat cccctcgagg gctggaggaa gaatgggtgc 4920 aggggtcccc aggccgggac aaatatgagg actcaggagc tgcttagatg taggcctggc 4980 acccaggggg cctcaccacc tggaaattcc ttgtttgagc accaatgttt caaggctggc 5040 tatgaagaca ggatgcatgt gatggcaaat aatataaaac tagctggtcc ccagctcagc 5100 accctcctcc agagccctgc gcccggccac caggtctgtg tccctcctgg ccaagctccc 5160 actcagaatg gcaatatcct aaatgcccca gatgtggtag gaacccagcc agagcacacg 5220 cacacacccc aagagggtca ggacctcata gagtcaagga cagctatgct gaaaatgtcc 5280 caaaaccctg acaaggaaga ggactcggct ctacagaagt tggatccaca ggacacccac 5340 tctcggagat tcaggaagtg cttcaaggag taggaaaccg tgagagaatt aaaaagccct 5400 cggacacttt tgcataaagg tctgcgtgat cctgcggcca aaaatgccac cgccaccaag 5460 aagtcttcta ggtattgccc tatgaggcaa ttttgttgga agatttaaca cctttccatc 5520 ctgctcccca aacattaatt gtttcgctgt gttctaaaac aggggtgcaa tcatcatcta 5580 cattttgtta aatgtggaca aaatcaatgg aataatttgt agctttgttt tgtatattcg 5640 agtgccaatt ccactatttt ttgctgctat ttactagaaa cttgtaccgc tcacctcttt 5700 ttgagatagg atctcttgct ctgttgccca ggctggaatg cagtggtgcg atctcagctc 5760 actgcagcct ccacctcctg ggctcaagca gtcctcccac ttcaacttcc tgagtagctg 5820 agactacagt tgtgcaccac catgcctgga taatttttgt gtttttgtag agacagggtt 5880 tcaccatgtt gcccaggctg gtctcaatct cctgagctca agcgatccac ctgcctcagc 5940 ctcccaaagt gctgggattg taagaatgag ccatcgtgcc tgaccttgtt ccccttttta 6000 agtaggttca tttttggtgg ctttttccta ttttccctgg actaatgaag acccctgagg 6060 tggtggtggt agattccctt cctgggtctc actctaggac agaacctata tgcggagcca 6120 agaatactgt gtctggaaat atcctttagt tcattgtaga gatgagaaca ctgaggctcc 6180 aagactagac catgcctctt gtctctcttg gtctgggctg ctgagacact cacccaggga 6240 aggaaatacc tagtctgtgc tagtttttca tagccgcaca catgacagat attgctaatg 6300 gatcacagca ctccctcccc taggtgtgac ctcggaatgc ttctcaactt ctctcgaggc 6360 agtggctacc aagggatctg agtcatggaa gtaaatcttg ctctaggagg gggctttcct 6420 cagggagaca cacagaactt gtcggtggag tgaaaaggag ctagttccag cacttggtgg 6480 agaaaacagg actgattttg gcaaaacctc aacacagagc ctctaagtag ctgtgtcatc 6540 tctgagttcg tatcctcgtg gttctttctc agagctctgg gaatttggag aggagctacc 6600 attcctagcc aggctcaaag ctaccaaggg acagggtttg tctccaaact tggggaggca 6660 acggcagccc ctgagtggca gggaggtggc accagggatg agagggtggg aaaatgccct 6720 gtcagtaccc cagccacagg gcagagagcc ccagggccat cccatgtctg gggttatgaa 6780 agcgtgcagg aggcagtgag aggtgcacaa agtagctctg tgactgcaaa catgtgactc 6840 aaaactgctt ctcctcccca tgtgtaagcc ggagacactg cttgccctag aggctgttga 6900 ggtgttgaga tgagatcatg cctgtgcctg catgctgccc gcacatggct atcactccac 6960 agatagcagc acagcatggt gaaggccgtg ctacggtttg acgtctgtgt ctccccagaa 7020 actcagttgc caatgtgatc atattaaaag gtggggactc taggagatga ttaagtcatg 7080 ggggcagagc ccccctgaat gaattattga tgctataaga gggctggacg aacccaacat 7140 tcagaccctt tttgcccttc tgccatgtga ggatgcagcg tgtgtcccct ctgggggtgg 7200 tgcagcgttc aatgtggcat cttaaaggca aagatggggg ctgggtgcgg taatcccagc 7260 acttttggga ggccgaggcg ggtggatcat ttgaggtcag gagttcaaga ccatcctggc 7320 caacatggtg aaaccccaac tctactaaaa atacgaaaat tagccatggc atgtgcctgt 7380 tatcccagct gctcgggagg ctgaggcatg agaattgctt gaacctggga ggcagaggtt 7440 gcagtgagcc aagattgcac cattgcactc cagcctgtgt gagaaagcaa ggctcagtct 7500 caaaaaaaaa aagcaaagat gggcccctca ccggaatcaa agctactgct ggtgccttga 7560 tcttggactt cccagcctcc agaactgcga gcagtaaatg tctattattt ataaattacc 7620 cagtctcacg tattcagtta cagcagcaca gacagactaa gacagacccc aagagcaaac 7680 tgatcccagc tctgcctctt ccgggtagtg tggccatgca cctgtctctt agcctctctg 7740 agcctcctca gccaaaaaac tgggcgtcat tacaacatgc gccttgtggg gctgttggga 7800 cagtggaatg ggatgacgct ccgagctttg catgcagcca gggcttgtta tctgcacccg 7860 ccagatgtgg agccatggtg ggtgacccaa aggcttttta gaaccaagat gaggatgctt 7920 gccactcaga aagggaaaat caaagcttga catctgtttc ctgaaggagc tatttgaggt 7980 cctgggtctt ctggtaaaaa ccagagaaaa atgcccatga gtcagcagag cagctgttga 8040 gttcctcccc tgcccctagg ccggcctcca cacaagtctg tgctggatcc cgcctcctca 8100 tggccccctc ctccaggaag ccctccagga tctcttagtc ctaggcagtg ccctttcagt 8160 cggactcctg gtcttcccag ttctgctttc taacgtcacc ccctctaacc acaacatctg 8220 gtcctctggg ggctgcctac caaggttcag tctccttttg gctcttcctc ctgtccccag 8280 cccagggacc accagtgcct gctaagaatc tagcgatttt tttgtttgtt tattgttgtt 8340 gttgttgctg ttgttttgag acagggtctc gctctgttgc ccaggctgga gtacagttgt 8400 caccttgact cactgtaacc tccacctccc aggctcaagc aatcctcccg cctcagcccc 8460 acaaaaagct gagactacaa gcatgcgctc ctatgcctgg ctaatttttt ttcttttgta 8520 gagacagggc ttcaccatgt tgcccaggct ggtcttgaga tcctgagctc aagcgaccta 8580 ccctcctcag cctcccaaag ggctgggatt ccagtaggag ccaccgcacc cggtgcagca 8640 attgttgaaa aacatttcca aggagagtca ttacataagg agaaactgag gttagatgga 8700 aggcagaact tcctcactca gagagttggg agccagccaa ggcagaggtc aggggaaatg 8760 aaggacgaag cctttcatcc ccccagtgac acccgaagac acaagcctgt gggctgggcc 8820 acctgagact catgtctacc gcttctgttt catggagccc tttggtggat ggattctgat 8880 tagtggagag ccggctactg gactcgtgtc agcctcaagg gcatggatgt catcttttct 8940 gaatctgtgg actctagccg tgtttttgag cccagaaagc cagcccggcc ctggctcagc 9000 tctgtgatgg gggccctccc cacccgcaga ccacggggca gccacttacc tccctggagc 9060 agcagcagga gcaagggggc ggcccagcca cgcggcatgc tgactcccac gggcctgcaa 9120 gagacgtaca gtggagggtg agccaggcgg gctcagggct cccccggcct ccccctcatc 9180 ctgtctcttc ccaggcagac ttgcccccga ttcttggaac tgtcttaaga aaaatgcaag 9240 atcccgaggg gcctgtggat gaccccaaga actcccacct cttagaaatg gcagattcac 9300 cccaagaatc ccttctccca ccttcttcct gcccccgccc catcacgtca caaatgcaaa 9360 gtgaatttgt tttattattt tattttaata tttttagaga tggggtcatg ttctgttgcc 9420 caggctggag tgcaatggcg caatcacggc tcactgcagc ctcaacctcc tgggctcaag 9480 cgatcctcct gcatcagcct cccgagtagc tgggaccaaa ggcacgtgcc accacgcctg 9540 gctaattttt aaaacatttt tgtagaggcg gcatcttgct atgttgccca ggctggtctc 9600 aaactcctgg gatcacatga tcctcctacc tcggcctctc aaagtgctgg gatgacaggc 9660 atgagtcacc acgcccagca caaggtgaat ttaaagaggg aggagactgc tgactatttt 9720 gcccttgctg agcctattga tgtagcattg tgtgtgaaat taaacttgtg tatttatcta 9780 tctaacaaag aaagagggag ggaaaaagag aggaagggag agagaaaagg agaaaaaact 9840 agtttggaaa gggggatctg ctgaggagca attttggatc ttccttccta taaacatcca 9900 tgttgtaaaa aaaagccagt aaattggctt ttttatgttt attttttaga caaagtctca 9960 ctctatcgcc caggctggag tgcagtggcc cgatcttggc tcactgcaac ctctacctcc 10020 caggttcaag cgattctcct gcctcagctt cccaagtagc tgggaataca ggtggccgcc 10080 accacgcctg gctaattttt gtatttttag tagagacagg gttttgccat gtggactagg 10140 ctggtctcga actcctggcc acaggtgatc cacccacctc agcctctcaa agtgctggaa 10200 ttacaggcat gagccactgc acccagccat caataaattg gcttcttact gaataataaa 10260 tccaaagtta acccacaaag ttaaccactg ctgaatactt aagataaggt tagcccaata 10320 atgaggctcc ttactccatc acaaagccca gattagccca ctcgggagca tgctaggaaa 10380 tccaggcctg accaaacaga gacaacctga cctcctaatt tgggccagga tggagaagga 10440 acctctttgg ggagcagagt gagtttatcc agaaagagtg ccctctactg tcttgggcag 10500 aggaatgccc caccagtctg gcgatgccag gcttctcctg ggacttgtct gtttcaccca 10560 ccagctctgt gtctgtttga ggaagtgcca gccagagggc agtggtccac ctgcccagac 10620 tcaggagccc ctcccagacc agcccctata gacaagaatg gagctaaagc taaacctaag 10680 agggatgccc catgcaggac ccaagtcaca ggcatggctt tctccagacg ccatccctga 10740 ctccacaagg ggaagatcct cctcctctgt gccggggttc ccaaagatcc tactctggcc 10800 ccatccatag ttccctctga cctccctcac agagccatca aagaaccctg gggcagggca 10860 caggctgggg ggccaggaag gttcagatga ccagctgtgt ggctccgggc cagtcccttg 10920 ccctctctga gccttgtaaa gtggaaacag caaggcctgc ttccaggctt gtgagaattc 10980 aaggaggtga tttgcataaa accctgcaga tggcccccat cacataaaca ctcaaacaac 11040 agctgtaatt gatggagaag gtagagcctc agaaacagag

ggggtctcag aatgatctag 11100 ccaagagttg ccaaaatatc tcatatccaa aattttggcc atatttgtat attgtgtgca 11160 ctgttgttta ttttaataca tattttaata tattttctat agataagcac acttaaaaat 11220 ttaactccag catcatccta agcaataaca tcactactca aataaaataa tgttgaggta 11280 gatgcagttt caacctgctt ttaggaatat gaatcctgtg tgattccaac ccctgcagag 11340 gtcctgttat cacactttga agatgtggcc tctgatcgca ggagggggag tctgtgtctg 11400 ccccctcact attctgtcca catccatcac cccagccacg gtgcgagtcc tgacatagct 11460 ctgtccccgt cactccctga tggttttatc ttgaccacag cacagcaaaa agtgcaaact 11520 cctttgccca gattcaatat tcctcatctc cagcttgacc aacatagcaa gaccttgtct 11580 ctaccaaaaa taaaaataga agtaaaaata agttagccgg gtgctgtgtg cctgtggtcc 11640 cagctattcg ggagactgag gaaggaggat cacttgagtg ccggaggtcc aggctgcagt 11700 gagccatgat tgcactattg cactccagcc tgaatgaaag agtgagaccc tgtctttaaa 11760 aaaacaaaca gacaaacaaa aaactcctca tccctcctaa tttcttacct ctgcctacat 11820 cacagagccc tagggatgtg gagggggtac ctgctgcttc tgcctgcccg gcgtctcttc 11880 ccaaagagct cctctatctg agctgcctct agtggtttgc ctgaggctga ctccacccca 11940 tgctccacaa agtgggctcc tgacccaggc ttggccaatc aaaacacccc atctgtctga 12000 ccacagtgat tgcatcaggg atggatgtca acccaggcaa ggccagtgaa agcagtcctg 12060 agattttttt tctagaactc tagaaaactg acactttttt tggtatcttc tcagagcttg 12120 tagccatctt tgtcacctct tagagacagc ctccttgaaa atgacagcca tactgagaaa 12180 tacaggtcag gagtccccat ggcatcatgg gaggtcctgg atccagccat atctgatgtc 12240 caccctgtgg atttctcatt cccatgagcc aaaaaccacc tatgttgctt aatcatgttg 12300 aattctattt ctgtcacttg ccaccaaaga tccctgacac agtggagttg agaaaagggc 12360 tcagaatttg aagccagaag atcttgaagc aaatagtagc tcttctctgg ctcttttatt 12420 ctctgaactc agttaccatc tctgaaatgg ggacaatagt gctcactgca cctggcagtg 12480 aagaagaggc tgtgagaaag cagatgtggt agtgcttcat ggactgtgaa gtgctgtacc 12540 agtgcacggg tttcctgcca ttactggctg actgccctga attactcatc gtctccccgg 12600 cagcccaggc ttcgtcatgc atctttgcct ctgctggagc agactcctgc cctccaacac 12660 ctttcccctc ccatcctctc aactcaactt tcatggtcca tgtgctgtat ccctgcagcc 12720 acactggagt gttccagaac tcctgccact agcctattgc gtttgaggtt ttcactggtg 12780 ctactattaa ttatccactt actattgatc tttaccttgc ctcctatttt ttactccaat 12840 aaacatgtct caaaaatatt atatcatgac catcagcaga taactggcat gaaataggca 12900 gtgtgcgcca tgctggtcaa gacacagact atggagctgg ctggtctcag ttcaaatccc 12960 agctctgcca cttataggct ttgtggcctt gggcagtttg cttaaccttt ctgggcctca 13020 tctgtaaaat gggagtaatg ataatcacag cacctgcctc ataggtcttt gggagaatta 13080 aattagtcaa catttataaa gcatttagaa cagtgcctgg catatagcag atattatata 13140 agtgttactt aaacaaaaca atcaacttac cataaaacta aatacaatac aacataaaaa 13200 ctaaatacaa gaaaaacaaa ggcagtaatt gaattttagt gacatgctac tgcctgcctg 13260 agagtcctag aaactggaga tgtggtttgt ctttgtttaa agttaagaaa gggaggccgg 13320 gcacactggg tcatgcctgt aatcccagca ctttgggagg ccgaggtggg aggatcattt 13380 gaggtcagga gttcgagacc agcctggcca acacggcgaa accccatctc tactaaaaat 13440 acaaaaatta gccgggtgtg atggtgtgtg cctataatcc cagctactca ggaggctgag 13500 acaggagaat tgcttgaacc caggaagagg cggaggttgc agtgagcaga gattacacca 13560 ctgtactcca gcctgggcaa cagagcaaga cactctgtct caaaaaaaaa caaaacaaaa 13620 cacaaaaaaa gagagagtag tgtttaagat gcgatagcac caaattgaga atcttgcctc 13680 ctccatcagc agggcaagtg atgaaagagc ctacgctttc actaggggat ccagggatat 13740 ttaacactgt ttaaaataaa ctgctcactt ctgggaaaca tcagcttaaa aacacagact 13800 ttctccatca ctccctcctc atgctcccac agcaccaaat ggggcctacc ctaattccac 13860 tgccttctct gccccgaatc catccccttt tgggtctttg cacgcaggct gacccctgag 13920 ggcaacagca ccttatcctc agatacagat cgggccacca tgtgtgtttc ctacaaggac 13980 agaaggtgtc ttaggaggct tcctgctaca ggagcccctt gatatggttt ggatatttgt 14040 cccacccaaa tctcatgttg aaatgtcatc cccaatgttg gaggtggggc ctggtgggag 14100 gtgattggat catgggggcg gatttctcat gaatggctta gtgccatccg cttggtgatg 14160 agtgagttca cctgagatct aattgttcag aagtgcgtgg cgcctccccc ttttgctcca 14220 gctgtcgcta tgtgatgtgt ctgcttctgc ttcaccttca gccttgagta aaggctccct 14280 gagggcctcc ttaccagaag ccgagaagat gctggcagta tgcttgtaca gcctgcagaa 14340 ccatgagcca attaaacctc ttttctttat aaattaccca gtctcaggta ttgctttatg 14400 gcaatgcaag aatggcctga cacaccgctg cagcacccaa cctctgcagt ctcctacgga 14460 ggagtgaagt gcacctgact tggagtgtgc caggtcccag ctgtgcgtct ttgggcaagt 14520 tctctaaccc ctctgattct ctgttttctc atccataaaa tggaggtcat caatctttgc 14580 tgcaggtttg tcatgagcat taaaagtgtc atcgaatttg aagtcctgta ggtgcattta 14640 tccaccagtt tcagtgctga gaaatgtttc ctggagttgc tgtaattcac cctaaaagtt 14700 gaccaaattt caagagattc tggggaaatg gagaagcttg gggcaggaaa ttagattgaa 14760 gtgagtgcta tggagaatat aaggtgctta acttcatggt caacgtggtg gctcatgcct 14820 gtaatcccaa tactttggga ggccgaggtg ggtggatcac ctgaggtcag gagttcgaga 14880 ccagcctggc caacatggtg aaacccatct ctactaaaaa tacaaaaatt agccaggtgt 14940 ggtggtgggc acctgtaatc ccagctactc tggaggctga ggcaggagaa ttgcttgaac 15000 ccgggaggtg gaggttgcag tgagccgaga ttgcaccact gcactccagc ctgggtgaca 15060 gagcgagact ccatctcaaa aaaaagaaaa agaagagaaa agaaatacaa agaaaaacca 15120 caatgacata ccattttaat tttgtcggat tggcaaaaac tgaaagtctg atgacatcac 15180 atattgctca ggatatgata taataggaac tctcatttat agccagagga aatgtaaatt 15240 attacaactg gtctggtaaa aagtttggca tgacctaata aaagcgaaca catgccctac 15300 ctaaaaccca gtaacttcac tcttatgtaa actggtaaag aaattcttcc tgcatgtcta 15360 tagcagcatt gattttaata gtgaaaaatt ggaaacaatc taaatagcca tcatcaggag 15420 agtgaaaaaa taaatcgcaa tactgtcact cagtgtaaac atgaatcatt tacagccaca 15480 cacaacaaag tgaatgactc tcacaaacaa aatttggagt gaaaaaagga agttagagaa 15540 cagcataagc ataataacac cattgacacc acactgaaaa ccatacaaaa tcatcctata 15600 tattgtttag gtaagcacac ccatcataaa aatatttaaa aattcatggg gataataaca 15660 tcaaattttg aatagtgtgg aaaaggaggg ggtgtgagaa ggaggaatgt tcaagggtct 15720 caacagtttg gtaaagcttg gtgaagctac catgggtgtt gattatattt tatgcccttt 15780 tgtatgttca aactatttca taccatttta aatgtaagtg aggcttagtt ttgtaaaata 15840 aaatatggat gagggacttc cacttctggc tataatgaag tagctgttgt cacactaatc 15900 ctcccactga gggcaactac taagcagaaa aaaaaaaaac tgcacaagag agtgccaact 15960 tatatagtac ttaagagagg aaaaatgcat tgaaattctg catgcaattt cccctcaaat 16020 catgtgcaaa gtcattgaaa ataagtagca tgccaagaaa cttagcaaaa aaacacctgc 16080 taataggtgg gaaaactaag aaggaatttt cacagtttta caagctgagg gaagaattag 16140 agttcagggc ctggcaaaaa aaagatgggc cctggtgagt acccagctat ttagaaaata 16200 ctattccttg ggaataagga aaaattagaa atagttcatt cttgactgca tcaaggtgta 16260 atctgcttat atgctatatg tctgctagaa gaaaatttaa attgatctag aaaaagttag 16320 cttcatccaa agcctctata ctttttatac ataatgtcca actttcagtt aaaattacaa 16380 ggtaaaccaa caagcagtat caaatgatca acaatgaaga taaaatagac aatagaaaag 16440 ttacttaggt gaattggaaa atggcattat cagatatgat ttaaaataac tatgattaat 16500 gtgttcaaaa attctatcag aaatttggaa cctatttaaa aagtagaaat cctggaactg 16560 aaaaatataa taactgaagt ttataaggac tgaagagatg gatttggcaa catactagta 16620 acatcagaag agaggattag taaactggaa gattggtcaa tagaaaatac caagattgaa 16680 gcatgcagag aaaaaaagtt caaaaataca gaaaagagca aaagagacat agaagacatt 16740 gtgaaatgat ctaacataca tgcatttgca attaagaagc agaagagaga gaaaacaagt 16800 aagaagaaat agcagtgatc ttaaaatttt tttccaaact aacaaaagac attaagccac 16860 agatttaaat agctctacaa accacaaata ggataaatac aaagaaaacc acatctaggc 16920 atatcataga aaaactattt taaaaaataa aagacaaaga tatgcagtat gcagaaaaaa 16980 aagacacttc accttcaagg aagaaacaat aagactggca ggggactttt taacaaaatg 17040 atggaagaca gcagacaatg gaacaacttc tttcaatgct gaaatttcaa aagccaattt 17100 agaattttat atccagaaaa aaatctctta aaaaataaaa gtaaggtcaa tatgatacag 17160 acagctataa aaatattaca gttaacatca tacttaatgg taaagaaatg aaagctttgc 17220 tcctaagatc atgaacaaga caagtatgtt cactcttgtg acttttataa aacattttac 17280 tagaggttct agccgaggca attaggtaag gaaaagaaat aaaagacttt cagattggaa 17340 aggaagaagt aaaccatttc tctttgcaga taatatgatc ttgtatatac aaaactaaag 17400 gaatttacac ctaaaacgaa caaaaaccaa ttagagctaa taaacaaatt tagcctagtt 17460 gtagaatata agattaatag acacaaataa tgatatttta tacacactgg caataaataa 17520 tctgaagatg aaattaagaa agtgatttca tttacaataa tacaaaacag aacaaaatgg 17580 ttaaagtaaa tttaacaaaa ttcattagtt atgttaatat gagatgaatt tgtttattag 17640 tgtgagactt gtgcaataaa acctacaaat gttattgaaa taaattatag aaaacctaaa 17700 taaatggagg atataccatg ttcatagatt ggaaaactta atgttatcaa gatgacgcct 17760 caaattgatc tacagattca aagtaatctc tataaaaaat cccagctggc tctttttttt 17820 tttttttttt ttttttgcag aaattgacaa actagtccta aaatttatag ggaaatgtaa 17880 gggactagaa tagccaaaaa ggttttgaga aagaacacag ttggaggagt caaacttcct 17940 tatttcaaag cttactacaa agctacactt atcaacactg tggtactgac acatagatgt 18000 acatatagat caatgggatt gaactgagag ttcagaaaga agtgcatata tttatgttta 18060 accgattctt gatgagggtg ccaaaacaat tcaatggggg aaagaatagt ctttaaaaca 18120 aatggtacca cacaaatgga tatccacaat gcaatataat gaagctagat ttctacttca 18180 caccgtatac aaaaatttac tcaaaataga ttaaagacta gatgtaaaaa gtaaaactat 18240 taaactctta gaataaacct aggtgtaaat cttcatgacc ttgtatgaga caataatagt 18300 tttttaaata tgacacaaaa gcataagtaa acaaagaaaa aatagatcta ttaaactgta 18360 tcaaaattta aaatgtttgt gcttcaaaaa atactatcaa gaaagtaaaa ctataacata 18420 cagcatggga aaaaaagcat ttgcaaatca cgtatttgtt aaaggtctgg tatttagaat 18480 atgcaaagaa ttcttgcaac tcaagatcta aaaggcaaga attaaaattt aaaatgggca 18540 aaagatttgg atagacattt ctccaaagaa gatatccaaa caacaaacac atgaaaaggt 18600 ggccattgac attactctct agagagatgc aaatcaagaa cataatgaga taccacttca 18660 catgcattag gatggtaata ataataatca atcacaagtg tgaatgaggg ggcggagggt 18720 gagaatgtaa aatggtgcag ccactgtgga aaacagtttt gcagttcttc aaaatgttaa 18780 atacagtcac gtgctgcata atgatatttc attcaatgac agactgcatg tacataggcg 18840 ggcccatagg actataatga aggtgaataa ttcctattgt ctagtgacgt catgacacaa 18900 cactttactc atgggcttgt gatgatgctg gtggaaacaa atttactgca ctgccagtca 18960 tataaaagta tagcatatgc agggctaggt gcggtggctc acgcctgtaa tcccagcact 19020 ttgggaggcc aaggcaggca gatcatttga ggtcaggact tcaagaccag tgtgacaaac 19080 atggtgaaac cccgtttcaa ctaaaaacac aaaaaactta gccaggcgtg gtgacacatg 19140 cctgtagtcc cagctactca ggaggccgag gcaggagaat cacttgaatg gggaggcaga 19200 ggtagcagtg agccgagatt gcgccactgc actccagcct ggacaacaga gtgagactcc 19260 gtctcaaaaa aaaaaaaaaa agatagcata tacaattatg tattgtacat aatgcttgat 19320 aataataata ataataataa taaactatgt tactggtgta tgtatttact atactatatc 19380 ttaaatcact attttagggt gtgctccttg tacttactta aaaaaaaatg ttggaggggc 19440 acagtggctt acgcctgtaa tcccagcact ttgggaggct gaggcaggca gatcacttga 19500 ggtcaggagt tggaaaccag cctggccaac atggtgaaac cccatctcta ctaaaaatac 19560 aaaaattagc caggcacggt ggtgtgtgct tgtaattcca gctattcgtg agactgaggc 19620 acaagaatcg catgaaccca ggaggtggag gctgcagtga gccaagattg tgccactgca 19680 ctccagcctg ggtgacagag caagactctt aaaaacaaac aaaaaatgtt aacttgtaaa 19740 acagcctcag gcaggtcctt caggatggat tccggaagaa ggcatcatta tcataggaga 19800 tgccagctcc atgtgtgttt ttgccactga agaccttcca gtgggacaaa atgtggaggt 19860 ggaagacagt catattggtg atcttgaccc tgtgtaggcc taggctaatg tgtctgcctg 19920 tgtcttagat tttttgtttt ttgttttctg ggtttttttg agatggagtt ttgctcttgt 19980 tgcctaggct ggagtgcaat ggcgtggtgt tggttttccc gggttcaagt gattctcctg 20040 tctcagcctc ccaagtagct gggattacag gcgcccgcca ccatgcttgg ccaattttgt 20100 atttttagta gagatggggc tttcaccatg ttggccaggc tggtctcgaa ttcctgattg 20160 caggcgatcc gccctccttg gcctcccaaa gtgctgggat tacgggcatg agccaccatg 20220 ccccgcgtgt cttaggtttt aacaaaaaaa atttttttaa gtaaaaaaat aaaaaataaa 20280 aaattttaaa atataatata taaaaaggct tttagaataa gaatataaag aaaaatattt 20340 tgtatagctg tacaatgtgt ttgtgtctta agtgttagta caagagtgca cgaaaacact 20400 taagtgttat tacaataaga taactgttat tacaaaagtg ttattcaaaa agttttaaaa 20460 agttaaaatg tttataaaga aaaaatgttg gcggggcgca gcggctcatg cctgtaatcc 20520 cagcactttg ggaggccaag gcgggcagat cacgaggtca ggtgatcaag actatcctgg 20580 ctaacatggt gaaaccccgt ctctactaaa aataccaaaa aaattagccg agcgtggtgg 20640 cgcgcacctg tagtcccagc tactctggag gctgagacag gagaattgcc tgaacctggg 20700 aggcggaggt tgcagtgagc tgagatcgcg acactgcact ccagcctggg tgatagagtg 20760 agactctgtc tcaaaaaatg aaataaaata gggccgggcg cggtagctta cgcctgtaat 20820 cccagcgctt tggaagggcg agacgggcag atcacaaggt caggagatcg agaccatcct 20880 ggttagcacg tgaaacccca tctctactaa aaatacaaaa aattagccgg gcgcagtggc 20940 aggcgcctgt agtcccagct acttgggagg ctgaggcagg agaatggcgt gaacccggga 21000 ggcggagctt gcagtgagcc gagatcacgc cactgcactc cagcctgggc gatagagcga 21060 acctccgtct taaataaaat aaaataacat aaataaaata aaataaaata aaataaaata 21120 aaataaaata aataggaaaa atgttacagt aagccaaagt taatttatta ttttaaaaat 21180 tcaaataaat ttagagtagc ctaagagttc agtgttgata aagtttacag gagtgtacag 21240 caatgtcctg ggccttcaca ttccctcagc acttaatcac tgacacccag agcaacttcg 21300 aatcctgcaa actgcattca tggtaagtgc cctacacagg tgtgccgttt tttcatattt 21360 tatattatgt tttcactgta ccttttctat gtttagatac acaaatactt acaattgtgt 21420 tacagttgcc tacagtattc agtaccgtat catgctgtgc aggtttgtag ccaattagca 21480 ataggctgta agatacagcc taggtgtgta gtaggctata ccacctaggt ttgtgtaagt 21540 gcactctatg acgttcacac aatgacaaaa ttacctaatg acacatttct cagaatgtat 21600 cctcgtcatt aagcaatgta tgactgtttc catatgacca agaaatttca cacctagtta 21660 tatagtgaaa agaattgacc agatgcagta gctcatgcct gtaatcccag catactggga 21720 ggccaaggcg ggagggttgc ttgagcccag gagtttgaga ctagcctggg taacatggca 21780 agacctcatc tctacaaaaa ttaaaatgaa aacattttct gggcatggtg gtatgtacct 21840 gtggccccag ctactcagga ggctaaggtg ggaggatcac atgagcccaa gagtttgagg 21900 ctacagtgag ctgttttcac accattgcag tccagcctgg gtgacagagt gtgaccctgt 21960 ctcaaaaaaa aaaaaaaaaa agaaaagaaa aaagaaagaa aaaggaaaga aaacaattgc 22020 ggatatatgt ccagaaagaa atgtacaaac cattgtagac aaatattcac agcaacatta 22080 ttcataatac caaaaaactg gaaacaagca aaatttctat cgactggtga atggataaag 22140 aaatgtagta tatccatgca gtagaatgtt atttagccat ataagggaat gaactactga 22200 tacctgctac aaaatggatg aactttaaaa attcatgcta aagtaaagac accagacaca 22260 aaaggcctca tattgtatga ctccatatgt atgaaatgtc cagcattggc agatccatag 22320 agacagaaag taggttagtg gttgccagga gctgaggctg aggggagggg ggatagagag 22380 taacagcaat ggatgtaggt ttgtttttgg agtgatgaaa atatagagtt agtggtgatg 22440 actgcacaac tttgttgaat tgcacatctt aagagtacat tttcagtttt gtgaattaca 22500 tctgaaatac agatttttga aaatgaaggc aaaataagga ttttttttca gacaaaaata 22560 agtaagagat tttgtcactg atagacctgc atcaagagat atacaacagt gagtttttca 22620 ggtaaaagaa aaataattcc agatgggaag caagttactg tagggtgaaa tgaagagcaa 22680 cagaaagtac tctaaatata tgaattaggc tgggtgcggt ggttcacgcc tgtaatccca 22740 gcactctggg aggcctaggt gggtggatga cctgaggtca ggagttcgag atcagcctgg 22800 acaacatggc aagacccagt ctctactaaa aacacaaaaa attagccagg tgtggtggcg 22860 caaacctgta gtgtaggcta ctagggaggc agaggcagga gaatcgcttg aatccggaag 22920 gtggagcttg cagtgagctg caactccagc ctgggtgaga aagtgagact ccagcctggg 22980 tgagaaagtg agactctgta tcaaaataaa taaataaata agtgaattac tatgattttt 23040 aatttcttaa catactaatg atgtaaacta tatgcagaat taaaatacac gaccatgaaa 23100 ccccaaaggt aagaagaggt gaataaagtt aaagaggact ttgtacaaga ggacttcaaa 23160 ttgtacaaga ggacttcaaa aagctcatgg aaacatggaa ttaaagataa caattaaaaa 23220 ataaactcta tttctcaaca taaattcagt caaggtcaag gcacttttgt aagtgatgat 23280 accaggcaca tagtccatct ccaaagaact ggggggtcct gggaatatag ccatgtcaat 23340 gcaggctttt ttacattatt aactaaagaa aaatgggtga tcgttaaaga ttgcttaaga 23400 ttaggagaca aaaagaagtc agaaggagcc aaacaggact ttaaggtgga tgcctaattt 23460 ctcatggaag ctgtaaaatt gcccttattt gatgagaaga atgagcagga gcattgctat 23520 ggtggaggac tctctggtga agctttcctg cgcgtttttc tgctaaagct ttggctaatt 23580 tccccaaaac actctcatga taagcagatg ttatcattct ttggctgtcc agaaagtcaa 23640 caagcaaaat gccttgagca tcccaaaaac ctgttgccat gacctttgct tttcttgacc 23700 ggtctgcttt tgtcttgact gaacctcgtc tacctctttg gagccactgc tttgtcttca 23760 ggatgaactg gtaaagccac gtttaatctc ctgttacaat tatgtgagga aatgcttctg 23820 aatcttgatc ctattagttt aaaatttgca tggaagtttc tgctgttgtc tgcagctgat 23880 ctgggcacag tggtttggca cccagtaagt gggaagtttg cccaacttta attttttggt 23940 cagaattgtg taacctgaac taattgagat gtctatggtg ttggctattg tttgggctgt 24000 taatagttgg tcctcttcaa ttagggcatg aacaagatga attttttcct aacaaattga 24060 tatggatgat cggccacgga gggctttatc tttatcattg atttcgccat ccttccaccc 24120 tagttgcacc ataaatttga tgtttgttcc tgcttcaatt ttagcagaat tcatgttgca 24180 atgatagggg cttttttgca aacagacgcc ttatccttct tagtgcctca aactagatcc 24240 tgttcagaca tgttatagca agttagtaca agtttatttt ggtgcagaaa aaagtttgaa 24300 ctttatgcat agttttttca taatttgaat tttccatgaa cttttggaag aattcatatt 24360 aggtagatcc taataataat aagtatgcat gttgtaatct ctaagataag cacaaaaaga 24420 atataaaaga aagtacaatg aacaaactaa taggaaaaat ggaataataa tttttaaaac 24480 ttgattttct aaaaactaga acaaagaagg aagaatagat aggtcaaaaa agaaaacaaa 24540 tagcaagatg gtagacataa tcccagacac atcagtgctt tcattgatgt ctgaaataaa 24600 aggcagtctt aataccaaaa cttgacatta aaggaaagta gaattacagg tcaatctttc 24660 ccataaataa tctaaaaaat atctgaaaca aaatattaac aagtcaaatc taataatatt 24720 tataaaaata gataacatat tattatcaca ttggtttatt tcaggaatgt aaggttagta 24780 tgacatttag aaaccataat caatttcacc acattaactc aataaacaag aaaatttata 24840 ttatcattta ataagttcag aaagcacatt taataaattg caacacccat tcatgatttt 24900 taaaacctgt caggaaagca gaaatagagg agtcttcttt aatctaatta cgatcacaca 24960 cacacacaca cacacacaca cacacacaca cactatacat agctaacatt atccttaatt 25020 gtgaaatatt gaacacttcc ttactgaggt cagggcaaaa acaaaaacat cttatactag 25080 aggtgctaat taacataata aggcaatgat aagaaacttt taatttataa agattagaaa 25140 actatgtgtt taagttatat aagaattgag gaaccagaaa agtccacgat actcaacaaa 25200 taaactatta gaataataat tgttattatt attcaaagaa gctgaagatg agatcaatat 25260 aaaaatctat tgtgtttcta acaatcagca aaattgacaa acccttagtt aaactaagaa 25320 aaaaagagag aagactcaaa ttaactaaaa tcagaaatga aagaggagac attgtaacta 25380 tgtcacagag ataaaaagga ttctaagaga ctactatgaa caattatatg ccaacgaatt 25440 ggatacttca aataaattag taaattccta gacatataca acctacccat actgatcatg 25500 acaaaataga aaatctaagc agacctaaac tagaaaggag attgaatcag taatcgaaaa 25560 cctcctagaa aaaaaaaaaa aaaggccaag accaaatggc ttcactagag aatgctacca 25620 aatacttaaa gaatcaacac caattcatcc ctaacacttc caaaaattga acagtaggga 25680 acacttccaa actcatttta tgatgcccca gtaccaaagc cagacaaaga tgcaagaaaa 25740 caaaaccata gaccagtact cttgatgaat actgatgcaa aaatcctcaa caaaatgcta 25800 gcaaaccaaa tttaacaaca cattaaaagg ctatacacca tgaccaagta ggatttatct 25860 gcagaattca aggatagtta caaaaatcag ataaatgtaa tacatgaaat taacacaaag 25920 aaggacaaaa atcacatggt catctcacct gatgcagaaa aagtatttga aaaaattcaa 25980 tactctttca tgataaagaa aacattcaag gaactagaaa tagaaataaa ttacctcaac 26040 ataacaaagt ttatatataa aaagcccaca gtgaatatat tcaacagtaa aaaactgaaa 26100 ccttttcctc taagatcagg aacaaggcat ggatgcccac

acttgccatt tctattcaac 26160 tggaagttct agccagagca attaggtaag aaaaagaaat aaaaggtatc taaattggaa 26220 aggaagaaat gcaattatct ctgttcacag attacgtgat tttatatgta ggaaacccta 26280 aatattaaaa aaatggttag aactaatacg tgaattaagc aaagttgcag gatacaaagt 26340 caacactcaa aactcagttg cattttttac attaacaatg gacaatccaa aaagaaactt 26400 aagcaaaaaa tttcatttac aatagcatca aaaagagaaa agtacttagg aataaactta 26460 gccaaggagg agaaagactt gtataaaaca tttttgaagg aaattaaaga acacagataa 26520 gtgaaaagac atcccatgat catggattgg aagacttaat attgttaaga tgacaatact 26580 actcacagaa aactgaattt aatataagcc ctatcaaaat ctcaatgaca gtttttgcag 26640 aaataaaaaa agaattcatc ctaaaattca tatggaactt caagggaccc taaatagcca 26700 aaataatctt gagaaagaag aacactctaa aaaaaaaaag aagaagaaga aggagaagga 26760 gaaggagaag gagaagaaag ctggaggcct cacatttcct gatttcaaaa tgtattaaaa 26820 agctacagta atcaatatag tatggtactg gcatacagat agatatacag accaatggaa 26880 aagaatagag agcccagaaa taaacccttg tgtatatagt caaatgatct acaaaggcac 26940 taagtctata caagagggaa aaggaagttt gttcaaggag atagtgctgg aagaaccaaa 27000 tatccatatg caaaagaatg atgttggatc cttattttac tccatatgca aacattaact 27060 caaaatgggt taaagaccca aacataagac ctgaaactat aaaactccta gaagaaaaca 27120 tatgggaaaa tcttcgtgaa attggaatgg gcaatgattt cttgtgtgtg ataacaaaag 27180 cacaggcaac aaaagtaaaa atagacatat gagactacat caaactttaa aacttctgtt 27240 cagcagagga aataatcaac ggaatcaaaa ggcaacctat ggaatgggac aaaatatttg 27300 caaaccatac aagagatatg gggttaatat ctagaatata caaagaactc ctacaactca 27360 ataacaacac aaaaacaaat aactcaattt aaaaatggcc gggtgtggtg gctcacacct 27420 gtaatcccag cattttggga ggccgaggcg ggcgaatcac ctgaggtcag gagttcaaga 27480 ccagcctggc caagatggcg aaatcccatc tctactaaca aatacaaaaa ttagccaggc 27540 atggtggcag gcacctgtaa ttccagttac tcaggaggct gaggcaggag aattgcttga 27600 acctgggagg cggaggttgc agtgagccaa gatcacacca ctgcactcca gcctgggcaa 27660 cagagcaaga ctctgtctca aaaaaaaaaa aaaaaaaaaa aaaaggacaa aggacatgaa 27720 tagacatttc cccaaagaag atcagcaaat gcccattaag catacgaaaa gatgctcaat 27780 atcactaatc atcagggaaa tacaaattaa aactacaatg agatatcatc ttgcacttgt 27840 taggatggtt attataaaaa aaaagaaaag aacaaatggt ggtgaagatg tagagaattg 27900 aaacccttgt gcacttttag tgggattgta aaatgatgcg aaactttaaa aaattttttt 27960 aattaaaaaa atagaaatac cagacatata tccaaaagaa ataaaaacag catctcaaag 28020 agatatttgc acacctgttt ctgcagcatt atttacaata gctaagatga gcaagtaacc 28080 taaatttatt gataggtgaa tggataaaga aaatgtagta catacataat ggaatattat 28140 tcagacttta aaaaggagga agtcctgtca tatgttacga catggatgct tcttgaggac 28200 attctgcaaa gtgaaataaa ccaattgcaa aaaaacaaat actgcattgt atcacttata 28260 tgatgtatgt aaagtaatca atctcataga aacagaaagt ggaatgctga ttgccaggga 28320 ctgggggagg gagaaatgaa aagttgtttt tcaatgggta tagagtttca cccatgcaat 28380 aggaaaaagt tctagagatc tcttgtaaac caatgtgcat acagtttaca atattgtact 28440 gcacatctaa aaattgttaa gggtaaaaaa gatctactgg ttaataaaat ccaaacccgg 28500 gacactatac tttgcactac aataccctct cttgttagat tagcaaagga aaggatccat 28560 aaacattaca aaagcaaaga aaggcaggaa gccctcaact ggttgtcagt tgaagtgggc 28620 tgggtttgat agctatttct tttaacatct ggaactttag ctttaaaaag tgttaataaa 28680 gttgtaaagt ataaaaactc attatatttc tgtacactac aaacagtcaa gaaattttaa 28740 aattatgcca tttacatcag catcaaaaaa tcaaatatcc gagaagaaat ctaatgaaag 28800 atgtacaaga cttttttttt ttttttgaga tggagtcttg ctctgtcact cagctctgtc 28860 actagaggca cagtctctgc tcactgcaac ctccgcctcc tgggttcaag caattctgcc 28920 tcagcctcct gagtagctgg gattacaagc atgcaccacc acactcagct aatttttgta 28980 tttttagtgg agacagggtt tcaccacatt ggccaggctg gtctcgaact cctgacctca 29040 aaagatccac ccaccttggc ctcccaaagt gctgggatta caggtgtcag ccactgcacc 29100 tagttgatgt acaagacttc tacacagaaa cctacaagac attactaaga aaaattagag 29160 aaaacataaa taagtggtgg tacatactat gttcatggat tggaaaattt aatactttaa 29220 ggatgctagt cccccccaaa ttgaagtaaa gtttcagtaa aatccaaagt ttcaagaaaa 29280 aaatctccag caagattttc tttggtggaa attgacaaga tagttctaaa atttatatag 29340 aaatacaaag gactgaaaat agtttagaca attttgaaga agaatgtcaa gatttataaa 29400 gtaatttaga aagtatggca ttgttgcaag gaaaaacaaa ctaaccaatg gaaaagaaca 29460 gaaacagtag aaataaactc atgtattcaa ccacttgatt ttcagtaagt caccagtgca 29520 aggcagtgga gagaaaacat gatgttttct taaatgatgg tatattaatt gaatattcat 29580 atggaaaaag taaaccttga cccctacctt acaccataca gaaaaataaa ctacagatag 29640 actgtagacc taaatgtgaa aggtaaaaca gtaaatcgtc tagaagaaaa cacagaataa 29700 tatttttatc actttattgc aggcaacact tcttaaacag cacatagaaa gttctggtca 29760 taaagaaaga tttataaata aacttcatga aaattaagat tttttattga tcaaaagttt 29820 ccattaagag aatggaaaag aaactgaaag aggaagaggg aaagaattat atatatgtaa 29880 atttataata aaatataaat ataaaaatat ttatttataa atattatata tatttataaa 29940 atataaatta tatataataa acatatataa tttattttta taaatatata tatggagaga 30000 gagagaaata aacacaattc caagtgctga aaaggataca gagtaactaa actctctgtc 30060 tctactcaat attctggtaa gagtgtaaat cgacacaacc actttggaaa acttctgctg 30120 agcagagact caacatttcc acttccaagt tatataacca aagagaaatg caagcatatg 30180 tgcccaagca gacatgtacc agcatgttca tagcagccat tatttacaac ctccccacac 30240 tggaaagaac ccaagttctt ggtttccctg tatctttttt ctgtcacacg attgaggaat 30300 gggatgcgtc tagttcctca agtcggccat tctctgcaat gtgacccagg tagggacatg 30360 gggctgaaag tgaaaggtgg gtagtgtggt aaaagtaaaa tggccctatt tcctataggg 30420 atcacaaaga aatgaggtgg gcgaagaggg actccccaga agtgccagga acaggagcca 30480 acaggtcacc agaagaggaa gtgttaaggc ggagggactt cccgagaggg aacacagaag 30540 aaagaactcc taaagttatc cagggtcttc aaggacttct gcccacctgc tctcctgctg 30600 gtcctcctct ctgaccttcc tgctatagct acagagctgt ggttcctcca actccattct 30660 tactccaacc gtgtcattcc atggctctct gttatccaca aaagaaaaac cccaaattcc 30720 ttgtctggta ttcaaggcat ttcgggatct agcccccagt attcatactg gacttcaggc 30780 tcatcagacc actgacagcc tcaccaacat gctccattca cctggcttgc acacctgtgc 30840 atgtgctgtt ttagccacgg aaactatctg ccctttttga ctgactgtta gcctcctact 30900 cagtcttcaa aatcccattc aaatggcagc tcctctgtca atctttgacc tctccaaaga 30960 gcatgagtca ctccttcctc tggggtccac cctccacata gtacatagct catggtgttt 31020 tcatctatat tctgggcacc atctaggcag ggacagtgtg tttctttgtt actgctgaat 31080 ccccagtaac taaccagtgc ctacagtggg tgctcaaaac atttgctaca tgagtgacta 31140 aatgaaagag aatgctttct atggaaacga attgtgaaca aacattatcc tcaatgttaa 31200 gacctatgta gaccgtacac tccaaagtta aatcttaccc tttttgaccc atttttctgg 31260 agcttagggg attgagagcc ttatgtattt ggtgaggata cgttttttgt tgttgtttgt 31320 tttgttttgt ttttgagatg gagtcttgct ctgttgccca tgctggagtg cagtggcacg 31380 atcttggctc actgcaacct ccgcctcccg ggttcaagtg agtctcctgc ctcagcctcc 31440 caagtagctg ggattacagg catgggctac caagcttggc taactttgta tttttagtag 31500 agacagggtt tcaccatgtt ggtcaggctg atctcaaact cctgacctca ggtgatccac 31560 cctccttggc ctcccaaaat actgggatta caggcatgag ccaccgcacc tggccgagga 31620 taaaagcttt aatgcttcaa tctttttttc ctgcatcagg gcctttggac atgcctttcc 31680 ctctgcctag aatgcttttc ccccagcttc tttcctttct catcctcagg tttcaagttc 31740 agtgttgcct cttcaaagag gcaaccctat ctgaactggg tctacgccac agatctctgt 31800 agcccacatc ccctagtttg tctccttcat agcaggtgta agatgaggaa tcgttttgta 31860 aatgtctctg tgtatgtgtt tgtaacttgt ttctgtactg aagcatgagc tctcagaggc 31920 tgggtaattg acctattttg ttcagcacag tatccctagt gcagagaagg ggacctgtag 31980 gtgccccata agtagctgaa gaagaatgtt aagtgaatga atgaatgagg ctaaaaatgg 32040 agacagcccc taagaggtcc tgctttcctc attagcttcc tcactgtccc cagaaactcc 32100 ccccagaccc acctcaggca acatcttctg cttccagagt gtgaataatc atgactaatg 32160 tcatcagatt agtaacaagg aactttcaga gaaggacagt gggctgccca gggtcacaca 32220 gcaagttctc atctcagtgg gttctcaggc tggggaagaa ctagcccagt ggccctgggt 32280 cagaaagcag actctgccat tcactcacag tgggaacttg gccgtatctc tgcctttctt 32340 tgagcctcag ctttcctcat ctgtggaatg ggggcagcag gcttagcctg gatgcggagt 32400 gtgaggcagg ttagtgtggt tagtattagg tgtgtggtta gtgttagttg cttttcatct 32460 tttttttttt cgccagtggc aaggggtggg ttatgggtgt ctgaaggcag gtatgttggg 32520 gacctcaggc ttacagttaa tcagtagaag acgaaggtga gaggaagggc cttctcctcc 32580 ccagctcgtg caacctctga ggctgtgcga cctctcagcc tatctctctc tcctagccag 32640 gcagcaaatc tctattagct ccagcaatct ttccctcctg gcacttcttc cagcctgtgc 32700 aacctctcct agctgtggag catgaggtcc cagtcgcagc acccgcaccc cccactcccc 32760 acccccgctg cctgtgcaat ctctatctcc ctgcaagccc tgccatcctc tgcagccttg 32820 cccaaacctt gcaacctctc agcctctgca gccagatctc actccagctc catttcctcc 32880 agcccaggag gggccgcttg tagagcagtg aaaggaggct ttgaactatg ccacgagatg 32940 gagcggagct ggccacccac cgtctctctc ccctcagttc ccagcagccc ccatcagccc 33000 gccctggccc tggccaaagt ggactggtgt tactgattca ttctggggaa agttggcaag 33060 ccctcgtcct agtcaagccc gcttggcaga tggggaaagg aggcccagag aggcccaggg 33120 ccttgtcctg ttcacacagt gggggatggc aggggcctgg gctaggaccc gggctggtgc 33180 acttgcccca actccccggg gacaattcag ggcaggggtc ggggtggcca gatccttcct 33240 gtgggctgca gagatcagca ggctgggttc ctgatgcaga tgtcctgggc agtcgggggc 33300 cctccttgtg gcccctgccc cccagcctgg ggaatattct caggtcgttt cagaagttcc 33360 aagtttactc agaggtctca tttgagctgt catgaaaagt ccctttggcc tggggccagg 33420 gcagggagtg ggggcggtgg cacctgtgag ccaggggtgg gggtgggttc ctcccattgc 33480 cccctctccc tccccttcac gtgtccgggg gctccggagg gcagggaagg agggcatgtg 33540 aggaaggggc tgcgagggca agaagaccct cttcctggag gaagcatctg ttcccctctt 33600 ttcactgctc ccttctagac ggttccctga gtgaccctag aaaaaccccc accctgctct 33660 gaacttcagt ttccccatct gtaaaagggg ggaagcagaa tcatcataag agctcttatt 33720 tactgaggtc tcatatatgc caatatgacc aataatattt attatttttt ttctgtattt 33780 tcatagctaa aagcaaagag ctcttacata tttaaagtaa aacataggcc ggccacggtg 33840 gctcatgtct ataatcccag cactttggga gactgagaca ggcagatcac ttgagcccag 33900 gagttcaaga ccagcctggc caacatggtg aaaccccgcc tccactaaaa atacaaaaat 33960 tagccaggtg tggtggcagg tgcctgtagt cccagctact cgggaggctg aggcaggaga 34020 atcgcttgaa cctgggagat gaaggttgca gtgagttgag attgcgccac cgcactccag 34080 cctgggcaac agagcaagac tctgtctcaa aaaaaaatta aaaaaaaaaa aaagtaaaac 34140 atgatagcgg gcacttctca tgagccatct gtgcccagct cttttgcctt ccccatcaca 34200 ctggctctct ctgtgtaccc cagagcttca ggacctaggg aggtgcctgg gccatggtag 34260 atgttccata aagaataggt tgagtaaatc ctcaccacag acctaagaca ggcattctta 34320 ttcctatttc cactcttaca gatgcagcaa ctgagctgca gaaaggttag gtggcttcct 34380 caaggtcaca caggtactaa gcggtggact tgaacccagg tctggccagt tccacagctc 34440 aaagtcacaa ggggtggtgt gctgggaagc cagttccagt tcccccatct aaccgcaggc 34500 accccttcct agttgtgtgg agaaggcccc tgtgcctgcc tgtgaggatg ctgaaggcgc 34560 tagcttccct ctctggctgt ggaaggaacg gataccccgt tcctttcatt catttattta 34620 tcaggcacct gggttgtcct gatgctacaa agaggtgggt tcctcccaga acttctggtg 34680 gctggctttc tcttgaatgg gcctcctcct ctggtatcca ccccctactc tactgcccac 34740 aagcccctta ctggctctaa tccccacctc ccagtttctt ctcctagcct gaaaccagga 34800 acgctgggca gtgggagcct gcagtctgag ctcagagctg tctttgcctt gctctcccca 34860 cctctcctgg tcctggattc tgcctcctcc tatcacaagg gcttttagct acctccactg 34920 tgggactgag taaggatccg gacaccctcc tggcagagga gcgggcagga caggggcatc 34980 gctgagccca cggtggggag caggtgagca gggcctctcc tctgggggtt gcctgtcctc 35040 gtgggaatga atactccgcc atccccatcc aatgctgctt cccagaagct acccaagccc 35100 atgagtcttg ttttcatgtt attctatcta gcaggtggtg gccttgtcct acagactggc 35160 atccaatccc agctccatgt catctggtca cagttaacct ggggcaaggt tgtccccact 35220 aagcctcact cttgtcatct gtgaaatggg ctgaatgagg ctgcccgctt cacctgttga 35280 ggactatggg agacgcttaa gtgcctacaa catggtgagt tggcactggg ggggccagag 35340 ccaggtggct ggccaggcag gaccacagca gcctgagtgt gtcgctcccc ctctgtcact 35400 cttggagtcg ggcccgtctc catgggctca aagcttctca tgccacaaac tctcttcctc 35460 aaaggaaaag ggccattagg agcacagctt tcaccctgtt cggccctgag gtccccacaa 35520 ctagctccac caggatgggg gtaccttggt ccccatttca tagatgagaa aaccatggct 35580 tctcaaggct agaagcagag agacttgccc aagttcaccg agtgaggaag gcgcgtggcc 35640 gggttttgac ctggcaggtt ttccccgact ccaccatgtg cttctgaaag ccccactctc 35700 cccccaggga atcacagatg tcctcctcgc agcagcctcc tggctttgaa tgtcccgcac 35760 ctggtcttct gccttcactg agcagagacg tcattcctgc ggcaagtcct aagttcccgc 35820 tccccagggc tcctccacca ggaatgttct tcctttccca tccttcaagg ctcagcctcc 35880 actgacctcc ccaccctcct ctcctccaca tgcacaggga actgggttgg cctcttacca 35940 tccgacccgt tccccgtgct tcatgagaaa gatgcagcga cagctcatgg ggtcacctgg 36000 cttgactgcc cgacctgcta cgtcccagtt gagtcaccga gggaaaatca ctctacctct 36060 cgcagcctca ttgcctcatc tgcgaaatgg gcacaccagc cattgttttg aatgactggg 36120 tggaatcaag gtggtaagac atagaaactg acacagaatg ggtcccatca ggctctacac 36180 ccatgcccgt cacctcccga ctcagccgcc cagaatcggg tgattgacat gttatgtttg 36240 gcttgaaact ggctttcctt ccccagcccg ctacattttg gaggctggtt ttcaagatag 36300 agggtcgtct cagccacatc cctgctgctg ggcccctaag ctgtgctgaa caagagaccc 36360 caacctcatt tctgcccagc ctcactgcat gaatcttctt tgcctcagtg tccccaattg 36420 tgacttgggc acagtcctcc gagccctccc actgtgacac tagatgcagc aggcagggaa 36480 cagtccctcc tgggcagaga aggcaccctg gtcccagcag aaaaccaggg ctggaattac 36540 cttctgtttc ggcttctttc tcagaaatcc attccgtgtg ctgggggcag tctgatgggc 36600 ttctgggggt ggcagctggg ttcagcgtgg gggtgacaga tgagggtctg agaaagcagg 36660 cagaggcctg gggtgagaat ccacacccag cccgagtcac caatcaggct gtcataagac 36720 ctgtcactct gatgacgggc agctgggcct ccaggcaggt gtcccacaga gcagcagaca 36780 gaggaaagag acggcaagga aggtaccagg cggccgcagc agggatgcac gggccgcaga 36840 cagacagaca cagctgcgtg gctgtcagcc acactcaggg agaggcaggc agagggccag 36900 acgccgagct tacggtcact cagcagagag acgccagtgg gtctgtctga ggccgctggc 36960 tgccacccag ccgcatgtgc agagtgggga ggaagaggag gatgtggtga gccgctcccc 37020 gccagccccg cccacccgca gctgggctcc tcctgggagg gagtgagaaa gaggtgggag 37080 ggggcctggc cccacagcag gcacccactg tcaccaaagg tcacctgcaa gtaagacatt 37140 tggggccatc ggggtgacac ttccctctta gggacctgtg acatcatccc cacaggcctc 37200 tcagtcataa cccatgaggc aggcagtgcc tcctggagcc accacattgc acagatatgg 37260 aaactgaggc cagaggtgtg aagatgtggc acagtttcac ccgtggaccc agagtcctca 37320 gtggccgact ccagcccctc agcacagaga gaaggaccag aaccttgccc aaggtcccac 37380 ggaagcccct ttttcagact ttgcaaacca gcctgtttct tattcctgag ccttacctgc 37440 ctaatagggt ctgagcttgg gaccctctgc tctggcttga cccccatgcc tctctgcctc 37500 catttccttt ggatccagtc tcctgctgca gagctccagg gagcaggacc ccttgggcac 37560 gtcacatcac ctctctgaat gcctcacttt gcccatctgt caaatggggg cacctagcta 37620 tgaggaagag ggcagacatt aacacatgag aattcaagaa gaaaatagag aaaagaaatt 37680 ctcaagagag ttttcatctt gctaacatgc cagaggccca tgagagggtc agatgtttct 37740 ggaagacatt caagaaacca ggagacaaaa tgtaacaagc ttaatgccta gtgcagtctc 37800 caaaggagag caagagaatt gctaccacca ccaccgctac caccaccacc actaccactg 37860 ccatcaccac cacctcgaat tactttggtg cctacatgat ctcactttat cccacaaagt 37920 gtgaggcaga caagattagt cccattttac agatggagag actgaggccc agaggagtga 37980 aacgtcacac aagccaagaa gaattagagc tgggatttga acctgggtgt gtttgattcc 38040 taagcttata ccaggctgtg aaaacggggc cataccagga tgaaatgtcc cacagttggg 38100 ggaggctatt ttatgctgtt ttgataacaa gatgcaattt gtagctctcc ctgctgactc 38160 atttttggga atcaacttgc tgttgacttt cgtgaccact ccagagaaaa aagagaaagc 38220 gtcttgctat tctttctttc tttttttttc tagctgtttt tgaggaactt ggtacaaatt 38280 cccctgtggt tgatttgagg gatcctggcc ctgtagagct cttcccagct accgaagtgt 38340 gtcgtgttct ctggagaacg aaccttggga agacaccagc ttttgttttt ctttctcttg 38400 atcttcgggg tgaattctct gataacatgc actgaaattt gtagtggttg aaagatcagg 38460 agaagagaca gtaaaaagag ctaatatttg ctgaaagctc attgtgtgcc agggttttaa 38520 ggactttaaa gatgcaccct ctcatttaaa cctcagaaat actcgagaag gctgtgtgca 38580 gcggtgcaca cctataattc catcattttg ggaggctgag gtgggaggat cgattgaggc 38640 taggagttca agaccagtct gggcaacata gcaagaccct gtctctataa aaaaatttaa 38700 aaattagaca gtcatggtgg tgcacgcctg tagtcccagc tactctggag gctgaggtgg 38760 gaggaccact tgagccaagg agctgaaggc tgcagtgagc tatgactgta gtgctgcact 38820 caagcctggg caacagagca agactctgtc ttcaaaaata aaatccccaa aatgaaaaag 38880 aatgagtcta gaagatagtt atcatcatca tcatcatcgt ccctgtttta cagatgagga 38940 aactgaggct cagagaggtt aagtgacttg tccaaggtca cacagctgat gagtgccaga 39000 gttgggattt gaactcagtc atttgggctc cagagcccgc actcttaact cactacactg 39060 ctctggggtg tgtatgaaaa caaggatatt caatttctcc tccctccctc ctgcctgcct 39120 tctttatcta cctgctttct acccacccac ctacctatcc acccattcat tcttctctgc 39180 acccacccac ccagccattc cttgcttcct ccccctaata tttattgagc aggctcagag 39240 aggttaagtg acttgtccaa aatcacgcag cttggctaca tcctggggct agcccacaga 39300 aatatacaca cagtgacagc ccccatggag cctatattct agtagggaga gttagacaaa 39360 aagaagtaac agacaaactc aatgtaaact gtactaaatg tcatagaaaa ccaatgagag 39420 atcaaggttg tgacatttgg gggtacacta ggtgctaaga agtgcccagc catgggaaga 39480 acattatcgg cagaaggatc tgcaagtgca aaggcactga ggtaggaaag aatgtggctg 39540 gctccaggaa cttgagaaca gcatggcgta atacagtgag caaatggagg aatagtagaa 39600 ggtgaaggca cggaaatcac acatctacac cacatgggac ctcacggttt tgggtgagga 39660 aggtagattg ttccaaagga aatgggaagt catcattgga gggtttcatc ttaagagtgg 39720 cttaatccag tttacatttt cagttcaggc caggagttgg ggaatgggtg ggatcagcag 39780 gggtcacagt cccttggtgc ttaaaaagta gatgcatttg gagctgccag gcccatgtcc 39840 actggggtaa acatgagtcc cctcagaaat agacatgtgg tcccttgagg ggttcctctg 39900 ggtctgtaaa tgtctcataa gtatggcctc tatacccttg gaaggaaggt gccataggat 39960 ctcagcggcc ctccaaccat gcctcagtgc tgatagctgg tcagcgggga ggaagtggtc 40020 acatttgcat ttcagccaag gggcctcctg ggaaaaaata attgacacac cccctccctc 40080 aacccatttc ctctttttta tttttatttt aagagcttta tttagctatg ttttacataa 40140 cataaaatca agccactgta aatgtacagt tcaatgattt ttagtgaatt tatcgaattg 40200 tgcaattata accacgatcc aattttaaaa caatcccatc actcctgaaa ggttccttgt 40260 gcctgcttat agtcaatctc cattcccatt cccagggcca ctcgtctact ttatgcctct 40320 actgattttg cttttttttt ttcctaggca tctcatatca atatgttgtc ttttgtgttt 40380 agtttctttc agcatcgcgt gtttggggtt caaccacgtt gtagcgtgtg ttggcaccac 40440 atatctaaaa atccattttt aggccaggcg cggtggctca tgcctgtaat ctcaacactt 40500 tgggaggcca aggtgggcgg atcacttgag atcaggagtt cgagaccagc ttggacaaca 40560 tggtaaaacc ccgcctctac taaaaaatac aataattagc ctggtgtggt tgcacacgcc 40620 gggagattga gaccagcctg gccaacatgg tgaaaccctg ttctactaaa aatacaaaaa 40680 ttagttaggt atggtgacgt gcacctgtag ccctagctac tcaggaggct gaggcaggag 40740 aattgcttga acacgggaag tggaggttgc agtgagcaga gctagtgcca ctgccctcca 40800 gcctgggaga cagagcgaaa ctccatctca aaaaaataaa aaataaaaat tgatttttat 40860 tgctgaagaa tattacattg catagatatg ccacattcat ttatcctttc atgagatgat 40920 ggacatttgg agtatgtcca ctttttcctt tactacaaac aagcatgcta tgaacacttg 40980 catgcgagtc tttgtgtaga catgcatttt caaatctttt ggatatatac ccagaaggag 41040 acttgttgag ttaaaggtag tttaacttat taacttttaa ataagtttaa cttattaaga 41100 aactgacaaa ctgtttttca cagcagctgc cctgtattat attcccacca gcaatgtctg 41160 aaggttccca tttctctacg tccttgacaa tatgtatttt

cttttttttt tttttctttt 41220 tttttttttt tttgagatgg agtctcggtc tgtcgcccag gctggagtgc agtggcgcga 41280 tctcggctca ctgcaagctc cgcctcccgg gttcacgcca ttctcctgcc tcagcttccc 41340 gagtagctgg gactacaggt gcctgccacc acgcctggct aattttttgt attttctttt 41400 gtagagacgg ggtttcaccg tgttagccag gatggtctcg atctcctgac ctcgtgatgc 41460 gcccgcctcg gcctcccaaa gtgctgggat tacaggcgtg agccaccgca cccagcagac 41520 aatatgtatt ttctgttggt ttttattata gccatcctag cggatgtgaa gtggtatctc 41580 attgtggttt tgatttgcat ttccctaatg gctaatgatg gtaagcatct tttcatgtgc 41640 ttggtagtca tttgcatatc ttgtttggag taatgtctat tgaagtccct tgccaaattt 41700 tatttatttt attttttttt tagacagagt cttgatctgt ctcccacact ggagtgcagt 41760 ggtgtgatct tggctcactg aaacctctac ctcctgggtt caagcgattc ttgtgcctcg 41820 gcctcccgag tagctgggct tacaaacgtg tgccaccata tccagctaat ttttgtgttt 41880 ttagtagaga cgggttttca ccatgttgac caggctggtc tcgaaccctt aacctcaggt 41940 gatccgccca cctcagtctc ccaaagtgct gggattacag gcataagcca ccgtgcccag 42000 ctccttgcca atttttaaat ttgattgttc atcatttgtt gttgaatttt aagagttctt 42060 tgtatattct ggaaactcac ctttatcaga tatatgaatt acaaatattt ttcccattct 42120 gtggctgtct tttcactttg ttgatagtgt gctttaatgc aaaaaaaatg aaaaaaaatt 42180 gactatagat tcatgggttt atttctgaac tctaagttca attccattga tctatatgtc 42240 tagccttgtg ccaggattat cacactgttt tgattactgt agctctgcag taagttttgt 42300 tttgttttgt ttttgttttt gtgttgtttt gagacagagt ttcgctcttg ttgcccaggc 42360 tggaatgcaa tggcacaatc ttggctctgc agtaaatttt aaaaatggga atttgagtcc 42420 tccaacttta ttgttttggc ttttctgggt cccttgaaat ttcacatgat ttttaggatc 42480 agtttttcca ttttttaaaa aaggccattg ggacttttat aggaattgca ttaaatctat 42540 aaattacttt gaggagtgca atgttaaatc ttccattaca ttaacataga atttaaaaaa 42600 ctttatttag gtcttttcta attttttcag caatatttta tagtgttcag tgtacaaatc 42660 ttgcacctcc ttggttaaat ttattcttaa gtattttatt atttttgatg ctattataaa 42720 tagaatttat ttaatttcct tttggggttg tttattgcta gtatatacaa acacaattta 42780 tttttgtatg ttgatcttat accctgtcac ttggctgaat ttattagctc taataatttc 42840 tttgtgcgtt ctttagaatt ttccatatat atgatcatgt catctgcgaa taacagtagt 42900 ttttactctt tccttttccc atttgaatgc cttttatttc tttttcttgc tgaattgttc 42960 tggttaaaac tttcagtgca atgttgaata ccagtggtga aagtgggcat cctctttcct 43020 gatcttggtg gaaaagtttt cagtctttca ctgatttagt gtgatgttag cttgttagct 43080 gcgggttttt ctttctttat ttctctttct ttctttctct ctttccttct ttctctgtct 43140 ctctcttttt tatttgagat ggagtctcac tctggtgcag gttggagtgc agtggcatga 43200 tcttagctca ttgcaacctc atgggttcaa gtgattctcc tgcctcagcc tcccaagtag 43260 ctgggattac aggtgtgtgc caccacatcc aactaatttt tgtattttta gtagagacag 43320 catttcacca tgttggccag gctggtctcg aactcctgat ctcaggtgat ctgcctgtct 43380 cggcctccca aagtgctggt attacaggca tgagccactg tgcccagcca ggtttttcat 43440 atatggtctt tatcatgttg agggagttct ctcttattcc tactttactg agtgttttta 43500 tcatgaaagg gtgttaaatt ttgtcaaatg cttttctgca tcaatttgga tgatcgtgtg 43560 ggttttctta ttctattaat gttatgtatt gtattgatta attttcttat gttgaaccac 43620 ccttgcattc ctgggataaa tcctttttgg tcacactgta taatcctttt aatgtgctgt 43680 agaattagtt tgcttgtatt tttgtgaaat tttatatcta tattcataag gaatattagt 43740 ctgtagtttt gttttcttgt ggttctcttg tctggatttc tctcctgttt tgtggaggag 43800 tttgagaaga actgatgtta attcttcttc caatgtttag tggaattcac cagtgtagcc 43860 atctgactct ggacttttct tcattgggag gtttttgatt caactcttta cttgttgtat 43920 gtttattaag attttctatt tcttcttgag tcagttttgg cagtttgtgt taccaaactt 43980 taatatttaa gttgacaact attattatct aatttgttgg catacagtca ttcacagtac 44040 tcccttatga tcctttttat ttctgtatga tttgtagtaa tattcccact tccatttctg 44100 attttagtga tttgtattgt ttcttttgct tagtccaact aaaggcttgt caattttgtc 44160 aatctttttg aagaaacgac tcttggtttc tttcatctct attgattttg tattctttat 44220 tttgcttgtt tgcactctaa tctttattat ttccttcctt ctgctacctt tgggtttcat 44280 ttgtccttct ttttttagtt cctttaaaag taaagttagg ttatgatttg tgatcttatt 44340 tgttgttttt ttttaatgaa ggcatgtagg gctacacatt tccctgttag cactgctttt 44400 gctacatccc ataattctgg gtgtgttttt ggtgctttca ttttcatttg tctctaagta 44460 ttttctaatt tccttgtgat ttcttcattg acatgatgac tgtttaaaaa tatgctattt 44520 aatttccaca taattgtaaa ttttccagtt ttccttctgc tattgatacc taacttcatt 44580 gtgatcagag aagacacttc atatgatttc aatcttttaa aatttatcaa gacttgtttt 44640 gtggcttaac atatagtcag tcctggagaa tgttccatgt gcactttgtg gtcactgtta 44700 ttgttgggtg gagtgttcta tgtgtgtcta ttagctctgg ttggtttata gttgtacaag 44760 tcccctactt tcttagtctt ctgtctagat ggtctccaac tattattgta gaactgtcta 44820 ttcctctatt cagttctgtc aatatttgct tcatatattt ttgggctctg atgtttagca 44880 catgtatggt cataattatt atgtcttctt ttttttgaga cagagtgttg ctctgtcaca 44940 caggctggag tgcaatggtg caatctcagc tcactgcaac ctccgcctcc caagttcaag 45000 caattctctt gcctcagcct ctcgagtagc tgggactaca ggtgtgcacc accgtgccct 45060 gctaattttt gtatttttag tagagacagg gtttcgccat gttggccagg ctggtctcaa 45120 actcctgacc tcaggtgatc cactcatctt ggcctcccaa agtgctggga tgacaggcat 45180 gagccaccac ctggccaatc ttcttgacaa attaacacat ttatcaacat gtaatgtcct 45240 tgtttgtctc ttgtaacaat tttttactta agactcattt tatctgacat tagtatagcc 45300 acctcagctc tctcttggtt tctattttca tagaatatct ttttccatcc tttcactttt 45360 aacttatttg tgtctttgga tataaagtga gtctcttgta gacagcatgt agttggatca 45420 tgttttttaa ttccttctgt caatccattc tgtcttctaa tcagtgaatt taatctactt 45480 acatttaaag tgattcatga taaaaggctt accactgcat tttgctattc atttcattta 45540 tatcaaattt tcttttgtta ttcaattccc tcattacctc cttctcttgt attgattgat 45600 tttcttctag tgcaccattt tgatagcctc ctcatttctt ttctgtatat ttttagttat 45660 tttcttagtg gttaccttga ggtttacact tcacaggaaa tttataacaa tatagtatga 45720 attgataaca acttggttta aatagcaata caaaaactct actcctatac agctccctct 45780 actctattct actcctatcc ctccccctct tttgttgtca ttatcacaaa ctacatcttt 45840 atgtggcatg tgctcattag catagattta taattatttt tatgtatgta tcttttaaat 45900 aatataggaa acaaaaagag gagtcataag ccaaaaatat aattatacta gtttttatat 45960 ttacctttac tagtgttctg tatttattca tatggctttg atttactgcc tagtgtcctt 46020 tcatctcagc ctgaaagact ccctttagtg tttttcatag ggaaagcctc ctagtgatga 46080 aatacttcac cttttttttt tgcaatcttt gaatgtttta atttcttaat cattttggaa 46140 agatagttgt gccagataaa gaattctcag ttggcagggt tttttccttc agcactttaa 46200 atatatcatc cttctgtctt ctggccttgt ggcttctgat gagactttgg tcattaatct 46260 tgttgaggac cccttgtatg tgacaagtta cttctctctt gctagtctcc agattctcga 46320 tttgtctttg tctttcaaca gtttgattat aatgtgtctt ggtgtagatt cctttggttt 46380 tatcctagtt ggagttcact gagcttcttg gatgtatata ctcatacctt tcatcaaatt 46440 ttggaaatgt ttggccttta ttcaaatatt ctttctggtc ctttctttct cctccttctg 46500 ggattcacat aatgcatatg tttatacctt tttttttttt ttgagatgga gtctcgctct 46560 gttgtctcac aggtctctca ggctctgttt atttttattc ctttttactt tatgctcctc 46620 agactggatt atttcaattt acctagcttc aagcttattg attctttctt ctgcctactc 46680 aaatctggtg ttggagctct ctaataaatt tttcattttg gctattatat ttttcaggtc 46740 caaaatttct atttggttat tttaaataat gtctagctct tattgatatt atctacttgt 46800 tcacacgtag ttctggttta acttagttct tcactcatgg tttcctttag ctttttgagc 46860 atatttaaga caattgattt aatgtctttg tctagcagct ccaatgtctg gactttttta 46920 gggacatttt ctatccattt ttcccccgtg aatgggctgg gtgttcctgt ttctttgtat 46980 acgttatagg tttttgttgg aaacttggca ttttgaatgt tacaatgtga taattctaga 47040 aatcacattc tcccttcccc tgtccacaag gcttgctttt gtcatttgct gtgagcttca 47100 gccattcatt tacttagtga cttttccaaa ctatttttgc aaagactata tccctttttg 47160 tgtatgttca ctgaaatctc cattctgtta tctcagcagt tggtcagtga tctgtaagag 47220 gtttccttat atacctgcag ccaaaaagaa aaacaaaata aaagatactc ccctagtctt 47280 tgtaaattgg ctctgagctg ggatactcct ttacacttag ccaggcagac tacaattctg 47340 tcttagcctt cacctctgct tgtgcagagc ccaattagcc agaggtgcaa gcctaggatc 47400 atctcatatc atttctgtgc atgacttcag ccctgggcat atgtgttata ttctagattc 47460 cctggtatac acggaagctt ttcaaagccc ttattccccc atatacattc ctccttagtc 47520 tttttcttct ccagcttttt ggtcggtctc tgctttttgc tttgtctgtc atcctttgcc 47580 ccacatgggt agtgtgtgtc tttaaagttt gtcaacagac attgcctggg aagctgctcc 47640 agcctagagt tcaggggaag tgggagggca aaacaaaggc aagcacctga gtggatcctt 47700 tagggagatg ccaaacagat caaaatgtag aaacacaatt ctttgagacc aaggcttgtg 47760 ttgttctttc tggtaccagc aagccatgcc aagaatgcag gctactgtac tcatagccac 47820 gactaagctg atgaatgtga gatggtaggc aagtaagcaa aaatgctaca gtattttctt 47880 accaagaatt agcagccttt tctggttgct aaagggtttt gattagattt caggttccaa 47940 aaaagtttct tttgacactt tcttccagct taatgtttgc tttagtggag gaactgatcc 48000 ttggagttcc ttactctgcc atttttaaat aatgtactta atggtgtttt ttttcttttg 48060 aagcataacc cttttaaatt ttattgaaac ccaatttatc aatttttttc ttttgtggat 48120 tggactttgg gtgccatatc taagaaatct tcacctatac caaagtagca aaaatctcct 48180 ctcatatttt cttctaaaag tttaatagtt ttagctcttg catttttatt ttttattatt 48240 tgtttatttt ctgagacagg gttttgctct gttgcccaga ctggagtcca gtggctcact 48300 gcaatctcag gtcactgcaa ccttggcctc ttgggttcaa gcgattctcc tacctcagcc 48360 tcctgagtag ctgggactat aggtgcatac caccatgtcc gcccaatttt tgtatttttg 48420 gtagatatgg ggtttctcca tgttgctcag gctggtcttg aactccgggc tcaagtgatc 48480 catctgcctt gacctcccaa agtgagccat tgagcccagc cttacctttt taaaattatg 48540 attttgactt aattttgtga atgctgtgag gtaatgatct aaatttatct tttttcatat 48600 aaatatctaa atgtttcagc acaatattta gaaaagattg ccctctccct tttgaattgc 48660 cctgacacct attttgataa ttgactataa atatatgggt taatttctgg accctgtatt 48720 ctgattcatt gctttatatg tctatccatt tactagcacc ttactgcctt gattgctgta 48780 gctttgtagt aagttttgaa atcaggaagt agaagtcttc caaatttgtt ctgctttttc 48840 agaattgttt tggtcattct gagtcctttg catttccatt ataaatatta aattccatat 48900 aagtttcagc ttgctaattt ctgcaaaaac tcctactgga attttgacaa ggattgtctt 48960 gaatcaatag atcagtttga ggaaatctac caccttaaaa acttggagac tttcaatcct 49020 ggatataggc actctctcca tttatttaga tctttgaaaa ttcttctcag acaaccgctt 49080 cctcatccat tcattttttt aaattaattt tttattgtaa taaatacaca tcacatgaca 49140 tgtatgatat ggtttggctg tgtccccacc caaatctcat ctgggattgt agctcccata 49200 gttcccacgt gttgtgggag ggagcctgtg ggagataact gaatcatggg ggtggttttc 49260 cctatactgt tcttgtggta gtgaatgagt ctcacgagat ctgatggttt tataaggaga 49320 aacccctttc acttggttct cattgtcttt tgtctgccgc catgtaagat gtgccctttg 49380 ctttctgcct ttcaccatga ttgtgaggcc tccccagcca catcaaactg tgagtccatt 49440 aaacctcttt ttctttataa attacccagt cttgggtatg tctttatcag cagcgtgaaa 49500 acagactaat acgatgtacc atcttatcca attttaagtg taagttcagt ggtattaact 49560 gatgcattcc tttctgaggt ccaacattgt ggggttcctg taatggacca gatggtctgc 49620 tgagagccac agccaagaaa gggcagggga tgtttcatgg ggagcaaacc acaaaggaag 49680 gagacgtcta gagggctgga ttgtgcgcaa catgtggctg aaactgggaa cccccttttc 49740 actcagttac cctgagacag caggtttatt ttaagagcga tggctgaata taaagaaaaa 49800 actaataaat agtgccctca gttatgagta acacagaaag aatatgttta tcctgcctta 49860 aaagctcatc ttggtttgta ttaggctctg gactaatagg ggtatcattt aagcagatga 49920 taagactaag aattttgtaa atttctttcc aaaattagga tctctaatgg gctgatcaga 49980 ctgccctagc cattgatcac atcacttgct gtttggcaca tgagttgtgt cttggttggg 50040 ttcccccaga ggcaaattct aagacaagga tttaaggata agtcatagat ttgggaaata 50100 atcctaggaa acacagggga atggagaagt ggggcaggca aagcgaggaa gccagtaaag 50160 ggcgctttaa ggagaaggcc actgctgttg gcaactggag ctgaatccca ctgggtacct 50220 ctctgggaga caatatagaa catgtctcca acttgtccca aaggagaaac aagaaatctg 50280 ggatatttct tttccaggac ttcatttgtt gttagttgag ggctgcttcc agggccacga 50340 ctctctgaca cttccagctt gccgattgca tgcacctgtt gccaaaaatg aaccctcacg 50400 cagggagttg caggtgtttg cagtacaaat gctttggtgt ataaagctga gaagttaagg 50460 gtaaggcttc aacatctgct acagggacca tgatagggac aattaattgt gtcccagctt 50520 ccatcttgtc cttctttctc atatcaggac catagtgtaa ttctggatgg caacagaccc 50580 agctaataga ccactttcca gtttcccttg cagctagtga tgggagtgtt cacatggacc 50640 ttctggaagc actctgtggc ccttccagta ttccccttcc tcctttctcc tgcctaaagt 50700 gtacacgtga tggctggagc tcaagcagcc atttagaact aaagaaaaag aataggataa 50760 tcacagagaa cttggccctg atatcatggg tcatgaacaa atgtcagcaa tcatttgtct 50820 ctagacttct cgttccataa aggaattagc tcttaagtgt ttaagccact gtggttggtc 50880 tctattagaa gttggaactt ctaaccatac aaggggattg agtgactatc aaaaggatga 50940 agaggcttat ctgtgtaccc aaactggtga agcaagaaat aaccaggtat gctgtgtgta 51000 taaagtactg actaggttga aataaatcat gaagagaggg ttggatgaat aatttataaa 51060 acatctgtga aacagaatac tctgtagcta tgccatgtga ggacataaca agaaggtggc 51120 tgtctgcaag ctagaaaggt tgttcagaag acaaccatca accaccttga cttagactac 51180 cagcctccag aagtgtggga aaataaactg ttgcctaagc cacacagtct atggtatttt 51240 gctatggcag cctgagctga ctaaaacaga tgtatctgta ttagtcagtg ttctccggag 51300 aaacagaact gtaaccaaca caattattaa taacaataat acaaaattgt ttacagttgt 51360 aagggacttg ccatctgcaa taagctatca aaaccacctt ttattttaat tttgtatttt 51420 tgagacaggg tctcactctg tcgcccaaac tggaatgcaa tggtgtgatc atggtgtgat 51480 cactgcagct taaacctctc aaactcaagc cattctccct cttcactctc ccgagtagct 51540 gggaccacag gtgtgcacca ccgcgcctgg ttaagtttta aaaagctttt tgtaggaatg 51600 aagtctcatt atattgccag ggctggtctc aaactcctgg gttcaagtgg tcctcctgcc 51660 tcagcctcct aaagtgttgg gattacaagc gtgagctgct gtgcccagcc ccaaactacc 51720 ttttaatatc cccattttag acataagaaa acaggtgcag gagactaagc aactggtcca 51780 caggcacata gccagtaagt ggcggggtca agatttgaac ctgggccgtc tagcctcaag 51840 ggttaatagg cccatttctg ggtggtggga atttttattt ttaatttttt atgttattct 51900 gtattatctg atttttaaaa tgatatctta cttctggaat ctgggaaaaa taaccaagct 51960 atttccattt tgaagagaaa aaaaaaagga catcacaaaa catcagtatg gtgggaaaag 52020 ggtgcagaga agggaaggta taaaaaaatc tatagagatg tgagccatgg tccccctgaa 52080 gcatgtgacg tcctttactt caagccagat attcaattac atgtatatga agattagaac 52140 cacagaacca acatgtcaca tggaaaatgc ttgcgagatt ttcccaaagt caagagtgtg 52200 tcacagccaa ggggaacctg aagcacccta gggttgcatt aatagaggta tggtgtccag 52260 aacaagggag gctattctca ctgcactggc cacaccttca gtctacgtcc agttctggac 52320 cttcattttt aaagaacatt ggcaaattga tgtatattca gaacaggcaa ccgagaaggt 52380 agagcagctg gtgtggagac agtaaaaaaa accatgagac agcagaagta tttggaaaga 52440 gaagactcag ggtgggggtc atgagcaggt ccagaccagc atagtcagag ttgatgaact 52500 ggagaagaca gtgagctccc tgtccctggt atgtaaacaa ctcctcactg tgggggtatg 52560 gttggattaa ctgagtgtgt ggttggacca agggcaatga agtctccttt catgctgagt 52620 gtctgtgtgt ctctgattct tagaaagaca taggagggct taccatggag ggagataatt 52680 atggaccgaa tgtgctgctt tcaacctctg ccaaattcat atgctgaagg cttaacccac 52740 aatgtggctg tatttggaga tgaggccgct aaggaagtaa ttaaggtgaa atgagctcag 52800 aagggtggga ccctgatctg ataatgttag tgcccttata agaagagaca ccagggagtt 52860 tgccctcaat ctccctcaca gcaagaaggc agctgtctgc cagccaggaa gagagccttc 52920 atcaggcatg gattcagctg cctttctttt ctgactgcag agctggggag atgggagttt 52980 accaaagttt tccagatgct gcacagagaa ttcaggtggt gcactgaaaa tgtgcctacc 53040 agagactggc acaaatgcgc gaaggaattt cctttgcatt ctgatcccta atgttattcc 53100 tggaccaact tgttgatcat ggactttcca gcctccagaa ctgtgagaaa taaacgtctg 53160 ttgtgcaaac cacccatcca ttgtgttata gcatcctgag cagctaagac agagggttac 53220 agcaaccact gagctgattg ggcacctctg caagctttct cttaggggac cctggggctg 53280 agccccagcc tccaagttca ggggtggggg tgcggtgggg tgagctgatt acagtcttag 53340 ggcagaggag gaattgccca tcttctccct cccctgccac ctccaaaatt ttcacagtta 53400 acaaaacaga tcttcttttc actgtcaaag gttttaaatt caatgggaag gcggatacac 53460 aggtctaaaa gcaaagcact tgagttagtg gcatgtacta agtataagag gtactgccgg 53520 gcatggtggc tcacgcctgt aatcccagca ctttggaagg ccgaggcctt aagctcttaa 53580 acacttaaga gctaattccc ttatggaatg agaagtctag agacaaatga ttgcttaggc 53640 cgagacctaa gatcaggaat ttgagaccat cctggccaac atggtgaaac cccatctgta 53700 ctaaaaatac aaaaattagc cgggcgtggt ggcacatgcc tgtattccca gctcctgggg 53760 aggctgaggc aggacaattg cttgaacccg ggaggtggag gttgcagtga tctgaaatcg 53820 tgccactaca ctccagcctg ggtgataaga gcaaaagtcc atctaaaaaa taaaggtact 53880 tataattgtt gctgtttact agactagtat gattattatt aacagcagta gcatattgtc 53940 tttgtttttg tttttgtttt tgttttgttt tgttttgttt tctagtcctg atttactctt 54000 ctcctggtca ctgtatcccg gctgtttctg agatatcatg ccccctccat tctcagttcc 54060 tttccctagg agcgtgcatg actctagcct aaaccaatcc actatattcc ctaagccaca 54120 ttaacagatc cagtcagagt ctaacagatc ccacgagact ctagagggat tgatcagggc 54180 aaagactccg cacttggagc cgggagaacg tattcatgtg ttccctgcag ctgctgtatg 54240 tctctgtggc cacacgggag aaactgcctc aggatggggt cgacctggaa gaagtgactc 54300 aagagatgaa ctgggccatc aggaatcagg atcccttgat ccagctatgc ctgaaagcca 54360 agacatttta gttaaatagg acaataaatt ccctcttggc ttcagccagt ttggattaag 54420 taagtttctt tcacttgaaa ctgaaagaat cttgacagat acattactgt gtgaccttaa 54480 agtgctataa tggtggttgg tacaaagtgc tatgacagca gacacgagcg acccctgatg 54540 atactggcca cagtgtggag ggagtgtcag ggaaggctgc atggaggagg ggtcacttgt 54600 tctggccttg gaaggatgaa gaacagttca gcgtgttgaa aatataggag agtgcatttc 54660 aaaacgttct gaaaatgtgg acaggaaacc aagaaatacg cttctaaatc acccaagaga 54720 caatgagaaa aattgcaatg gaaaattaga aaacattttg aactgaatgg ttttaaaaaa 54780 acaaccacat gtaaacttgt gggttgcagc taaaaaagta ctcagaggga aatttgtagc 54840 cttaacagtc acagattagc aaagaagaaa agctaaaaat taatgttcca aacgtctctc 54900 tcgagaagtt agaaaatgac agtgaaataa acccaaaaga aaggaaagga aaattataaa 54960 gatcagaatt ggctgaaaga ggaaggaagc acagcagagg gccaaccaag cctggagtcg 55020 gttctctggt aagccaaaga catgagggcg tgacttcccc gcctgttcgg ggaatgatga 55080 gggccctggt gtgggtgggg agggcagagg gggacaccag agcaaggagg aagtgtcgag 55140 gccatagcct gctggcgaag acagtgccag agcctggaga agtttccctg tagagcagcg 55200 ggagcagaac cccaccccat cgacggccag agccatgaga ggatctcagg aggggagtga 55260 tgggagcaag ctacctttgg tccctatcca actgggctgg gaacttgtag ggtcctggga 55320 gactgtctgc agatgagggt atgtggcccg tgtgcctagg tctgattgga aaggagaaga 55380 taagccacag aggataccaa gtttagtttt ccaggaaccg gcctctgtcg ctggggaaac 55440 tgccaccaca gctgctggct gtccagtgcg accttggcac aggctggctt ccctttcagg 55500 gaggggcacc cagacccatt gagtggtgtc tgagaaggcc ctggagggct ggggggaggc 55560 tgccgagcta cggcatctcc agcccttcaa tagaatattt ggcagggttt ctccaccttg 55620 atagtgttga catttgggct ggatcattta cggtgggggc catcctgtgc atggtgggat 55680 ggtgaccagc atccctggcc tcttcccact agatgccttt cccccagttg tgaccaccaa 55740 aggtgtcccc tggagggcaa aattgccccc aggttgagaa cctctgattt catgttgtag 55800 ggggaggagc actggaccca gagtcctaca gaagatgtcg gcagtgccaa actcatccgc 55860 tagggctgca tgaccccagg caagatcctt cctctctctg agcctctaca acccctacct 55920 gccaattgtg gtgagaatac ttacccctgc ccccaggata attaggagga tgctgtgcct 55980 ggaagagcag gctgccacaa acagcagagg ttaggtggga atcattgccc ctgtacctgg 56040 agcgaggcta aagagctgaa cggcgtgtgg ggcggggtcc ccatggtgac aatagcatga 56100 tctttcttgc ttccttgctt tctctcacca ccccaatccc acccgttggg aattcctgtt 56160 ggttccactt tcaaaacatg cccagaatcc acagctgtgg tcgccgcctg ctctggacat 56220 tggaaaatta gggcctcccc taccaaaaac atccccttct

aactgcagcc agaatgatca 56280 ttttgagaca atcatctgat caagtcactt ccacatccaa agccttccta tggttggtca 56340 aacaaaccct aggtcctgtg tgtggccagt aaggccctcc gtggctgatc cccattgcct 56400 ctgaatttgt ctctttcctc tcccccagtt ttctgactct ggcctcctgg ctgcaccccc 56460 ttcacgccat tcccttccca gggccttcgc acagcccgct ccctcctcct agaacaatct 56520 ctttctccct gtccctccag atatctgcgt ggctcattcc cctagttcac tcaagcctct 56580 attcagacct ccttggtgag gtctttcctg gcacctccta aacagctctc cacccgctaa 56640 tttcccctgg cccctcccct gcctgtctgt tctgcatagc actcaagccc tctgacgtca 56700 cactgtgggc tcacctgcac atcttttggc tctctgtcct cactaggtgt cacttcatga 56760 gagcagggac tctgttctgc tcactgcaga accttcctca gtccctttag ggcagtgcct 56820 tgcacacagt aggtgctcac taaaaagtca gccctgcttc agccccactt aggattcact 56880 ctgggaaaaa gaatcacccc tgcccccctg agtcaggttg gcctacaaat ggcaaggcct 56940 cttcagtctc aaatctcaca tgtggctcac acagctctgc ctttccctcc ggctgaatta 57000 gcctctcacc tcaccctccc ttcttgcatg atgtgcttca gccggaatgg gttgtttcct 57060 cctggctccc acatgggccg tgtcctctac ctggtgcatt ctttctgttg ccatctgttg 57120 gaaaactagt tccagtcctt cagattttaa cttagatgtc atctcctcca ggaagctttc 57180 cctgattgcc caagtctggg tcaattgtcc ctcctctgag cccaagcttc cgagtcttgt 57240 tatgaacaca ctgcattgtt gttgcttatg cacttgatcc caatcccatg caggtagggg 57300 ccagtcttgt atcccccagg gcctaggaca gtgctgggac tcaagaaacg tgattggagt 57360 gaatcagtgg atgaatgaat gctctgaatg acaccacctc aaaggacctg gagcattctc 57420 cttaatcact cttgtgggtg ccaatctcct tgggcgcctt tagaagcact atattttctt 57480 cctgagtact gcagctcttt ttaggcccct gaaaggcacc tggctctttc agttttatgg 57540 gcatttgtga aagaactcag cccaacagac ctctgatggg cacacctaga accacctcca 57600 aatggattca ccatggcaca tgctgggacc accttctacc ttggagagag cttcccaccc 57660 attctctcat ttctgcctcc cctaaacagt gagctggcct ggaaaagtct ggcctctcca 57720 ggtggggagg atggagtcag gtgactatgt cctctttgat gagtgggcag tgaggtcccc 57780 caagaagaag gggcaatgct gagtagagac cccttcccca ccctccagtc atctgagacc 57840 atgacttggt tatcttcttg gaacccacat ggtaatcttg gcacccatca gagactgtcc 57900 ttgtctctct agggctggga tgggggacat gctggaggac ccagatgtgc acaccacagg 57960 tgcccacaac acacaaatga aaacagcagc agtagcaggt gttgagtact tactcggtgc 58020 cagatgctag tatcttattt attccactaa acaactccaa gtctgtgttg ttgctggagg 58080 cttagggtcc ttgggccaca tgtggggctg aaaataatcc tgatctctgt ccgtctgtga 58140 taagagctag taataataat aatgtctgcc ttctgttgtc tccctgtatt cctgtattag 58200 tctgttttca cgctgctggt aaagacatac ctgagactgg gcaatttaca aaagaaaggt 58260 ttaattagac ttaacagttc aatatggctg gggaggcctc ataatcatgg ccaaaggcaa 58320 ggaggagcaa gttacatctt atgtaaatgg aggcaggcag agagcttgtg caggcaaact 58380 cccgttttta aagccatcag atgtcatgag acttattcac tatcatgaga acagcacagg 58440 aaagacctgt ccccatgatt caattatctc ccactgggtc cctcccacaa cacatgggaa 58500 ttatgggagc tacaagatga gatttgggtg gggacacaga gccaaaccat atcacccccc 58560 aacaagagaa ggcaggcatt attactagcg catatcacag atggggaaac caagccacag 58620 ggaatttaag taacttgttc attgtcacac agataagaag acatgaacct gggattcaaa 58680 cctggacagt tcggctgcag agcttgagct gttaacgatg acactctcat ctttttcaca 58740 aatgcccaca caagtcccct tctgtacata tcaacatatc tcacttacat atatgtgcaa 58800 atcagcatat gaaatcccgc tcttgctcat gcactcatgg acacatacaa cccaatcaca 58860 aagacataga ttctcctgta gcatgaaatt catcatacat cgtcaatagg cagcgagaat 58920 tcacccgagt cacgcacatt tacacacagg tgtgcatgtc cctgttaata tgcctgcgtg 58980 aaaagcatgc agaaccgcca ctcacaagtt cacacatgtg caagtacaca cacattgtca 59040 cataggcaca gacatatata tgtgtgcaca cttgcatata aacatttctt gagcacctac 59100 tatgtgccag gcactgtgcc ggttctggga atataaaagt gaagaaaacc gagtttctac 59160 tctctcagag tgtgtaatcc aggggagggg gaggcatgcg tgagcaaatc agatagaaag 59220 tgcagtggct catgcctgta atcccaacac ttttagaggc cgaggcaggc acatcacctg 59280 agttcaggag ttcaagacca gcctggccaa catagtgaag ccacgtctct actaaaaata 59340 caaaaattag ctgggcaagg tggcgcatgc tgtaatctca actacttggg aggctgaggc 59400 gggagaatca tctaagcccg ggaggcagag gatgttgcag tgagccaaga tcgtgccact 59460 gcactcaagc ctgggtgaca cagcaagact ccatctcaaa aaaaaaaaaa aaaagaaagc 59520 tcatagttat ggagtgctta ttatgcataa gttgcagagc tgaatctgcc tcatgcatta 59580 ggggtccgta aacaccacag cccccatgct tacatgcaca catatgaata taaacctccc 59640 tccctgcaga agccacatgg cacaacctgg gcccttgaca ggagtggatg ggtgtagggg 59700 acggggatca gcattatttt cagcccccct cgtggcccaa ggacctcaag cctccagcaa 59760 cttccctgag atgagaccat ggcctgtgcg acaaaatgca catgcaattt gcagacatgt 59820 gtacatctct ccacgtgctt gtatgtgtgc aggtccctca gcagtccccc attctcctca 59880 ggacctcccg ccctatcaag agcccctgtc cctaagtcac caacctcctc cccctaccct 59940 catcttgcac cttagggacc tgtgggacgg agacatttgg atcttccaag acgctgcaca 60000 cacaggcgga ggatgggctg caatgactcg gcttccaggc agctgtggtg agtttctgca 60060 aagactccct tcctcaaaga ggctctgctt tactgaaagt tcaggttgcc gtggtttccc 60120 cgcattcaca tgggggaaag gtcagactct cacaaaaata tttaccaaga actgtctgtc 60180 agtcaatcaa caaacattta cagacagctg ctgctcacat gctggggaga gaggggggat 60240 gaataagcac aggccttgtc ctacataaaa gctcaggggc catgagggag ctgtgattga 60300 cccagcaagg gacacagtgg ccaggaagag ggaactgctt gtccaaaacc tggcatcaaa 60360 agagtgtgcc ctggtgagac caaaacaacc tttttttttt ttttttaatt gagacggagt 60420 cttgctctgt cgcccaggct ggagtgcagg ggcgcgatct cggctcactg caagctccgc 60480 ctcccaggtt cactccattc tcttgcctca gcctcctgag tagctgggac tacaggcacc 60540 cgccaccatg cccggctaat ttttttgtat ttttagtaga gacggggttt caccgtgtta 60600 gccaggatgg tcacgatctc ccgactttgt gatctgcctg cctcggcctc ccaaagtgct 60660 ggaattacag gagtgagcca ctgtacccag ctcaaaacaa ccatttttta tggcccctaa 60720 aatcacaacc agtggtgtgc tggtaagcca acaaaaacaa caggaaacaa accaaaaacc 60780 caatatgtgt gtttgctgca aaatcctgaa acacggtgtt tgctgctttg catggtgtag 60840 atagtctcat tgtggccaat ttcaggctac caaggtgatg tcaggggctg cggaggcaaa 60900 gatgaccaca ggagctggtg caagctggac tcaacacact gtttcttccc acagacacca 60960 ggaggacaga gccagagctt tggtgcctgg aacagagtaa ggagtctcag ttagaagtgc 61020 cagggacaag acccaaacct cttgttcatc ttttgatgca tattccgacc tgttctttca 61080 ttcactcagc acttaccctg tgccaggtac tgtgcacctg aaggtgctga gaccattgct 61140 gcctgtgctg atggggaagt actggtgtta aggagcccag aagagaggca ttaaccaatg 61200 cctggggagc tagaggactt cctggaggag gtagtatcta agctgaaatg gaagggagta 61260 aagcagcaag tgaggggtag aaaagagtta tttgcaagca gaggaaatag gatgtgcaac 61320 agcttagtga tctcatgcac cactgggttc ccccagcaaa agcatcctgt tgagcagatt 61380 ttgtaaatga accagcacca tcctaggtgc tttacaagta ttcactcatt taatcctcag 61440 acgatcctat ggggtattta atattcttag ctcctttgta cagatgggac cactgggaca 61500 gatggaagtt atgaataagt caatatgccc aagatcactg gtctgccctt cctctccacg 61560 atgctacctt gaagcatctg aagtaggtgc agcttgggga aattttaggg ggtacccaga 61620 tatgggaaac aactgtaaat ttaattctgg gtttaatgaa atgtcagagt gtgggaagtg 61680 atgggttcag tccctcccct gctccctgcc agcccttcac acacaaagac aagagtattg 61740 tccatcatgg acacattgac cgaggctgtc ggtcagccag tcaacagaga tttgtcagaa 61800 tctttatgtc ttggtctggc ctaggactgg gatgtttgca gtgaacctct gctcacattt 61860 ttgttttgtt tttccttttt tttttttttt tttgagacag agtcttgccc tgttgcccag 61920 gctggagtgc aatggcacaa tctcggctaa ctgcaacctc cgcctcccgg gttcaagcga 61980 ttctcctccc tcagcctcct gtgtagctgg gattacaggg gcatgccacc acgcccagct 62040 aattttttgt gtctttagta gagacggggt ttcacaatgt taaccaggct ggtcttgaac 62100 tcctgacctt gtgatctgcc caccttggcc tcccaaagtg ctgggattac aggcgtgagc 62160 caccccatct ggccttgttt tgtttttcta ctgaacactt aatctgggct gggctctagg 62220 ctgaggattg gggcacagaa gaagtaatga atctgtgtcc ctgtcatcaa ggggcatgat 62280 aggagacata caggagacaa ggttttctgt atgtttgaaa tattccatga tttaatataa 62340 aaaataattt ttatattaaa ttttagacac aaggtcttgc tctgttgccc aggctggagt 62400 gcagtggcac gattatgact cactgcagcc tcaaactcct gggcttaaac gatcctcctg 62460 cctcagcctc ctgagtagct gggactacag gcgcaagtag tcaattgcaa taaatgacga 62520 ttacaataag tgatcgaaga ggcccaagga caaaggagag aacaattctg ccccccatcg 62580 aggggaggac tcacggggaa aggcctgctg ccactgggcc tgagaagatg atattttaca 62640 gcagttgcca gggaatcagg ggaggacgat ttggacctac agatatggat aaggattttg 62700 gttttttttt agagaagagg caatggtgga aatgttctgg aattacatag tggagatgtg 62760 tgtactacaa cattgtgaat acactgaaac ccactgagtt atatattttt aagtggttga 62820 aatagtgaat tttacgtcat ataaatttta tctcaataaa aaataagctt atgactgggt 62880 gcagtggctc gcacctataa tcccagcact ttgggaggcc aaggcgagtg ggtcacctga 62940 ggtcaggagt ttgagaccaa cctggacaac atggcaaaat gccatctcta ctaaaaaata 63000 caaaaattag ctgagcgtgg tggtgggtgc cagtaatccc agctactaag aaagctgagg 63060 caggagaatt ccttgaacct gggaagcaga ggttgcagtg agccaagata gtgccattgc 63120 actccagccc aggcaacaag aatgaaactc catctcaaat aaatacataa ataaataagc 63180 ttatgtttca accacaggga gctttctggt tgtaagtgac aaaaatgcct gtttaaaaac 63240 aaagtttagg ccgggcatgg tggctcacac ctgtaatccc agagctttgg gagactgagg 63300 caggagaatc acttgagact aggagtttga aatcagcctg ggcaacattc agcctgggca 63360 acatagcgag acaccatttc tacaaaaaat acaagtatta gccaggcgtg gtcacttgtg 63420 cctgtagtct cagctactca ggaggctgag gcaggaggat cgtttaagcc caggagtttg 63480 aggctgcagt gagccataat cgtgccactg cactccagcc tgggcaacag agcaagacct 63540 tgtgtctaaa ataataataa tacattttaa aaataaataa gaagtttatg aatatagttt 63600 caggcatgga tagatcaagg ttctcaaata atttcttcag gttggaaggt tctactctgg 63660 gatctgcttt cttttgtatt ggctttagtc tcattctcaa gcaagctgag gaagcctcat 63720 tggtggccaa agaaaagcag ggacctcaga agccaagacc taccacctac cagctttcaa 63780 agcccaaagg aaagaaggca tgttttccta atggttccaa tgatgctacc aggaaaagct 63840 ctcattggtc agttttaatc atatgcctgt ccctgaacca atcttcagat tgggaaagga 63900 atgctctgat tggccaagcc tgagtcatgt gccctgttgg aaagctagga gttagatcag 63960 ttctctggca gttagatcag ttctctggaa ctgcgagtgg gaagacgctg tcccggcaag 64020 gaccaccagg tggtgatgtc agaagaaggg agagtagata cagggcaggc aggatagccg 64080 ggttcgtccg gaagagtggt agggaggggc gtgcctgcca gagggaaagg catgagaaaa 64140 ggcttggagg ttgaaattgc atggtgtggg ctgggaactg tggctgggtg gtggcgtgta 64200 gtcagatgat ggtgtctaga ggggtcaggt catgtgtctg aagacatgag aagcctcact 64260 tctcactttg aaccagggcc aggtacaaag aggtttggac agggcctgta cgccagtgtt 64320 gcccaaactc tcatccctat gcacacaagt catttctgcc gtatttacat tttttctcca 64380 tcattacatt ttcttcatat tggcctcctc gctttgttta aatcacttta ttgtcaaatg 64440 aaacattgtc accatcacac atgaaagatc aggatccttt actgtaaata gaagggaact 64500 gtaaaaacaa gcccattgaa accaaaaccg agtgattaac ttccagctag atgctgcagc 64560 ctgtggtagg ctaatctgga gaagagaaag agaactgaaa aaggaattgt tttctcagct 64620 cactatgccc acatgccacc taaaatcatc ttagggaccc cagggagcag ggaaggacgc 64680 agtcacaaag atggtcctgg tcctggcatg ggggagtgtt tgataaagac actaccggca 64740 taaacccagg ggttacaata gggaggctga catggctgaa cagaggagct ggagagcagg 64800 gtcccaggga cagactcaca gtaccggaag ttctgcaggg aggaaggagg tagtcaattg 64860 aggagcaccc gctctcttgc tttagcaatc aaaaccagga accagaagga ggaggtttgg 64920 cgaggatgag aaagagctcg tttggatata ctgaatctga gatgtgggga tgtctctgtg 64980 gagatccatt catctgttca agaagtattt atttatttac ttatttttgt gtatttattt 65040 agagacaggg tctctgtcgc ccaggctgga gtgcagtggc gtgatcgtgg ctcactccaa 65100 cctcgacctc ctgggctcaa gcgatcctct cacctaagcc tccccagtag atgagactac 65160 agttatgcac cgccatgcct ggctactttt tgtatttttt gtagagatgg agttttgcca 65220 tgttgcccag gctggtctca aactcctggg ctcaagcagt ccacctgcct tggcctccca 65280 aagtgctggg atgacaggcg tgagccatgg tgcccagcca agaagtattt attgagcacc 65340 tactatgtgc caggcactgt tctaggccct gggataaggc aatgagccaa ccaaactgaa 65400 atctctgccc accatgagtc tactttcttt gggggagata tgggagacct ggtgagaaat 65460 aagaaataaa agacttaata atatatacaa tatataataa catatgtcac acgagatagg 65520 aatggctact agctaggaca tgcaaaatgt ttgggtggga gatgtggaaa ctgagacata 65580 cccaactgca gttgaatgta taactctgga gttcaggcaa gagatctcag ctggggagat 65640 atatttagaa gacatcattc attcattcat tcattcactt agtaaatgtt tattgagtat 65700 gctctatgtg ccaagcacag tgctgggctg tggaccacac agttcctgat ccatgcgact 65760 ccagtacaga tttagtgaag gttgtaatta actctgtgga aagaggagag gagatgggga 65820 gggccccaga tgggccctgc ggacccctaa cgtgtaagag agggcagagg agaagacctt 65880 ggaaaaagag actggagcca ggtgcggtgg ctcatgcctg taatcccagc actttgggag 65940 accgaggcag gcgaatcaca agatcaagag ctcaagacca gcatggccaa catggtgaaa 66000 ccccgtctct actaaaaata caaaaaaatt atccaggtgt ggtggtggtg ggtgcctgta 66060 atcccagcta cttgggaggc tgaggcagga gaatcgtttg aacctgggag atggaggttg 66120 tagtgagctg agactgcgcc actgcactcc agcctgggtg atagagcaag actctgcctc 66180 agaaaaaaag aaaaagagac tggacaggaa caaacagcca gtgtcagaga accaagaggg 66240 gccgagtttc aggagtgagg gagatgctag tgccacagag tagtagggat tgaaactgag 66300 acttggcttg ggtggtggag ctggcttcag tgaaagtagt ttagtagcag agtcaggccg 66360 actctgcaca gagagggtag aggagtgggt gcagaggaag gaagtgaagc ccaagagagg 66420 agacagcttt tgagcacaag cactgtggct gggggagaga aggcgagaga gaaataggag 66480 ccggggtgga gggaagcttg ggctgtttga agatggaaag acttgagtag atatgtgtgg 66540 caatgaggaa aggccagcag agggaggcag gtgaagacac aggggagtga tggtgtttta 66600 gggtccagag ggagaagaga gctcagcctg ggagagaggg aatgaccccg catcaggaat 66660 ggctgcatca ttggaggggc actgtgcaaa ataaaaatgc aggtcttctt attcaaaaag 66720 ggctgataat ttggccaggc gcggtggttc acgcttgtaa tcccagcact ttgggaggcc 66780 aaggcgggcg gatcacttga gtccagcagt ttgagaccag cctggccaac atggtgaaac 66840 cccgtctcta ctaaaaataa aaaattagct gggcatggtg gctcatgcct gcaatcccag 66900 ctacttggga ggctgaggca ggagaattgc ttgtactcgg gaggcagagg ttgcagtgag 66960 ccgagattga gccactgcac tccagcctag gggacaagag cgagactccg tctcaaaaag 67020 aaaaaacaaa agggctgaga aatcaagatg gtcactcttg gttcctggat tagaagtggg 67080 ctgagttgtg gcttctggct gcccagcaga tacctctggg gtaggaatgg ctgtcaccat 67140 gctccacccc aaggtcccgt gaggtgagct actgaccttg atgatccttt gcacctgtgc 67200 ccaggcctcc ttcaagggca gaaggcagca atggtcatgg atggtggtgg aatggggagg 67260 atgagtaggg cctggtgtgc tggggacaga gcagcaggtg gccaggagcc tgtaccaggg 67320 aagcaggcag gtggcagatg tcccagggaa gcaggcagga ggtgggaccc tgtgtgagct 67380 gaggctccaa gccctcaatg ggtgctcctt ggtcccatta aacctcccta taaaacacaa 67440 cttcagagat aaaattttta agaatttctt tttctttttt tatttttttt tgaaacaaag 67500 tctcgctctg ttgctcaggc tggagtgcag tggcacagtc ttggctcact gcaacctcca 67560 cctcccgggt tcaagtgatt ctcctgcctc agcctccctg agtagctggg attacaggca 67620 tgcaccacca tgcctggcta atttttgtat ttttagtaga gacggggttt caccatattg 67680 gccaggctgg tctcaaactc ttgacttcga gtgatccgcc caccttggcc tcccaaagtg 67740 ctgggattac aggtgtgggc cactgcgcct ggccaagaat ttcaaaactg ggacttcaga 67800 gcatgacact gaagagtagg ggtcctccga gcaggggggt acgggctgct gcctggcgca 67860 cagctgtgga gccagcctgc cctacgtcca ctctaacaag aaagggtgga tgccgatatg 67920 gaatgggcag gaatttggca ggatcctctg gaagtgaaga ttgagtgcca ataaagaggg 67980 agaggatggg gattggggaa ggagcttgag gaagagtcca gagcctaaag ttggatgcgt 68040 gaatgcacct gcacgttcat ttcccattgc ttctctgaca aatttccaca agtttagtgg 68100 cctaaaactg aaatttatga tcttgcaatt ccagaggtca gaagtctgaa actggtctca 68160 ctgagctcaa aatcagggtg ttaacaaggc tgtgttcatt ccagaaactc tagggaagtc 68220 tgttccttga cttttccagt ttcaagtggt cacctgcatt ccttagccca tggcccttcc 68280 ctccatcttc aaagccagca gcataatatt ttgaaatctc tctctgactt ggcttctcct 68340 gcatctctca cttatgaaga tccttgggat tatattgggc ctgtctggat aattaaggat 68400 aatctccctc tacatctcag gttcagttga ttagcaaccc tgcttccatc tgtaacctta 68460 atccccccct tgccatgtaa tgtaacatgt tcacaggttc cagggattgg gacatggaca 68520 tcttgggggg ttcattattg tacctactac aatttttaag gttgctttat gactgcaaaa 68580 gaaaactcac agtggtttaa agaaaagcac acttacagtt tatttaagaa tttaaggttg 68640 gtttgatcac ttggtgatgt aatcaaagcc ccaggcaggc cttcttcttc ttccactcca 68700 ccatattcag aatgcctcct ttcatctcca ggcttgtctc ctcatgattg caaaagcacc 68760 atccaaagca gaaaggaagg tggcaaagga gccttctcct ccatgctgct ttttgattag 68820 gaaggagaac tgctgacttg ctctgtggtt ccatggcaga gaggaaggtc aggcagccca 68880 gctatgttga ccaactcaag ctaaatttgc tgctagaaat caggaaatca tggatggtgg 68940 tggaaggggg aggatgcgta gggcctggtg tgcccagggc agagcagcag gtggccaaga 69000 gcctgtccca ggaaatgcca gttcctgggt gggatcacgc taaggtgggt catcagtata 69060 taggagtggt ggttcagggc agccagaatc agagatcagc ctatagctgg gatcaaagat 69120 ccatctgtgg tgagggtcag gggtcagcat gtggctgggg ccagcctgca accaggatca 69180 tgagtaagtc agtaacatgg gtcgggactc agttcctggc ttgagtcaga gctgttctgt 69240 ggctgaggac aggaggtgga ctgtggccag gatcagaagt cagtctgtgg ctggatcagg 69300 gtcattctgt ggttggagtc agatatcata atggagttgg gatcaatcag tttagtcttc 69360 tctgtctgtc tttctctctg catctctgct tctctctgtc tcattctcct ttgctgtaaa 69420 cacctgttgg aagctcagca acttgggtcc atggggaaga gggaagttga gtctgggcaa 69480 ggtcctataa gtcccggtat ggaagaacat aagggaaaaa gagaaaaatg tcaggaatat 69540 aaagatattc cctagaaaaa tgtcaggaat ataaatatat gtatgtatga tgagaaatga 69600 atgatataaa accgcaaata cgcaaacact gactctgcat gtaaacaaca cacagaagtg 69660 gcacaaacag atacaaacac aagtgggttg aatgatggga ttgggacaat tttttgtcct 69720 ggattactgt tgctccattc tgcccttttt gtaataaagt caaattaaat ttagaatgtc 69780 attatttcct ctaattatgt tttataccct cacctcccac aaaatccaca gacatattca 69840 gtgtaatatt aaggataatg ttagcagact aggcatggtg gctcacatct gtaatcctag 69900 cactttggga ggccgaggtg gatggattgc ttgaacccta ggagtctggg actagcctag 69960 gcaacatagt gattccctgt ctctacaaaa aattaaaaaa caaattagcc aggcttagtg 70020 gtgtgtgcac ctgtggtcac agctacttga gaggctgaag tgggagaatc acatgagccc 70080 aggtgttcaa ggctgcagtg agccgggatt gggccactgc gctccagcct gggtgacaga 70140 gaaagaccct gtctcaaaaa aaaaaaaaaa agaagaagaa gaagaatgtt agcaaatact 70200 tttaaggcat taactatttg caggcatagt tctttttttt tgagacagag tcccacttgt 70260 tgcccaggct ggagtgcagt ggcacaatct ccgcttgctg caacctctgc ctcctgggtt 70320 caggcaattc tcctgcctca tcctcccgag tagctgggat tacaggcacc cgccaccatg 70380 tctggctgat ttttgtattt ttaatagaga cagggtttca ccatgttggc caggctggtc 70440 tcaaacgcct acctcaggca atctgcccac cttggcctcc catagtgctg ggattacagg 70500 cgtgagccac tgtgcccagc cgcaggcata gttctaagga cctcacaggt actaacatat 70560 ttaattctca caatagctac atgaggcaga tattataatg attcccattt tacagataac 70620 gaatcaagca cagagaggtt acacgctggc ctggcgccac acagcatatc taatggacat 70680 taggctcttt tgtctggtgc tgggggaatt tctggaaaat gcagagatgg aagagaggtg 70740 ggatcggggc agggcgggtg gggaatctcc caggccaagt tttctgggca cggcctggac 70800 ctggagctgc tgggatggag caggtgctga tgctgatgtg gtcaccagag ggcagcaggg 70860 accagggagc tgacttcctc ttcacacccc ctccacccgg ggcggtgccc ctgacttgtc 70920 aataaacgtc gtggaaacag gtgggatttg gtggaagaac tgacacggtc ccgccaccat 70980 cacaggcaag ttaagcttga gagcagcctg gtgtcacctg gcctccaccc accacgtcat 71040 ctgctgctca ccccaccact gtggcgtgca gagtactgct cttgtgtgcc ttttaccaag 71100 gaggaaattt aggcccagag tgggcaaaca cttttcccat ggtcacacag cttggaattg 71160 gaaaggctgg acctaaccag gcagggtgag cgcagagccc ttgttcctct gagtctctgt 71220 tctgagctgg aaccagggaa gatgatgagc ccatggtata tacagggcgg ttagctagga 71280 ctctggtttg aagcctggct atgccactcg gtaacagcac

atcgtgggca agtcccatgc 71340 agcctcatcg gctgccggga ggatccagtg acgtcaggcc tccctcgtgg tgggagctca 71400 aacacttgct gggttttttg ttttttcggt gttttgcttt ttgtttttta attaataact 71460 ttcgtgggaa aaataaagtc agaaaggttg ttcaaccgga tgttcacgga gatactacgc 71520 cttcaaaaac aagcgctcat ttcgtgatcg cccacacagt gccagatgct ggggcgggga 71580 tgggatttcc ggtggttttt cttcactttc ctctttatat cttcaggctt tctctgaata 71640 ggaaaaggtc tgcacgtgct gatagagtaa tcagaaaaat gcaataagcc actttttgta 71700 aaaaaaaaaa aaaaattcag atgcactcac tgccttcctc ccccctttcc ccctcccctc 71760 atcccccaca aattctgtgt aaaggcaggg ctgagggagt acccccaaat caacatctct 71820 gaaagctccc ccgcaggggc ccaggccact cagcccactc tggcgctgct acccaggggc 71880 gtcccctcct gccctctggt ccagccccgg aggctgaaac tcccgcagcc ggcatctcac 71940 tgggcctatg accctggact tttcctactt cctgcagggt gggggccagc ctgagttggg 72000 cccacctgat ccaccaggtc ttctcatccc tctgtagctt cccaaagccc agccagttca 72060 gtaatagttg ctgaggccaa caacttggat ttcttttgcc taagatcctt tgttgggctc 72120 ccagtcccat tgcatctcta gcagtcaaat atattttcta tttgctgcct aatttttaaa 72180 ttagatttct ctgagacaat gctcaggcaa agtggattcg agccgacatc tgcgtaggtg 72240 tcaggatgac ataagctgat atatcacggc ggctgggagc tcttaggtca gacccgggtt 72300 caaatcctgg ctccaggatt tcctagctgt gtggttctag gctaatcctc aacctctctg 72360 gacctgtttc cttatctgta aaattgggat aatcattgag cctcccaact agagctgttt 72420 ggagacgtaa atgacacagt ggctggacgg cacttagcat attgcttggc cctaaacaga 72480 taactatttt tatttaaaat tatattttat tataaagagc ttttggaggc aagaagtcca 72540 gaaggctagt tcaggctctg ccattctctg ggtgactttg ggcaagtccc ttttcctctc 72600 taaaactgtt tctccatctg taaactgtgt gatttgggca gctaaaggga ttgattttgg 72660 cttagaacaa ggtttttttt gttttttttt ttttttcctg agacagggtc tcactctgtc 72720 atccaagctg gagtgcaatg gcacaatctc agctcactgc aacctccgcg tcccaggttg 72780 aagcaattct accaactcag cctcccaagt agctgagatt acaggcaccc accaccacac 72840 ccggctaatt ttttattttt tggtagagat agggtttcac catattggcc aggctggtct 72900 tgaactcctc acctcaagtg atacacccgc ctcagcctcc caaattgctg ggattacagc 72960 catgagccat catgcctggc gcaaggaggt gtttctaaca aacatcaggc tgcatgtgag 73020 gtagagagct ccccatcatt tccagtgtag tatgtaggct cctagagcag gaaggaatct 73080 aaaatggcca agcccctcaa gtgtgaaatc agggcagcta ctcagagaag caaaagtgat 73140 acagacatag gagcttcagg ttcccagcct tcccatctaa ggcccacacc atccctctgc 73200 ctgcctcgga tgtgaagcgc agatcactac agagagcctg ggtgatggag acactgtctg 73260 gattccacta tagatctgtt gctaactagc tgtgtgacct tggcagtgta tcatcagcca 73320 ctgttggtgt cctgttcctg tcttcttggt gtgccagagt ctctcaacac ctccaactct 73380 cagttgaaga attttttttt tcctggagta gctggaagtg ctggagaatt aatgctcctg 73440 gaagcagctc tcagccaatg acagatggga gctggtggac aaatacccca gctccctctc 73500 ctttgcaggc ataactgggc atgtgccatg catggtcttc cagagatccc cggcaggtaa 73560 gcatcagtta ctcatgctgg taatctgttg aataactcac ccttttgtca gaggtttatt 73620 taaactggag ctttgaagag gggctgggta aaatgaggct gagacttact gggctgcatt 73680 cctgggaggt tagggcattc ttagtcatag aatgagacag gaggtcgtca ccgatacaag 73740 tcataaagac cttgctgata aaacaggttg cagtaaagaa gatggctcta acccaccaaa 73800 accaagatgg caacaagagt gacctctggt tgtcctcact gctacactcc caacagcgcc 73860 atgacagttt acaaatgcca tggcaacatc aggaagttac cctatatggt ctaaaaggga 73920 ggcataataa tccacccctt gtttggaata taattaatca ataaccataa aaataggcag 73980 ccagcaaccc tcagggctgc tctgtctacg gagtagccat tcttttattc ctttactttc 74040 ctcattaact tgctttcact gactctatgg actcgctctg aattctttct tgcatgagat 74100 tcaagaaccc tcttttgggg tctgcatctg gacccctttc aggtaacact ttcttggttt 74160 gtgtccttcc ctgcctcagt tccccactac cttagcaggg tttcccggga ttacttccta 74220 aatcaactat ttgaatcctt gtatggggaa cccaaaccca gacggcaagt ttcttaacct 74280 cttgcatcct ggttcctctg gatgggtgaa ttccaggagt taacagagac acagcactca 74340 cagcagtgtc tggaggtatg tttggttaat acataaaaaa taaagagaca aagaaacttt 74400 atttatacaa aactccaccc cttctgttcc actctcctca gcaaacacag ataacaggtg 74460 atgaaactaa aacacacaga cgagcattac tcaacccaag gttcccgcct tccctagcac 74520 ctgaggtctg ggccaacatg cagggtaact ggtgccttat gcctgctgtc tggattgccc 74580 ggcccacagg gtggctgagc atatttattc tgggggttcc atgcatacga ggagccccca 74640 gccatacagc tgggcatggg tgtttggcag caaattgtcc ctgctttagt cacagcaatt 74700 tttcatgtcc tctgttgctc cccttaactg aacacccctt gacagccggg attctgcctc 74760 cagcatcaac ctaaggaagc tctgttaaat gaatgagcac ctctaggcaa tgaccaccct 74820 ccctgaagtt caatttctca tcgatttccc aaggccgccc aagtcattcc cttgatggct 74880 gtgttagttt ctaggctcgg cttctagttc agtgagacag aggcaggtgg gcctccatgt 74940 gggccatgat ctggtgggca aaacatgcct tgggcagtta caacagggac agtctgctgc 75000 agaagctgtg gagggagcag ccaacaactc gttcacaagt cgaggtgccc aagggcctca 75060 gttttcctgc aggggcctgc cgtggggcac ggcgacccag tgccctctac tctcatggga 75120 tgtggcgagc ccagcccaat gctgggcggg gtccagtctc tgcagcccag tgttaggcca 75180 acgtcagtgg ggccaatcac cttcatacca tggttcttca agtcttttgg aaatctgcca 75240 gcctggctgc cttccaggag gtggcatttc ccaggcatgg ataagcccta gtcctcatct 75300 gcagactcag caacaagagg acatgcacct aagagaccct catgtatgtg ggtcccacgg 75360 agacaaagtt cacgattttg ggggtctggc ttgagctctg agcattgcca ggggcaggat 75420 ggaaggatga tgaggattta ctcttctctg agatgcccga gggtgccagg gaggccggac 75480 acagactggc ctccagtgga accccacctg gagaggggtc tggggccctc aggggggttg 75540 tagggggcga ggacctgtct ccacagcagc agccacagca aggactggcc atgacagggg 75600 tctggccacc atcctcctgg ccatgacact gtttcaggtg gccgcacagg tggcaggtaa 75660 gggctgagta gacaatgcca ctgcccaggc tgtccacaag ggggtctgtg gcctgctcct 75720 ggggaagtgg gggctttggc atgtcctcta ccttttcccc cggctccaga cccaggtgct 75780 ctggggagct gcttgggaga tgtgagctct gcggactgcg aggtggctcc ctgtccagtc 75840 caaaggtgaa caaggggaca gggactgggg cagggtcccc agggcagcca ggaatgaggt 75900 cttggaaagg cttatacccc tcttccccac tgctagcccc aaacccacat ttctctgggg 75960 acacagcact gctggcaagc aggcttgaga aggccttgta accagcctct cctgggggac 76020 ccaagcccac caccgcactg gcctgggtgc caccctgctc caccgcatgt acaaactcct 76080 gatagccact ggtgggggcc gagacggggg ctgcagctgc cccatgctgg aggacatttc 76140 ggcggaggat ctgctcccag gtttctggct caggttgggg cacagtggtt ggctcagaga 76200 gctgggggac acagggcatc tcgggttcta cttcctccag gtgtctggcc agcagtgggt 76260 ctggacccag ctctctggga cacggtgact ggctcaggga gttgctgaag ctgcggtaag 76320 cagggttgcc tgcgatgacg aggggcgtct ctgtgcaagt caggttgtct ggactctggg 76380 tcgggctggc aggaggactt ggctccaggt ggagaggctg ctccttgccc cagggaggtg 76440 cctccttggg ccctgcactt gggaactcat cccagggcat gtgagcactc gtacttcccg 76500 aaggtggaag aaggcatgac tcccccatgt cctgctggca aaagccccca ttctcctctc 76560 cgagcaggtc caggaacagg ctctctgtta gccgggccac aatgccctcc cttccctcct 76620 ggaagtcatc cctgctgctc tcaggcgatg cacagaagct ccctttttct tcctctacct 76680 cctcctcctc ctcacactcc accggggcct caaacaactc cacacatcgc accacgctga 76740 tgctctctgg ccagaggact gtcttgctga tctccactgg gcaccatgct gattttccag 76800 agccctggaa aggcatctct ttggcagcct tgtgaggatc ttcatccctt ttcatgttgt 76860 gctccagaaa acagggcaag agcttggtaa gacaattctt ccagtgtctg caaaagcaaa 76920 ggttggtcag ggttcagttt cgacacttga aaaactccta ttcattcttc aaagcccagc 76980 ctgaaatcac catgtcctga tggctgtcta cttcagacat gccttctaac gttataatca 77040 gtaattgatc agtctatgtt gctagaatga gaaatccctg agggtagggt ctgtgtcttt 77100 gggatttcgg gaacccagca catgtcctgg gcaccaaata caagctcaga tctgttgagt 77160 aagaaaaagt ctttatggat gatgataatt aaaagcaaca ccattatccc aagccatgac 77220 caattagaaa ggatatggaa gccgggtgcg gtggctcacg cctgtaatcc cagcacttta 77280 ggaggctgag gagggaggat cgcttgagct caggagtttg agaacagcct gggcaacaga 77340 gcccaagccg tctcaaacat ctctacaaga attaaaaaaa aggaaaatta gctgggcatg 77400 ttggtgtatg actgtagtcc cagctactta gaaggctgag gctagagaat cacttgagcc 77460 cacgagtttg aggctgcagt gaatcactgc actccagcct gagggacaca gcaagaccct 77520 gcgtccaaaa aaaaaaaaaa aaaaaaaaga tgcgggatcc tagggttgaa gcagggtcct 77580 ggaccccctt tagtttctgg agtatggaaa ggtccagcag gccccaggga agaagctaag 77640 gaggccacag tggcggggca acacagcgag agcttgtctt tacaaaaact ttttttggta 77700 ttattttttt tgagacaaga gtctcactct gtcgcccagg ctggagtgca gtggcacaat 77760 ctgcgctcac tgcaacctct gcctcccggg ttcaagtgat tctcatgcct cagcctcctg 77820 agtagctgga attacaggca aaaacctttt aaaatagcca ggcgtggtaa tgcacatctg 77880 tagtatcagc tactcgggag gctaagatgg gaggatcact tgagctcagg agtttgaagg 77940 tgcagtgagc tatgatcgca ccactgcaca ccagcctggg ccacagagtg agatcctgta 78000 tattaaaaaa aaggagggga gtgttcagga tagtggctat ttgccgacaa gtatacaatt 78060 aggtcaagat tttaacaaca ggcacggctg ctctggttgt taactataac agcactccca 78120 aggggtatca ccatgtacct gacacaccag tgaaactcag ggggcttggc cagcagtcca 78180 cgtttccaga acacccctcc tcctgccgtc ccttgaaggc taatgtcaca gggcaattac 78240 cgaatgcagg gaagagaagg cagacatgga gctaggggct tgcctctcca attctatcac 78300 cttcccaatg catgcaggct gtgacctaag ttcagataca tacgggcact tggctggttc 78360 ctggcctcgg gaccgcttct cccactgtga cccctaaaat acagagacag ccaatgaaag 78420 agcagagtgg ccttgcagcc cggcctggag tacggcaatg ctcaggaagg ggtacaagcc 78480 tcgacatcac acagatcaga agtccaacag gtcacttacg ctttctgagc ctcagattcc 78540 tgggaattaa ttacattgtc ctaatattta cagggactgt ggtcgatggc aaaagtggcc 78600 acagaggagt gctctgaatc tcctcccatc ccttgaatct acactggccc tgggacttgc 78660 ttgggcccat agaatgtgat ggaagtgata tcatgccagt gctgcgccta ggcctcaaga 78720 agccttggga cttcccaccc ttgaaaactg agccactctg taaacaagcc tgggccggcc 78780 tgctggagag tgagaggcac atggctcagg cacctgcact gtcccaggta aaagtcagcc 78840 aactaggctg ggtgcagtgg ctcacgcctg taatcccagc actttgggag gctgaggtgg 78900 gtggatcatc tgatattagg agttccagac cagcctgacc aacatgaaga aaccccatct 78960 ctattaaaaa tacaaaatta gcctggcgtg gtggcacatg cctgtaatcc cagctacttg 79020 ggaggctgag gcaagaaaat tgcttgaacc caggaggcag aggttgcagt gagccaagat 79080 catgccattg cactcctggg caataagagc aaaactccgt ctcagaaaaa gaaaaaaaaa 79140 aaaaaagcca actgtcagac acggagggag gctacgggga cgggtcagct caattcagat 79200 ctgcccagcc cagcccagcc cagcctcccc cgccaccctc actgctgagc ccagccccaa 79260 atgccagcct gcagaagcat aagctaaata aagttgttgt aaccctctac atttcggggt 79320 catttgttac acagcaaaca ctaactgata cataaatggc acttggtatt tgcaagaggc 79380 tggaatatgc actttatatg tattgtctca ataaattcca atatctgtcc taccaggtaa 79440 acactgttat aacccacaag aggaaactga ggctcagaga attaagcaac ttgctcaagt 79500 ccacactgcc agtgagtggc agagtcagga ttcaaagcca acactagatt ccagagtccc 79560 tggaaaagca aggaccctca gaccttctct tctagacagg acctataatc cccctttctc 79620 ctacctagag aactctcatc ttgctcaggc aaaccccacc cactcaccca caggaccatc 79680 gccttgggga agctgactcc acccctggct taggggtggg atacatttca ggccaatgag 79740 aacccggaag ggatttgcta gtgcttcggg ggcagtttcc tatgcttaag aagaagacac 79800 agaaagcctg tcaccctctc cttcctccac tccttctggc ctctagccct ggtactggca 79860 cctccccagg gaatgacaca gtgatgatga cccaagggtc attctcccaa ggggagggaa 79920 aaaaacagga aaccaacccc agagaagccc accccaggct gggattgcca aggggaaggg 79980 cgggaaagcc tctggtctgc taatgacctc attcggcttc tgcctctgtc tcccctgcaa 80040 ccctcctgac tgtctatttg ccattccaga agccagtgaa ggtccttaac agcccaggtg 80100 ttctgaacca cacttcttca tgtacacagt cccacatgtc ctcatccacg cctactccta 80160 cctgagcatc ctggattatt atagccacga ggcggctgcg ggctgggttg ggaatctgat 80220 cccaccattc tttcttaatc ctgaaaaaca aaaggaaaag gtgttgatct ctgaattatt 80280 gacaggtgaa cgctcctccc ataaaagtgg tcgttatcac caatacttat ttggcatcag 80340 agatcaggct gattagtttc tggaggtcaa accactgact catcattcta acctggtgac 80400 atggattcta ttgctaaccc tattctacag atgaggagag tgaggcccaa ggaggtggag 80460 tgacagctgg taggtggcag agccaggttc acactcccgc attctgttac ccagcacaga 80520 gttcctgagg aggccaccta tgtcctcagt gggaatgaga ccccagggga ttaggaaaga 80580 ctcaaactgc agagagagga ttctaggttc ccctcaacaa tccctttttc agagcagctg 80640 agggcaaatg accagggaat ggcctgggat ggcctgggaa gctgggggac ctacggagaa 80700 acacaaccct tctctcttcc tggcctgcct ggggccatgg tgcatcatca ctgggaggta 80760 aggtcctcag accgcttctg aattagcctc tgcccatggt ttcaataaat aaacacatat 80820 ttattgagca ccttctatgt ttgactcact gtgcctggcc ctgaggctac aacagtgaac 80880 gagacaaaga tgcctgcctt ggtgaaactg accttcaaga gggaaagatg gatgataaat 80940 ggagacagaa atgcttatat gaggccaggc atggcggctc acgcctgtaa tcccagcact 81000 ttgggaggcc aaggcaggtg tatcacctga ggtcagaagt tcgagaccag cctggccaat 81060 atggtgaaac accgtctcta ctaaaaatgc aaaacttagc caggcatggt ggcaggcgcc 81120 tgcaatccca gctactcagg aggctgaggc aggagaatcg cttgaacccg gaaggcggag 81180 gttgcagtga gctgagatcg cgccactgca ctccagcctg ggggacaaca tgagactcca 81240 tctcaaagaa aaaaaaaaaa aaacgaaaga aatgcttata tgaggtcatt cattcatggt 81300 aataagtgcc atgaataaag ataaaggggt agaagttcaa gatcagcctg ggctacataa 81360 gtgacagcca gtctctacaa aatgtaagaa aattggccag gcacagtggc tcatgcctgt 81420 acccagcacc ttgggagaac gaggtgggtg gatcacttga ggtcaggagt tcaagaccat 81480 cctggccaac atggtgaaac cccatctcta ctaaaaatac aaaaattagc tgggtgtcgt 81540 ggcacacgcc tgtaatccca gctagtcggg aggctaaggc aggagaatcg cttgaacccg 81600 ggaggcagag attgcagtga gtcgagatcg caccactgca ctctagcttg gtttttgaga 81660 ctccactctg tcccaaaaaa taaaataaaa ttaaattaaa ataagaaaat tagccaggtg 81720 tggtggcaag cgcctgttgt cccagctgct ctggaagttg aaatgggagg atctctttag 81780 cccaggaatt ggaggctgca gtgagccgta accgcgccac tgcacttcag cctggctaac 81840 agagtgagac cctgtctaaa aaacagaaca aaacaagaaa ccaaagacaa agaacagcaa 81900 agaggctggc atgctgggat atagtgatgg ggagtgggaa gaggtacagg tgatggggaa 81960 actgaggctc agagcaaggc gatcagtcca aggttgctcg gaggcatcag tgacaagata 82020 tggaggggaa tttgcattcc ctgcctacct agtgtgtgct gggccaagag tgggcacttt 82080 catcaatcat gttaataatc cttagccacc tctaatcctg acttagtgag gttgagcttg 82140 ctagtcccat ttcagagatg ggaggactga ggctctgaga caccatgatt ccaagggctg 82200 gcaagagctc tctggaaacg aggctccttc caccacctcc cagccactcg ccactcatct 82260 cctccctcag cctttactgt gatttggcct cttctggctg atgaaatatg ttctcaccat 82320 cactgtcact ctgcaaagca ggaaccccag gctcaggggt ttcagcaact tgtcggccgc 82380 cagctaccaa gctccaaggc caggattcaa acccaggtct cctgactcca tcagggcaat 82440 cgtcgcaaat tcctctggct cctaccacat gcccccaaat gcccccagtt gcccaggcac 82500 ccagtgttgt aaggggatgt gggatgactc catcttgctt atccctggcc agctcattct 82560 ccccagtcag ggcccccctc aaattttccc tccttggaga gccttatcct ggtccccctt 82620 gcaaaaagca ccccagaaga tctctggcct ccaccctggt ctagtgcatt cagagagtgg 82680 gtcactatgg aaaaccacct gtttgtgatt gatatgtatt ttcagtctcc ccagtaggat 82740 gccggtttta gaaatgtgga gtcctggttg ggtgcagtgg ctcacacctg taatcccagc 82800 actttgggag gctgaggcag gcagatcatg aggtcaggag ttcgagacca gcctgaccaa 82860 catggtgaaa ccccatctct accaaaaata caaaaaatta gccgggcata gtggcatgcg 82920 cctgtaatcc cagctactct ggaggctgag gcagaagaat tgcttgaacc cgggaggcgg 82980 aggttactgt taggcaagat tgtgccacta tactccagcc tgggtgacag agtgagactc 83040 cgtctcaaaa aaaaaaaaaa aagaaaaaaa gaaaaaagtg gggtccttgt cgatatgttc 83100 tcagctctac tctcaaggtg tgtagtagct gctcagtaaa tacgtattat tgactgagag 83160 gactgcaaag gggcagacta gaggggcaag tccctgccac cctgcactcc agagggactg 83220 ctcggcagca ctgggcccag gactcacttg gtgatgctga catagcacaa caggcagacg 83280 gccaggatga caatgcagga aacgctgacg cccagcagga ggtgctgctc gaagggctcc 83340 ctgtaggctg gaagtgggag gggaagccat gagctgaggt cagccatggt acagcctggg 83400 ctccacttcc caggactgga gacccagggt caggaccctc gtccgagacc ctccacgatc 83460 tgaccgccga ccacccctcc acctcatctc cccaccactc tccctcttgc acattgagtc 83520 ctcgtcacac cggccacctc attgccccct gctcctccaa agctcagtgc cacctccaga 83580 tcttaaactt gctgctcttt ctttccggaa gcacagaagt caaatgcacc aacttaggag 83640 ccggatggct ccttgctgtg tgacacagga caagtccctt atcttctctg tgcctccgca 83700 tcctcatctg tactatgggg ttaatgacag aattgatcgt gcagggctgt tgcgaggatg 83760 acatgatgaa tcaatagata atgcactcag aacagagtgg cctgtccaca tcattagctg 83820 ccatcatctc aacacggttg acttctcctt gttcactttc atcgtcacat tctccaaaaa 83880 gttttccccg acaccctccc taccgttgcc ccctgcccat cgctccatcc ccttcctctg 83940 cttttatttt cttcccagca ctcactgtca tccagatcca tgtgatttct ttctttcttt 84000 atatgtttgc tttttgtctc tccctccaga atgtcagctc cacaagagtg gggatttcta 84060 tctgctttgt tcatcgccga ttccccagtg tctagcacag ggctagccat gtaggaggaa 84120 ctcagcaaat atttgctgaa ataatgaatg agggtcccat acattgttat ctgtttcata 84180 caagctagtt cccttcccca tgccaatggc agatgagaat caattcaaaa ctttcccaag 84240 cacctggttc tgtgctgggt tctggagata cagccatgga gaccctgcct atggagctca 84300 cagcccaggg caggagacac gtggctcaac tgatatccac agtgcaggat gagcaatgct 84360 gagagggctg tgcggtggca gggtcagggt actgccagct gccactgcac ctaccacgtt 84420 gaagaatctt ctttcattga gttttcacga cagcccttac tcctcccatt atacagatgg 84480 ggaaactgaa gcagtcttct tcactcaaca gctttttcct ctctcccttg tcagggagcc 84540 atgcccaccc taaggcttga gccaatcacg attatcctgt tcctttttcc aggcatttga 84600 gttcccagtc ttccttgcaa tgagcaagta ataacagtaa ccatgtaaca tgtaacagtc 84660 ttggccaatg agatgtaagg ggaattttgc tagggaagct tctagaaaga tgtttgcttc 84720 ctgattaaaa aagagatgca caaggtacac tctctcgtag cagcctttcc tttccttcct 84780 gctttatgta agacagtgtg caacggccaa cctgagggca tgggtggcaa gtctaaggat 84840 acaatgccac catgtgaagg acggcagaag ggaagacagc agaagggaag agatatcact 84900 tggctgctgt acaaaaccca gtaggtccat ttgtaagact tcattttatg tgagataatc 84960 aatatttcta ctgcgaaaga cctgtgggat acctcactga aaagagttgt ttctgggatg 85020 taggttccaa gtgagaaagt tcccagcctc ctttgtggct atgtgtagtc atgtgaccat 85080 tttcaccaat agaatttgag aggaagtgat gcggcccact tccgggctgt ctgctttcaa 85140 gaagcagatg tgatctccat aatcacgcct cttctgctgg ctggatacag acaatggtgg 85200 ggccctagag cagggattct tcacttgggg ttgatggacc aacatggctg caggtagaat 85260 tcaggggtct gagaacttgg atgggaaaga aattacatct ttactttccc taacctctca 85320 cacctagcat ctccttccat taagaatgga ggaaggggcc aggaacactg gctcccactt 85380 gtaatcccag cactttggga agccaagatg ggaagatcac tggagcccag gaggtcaaaa 85440 gcagcctggg caacatagtg ataccctgtc tctatttatt ttaaaatttt aaaaataaaa 85500 taaaatacaa ttttaaaaaa agaatggagg gaagagaccc cagtaatgac agcatcagtg 85560 atcctgtccc cagcaaaaca aaactcacag ctattctcat atcacatcac aatttcagta 85620 tcataaagta ttatttatgc ttatctactt tgaaattaca gtatttatgc gagctacccc 85680 tagaacttat tatttataga tcttaataaa gaagcacata tatctctaga ttaaaaatgt 85740 gtttaaatgt tttataactg catttcaata caattagttt cctctgtagt tctatttatt 85800 ttattttatt tcattgtatt tatttatttt tagagatggg ctcttgctct gtcacccagg 85860 tgggagtaaa gtggtacgat cacggctcac tgcagcctca aactcctggg ctcaagtgat 85920 cctcccacct cagccttccc agtacctggg actacaggtg catgccacca tgcctggcta 85980 atttttgtac agatgggggt ttcactatat tgcccaggct gggattttat gtatttaaaa 86040 atattctgaa atgaggtcca taggcttcac caggctgcca aaggaaacca ggacacaaaa 86100 gtaggtaaga atctttgtcc tggagaatga tggcaccaca agatggaaag aacctcactc 86160 cccagatttc tacagggagg gaaaccacct acaaaccagc aacattcacg ttagcttgtt 86220 acatgagcaa gaaatatatt cctatagtgt taagccatgg aaattgtggg cttgtgtgtt 86280 actgcagcct agcctgccta cctggcagat acaccacagc aggtgtatct aatggcagga 86340 ttgccattag aggcatccta tgatataaaa gcccagagac

tcagccaggg gaagtgtctg 86400 gccacagggg aagaatggag agtggagatt accattgctt aggcctcttg atactcacag 86460 ttgtgccact tggtgctggg gctccactca ctccaggtgg tgttatagca ctgagcccag 86520 gccctcaccc gtgccctgta ggaaatccca gacttcaggg tgctggctgc gatgcggagg 86580 gagggttcta ggtaggtcac gttatagatt ctgaactggt agagaaaagg gagatgttag 86640 ggcaccagcc gggggcgagg ggcaccagtg cccaccctgg ttgctgtcgt tagctgaccc 86700 caccatgttt aagagccacc agcctgggta tttggttgtt cgtcctgttc aggccattgc 86760 ttgccacctc tgtaactgtg accaggctag agcaacatgc gtcaggctcc tcgactaatt 86820 agctgtatgt tacttaggca agttgtttaa cctttctgag ttttcgtgag ataattcaat 86880 ctcatctgta aaacaagggt aagagtccct actaccactt cctggctgtg tgaccttaag 86940 taagtgacat aacctctctg gacctcaatg tcctcatctg caaaatgaag ataataaaag 87000 tgcctatctc ccttcccgca gggttgtggt gataaggaaa tgggttaata tacgtaagta 87060 tttcaaatag ggtttggtat acaaaaagca ctgggtgatc tttctaaagg actggcaggg 87120 tcgaggcatc tctctctgtc cttcatgggc ttcctagtgt ctccatggtc aagtccaaac 87180 tcactggcct agaatccaat gctctgccca acctagcctc tactgagaag tgacttccaa 87240 gatctattgt taagtaaaca agggaagatg cagatgtgta tgtgttctct gcttcttttt 87300 atgttttttg tttgtttgtt tgttttttaa agagagatgg ggctgggcat ggtggctcat 87360 gcctgtaatc ccagcacttt gggaggccga agtgggtgga tcacttgagg tgaggagttc 87420 aagaccagcc tggccaacat ggtgaaactc tgtctctact aaaaatacaa aaattagctg 87480 ggtgtggtgg cgggcacctg taatcccagc tactcaggag gctgaggcag aagaatcatt 87540 tgaacctggg aggcagagat tgtggtaagc tgagatcatg ccactgcact ccagcctggg 87600 tgacagagtg acgacttcgt ctcggaaaaa aaaaaaaaaa aaagagagag agagagatgg 87660 cacagtgatc actgcagaca aacctaagat ggccccgatg atccccactt tgaatcacat 87720 ctttgtgtca tctggtcttc tcgagtgtgg gcaggacctg ctacttgttt ctaacccaca 87780 gaatatggta aaggtgatgg gatatataga ggatttcatg gatgtgatta tattacagaa 87840 gactgtaata tcctacttgc taggcaagtc tctctgcctt gatggctttg caaagggcac 87900 atgggaagga attcagggtt aactacaacc aacagccagc acgaaactga ggctctcagt 87960 tcaacagcct gccagggact gaatgtcacc aatattgtat gagcttagaa gtgacccttc 88020 cccaattgag cctcagatga aactgcagcc ccatcccaca ccttgattat agcctcacga 88080 gactctaaag cagagaacct ggctaagcta tgtccagact cccaaccaca gaaactatga 88140 gataataaat gggtgttgtt taaagctgtg aagctggtag taccattatt atgcagcaat 88200 agaaaactaa gacagcgatg actgaacaca cacatttgct tgtatagaca taagctctct 88260 ctaaaggatg cacatgccct ctgcctgccc taaccttgcc tccctttccc ttctggcaag 88320 agacagacag cagtggtcag tctctgggaa gaaaaacagg aggctggagt cgggagagaa 88380 acatgtttcc attctaagtc cttttgtacc atttgtattt ttattttgtg taactattac 88440 ttttaccctc caaaaaagct ttaaaaagtg atacatgaac gctatattca tgactgaccc 88500 tgatggcaag tgagcaacct gagttctcat cctgtcaccc ctctccctgc gagatccagg 88560 gctatgcacc tccctctcct cttcctcagc ccccggcagt ttcttcctgg ttactggcag 88620 ctccatttct gcttgtggtt ggaagcacat ttggggaaac ctggtgcaaa gtgagccaag 88680 gggaggtttt ttcacaagaa agaagcaaag ttccaaaaga ttgctggagc aactacagag 88740 agttcagaga ttttcctatt acagagggag ctgagggtgg ggtgtgagtt gtggggagac 88800 tggggaaggg agagaaaatt ctcactctct tttgtctttc tttctaaaca tgagcaaaca 88860 agaagcaaca gctagatgta agttaccgaa gaaatcttac accctggaag ggagctggga 88920 actgggaaat gttttactaa gaaacaaaag agaaatctga gcttccaaag aactacttgc 88980 ggtttgtctg ttttttaaca agaaggacag gtgtttggga gaccatctct gagagaaatg 89040 tccatgggag gaaaaagagc acaggcttgg gagttcaaat actgggtgca aatctcagat 89100 ctgggctgga tgcggtggct caagcctgta atcccagcac cttgggaggc tgagccaggg 89160 ggatcacttg aagccaggag atccagacca gcccggacaa catggcgaga ccctgtcgct 89220 actaaaaata caaaaaatta gcctggcgtg gtggcctgtg cctgtagtcc cagctacctg 89280 ggaggcagag gtgggagaat cacctgatcc caggaagtca aggatgcagt gagccatgat 89340 cacgcccctg cattccagcc tgggtgatag agtgagactc tgtctcaaca acaacaacaa 89400 caacaaaaac tcagccctgc catttccagc tgtgtgaccc tgagcaatgt gcctggcaca 89460 tagtaggcat ataaaatatg gaaaccatta tgaggcaggc attcagggct ctaactaaac 89520 ttttggcttt gtttggtgct ttatcgctac acatattccc acatccccca accccataat 89580 cctcaaacat tccatgaatt ttcacaccca taaacctttt atcatgcagt tccctctgcc 89640 tggtctaccg ttcccttctt tccccatcta gaaaactcct atgcatcctt caatgcccaa 89700 gccaaaactc cccctccact atggaacctt gcctagaaat tatctagggg aatgaatgag 89760 tagctctagc cccaaaattc tatagtaagt tcctaatgtt tctttctgga atttatccca 89820 ctcagtatga gccacagttg gtgcctatag tgctgtggca gatgcaggtg gacacccgcc 89880 cagcttttat gccatcgggt ttaggtgccc actgcttagg acagaactcc actgcctgca 89940 actccagggc caaccctcat tagtctaagc caattgtgcc aatcccatca cctggcctat 90000 cattcatcag aggggagcat gtggccaagc tctggccaat cactaaattc tttatgttca 90060 gctctgttct tgggaaggga gagatggggt atctcagttg ccactgttcc acccacaaga 90120 gaagctggac ttcattcttc tgcctcagac ctggtatctg gaggcctgtg gcatccctga 90180 gcaccaacat acccctaagc catagcccag gcatgctgac caatccaatc ccatgactgg 90240 catctggtgc cacgtcttaa acctggctgg cattttagcc agcacagtcc tgcttcctct 90300 tccccagttt cccctgggga agccactcct gccttatttt cagttcatct ggtttggatg 90360 gagctgagcc cacctgcagc cctagaggca ggtatgtgac tcctccctgg acaagcaggg 90420 ccctccacca gcaccctctc tccacccaca gccagtgatt ggttcagagg ctggcacagc 90480 atcgaagcca ggccactgag cagcagcctt aggactctta ttagaaccgt caagaaaaaa 90540 aacccttttc ccactgtggc tttgggatgt aggcctggag caactagtag gcatatctgc 90600 cactacatga gggaagagcc tgcctaaaaa cgaagccaat acagaataaa gcagggcgca 90660 gagctggaca ggttcctaga agctggatcc agccatgcct gaaggcagat ctagaaatat 90720 gaaaatatta aataacatat ctttggactc tgaaggtggt acctttctct tccccccagc 90780 ccatccttgg gactctctgt aggtcttgca caggcctggg gcatgagaag gagatttatt 90840 gaccaaacaa aacttgcccc actctctagc atctgtgcac tccttccccc catgcccccc 90900 gctttctgag cctctgctgt gtgagccaac tgccccattc ctgctctttc cccatacctg 90960 gcccagtgcc aaaggggaaa ccagacctga gagatataaa tcgacacccc tggcttggcc 91020 agattgagag gaagcagtga cagacaccag catgggagct taaggtggcg ggtgcctgct 91080 ccctgagagc caaccaggcc cagcctgggc ccctgggatg cacagagccg gccctcacaa 91140 acctcaagga acaaatgaca gaccagtgtg tgggacagcg ggcagatggg gatctctgtg 91200 gcctggggca ctcaagagaa agggcacatg gcttttagga cacatgtggg ccaaacaggg 91260 gagctcatct gtatcacaca ccttgtactg tcatagccct gtgggcgggt gctcttgtca 91320 ctcctgtgtt acagatgagg aaactgaggc acaaagaggt tatgggatat gcccaacatc 91380 acacagctgg cgagtggcac agcaggcatt ggattgcagc cctgtgggac cccaggtccc 91440 atatgtccag agattgtccc tccaatggga atgtgaggaa ggtggattca gggtggggaa 91500 acatgggtgg caggaaatga cctccctgca caccagcaga aaaatagggg gactcagcca 91560 cccactgtgg acggcccctg ccattactga cctgatagtt tcctgtaaat cctctcgccc 91620 tttttaaaga tgtcaataca tgtacttaag aaggaactcc actggccagg tgcggtggct 91680 cacacctgta atcccagcac tttgggaggc caaggcgagt ggatcatctg aggtcaggag 91740 tttgagacca gcctggccaa cgcagtgaaa ccccatctct actaaatata caaaaattag 91800 ctgggcacgg tggcaggtgc ctgtagtccc ggctgctcag gaggctgagg caggagaatc 91860 gctagaaccc aggaggcgga ggttgcagtg agccaagatc acgccactgt gttccaccct 91920 gggtgacaga gtgagactct gtctcaaaac aaacaacaac aaaaactcca ccaacactac 91980 tgtgaaggga atgtcaggac cagcttccat ggggaacaac caacaaactg atcttgtttt 92040 cttgggaaag tctaagcctg gggtgtggcc tctgctctct tcattaagag gaattagtaa 92100 acgagtcagc tgtatgggga cttcctcctt aacccgttgg cctcaaaagg gaactgacac 92160 gggtcactgt ctcttgagtc agtattattc tcttcaataa tagggttatt caagtgagct 92220 gtcaatgtgc cacctacgat cacttgtgtc ccccaaactg ggaaacacag atgtggccca 92280 cagccctacc tttacagagg aggagcggca tgcagagata actccccttc tctgatgtga 92340 ggggtggggg gccaggaagc aaagggactt gcctagggca gagccaggtg agaagccagg 92400 tctctagctc ctgggatttg ctatccagcc ccagggcttg cctcccaggc tcctggctcc 92460 agggactggg cctctgctgg tcacccaccc tgttccccag aggtcacaga aaaacgtcaa 92520 agcatgccca ctcacatctg ccgggtcgtt ttcactccaa atgttgactg cataggtgag 92580 atgattatac aggtaattgt cagggggata cgggttgctc caggtcagca gcagagtgtc 92640 ggagacattg gtgtgaactg tcaggtttcc tggggccctg ggtttcactg cgggagaagc 92700 ggcctctatg tgaggatcta tgcagtgact cccccatcac ccagcccagc cccggcctac 92760 aaaccacagc caccacccag atttttggtc gacaaaacct ccatagcata cttctggagc 92820 accacacaga ttcagagcca tagcaaagaa gattataata acaataatgt tgctaaaatt 92880 ctacatagaa ctttagtgtt tattaattac cctttttttc ttcttttttg agacagggtc 92940 ccactctgtt gcccaggctg gagtgcagtg gcacaatccc agctcactgc agcctcaata 93000 gcccagggct caagtgatct gcctgcgctg ggctcccaaa gtgctgggat tccaggcatg 93060 agccaccgcg cctggtcttt actaaccacc tttctacagt gaaggcaaag tgtatgagcc 93120 aaagtaaccc aaacttcagc cttttaaaaa atctttttat ttttatttat ttttattttt 93180 ttgagacgta atctcactct gtcgcccagg ctggagggca gtagcatgat ctaagctcac 93240 tgcaacctcc ttctcctggg ttcaagccat tctcctacct cagcctccca agtagctggg 93300 attacaggcg cctgccacca tgccagacta attttttgta tttttagtag agacggggtt 93360 ttaccatgtt gtccaggctg gtctcgaact cctgacctcc ggtgatccgc tgggctcggc 93420 ctcccaaagt gctgggatta taggcatgaa ccaccacacc cggcctcaaa cttcagcttt 93480 tacagctgca tccttcacac ctttggccac atctctgtac tttatgatga tttactgagt 93540 atttttcttt attttgattc atctgtttta aaacttggtc tcctttgaag caatcatatc 93600 catgagacct caggttttta ttcctgtgat gcatgtcata tcatgcatca ctggaataca 93660 acgcatgttt taaactcctg tcaccacagc ccagcaagct caagggggat tcctgcaggg 93720 actccgggta ccagaagaag gtcagccttc cacggtcaag accatccaaa gccgcagcag 93780 ctgaccagtg gacacagaat ggaaggaatg ttcccagttc ggttttcaat cagcagcagg 93840 catgccttcc agcaagcaca cctcatctca atacgccttc ctgggcactt aagttcaggg 93900 gtggaaacag ctgggctgaa gggggcttgg tggtctactg cgaccagctc cgtttcaaag 93960 gtgggaggac tgaggcccag gcaggggcag gaatcccatc agatttggga aaatacaggc 94020 ggcttcctcc tgctgttgct atgaccccac ctcctcccct gcccacccca ccctcagccc 94080 aggcagctgt gggaacacac ccagccaccc ctgccgcact ccgccctgct caccatgctc 94140 gctgggcttg aaggagccct tccacagcag ctgctgccca gcccacaggt ccagtgtata 94200 gttatccgca ctgaccacgt catccatgag caggtggcac acgcaccccg cgcctccgtt 94260 gttctcaggg atacacgtgt gggctctgca gacacagggg ctgggttagg ctggtcacct 94320 gtaggctggc tgggccccac agctgtgtcc gggacatgcc tccaggaacc gcatcagact 94380 cttccagctt cccgatcacg gatgtgagga cagatctggg ggcttgggag cggccagggt 94440 gtgctggggc agccgactag ggttgcctct cttgctggcc acagcctctt accacatgca 94500 ccaaggatgc tcctcacacc tgcctagctg ccggccctgg tcctgggact tagcttgttc 94560 ctgaaagccc atgaaaggga agaaccggtg gcactaatta atttagaaca acataagctc 94620 tgcatcgcac agtttggcca tcgtaagata attccaatgt tccaccttaa cagccaaccc 94680 acatctcacg ggctaggcca ggggtctgca aactttttct gtaacaggcc agatagttaa 94740 tattttggac ttcaaaggcc acacaatttc cttttttttt tttttttttg gttatttgta 94800 taaatttagg gggtacgagt tttttgtttt ttgtttttgt ttttttttcc tgagacaaag 94860 tctcactgtc acccaggcta gagtacagtg atgtgatctc ggctcactgc aacctccacc 94920 tcctgggctc aagcaattct cccgcctcag cctccctagt agctgggatt acaggtgtgc 94980 gccactacac ccagttaatt gttgtatttt tagtagagat ggagcttcac catgttggcc 95040 aggccggtct cgaactcctg acctcaagtg atctgcccac ctcggcctcc caaagggctg 95100 ggattacagc cgcgagccac cacagtgccc ggccttacaa gtgcaatttt gatggcaagc 95160 ctcagatctg ttctcactgg atatgctgcc tagtggtgaa gtctggggtt ctggtgggtc 95220 cgtcacccaa atggtgtaca ctatgcccat taagtaactt ctcttccttc acccctctcc 95280 caccctccca agtcttcaat gtctgttatt ccatggtaca tggaataatg cccatgtgta 95340 cacattattt aactcttact tataagtgag aacatgcagt atttgacttt ccatttctga 95400 attatttcaa ttaagataaa ggaggaggcc gggctcagtg gctcatgcct gtaatcccag 95460 cactttggga cgctgaggcg ggtggatcac ctgagttcag gagttcgaga ccagcttggc 95520 caacatgatg aaaaccggtc tctactaaaa atacgaaaaa ttagccgggt gtggtggcag 95580 gcaccattaa tcccaactac tcgggaggct gagacaggag aatcgcttga acccgggagg 95640 cagaggttgc agtgagccga gattgtgcca ctgcactcca gcctgggcaa caagagcgaa 95700 actctgtctc aaagaaaaca aacaaacaaa caaacaaaca aaaaacataa aggagggcca 95760 tgcagtttct gttacaacta cagcagccag tgtgttccaa taaaacttta tttacaaaaa 95820 caagcagtgg gccggatttg gctcacgggc catagtttgc caatccctgg actaagccaa 95880 tggaaacaaa gcacagccca tctaccttag aataatcatg gaataccacc tttactggtg 95940 aacgctgcat gctggtgtgc cgtatatccc cagtctgccc cgtaactgtg tgtcttgtgt 96000 acgtgattat gtcctgggca ccagtgtaac gaagcatcct gtgcaccatg gagtagccca 96060 gggaaccacc tgactcctat gccatgtgtc ctggtccatc caccatcaca gaggactcca 96120 ctctcaacac cgtactcctc aatcaagggt ctagtccagt ggttctcaaa gtgtggtctg 96180 ggcccaccag gtaaaactct tttcatcata atacagggac gttatttgcc ttttcactct 96240 ctctcactgg cgtacagtgg aattttctag aggctacatg acctgtgatc atgtaattga 96300 tctgaagatt tgttgaatgc gggcttgcat gttgttggga cttaaaattt ctccattttg 96360 ggccggatac agcggctcac gcttgtaatc ccagcacttt gggaggcgga ggcaggagga 96420 tcactggagc ccaggagttt gagagcagcc tgggcaacac agtgagacct ccatttcagt 96480 aagaaaaaaa atttttaatt gttttaattt tttaaaaatt acaccatttc ttcattgcaa 96540 gttctaatta ggtaaacata gaatgatgtg ttacctacac caaagccctt tgggtcctca 96600 ctcattttta agtgtgaaaa agggtcctga gaccaaaaag tgtgagaact gctggtctag 96660 tccagtattc cagccgtatc catgtgccac gtggaccaac actcccacca tgcgctccac 96720 tgtatcttac ataacatgtc cctggagatg ggacctcccc tctaacacca tataaacagc 96780 agcaaaccat actcagcatc ctgcactcca gcgcaccacc tggacccagc tcactcttgg 96840 ggcaaaccct ttgggcacaa cctccccaaa cctccagctc cagcgcaggc ttactcggag 96900 agcagaaaaa ccagctggta caacaggcgg agctcggtgc tgcaattggt gggaccattc 96960 atcttccact cgcaagtaga gatgctcatg tagtcggaga cgcaggtggg ctcctgcaag 97020 accttcatgt tccctgggga gacacaggag cagcctgagc ggcccaggaa ctcaatgcgt 97080 gagggggact tgcacccaaa caggaccaag gggaatagca caaaagcaca gtgcaggcaa 97140 agttgagggc cagcgcccac agagctgggg ccaggaccag ggctgctgga gcttcccagc 97200 tagtgtcggg atgactgtgg aagctgaaga aatacccatg ggtgcggggg ccattagaca 97260 agaacgaacc aagtgcacct ttatctcaca caaacttgac aacttgttct tggccaaatt 97320 aaagggaaaa tatgtagcag tttaaggagt gccattccat cacaccaagc ctctgccctc 97380 gagctagggg aagtgccact tccccatttg cccactatct gcgcccctct ttgtctccac 97440 ccctttctgc cctctctgtg ctcaggaggc tgataccaca gtcttcatca ccgggtaccc 97500 ttggccttta gtttccggtt gggtttggcc aatgggagag aggagatgga tgggctggag 97560 gagacatcgg ggtccttctc ctttcagcgc cctcctagct tgtcagtggc catggcagga 97620 ctggaattgg caggatgtac actgagtttg ctaaaccacc tgagcatgag accactcagg 97680 cgcctccatg ctcccatgct cagtgccacg cccatctcag ctgacaacaa cttgcataag 97740 ggcttaccat gttaaggagc tgtcttagat agcaaataaa tattccaagc ttatctcatg 97800 tccaaaacta actcctgatt cccgcccctc cccaaagtac cttctcttct ggcatccttc 97860 tgcatttcag aaatgacaac cgcgtccttc ccagtagttt aggtcagaaa tcttggagtc 97920 atcctggcca cttctctttc tctgtctttc acctcaattc caatctgtcc tcaaatctac 97980 ctcccagagg gatccggagc ctctcatctc ctccactcca gacctccctc cagacctcct 98040 ggcttccacc ctcttccttt ctccacacgg catcctgaag agtcctttgg aacataatgg 98100 atgatatgga tcctctgctc aaacactgca aggtcttttt tgattcagcc actctgtcac 98160 cttgctgacc ttcactgctc cctcctgctg ctcactctcc tccaccacat tccctcctgt 98220 gctcaaccct cccgccaggc tcagtgcagg gcctctctgc caagaatgag ccttccccag 98280 acatctgcac ggttcccgct tcctccaagt cagctcaatg aggccttccc tgacttccct 98340 actttcaact gcaatccccc ccactccacc cagcttcccc aaccccctct tatgctgttc 98400 tgtgttcttt ccccatcacc ttctcttttt ttttgacagg gtctcacttt ctcacgcagg 98460 ctggagtgga gtggcgcgat cacagttcac tgcagccttg acctcctgag ttcaagggat 98520 cctcccacct cggcctcctg agtagctggg gctacaggcg tgcactgcca cacctggcta 98580 atattttatt atttgtagag acaaggtctc cctatgttgc ccaagctgac ctcgaactcc 98640 tgggctcaag cgatcctcct gcctcggcct cctgaagtgc tgggattaca ggtgtgagcc 98700 actgcacccg gctcccatca ctttctaaca atctttagaa tttactgata atcagggcca 98760 cttgtcattg gctgtcccct cccactgcag tgtaatctcc ataaggccag gtgacatact 98820 gcaggaccct ggcacgccat taatagccaa caaatgaatg attgagaagt ggtgacttac 98880 cagagcttgc cacctgcagc aggaccaggc agctcacagg gaacaggagc ccagagcaaa 98940 gccaccccat tgggagatgc caaggcacct gcagagagag aggaagcagg ttagtgctga 99000 cctgtgcttc cagcagagct catcttattc aatatgcaac cctcccctgc cctggccatt 99060 tatttgtttg tttgtttgct tgtttgtttg tttttccgag acagagtctc gctctgtctc 99120 caaggctgga gtgcagtggt gcaatctcag ctcactgcaa cctccacctc ccgggttcaa 99180 gcgattcttc tgcctcagcc tcccgagtag ctgggattac aggtgcccac caccacacct 99240 ggctaatttt tgtattttta gtagagacgg ggtttcacca tgttggccag gctggtcttg 99300 aacttctgac ctcatgatct gcccctctgg tcttctcgaa gtgctgggat tacaggcctg 99360 agccactgcg gctgcccagc catgccctgg ccatttgtgt ctctgccgac aacaggtcct 99420 ccacgttcac agagctctct ctataatcat gaaacaacac ccagacccac aaagtccaga 99480 gggtgagcta cacaacaact gtcctgggag atgcttctgg atgtggcctt ccttggacaa 99540 ccctctacac atgtttctct gattatttcc tgaggtttaa atacctgctc tgtcactgac 99600 tagatgtgaa ccttgagcaa gctgtgcttc catgcctcag tttccttatc tataaagtgg 99660 gactaagaat agccccatct gattgaggtg ttgtggggat taaatgagct aacacagaca 99720 gagccctcca acagtgaccg gaacacttta aagtctcaag taacgttaca agtcagctta 99780 gtccatatag gctgctataa caaaaatacc ttagagcagg taatttacaa agaacagaaa 99840 cgtattgctc tattgggtgc aatggtgtgc atctctagtc ccagctactc gggaaggtgg 99900 ggtggatcac ttggagtcca ggggtttgag accagcctag ggaatatagt gagacctcat 99960 ggtgaatttt tgaaaaattg ttttaaattt tattattcag gccagacaca gtagctcaca 100020 cctgtatttc cagcactttg gggagccaag gtgggtggat cacctgaggt caggagttcg 100080 agatcagcct ggccaacatg gagaaacccc gtctgtacta aaaatacaaa aactagccga 100140 gtgtggtggc gggcacctgt aattccagct actctggagg ctgaggcagg agaatcgctt 100200 gaactggggt aggcagaggt tgcagtgagt cgagatcgcg tcattgtact ccagcctggg 100260 cgacaagagc aagactccat ctcaaaataa ataaataata aataaataaa ttttctaatg 100320 catagttctg gagactggga agtccaagat cacgacacta ggagatccag atctggtgag 100380 agcccattcc tcacggacga tgccatgctg ctgtgtcttc atatgacaga aggggcaaac 100440 acacttcctt cagcccgttc atgagggcac taaccccatt catgaccaac cacagagccc 100500 tcatgactta atcacttctc caaaggcccc atctcttaat atgactacaa tgggatgaag 100560 tttcaacata tgaatttggg ggaacacatt cagcccatag caccagctat tatcattaag 100620 aaaaaaatgt tcagaagcca ggtgtggtgg ctcacacctg taatcccagc actttgggag 100680 gctaaggagg gaggatggct tgagcccagg agttcaagac cagcctgggc aacagagtga 100740 gaccttatct ctacaaaaat taaacaaaat cagccaggca cggtggtgca tgcctgtaat 100800 cctagctact tgggaggttg aggtgggaag atcgtttgag cctgggatgt caaggctgca 100860 gtaagccaag actgtaccac tgcactccag caagaccctg tcataaaaaa aaaagaaaag 100920 aaaagaaaac agaaagtttg ggaggctgag gtgggcggat cacgaggtca ggagtttgag 100980 accagcctgg ccaacatagt gaaaccctgt ctctactaaa aatacaaaaa ttagccaagt 101040 gtggtggcgc acacctgtag tcccagctac acgggaggcc gaggtgggag aattgcttga 101100 acccaggagg cggaggttgc agcgagccga gaccatgcca ttgcactcca gcctgggtga 101160 cagagtgaga ctctgtctca aaaaaaaaaa aaaaggaaaa gaaaaaaatg ctaagaagca 101220 gaatctcacg gagtcaagtg tgtacttttc cccagcactc ctggtacata acgtcaaata 101280 ccctccaaaa agtgtgtact ggggccgggc gtggtggctc acatctgtaa tcccagcact 101340 ttaggagttc acaaccagcc tggccaacat ggtgaaacct gtctctacca aaaatacaaa 101400 aaaaaattag ccgggcggta gtggcgcgtg cctgtagtcc

cagctactca ggaggctgag 101460 gcaggaaaat cgcttgagcc taggaggtgg aggttgtggt gagccgagat ggcaccactg 101520 cactctaggc tgggcgacac agtgagaccc tgttgcaaaa agagaaaaaa aaaaaaaaga 101580 aaaagaaaaa aagaaaacag cacgcgctgg gttatattcc ctgcaacatt tcaagtgccc 101640 atccttcact tcccgcttgg gtctctctcc tgggatctta ttcaaaaaag cagctcccag 101700 gctttgttct gctagtgagt agttctttgc cattaacagc atttctcttt tggtgctctg 101760 gctgtgtttc tgtggcacag gaatggttcc aaggctaacc tcacaggctt gcagcaaaca 101820 ggaacggagg tgttgcctct gaagtactgt gtgcagtgtc tggggcatca taagaactta 101880 aggccacctc aggagctcta ggcccctgct ggatgctctc cgagtgactt gggcacactt 101940 gaaggccctc cccccagcag ggctccacct ccctctccac ctgggcccag cacccatgca 102000 tcctacgcct gccacctaca cttcttcttt tttgtttttt aacattttat tttattttat 102060 ttcattttat ttttttgaga cagagtttca ctcttgttgc ccaggctgga gtgcaatggc 102120 gcgatctcgg ctcactgcaa cctccgcctc ccaggttcaa gcgattctcc tgcctcagcc 102180 ttctgagtag ctgggattac aggcatgggc cacacaccca actaattttg tatttttagt 102240 agagacaggg tttctccatg ctggtcaggc tggtcttgaa ctcccgaccc caggtgatcc 102300 gcctgcctca gcctcccaaa gtgctgggat tacaggcgtg agccactgag cccggctgcc 102360 acctgcactt cttaaacaca ccaaactcag tcccacctcc atgccttttc tctctctgtc 102420 ccctccacct ggagggacct tcccttcctc cctttccttt tttttttttt tttttttgag 102480 acagagttta ctctgttacc caagctagag tgtagtggca caacctcgac tccctgcaac 102540 ctccacctcc cgggctcaag caatcttccc acctcagcct cctgagtagc tgggaccaca 102600 gatgtgcacc atgaggcctg gctaagtgtt tgtatttttg atagagatgg ggtctcatta 102660 tgttgcccag gctggtctcg aactcctgag ttcaagcgat ctgcccacct tggtctccca 102720 aagtgctggg attacaggcg tgagccactg tacccggccc ttcctccctt acgcgtgctt 102780 ttcctctctt cccctaattt atgatggtgc aaaatcaatg ggtgttcagt agaagcagta 102840 cttggagcac ccatacgacc attctgtttt tcactctcag tattccataa actacacaag 102900 ctattcaaca ctttataaca aaacaggctt tgtgttagat gactttgccc aactgtaggc 102960 taatgcaagt gttccgagct aagttaagct ataacgttca gtaggttagg agtatgaagt 103020 acatttctga cttaataata ttttcaactt atgatgagtt tatggggaca aaaccccatc 103080 attaagaaac atctgtgtat gagacacata aacacactca catatctcca taatgaataa 103140 attaccccaa tacctcagtg tgcatgtcag aaaaacaagg acattctctg cataatcaca 103200 atacaaccat cgaaatcagg aaattaacag tgatgcatca ccactatctc atccacggcc 103260 cctatttatg ttttgctaat tgcctcaata atgcccttta tacgtccagg atgcaatcca 103320 agatcattca tcacatttga ttatcatgcc ttcctagtga ccaggtcttg gcttctaaat 103380 actattctgt actgaaagga accagagcgc cttagagaaa tagctgattc caggagaagg 103440 aaaagtacca gataggcttg gaattcttgt gccagacaat aaggaagtac tcagcagcta 103500 gcctgacctg gacaaaacag gcacattttg agcatcaaaa tgaatattat ggtatccact 103560 tgtaaccctt ggagttaaag tccaggagtc tgtattgaca tagatcaata aatggaggag 103620 gccgggtgca gtgtggtgtg tgcctgtaat cccaacttct tgggaggctg aggcaggagg 103680 atggcttgag cctgggagtt ggaggctgca gtgagctatg atcgcaccac cgcactccag 103740 cctgggcaac agagtcagac ttcatctcta ataaataaat aaacagggaa gaagggaaag 103800 ctctatttta ctatgacagc ctcaaagtaa ctccctgcaa aacttgtatt gtatattaac 103860 taatgagtga aaatacatat caatgaacgt tacaatacag aaacctggca gacaccacct 103920 tcaccaagtg atctaaactc accaacaaga atgggacaaa tccccaacat cacctccctg 103980 aaacaattcc tgagaagagc acagcatgtc ttttcctggg ttcctgccaa ggatgcatac 104040 atcatttgga aacgtcagac aaatgtacac caagggccag tctatataca agaactggcc 104100 gctgctcctc acgaagtgcc gggggtgcaa aggacaagcc agcgtgggac atctgtccct 104160 ttttaaattg tctgtcatcc tgacagccag ggtctgtccc attcactgcc gagtccccag 104220 tatctacaac ggtgcctggc acataggacg tgttcagtaa ctgtttgttg attaaattca 104280 ttttggacat gatgtggggc ccctaactgg ctcagcatga ggctgcaaag ctctctctag 104340 gggaaggctc tgtatcggct caaaggttga actcgtcttc cttgctgaga gttacaaaag 104400 taaagtctca agtaaaggag gggcgggaat ggggtggtgg gttcctgaca ctgatttcct 104460 tggcaagtcc tagtaacttt tactctcaca actacacaga tgcccacagt atcctgtggc 104520 agagatggca aaggtgtcac tttagcctca attatgaata aagagaaaca gagaggccaa 104580 gacgcctgtc caaagtcaca cagcagaaga ctttcatggt aaagtcctcc agggaaacag 104640 gggagtcaac aaccctgtca agattcacta aaacatccca gcatggcgtc aggattgata 104700 tccttaatct gttgtggaag gcacgcccag acttacgcat gcccatgtgc acacacacgt 104760 gccgcagcta gcttctcagg gatgtgtggg tgctacattt caggagagat cacagatgca 104820 gtgggagaag aggcttccct gaacatcaga ggcaaaaaga acaaagatgc aggtgagtaa 104880 aagaattaaa aagataagtg ccaggttatc gcgttcaagt acacggcaag aggagatgat 104940 ttcaagttcc cattctaaat caagagactc ctggctgggc gcggtggctc acgcctgtaa 105000 tcccagcact tcgggaggct gaggcaggca gatcatgagg tcaggagatc gagaccatcc 105060 tggctaacat ggtgaaactc agtctctact aaaaatacaa aaaattagct gggcgtggtg 105120 gcgggtgcct gtagtcccag ctactcagga ggctgaggca ggagaatggc ttgaacccgg 105180 gaggcggagc ttgcagtgag ccaagattga gccactgcac tccagcctgg gtgacagagc 105240 aagactgtct cagaaaaaaa aaaaagagat tcctgaagtg gtggagttgg aaggggtctc 105300 agcggtcacc tgccaagctc aactgccaac ctggacaagg atgtcctgcg aacaccccac 105360 ttctgacatg ctgctaggca gtttcagccc atatgccacc acagatggag caatcactct 105420 caggtctcag ttaagcagag ccattggaaa cagcctcctt atgttgagtc aaggctttgt 105480 gtgaacaaaa tccctgcttg cccagcactt accttgtact tgacactttg ctaggtgctt 105540 tagaagtatt acctcaccga atccacacaa gaatcctgaa aggtaggctg ttatcccaac 105600 tgtctagcta aggaaactga ggccagagag gccaaataac ccatccacca ggtcccacag 105660 ctggtaaaag gtcagagcta gaatcagaac ccacgtttgt ttgcttccga gcccaccgtg 105720 gattccagta taccagttcc ttcaatctca tctcctaaaa ctgcaggctg aacccaataa 105780 gatgaaataa aatgagccgg gcagagttgg ccatcctgct tgtcaatttc aaacaacagt 105840 tccgcaagtg taggataaga aaatctgact gagcagcaga aaagtcctga ggattttatc 105900 atctgcaaaa tctccaaaag ccaatggtga agcagcaact taaaaaaaaa aaaagagtca 105960 cccgaggtca tattaacaga ggtatccatt tatagactgt tacatctcac aacagcagaa 106020 tatacattct cctcaggttc atgtggaata atcaccaaga cagaccacat tctgggacat 106080 aaaacaacct taagagattg aaaacattag aaatcacaca acatatgctc ttaggccaca 106140 atggattcaa attagaagtc actagcagga agaaagccag aaaatttcca agtatttgga 106200 gatttaaaaa cacacttccg aataacatat ggatcaaaga agaaatcaca agagaaacta 106260 aaaatatttt taactaaata aaaatgaaaa tatagcctat caaaatcggt gagacgtagc 106320 aaaagcagtg cttagaggga aatttatatt attgaatgta tatcttagaa agaaagttct 106380 aaaatcaata atctgtttcc atctagagta ctagaaacag aaaagcaata tatgtctaaa 106440 acaagcagaa gaaaaaaata caaaattgga gcatacatca atgaaattaa aaatagaaaa 106500 ttaatcgagg aaaatcaatg aaatcaaaaa ttggttcttc gaaaagatca atgaaattga 106560 taaatctcta cctaggcgaa tcaaaaaaat gagagagaag gcactaatta ataataatgt 106620 gatatggcct ggctctgtgt ccccacccaa atctcacctt gaattgtaat atgaattgta 106680 atccccacgt gttgggggaa ggacctcatg ggaggtgatt agattacggg agcagttccc 106740 ccatgctgtt ttcgtgacag tgagtgagtt ctcctgagat ctgatggttt tataaagggc 106800 tttccccccc tttccttggc gtgtctcctt cctgacgtca tgtgaaaaag gatgtctttg 106860 cttccctttc cgccatgata atatgtttcc tgaagcctcc ccagccctgc agaactgtga 106920 gtcaaacctc tttcctttat aaattaccca gtcttgggca tgtccttaga gcagcatgag 106980 aacaaactaa tactattgta aattaaagaa aagttattag aggtggcaca tgcctgtagc 107040 cccagctact gggaaggctg aggcaggagg atcacttgag cccaggaggt taaggctgca 107100 gtgggctgtg ttcatgccac tgaactccag cctgggcaac agagaaagat cctgtctcaa 107160 aaaataaaaa ataaaatttt taaattaaaa aaaagtggca ctatataatg attcgggagt 107220 taattctcca agaagatgta aataacaatc cttaatgtgt aggcaactaa caacaaaggg 107280 tcaaaatatg taaggcaaaa actgagagca gcaaggagaa atagagaaat ctgtgtgatt 107340 attctgaata atactattat tggataataa tagtattatt actattactt caactctttt 107400 ggtaatcgac agatccagca ggcagaaatc agtaaggaca cagttgactt gaacagtgct 107460 gtcaatcaac tggatctaag tgcagtgtgc aaccacaact tggcatccct ttggaaggaa 107520 tactgctgtc atgtatgaca acaaccaaca cagagagggc ctgataatcc caggaggcct 107580 agaaagtctt tacctggaaa gctctgccct cctatactgg gcagcctcac cattcactta 107640 aacacacaga gtttaccaca tgccaagttt tcttttacat cttttacata caagaatgca 107700 tttaataatc tatgaaatag gtaatattat tttctccatt ttcagatgag gaaaccaagg 107760 cacagaaagg ctgagtgatt tgtctgaggt tgcacagctg tgacagtcat gggtcttaaa 107820 tccagcactc aagctccaaa gctttggttc ttaactacta tctgtctaga agtcagagac 107880 cttagtgttt tggggatcaa tgacctatct gagcatctaa tgtaaacccc aaaccctgtt 107940 tctggcaagg tgcatacact taattctgca ttccatttta ggggttcaca gaatccctga 108000 agcccaagtg aggacctcag gtcaaaggcc tgtcctctaa gcagtgggca gcttttgagg 108060 gtctttgagc aggggagtgg caaatttttg agaagtctgg ggggatgtgg gcagttagca 108120 gttgctgcag atgaggctca ccctgcctct tctccacatc acagccagag ccattttaaa 108180 atcagaaatg agaccagggc cagctcacac atccaatgcc ccctggctcc taaagggctt 108240 cttcacaggg gccacagatc cccagggggt tcacagctca ttagagaagc atctctgact 108300 tagaataaaa tccaagccct agagtgaaaa aagccagttc ccagagtaca caccccgtga 108360 ctccatttat gtaacatact tgaaatcaca taaccatgga aatggaaaac acatcagtgg 108420 gtgccagggg ctgagggggt gggtcaggaa gggagttgga tgtggctata aaaggagaat 108480 ttgagccatc cttgtgatga tagaactgtg ctggatcctg gctgcataca caaacctatg 108540 ccgtgataaa attgcatagc actaaaccac acatgcgtac gcaaacacaa aaatgagtac 108600 aagtaaatcg ggaaatccca ataaggtcag tagcttatat cagtgtcagt atcccagctg 108660 tgatactgta ttacagattt ctacaatctt gccctcgggg ggaacagcat agaagcaggg 108720 gtctctctgt gatttctttt tctttttctt ttcttttttt tttttttgag atggagtctc 108780 actctgttgc ccaggctgga gtgcagtggt gtgatctcgg ctcactgcaa cctctacctc 108840 ctaggctcaa gtgattctcc tgcctcagcc tccccagtag ctgggactac aggcaagcca 108900 ccatgcccaa ctaatttttg tatttttagt agagatgggg tttcatcata ctgaccaggc 108960 tggtctcgaa ctcctgatct caagtgatct gcccgcctca gcctccaaaa gtgctgggat 109020 tataggcatg agccaccacg cccggcttct ctatgatttc ttccagctgt gtgtaaatct 109080 ttaattatct gaaaataaaa agtttaattt aagaaaatga agaaaagagc caattcccct 109140 gcttatccct ctaacattcc cctcctatcc cctctccctc ttctgcaatc ctccattggc 109200 ctcagccctg ggatacactc ccagctctcc cttttccaag catttgcacc agaaacactt 109260 cagaagcctt ctctgacccc cctcaactaa tgcgggccct ccctgtgact tcttatcaca 109320 gcaccctgtt tatgtctgtc ccagtactta tcactatctc taatcagatt aatgaggtta 109380 catgtatgaa tgtttatttt atttcaagat gcagatagca aagatagctt tgactgcaaa 109440 atgttttcaa cttccctaga tttttttttt tttttttgag acggaggttt gctctgttcc 109500 ccaagctgga gagcagtggc gtgatgtcgg ctcactgcaa cctccacctc ccgggttcaa 109560 gcgattcttg tgtctcagcc tctcgagtag ctggaattac aggtgtgtgc caccacaccc 109620 aactaatttt ttgtgttttt agtagagaca gggtttcccc atgttggcca ggctggtctt 109680 aaactcctga cctcaagtga tccacccgcc ttggcctcca aagtgctggg attacaggca 109740 tgagccactg tgtctggcct agattaaaat atgccataag aaaaagacag aactcaagtg 109800 tgttagtcca tttttgcatt gctataaaga aatatctgag actgagtaat ttacaaagaa 109860 aagaggttta attggcttat ggctctgcag gctgtacagg aagcatagca cccgtatcta 109920 cttggcttct ggggaggcct cagggagctt ttattcacgg ggaaggcaag aggggagcag 109980 gcgtacatgg tgacagccag ggcaagagag aggagggagg tgccgcacac ttctaaacaa 110040 ccagatcttg tgagaactca ctcactatgg caaggacaga accaaaccat gagggacccg 110100 tccccatgac ccaacatctc ccaccaggac ccacctccaa cactggggat cacatttcaa 110160 caagagattt agagaggatg acatcccaac tatatcagca agcaaaagac tagaaaaagg 110220 tagcaatcta cattatgaac ggttaatggc cttaatatat gaagagttct tacaaatcaa 110280 taagacaaac atccctttaa gaaaaatagg caaagaacat aaatggttca caaaagacat 110340 attcctccat cagatgtgaa atagccacct caatatactg ctagtttttc aagttgggag 110400 tataataggt aacctcaaat ttaaattatg tatttgtcta taaatgcatt gtttgcctcc 110460 ctctgagaga ccacaagccc catgagggca gagctttatg tctgctgtgc tcactgcttt 110520 atccccagtg cctgaaacag tgcctgacac aggcatagca tgtgtacaat aaacatatat 110580 taaatgaatg aatggtttaa aagtcagcaa taataggctg ggcacagtgg ctcacgcctg 110640 taatcccagc actttgggag gccgaggcag gtggatcacc tgaggtcagg agttcaccaa 110700 catggtgaaa ccacatctct acaaaaatac aaaaattagc cgggcatgat ggtgggtgcc 110760 tgtattccca gctactcggg aggctgagag gctgaggcag gagaatcgct tgaacccaag 110820 aggcagaggc tgcaatgagt tgagatcaca ccattgcact ccagcccggg caatagagca 110880 aagttctgtc tcaaaaaaaa aaaaattcag caataataaa ttcaacaatc tttttttttt 110940 ttttttttga gatggagcct agctttgtca ccagtctgga gtgcagtggc acgatatcag 111000 ctcactgcaa cctccgcgtg ggttcaagtg attctcctgc ctcagcctcc tgagtagctg 111060 ggactacagg agcacgccat cacacctggc taatttttgt atttttagta gaaacgggat 111120 ttcaccatgt tggccaggat ggtctcgatc tcttgacctc ctgatccgcc tgcctcagcc 111180 tcccatagtg ctgggattac aggcatgagc taccgcacct ggcctcaata atcattttta 111240 aacatgaacc aggtcccaag cttttgtcca tagttaactg tatatgcagt acttattttg 111300 tgcctggccc tgttctaagt acttacaata tattaatcaa tttggttctc ccaacgaccc 111360 tatgaggtag gtgccattaa cattcccatt ttactgatga gaagactgag gcacaaaaag 111420 gctagagcac ttgctggtgg tcacagaacc ataacctagg cattgtcact ccaaaatcca 111480 cgtgtttaag gactgtggtt gctgaccctt gaaatgcagt ccccaaaacc ctggaacttt 111540 gtccggggag ccccactggc ctgagaccca ttggcctgag acccagagac agcaagtgac 111600 aaggccaagg tcacagagtt aggaagcagc aaagtcagtg aaggccgcag ccaggaaatg 111660 agcagaagca tgggcctggg aaaacacgcc cgccaccagg agaacacact tgactcactt 111720 tttttaaaaa actcggggga gattaagtga ttaagccttg aaatgtcaga agccagcagc 111780 agcctggaag cgacccagca ggactcttcc cgaacccaca gcgggtggca ggggggcagg 111840 gggcggagga ctgggggcgc acagggggcg gtacaggcct tctgctcttc ttccctctgg 111900 gcaggaaacc tggcttggag gaggggctgg ttctgcagac aggatgcaag cacctctgtg 111960 acaggtggca tgtggagagg aatccaggga cagcaggggc ctcgggtggg ggctggggct 112020 ttggagccgc tgacccctga aactccacga cagaaactgg cctgaaacct cagctcctgt 112080 ggcgcaatca gttagtcatg ctacttataa gaaaccagcc cagagcttcc tctggcttct 112140 gggacagctg ggctgggctg gcaggaagct aggcatgcac ttcttcctgg gacagtaacc 112200 tacctagaac tggggtttcc agaaatggca gccacgtcgg tggccccact gggatagtaa 112260 ggcaggccag gccagggccc cagaatggcc cgggatctgc ctggttggct ccgtccccag 112320 cccagccagt gggagggagg ggaggcaaca gactggctct gcagccacca ggaacacaaa 112380 tcggccccct ccaccaacag cacccaccag cctccctgtg ggcctgctgt ggctcaacca 112440 tcctcaccca cttgctcaag tacctcactt attctccttc tcttttttgc tttagagaca 112500 gggtctcatt ctgtggctag aatgcagtcg caccgtcacg gctcagtgca gccttgaact 112560 cctgggctca agtgatcctc ctgccttagc ctcccaagta gttgtgacta caggcacaca 112620 ccaccatgct cgaataattt ttaaattttt gtagagatgg ggatctctct ctgtggccca 112680 agctggagtg ctatggcaca atcatagctc actgcaacct caaactcctg gcctcaagca 112740 aacctcctgc ctcggccttc caacatgcta caggcatgag ctgccgtgtc cagccatctc 112800 ctcatttatt cttgctgtgg gtgttttcca agtactgcct atgtgctggc tattacagat 112860 gcaaacaggc cattcattca ttcattcctt tcttctttta acaaatattt gagcaaggaa 112920 gaagttgcca gagcgttttg agcaaagaag tgacatgatg agactttaaa aaagacctct 112980 cttgcttggg tggtgggggc acgggtagaa gtgggtagca gtgaggaggc taccacactg 113040 gccccagaga gaggtgccag ggctcacacc aggctgcgga gatggggagg ttgagaaggg 113100 gctgaattct agatctgtgt tgaagacaga gcccacagga tttgctgaca ggatggatgc 113160 tgtggggcgg gagagagaac aggaaggcta ataggagtca agaatgattc cacagccatt 113220 ggccttttcc aagatggcgg cctgtggggg tagcccgtgt gacaagccag cctgctgggc 113280 aagctgcagg acttcagatt agcagaggag gctgttagca ggcgctcagg gaagaccctc 113340 cctcatccta gagggtcagg tttctttatc ggggatgcac cagagttctc tcggggaggg 113400 gcccgacata cagcaggcac tcaataagtg agcatgtgta ttttatttta ttcttatttt 113460 aacaggtata cagtgtttcc ttggtgccag acactggcct aggtgcttta agccctccct 113520 atgagataag tataattatt aaacccattt tatgattctg aaggttcttt tttttttttt 113580 gagacagagt ctcactgtgt agtccaggct ggagtgcaat ggtgggatct cagttcactg 113640 caacctctgt ctcccaggtt caagtagttc tcctgcctca gtctcccgag tagctgggac 113700 tacaggtgcc cacaacaacg cccagctaat ttttttgtat ttttagcaga cacagggttt 113760 tgccatgttg gccaagctgg tctcaaactc ctgacctcag gtgatccacc cacctcagcc 113820 tcccaaagtg ctgggattac aggcgtgagc cactgcgtct ggcagattct gaaggttctt 113880 atgagagggc ggggtgaggg agggaatgag ggaagaccca gcgaataggc aggcaggagg 113940 gagggcacgg cccacttccc accctacaag cggggccctt ggctggagca agggggagaa 114000 ggactggctg ggatggcagc tggaaggttg gcaggccagg gacaacatcg tctgccaagc 114060 catggcagta gactcaaact ctgtctctag ggcagtgggg agccagtgaa ggctcttaag 114120 caggagagga gcgggctgag ctgagagctg gcgaagaggg ggcaggtttg agcctgtttg 114180 ggaggaggag gcagcaggga cttggtggcg cttggtgggg aaagcaggct ggggagagcg 114240 gggacagagg gaaagtggcc tcaggctggg gtggtgggga ggatcccagg gacccttccc 114300 acgatggaga gccgagaaga gaagcagatt tgctggggaa gaccacgcgc tccggcgcac 114360 tgtggacact ttgagtccac acagtcaaca ggcaactcca tccgggatcc tggaagccag 114420 gcctgtcttc agaagattcc agaggcccaa aggaggaaac agagcctgag gaaactgtgt 114480 ccagaagcca cctacccagg aacagcagcc agcctggcct ccatctttct gcaacctgtt 114540 atcttgcttt atttctctgc aatctttgtg gcatattgca cactcgggcc ttatgtgttt 114600 ggtgctcata tcccctactg caaggccagc tcacacaagg ccagtgtcga tcccgctcac 114660 agccatctcc ccagagccta gcacggtgcc tggcacacag caggtgctcc atgaatattt 114720 gctgaatgga agaacgacag gatgaattcc cctggctgtg agcagggagg cctggttggt 114780 catccccaat tttccacctc tccctgctgg gcaaacacta gcaaatcctg gcttggagtc 114840 cccttaaatg tcctaatgca ggcacagatc tcagcccacc tcggggccct gacctccaga 114900 gagctggtta tcgactatca ctcactgtcc tgagcagcgg acaatggcct cttggcgcca 114960 gggtgctgat tagattttga ctgtcttagt gacagctatg gctcctagct ggcctccttc 115020 ttgaggtctg cagaggtcac tgtgactggc cacgtgtccc ctcttccttc agccatgaga 115080 ggccatgccc tgggcccttc tgcccttggt ttcgcccctc cccaactggg tgcaactttg 115140 ctcaccagcc ttggcattgg ttgtaccagt ctcactaaca aaaaaaagca cttgaaggct 115200 gggcgcggtg gctcctacct gtaatcacag ccatttggga ggctgaggca ggtagatcac 115260 ttgagtccag gagttcaaga ccagcctggg taacatggca agaccccgtc tctaccaaaa 115320 atacaaaaaa aaattgccag gcatggtggc acatgcctgt agtcccagct actaagggag 115380 atgaggcagg aggatggctt gagcccacga ggtcgcagct gcagtgaact gtgatcatgc 115440 cactgcactc cagcctgggc aacagagtga gaccctatct ctaaaaaaaa aaaaagaagc 115500 ccttgagcct cactcactca cttaaccatg gacccaatcc aaggagcatt gcagatacaa 115560 ttaatatcat ctccattgca gagatggaga agccgaggcc cagagagggt caggaacctg 115620 cctaaggatg cacagctagt gagtgggctg ggatttgaac ccaggccttt ggctctggag 115680 tgcacagcac agtgcctggc acagaagcag tcataaactt gcttttttat acaccactct 115740 tttttgcttc tcctggaaag ctcactatgg aagatacagt aaaatattct ccctgcaagc 115800 acagatggag gaatgaggct gagggaggtt cagaatttgc ccgaggtgat ggtgacggag 115860 caggaagtaa ccaggtagga atttagaatc tggcctttgt caccatttgc tcggccctca 115920 gcagcagtga acttgtctca acaatgacca ggactttgcc ccctgggcaa acccaaagcc 115980 aggacaacac atgggcacag gtaagagctt cccgttcatt ttctgcttgc agactggcat 116040 atggaaggca gtgggtgggg actacccatg gagctcttcc tgttgtctca ccccatacca 116100 ttcctggatg ggtggggaca cacagacatg ggggccacct ttgtgggcag ggacccccaa 116160 gcatctgaga cacagaagca taactctggg cctaagtgca ggctccaacc tgactcggtg 116220 ggagaccgtg gcaggctgga gtccactcag agcctcagtc tcctcatctg tgaaatgaga 116280 ataagatcac ttgcttcatg gctggttgca aggctgctgt tgtttttaac tcatctggct 116340 tattagtaca ggagtactga catcaataaa atagaattca tgacaatgat gtcttgagta 116400 agtattagaa gagagaaggt ggatggtttt tttgtttttg tttttttgag aatgtctcac 116460 tctgtcacct agactggagt gcagtggagc aatcatagct

cactgcggcc ttgaattcct 116520 ggtctcaagt gattctccca cctcgccctc ccaagtagct ggaactacag gtgcacgcca 116580 ccacacccag ctaattttta gtttttggta gagatggggt ctcactatgc tgcccaggct 116640 ggaagttggc tcttttatac tgattaaagg aacaattcag ccagctgcgg tggctcatgc 116700 ctgtaatccc agcactttgg gaggctgagg tgggcagact gcttgaacct aggagtttga 116760 gactggcccg ggaaacatag cgaaatccca tctctacaaa aaacacaaac atcagctagg 116820 cgtggtagtg cacatctgta gtcccagcta ctcgggaaaa tgcaacaatt caccaaaaag 116880 gtataaaaat cctgaaccct tattctactc atatggctat gcacttaaca acatagcttc 116940 aaaaataaag caaaaaactg acagactcaa aaggagaact taacaaagcc tctattatgg 117000 tgaattgtaa ttgtattacg gtgggaggta aatcaggtaa aaaaaaaaaa aaccacatag 117060 tgacataaca aatatagata atataatagc tagcttgatc aaatagatac atttctagaa 117120 ccttataccc aacagaaaat atacattctt ttcaaacaca agaggaaaaa aatgaccaga 117180 aaatcctata aagatctcag caaattctct accactgata ttataggaac cacatatatt 117240 gaccacaaca accatgatga ccaaatttag aaatcaacaa taaaaagata acttaaaaaa 117300 ttctatctgt ctcaaaaatt taaaaagcat ataattaaat aagtcttaaa atgggaacca 117360 aaagggaaat tctaaactgg gcagtgaata acattgtgta acaagttgtg gaaagaaacc 117420 aaagcagtac tcaaagaaaa aatgatagca tgttagcctg aagtagattt attagagagc 117480 atgaaaaact ggaacctctt agctttagac aaggactctc agaatagaag aaacttggat 117540 taacagcagc tcatatagaa gagaaacatg gagattttag ctgacaagcc caatatgcac 117600 tccaagtggg atatggctgc tacacagtat gattccaggt tgcatcaaca gaagtataag 117660 ctctagatcc aaggaggtaa tagtctcctt cctctttgac caaacaaaca gtctttgcgt 117720 gggactccaa ccagttgact catgtgccaa ggcaggccaa acttataatt tcctgagtca 117780 gggagaatac ctgaaataac tacagttgtg tagtctgaat aagagaagta tccgagtttg 117840 gagggagatg gtcactacct ccaagaggca gaaacagact cagaaagtgg aagctcctag 117900 aaagctaatt tgggctgggc acggtggctc acacctgtaa taccagcact ttgggaggct 117960 gaggtgggcg gatcacccta ggtcaggagt ttgagaccag tctggccagc atggtgaaac 118020 cccattgcta ataaaaatac aaaaattagc caggcgtggt ggtgtgtgcc tgaaatccca 118080 gctactcggg aggttgaggc aggagaatca cttgaacccg ggaggcagag attgcagtga 118140 gccgagattg cgccactgca ctccagcctg ggcagcagag tgagactcca tctcaaaaaa 118200 aaaaaaaaga aagaaagaaa gctaatttgg ctaaatctca ggaagatttt ctcccagtta 118260 gagctgccca cctgtggaaa tgtggcttct ggaagaaata tgctagaggc aagaccgtcg 118320 ggaggctgtg gccctgccag cttggagacc tgcccagccc agagagctgg tgcctctcct 118380 ccctagaatt cagtcttccc ttttaaaaag gcagaatact tccctggccg cttcaggaaa 118440 acataggcct tctctgagtt tttcaaaaat ggttctgggc gactatgctc actgatttcc 118500 aaccaagaaa tgtaatcctt ttaaactctc ctctttgggc tggatagctc agaaccgtgg 118560 ttaaatgcac aggcttctgg gtccgtagcc ctgggtttga atctggactc tgccacttag 118620 gagccgtgtc acccaggtaa gtctctagcc tgagcctcag gttcctcctc agggaaatga 118680 gcaggacacc cactcatctc cattcctgca cccagcacag agcgtgatgt ggactccagc 118740 ctccactgcc agaggccgtg gttggggcca tggcttcggg atgattcaag cacattacat 118800 gtattacgta ctttatttct attattatta tatggtaaca tataatgaaa taattataca 118860 actcaccata atgcagaatc agtgggagcc ctgagcttat tttcctgcaa ttagatggtc 118920 ccatccaagg gcgatgggag acagtgacag atcatcaggc attagagtct cataagaagc 118980 acgcacccag ggccgcgtgc agtggctcat gcctgtaatc ccagcacttt gggagactga 119040 ggcaggtgga tcacctgagg ccaggagttc gacaccagcc tggccaacac agtgaaaccc 119100 tgtctctact aaaaatacaa aaattagctg ggtgtgatag cacaggcctg taatcccagc 119160 caccaggctg aggcaggaga atcacttgaa catggggagg tggagattgc agtgagccaa 119220 gatcgcacca ttgcactcca gtctgggcga caagagcgaa actccgtctt aaaaaaaaaa 119280 gaaaaagtac gcaacccaga tccctcacat gtgcagttca caaaagggtt tgtgctcctg 119340 tgagaatcat ctaatgcctc agctgacctg acaggtggcc gagcttgggt ggtaatgcga 119400 gcgatgggga gcagctgtaa atacagataa aacttcgctc actcacctgc cactcacctc 119460 ctgctgtgtg gccctgctgt gtggggaccc ctggtctgag acactgagga aggtgctcat 119520 tcataaaata atatgtaaaa tgattggact gtctgtaaaa tggggcttgg aggaaccagt 119580 cctgcgctgc ccacattcac agtatttctg ggcagatgga tggcagggag tacaccgaca 119640 atgtaaaaaa gtggatcgat ctaaatggca ggtgtgtctt cagcactcag gcatagattt 119700 tattgtgttt aaacaccatg gtgaggactg aggactctgg agcttctagg ttcacagccc 119760 tgctctgcca ccttcaggtg tgtgaccctg gacaagtcac tgaacctctc tgtgcctctg 119820 ttacttcatc tgtaacattt gcatgagagt agctcctatc tggtagggtt attacacaag 119880 ttaaatgaga caatgcaggt atctggcccc agctccctcc ctgacctctg ctcctagacc 119940 tctttcttgt ttggttccat ccaaccacac cagcttcctt gctgtgggtc aaacacacca 120000 agctcattca tgcctctggg cctttgcact ctctgttccc actgccagga tcactctttc 120060 cccagatatc cacatgcctt ggtatttcag gtctcagttc aaaagccact tcctcaaagt 120120 caccaagccc agaatgcttt cagctccatg agggccctca tttctgggtc actcattccc 120180 gtgtccccag cccctggcac tttggctaag caagcgttaa gcacctagcc ccgggctggc 120240 actcgagata tgtgagcgcg tgttacgttc aactttgctt tgcaggcgag aggctctaag 120300 acccagaaca cagcccctcc ccaccaggaa ctcaattcca ccaaaccctt ggaaggcccc 120360 acggctcctc ttcccctccc gggacccact tcctccaact ccagatgcaa gcccagctgg 120420 gcaccatcct cggccgagag gttacagcac ccaggctggc tttagcgcag acctcagatt 120480 tcttaagcaa acgagaagag gtgaggaggc ctgtgggggc acctaaagag agaatgagtc 120540 agtcagagct agacttgaaa cgtaagagac atcagcactt cttggaaagg caagaacgga 120600 cgcatttaga cagaactgtg catgcagaat gaggggtaag cacaggccag tcaggaagtg 120660 gagtcttccg tcctctgtga gtccctccac ctggccctct cttcctcatt tccctctcaa 120720 atgtcacctc ctcagggaag ccctccctga ttccacttgc acccagcttt gctttccttc 120780 ctagcacagt ctccacctga taggatattg caggtccatg ttctctcctg taaaatcctg 120840 ggggccagac atagttcaga attcaggatg actcaggctg tagaaaggtc accttgtata 120900 aaatacccac cgacatgccc atcatgcagg gcgggaaagc acccaaagat caattgcact 120960 aatatctctg cagaaagaca tctgagcatc acaccgaaga ggatgaagac tctaaaaagc 121020 ctcaagttta gcttcaaaat gagttatgaa aaactttcgg ttttcagaaa tcgtggattt 121080 cagaataaca gataaagctg ccgatctgaa catatgtatt ggttgtacat tgcctgtatg 121140 ctcaactaga atctaaaccc caggagagca aaggacttgg tccgatgctc tggcacatat 121200 aggtgctcaa taaattcctg aggaagaaat gcaggaggaa gccaggtgct ggatacgcag 121260 aagggactcg gtccaaaatg cagagaaaaa cagcaatcac tgagcgcctt ccaagggcaa 121320 aatgtctata caaactatct catgaatgct ctccctgcaa atacccatga caaaacaaag 121380 ggaggctcag agaggttaaa caacttcctc cagattacac agtggagagc tgggaagtag 121440 ccagctccga tttaaacctg ggatcccctc ttcaagctca ggcccttctc tctgccttaa 121500 gctcagagac gcagagcaat ttgctggata tcataaaaca gtggtgatgc acccggtctg 121560 acagtggatc ccccagcctc tgtagcacct caagcctctg ccggggcgct ccgatccaca 121620 ttcaccaagg gcaggtctgt gcaacctcca gacaagctcc tggcctctct tctagcccag 121680 ccatggtaag tctagctaaa cgtggctggg catgtctcca ctgtgccagc caggaggaaa 121740 actcagaaat gtctttcttg gtcaccccag tgaactggta caagcaggaa agaatacatg 121800 ggaagaaaag taggatggag tctgtccctc agatgtggac ctgaggcaag gctactgcat 121860 tcatctccgg gtctcagttc tcacctgcaa aatgagtgga ctaaatccca accaataaag 121920 ctcaaaggag gcaacagtaa agatttattc atttaggcca ggcacgtttg ctcgcatcta 121980 taaatccaac actttaggag gctgaggcag gaggactgct tgagaccagc agttcaagac 122040 tagcctaggc aacatagcaa gatcctgtct ctaaaaatat aaaaacaatt agccagccgt 122100 ggtggcacac acctgtagtc ccagctactc aggaggctga ggtggaagga ttacttgaac 122160 ccaggtattg gatgctacag tgagctatga ttgtgccact gtgctccagc ctgggaaata 122220 gagagagacc atggctctaa aaaaaaaaaa aaaaggtatt catttaacaa atacatagtt 122280 tgtactacat accaggtact gttttgtatg cttgacatat attaactcat ttagtcatca 122340 ctacaatggc atagtcacta taatctcata aatgcagaaa ctgaggctct gggaagtcag 122400 ttacacagta agcagcaaag ccagcatttg cactgggtga tctggctcaa gtccatactg 122460 tcactcccca gctatacgtt ttgacaggta acaacagaca tatacattga ttgtggaccc 122520 actgggtgtt ggacattgtg tgaagtagct tataaaacat gtcatcaacc ctcaagaaag 122580 ttcactttta caaatgtgag gaagaacagt acttaggcgg cttctgtcct ggcctcactc 122640 ccaggtacca gtgagggcag tcgagaggtg gggcatctaa tcccaaacgc tctcctccat 122700 cgtcccagat ggggtgggtg gaagagggca gggagatggg ggaggtagaa aaggttctct 122760 cttctcatgt atttatgtat ttgtttattt tgagacagag tctcgctctg tcacccaggc 122820 tacagtgcag tggcacaatc atagttcact gcagccttga ccacctgggc tcaagccatc 122880 ctcccacctc agcctcccaa atagctgggg ctagaggcac ctgctaccac gcctggctaa 122940 gtttttaaat tttttttgta ggaacaggat ctcactatgt tgcccaggct ggtctcaaac 123000 tcctgggttc aagcgatcct ctctcagcct cccaaagtgc tgggattaca ggtgtgagtc 123060 actgcgccca gcttgtcact tctttgacaa ttcatttcag ctcctttttg aactgggagc 123120 attagaaaac tgaacacccc cacccttgcc ctgggcattt tgcaaccggg attgtattgt 123180 attaaacata tcagtcattg agtcacttcc tagcatttgg aaacccttag gttatctcgg 123240 acctgccggt aactgcatgt gttgtaactg gggtgagtga ctgcaggttt tcctccctgc 123300 tgcctcccca gcgtctccct ccagattcag atccagaggc ccagggtggg gcccaggaat 123360 ctgcactgtt gcttttacaa ctcaagccaa ggtgaataca ctggcccaag ttccttggca 123420 tggcattcaa ggctttccac aatgcagccc agcccaactt tccatccatc tctggcttct 123480 gcttactccc taggttctgt acacagtgcc ctttcacact tcccagcctc cgcccactct 123540 gtgcctccct ttacctccca tcctctacct gccagactcc cctgctctac tcacaaggct 123600 agctcagggt ccctgggtgc tgggaagcct ggcccagtgc cctttgaaga gctgatcccc 123660 tctggcttgt ggtcctgcag ccgctggtgc atccgcaccc tcccagccct cacgtctgtc 123720 ttcctgcaga gggggtgctc cttgaggcaa gcaccagtgc ccagaacacc acccagggcc 123780 tcgcaggagc tgaatgagtg ctagtgagtg aatgagtggc ttagcaggtc acctcccatt 123840 catgggctgg gctagccgag ttctaagaaa cccctgcagg accctggaat gccctaatgt 123900 gcaggacacc ctggccaaac ctacacctga gcctgtttcc ccattggact ttgggatttt 123960 gataaaggca ctgaggtccc agcaaatgcc ttgggctcct cataggctac tgggggcagg 124020 gaagtgcagt gggtcatgcc tcccagaggg ctggggaagg ggagagacta gaactggagc 124080 agggttagcg ctggcctggg ggcattcctt gaaccccaga agcagacaaa ctagtggtcc 124140 cctatcttgc ttaagtctaa gaaatccaga ttttgtttaa aaaaaaaatc agggctcagt 124200 ggctcacacc tgtaatccca gcattttggg aggccgaggt gggaggatca cttgaggcca 124260 gcctggacaa catagtgaga caccatcttt ataaaataaa tttttaaaaa taaaataaag 124320 atcagattcc tgggtttact gaatctgaat ctcagggcaa ggacctggga atctgcattt 124380 taagcaaatc tcccggagat tctggttatt gtgggggctc ctcaaacgtg attgagagat 124440 ggagagcagc tggctctggc gtgggattgc ctgggctcta gtcctgctgc cccacggact 124500 cccaagtggg cggcacctta gggcaagtta ctaaagcggc tggggtcttg gtgacctcat 124560 ctgtacaatg tccatgacgc acagcaccgg ccacagagaa gttagtggga ggtttcactt 124620 ggcatccttt cagtaaaaca agtcccagtc aatgatcgcg attattgggt ccatctgata 124680 ttcggaggcg gagaccgccc cgattcactg acctactact caaatttaca gttctcagaa 124740 ctttccccac cctttctctc tccgaagtcc tcactgactt cctgagcgca gggctgggcg 124800 ccactgcccg gagaagaggg gctgcgtggc ctccgacagc ctaaccccaa gccccccacc 124860 cccgcaggcc gtcccggcgg aggtctcact gccagcaccg gctttgggga gcgctgggtg 124920 tccacgtacc ccgggcctcg cgcctccgct gccgttcggc ggcaaggacg gaaccgggcg 124980 ctgctccgcg cgcaccgaga gtcgcgggct cgggccccgc tcggagtcgg ggtggtcccc 125040 gtggtccccg ccggcggccg agcccttccc gaacgccctc agccggcgtg cccgggcgat 125100 accttcgcaa cccgatccgc gcgcgggccc cggatcctac ctgcgcgccc ctgctccatt 125160 cgccgtccgg gcgcccaagc gggaagtggg ggacccctgg gtggcccttc cctggcccgc 125220 gccgcgcggg gctttcctgg gcgcggagac ccagcccgcc ccggcttcct gctccccgcc 125280 ccggcgcccg gcagatttgc atgcgccgcc ccgcccggcg cgcccggccc gggtcccggc 125340 cacctcgcgc ccctgtcgct gtccccgccg ccgccgccgg acagtccgag ccgccgtcct 125400 ctcctggggg cgaccccagc acctcgctgg gtgtcttgcc gcgcgcttcc tcccacgctg 125460 cagcccgagg tcacgcccag cggagaactc tgcagtggat ccccttggtc tcggaataaa 125520 acgcccacgc cgaccaggcc tccaggacct gctcctgcct ggcctccgcg gcctcttttg 125580 cgccggggca gttgcgtttg tggggggagg gctgggcgac tggaatcctc ctccccttcc 125640 taacagcccg gcttccaagt gagagtggtt gcacattctt caataaatgt ttacccaact 125700 cctactccag ccacatttgg atagtaaaac agcccttcaa tcagtaacta gtacaatctg 125760 tcgggcactg caaatgttaa ctcgttcaat cctggcaact accctataag ggagtgatac 125820 tatcaccggt tttacaaatc aggaaactga ggcacagagt ccttaagaaa ttcaaaatgg 125880 gttgggcaaa gtggctcatg cctgtaatcc cagcactttg ggaagctgag gcgggaggat 125940 cacttgaacc ccatagttcg agaccagcct gggcaaccct gttagcgaga cccctatcta 126000 caaaaattga catttataaa attttaaaaa agaaaagaaa gaaactcaga atcactctgc 126060 aggtaagcag gagcaggggt ggagctggga ttcgaacaca ggccccatag accattaacg 126120 tctggacacc ataagaaccc ggatgcagcc agatgactgg ggggaaaaca ctcaacaaag 126180 tatttcaggt caggagggcc caagaagaaa gtagggacct cttagaccag tggggcaggt 126240 ggtctggaag aggctttttg agtagaggcc atttgagctg agactctgaa gagcctatgg 126300 gggaagactt gtgtagggag agggaagagc tagtgcaaag gccctgtggg aaccagctca 126360 gcctgcagga ggaattgctg aaggccggca tggctgtgga gccctgagtg aggaaagagc 126420 ggagacaggg gaggtgggag atatttggac tcacactggg cagtgctgag aacccagaga 126480 gtaatttgac cccctgctgc cctcaaggag ctctcagttt gagggtggaa gccaaggact 126540 gcccaatggg atcaagtgct gtggtggagg gtgagggttg gcttcccagg gcaggggcca 126600 ctgcacgtca atcattcttt tttttttttt gaagatgggg tctcattcca ttgcctaggc 126660 tggagtgcag tgggtgatct cagctcactg caacctccac ttcctgagtt caagcaattc 126720 tcctgcctca gcctcctgag tagctggaat tacaagcgtg caccaccacg tccggctcat 126780 ttttgtattt ttagtagaca tgaggtttca ctatgttggc caggctggta tcaaactcct 126840 gacctcaggt gatctgcctg cctctgcctc ccgaagtgct gggattacag gtgtgagcca 126900 ccacgcctgg cccatatgtc aatcattctt tcatgtactt ggtctgtctt ggtcctactg 126960 tgtgtctaac cttgtgacaa gcgctgatga cgcagacctg aagacatgaa acagacacaa 127020 tccctgccct ctgagacctt acaaccaagt gagcaagaca gccaccaacc cacccccaga 127080 agtgacagta aagataagtc agttccctgg ggtcagggat ggcttcctgg aggaggtgac 127140 ctctgaccag tgcctgaatc aatgcagaaa aacatgcagg tggaagggac agcatgagca 127200 aatacctgga ggtggcgtgg gagtgtggaa aaactcccat ccttcggcat ctgtttgtgg 127260 atgcctccat atgccaagtc gtgagctggg ggctagggag agcaagaaat agatccagcc 127320 cctgcttcct agccctgagg acctggttgg ggcactgaca agtgagcaga cgtaagcaat 127380 tgaagtgaac agatgtaagc aattgagtgt aataaacccc gtactggggg cagcccaggc 127440 agacgctaag aggactaggg agagaaggcc acaggaaggg ctttcctgga agcttgggtt 127500 tcaggaggaa taggaatcct ctggattagg tgtgacttcc actcccgtcc acactcatac 127560 tccctcacac tcattactca cacatccgca cacccacacg gttgatgctt catcccaggc 127620 agttcagccc tgcaagctgc ctttgtcctg tctgtgtttc tgagtttgtg ccccatgtca 127680 tcgggagacg acagcaaaag tctacctttg aagtcctttg ggaatgtatg gcttgggcta 127740 tatcccctcc tggactccca aactgggctc ccatcctgtt ttcccagtca agtcatcctt 127800 cgtcggtttc ctcgcttgcc tctagactgc cggcattggc ataccaactc tgctacgtat 127860 tgactatgtg actttgggca aagtagatca tcctccctgt gcttcagttt ccccatctgt 127920 aaaatgggga taataataga cccacctcat agggctatga aatggtgaaa tggcaaggta 127980 cgtatcgcag tgcctagaat agtgcctggc gtggattagg cattcaaaat gtcaattgct 128040 gttcttatca ggatgaaaat gagaaagaag caaagtcgaa gcctgttgcc ttggtctgtt 128100 gcttggttgt gaaatggaga tgaacgtggc attgagaaga caggcaaaga aagcacagct 128160 gtggtcagag gctaagacat tcaggaaatc tgggggaaga gacttcggga attctttgta 128220 ctcttctccc aactttcccg ccagtctgaa attatgtcaa attacaaagt taaagaaaac 128280 agggcattgt tctcgggtgc aagagagata ataaatgtct tcttgcatac agtcccactg 128340 ttcaagttcc tgcccaagat gcccagactt tatctgtgac tgctccgtca gtcctgggag 128400 caggtccaaa ggcaccaaga tagtaaaacc tctgttcccc catctagggg ttcccaagtc 128460 ttgcctcagt ataagctcaa tcctcattat atcctaacac gcttacttgt ttccaagcct 128520 ttcggtcttt aaaactaata gagtttattt tttagaacca ttgtagatta tggaaaaatt 128580 gagccgatag tactgagagt tcccatacat ccctctcccc catgcaaaat ttcctctttt 128640 atcaacatct tgcattagtg tgtagtgcct tccttataat gctgatacat tattataaac 128700 taaagtccat agcatacatt gtttcactgt ttctcggggt tttgttttgt tttgttttgt 128760 tttgtgtttg tttgcttgtt tggaaacaag gtctcactct gttgcccagg ctggagcagc 128820 ctggaactcc tggactcagg ggatcctcct gcttctgcct tctgagtagc ttgggctaca 128880 agaacatgcc accacactag gctttttttt ttttatactt ttagagacag tcttgctaca 128940 ttgcccaagc tggtctcaga ctcctaagct gaagttatcc ttccatctca gcctcctaaa 129000 gtgctgggat tacaagcatg agccactgtg cctaacctct gcagcacagt tctgagtctt 129060 ttttttttct ttgagacaga gtcttgttct gtcacccagg ctggggtaca gtggcgtgat 129120 ctcaactcac tgcaacctgt gcctcccagg ttcaagcgat tctcactcct cagccttcca 129180 agtagctggg actacaggtg cacgcctggc taatttttgt atttttagta gagacggagt 129240 ttccccatac tggccaagct ggttttgaac tcctgacctc aagtgatcca ccagcctcgg 129300 cctcccaaag tgttgggatt acaggcatga gccaatgcac ccagccaagt tctgggtctt 129360 aacaatgtat aatgccatgg atccacacat gtagcaacaa aaagaatagt ttcccagcct 129420 gggcaacata gcaagacccg gtctctacca aaaaacacaa acaaacaaca acaacaaaaa 129480 acaggccagg catggtggtg cactgtagtc ccagctactc agaggctgag gcaggaggat 129540 cccttgagcc caggagttcg aggctgcagt gaactatgat catgccactg cactccagcc 129600 tgggcaacag aatgagaccc tgtctctaaa aaaaaaaaga aaaaaataaa gaatagtttt 129660 acttccctaa aaatccccaa ttctctacct attcttcctg gattctggaa accactgatt 129720 tttttactgt ctccatagtt ttgccttttt cagaatgtca cgtaattgaa atcagaaagt 129780 atgtagtctc ttaatactga cttcttttac ttagcaatat gcttttcagg ttcctcaatg 129840 tcttttccag gcttcacacc tcatttcctt ttgtggctga atagtgccct attgtatgga 129900 tgtaccacag tttatccatt ctacctattg aaagacatct ttgttgcttc caatttttgg 129960 caattatgaa taaagctgct atcaacactc atgtgcaagt ttttgggtgg acataagctt 130020 89 9900 DNA Homo sapiens 89 gaattcaata aaaaacaagc agggcgcgtg gtggggcact gactaggagg gctgatttgt 60 aagttggtaa gactgtagct ctttttccta attagctgag gatgtgttta ggttccattc 120 aaaaagtggg cattcctggc caggcatggt ggctcacacc tgtaatctca gagctttggg 180 agactgaggt aggaggatca cttgagccca ggaatttgag atgagcctag gcaacatagt 240 gagactctta tctctatcaa aaaataaaaa taaaaatgag ccaggcatgg tgcggtggac 300 cacgcaccta ctgctagggg ggctgaggtg ggaggatcat tgagcctggg aggttgaggc 360 tgcagtgatc cctgatcaaa cattgcattt cagcctgggt gacagagtga gaccctgtct 420 cagaaaaaaa aaaaaaaagt cattcctgaa acctcagaat agacctacct tgccaagggc 480 ttccttatgg gtaaggacct tatggacctg ctgggaccca aactaggcct cacctgatac 540 gacctgtcct tctcaaaaca ctaaacttgg gagaacattg tcccccagtg ctggggtagg 600 agagtctgcc tgttattctg cctctatgca gagaaggagc cccagatcat cttttccatg 660 acaggacagt ttccaagatg ccacctgtac ttggaagaag ccaggttaaa atacttttca 720 agtaaaactt tcttgatatt actctatctt tccccaggag gactgcatta caacaaattc 780 ggacacctgt ggcctctccc ttctatgcaa agcaaaaagc cagcagcagc cccaagctga 840 taagattaat ctaaagagca aattatggtg taatttccta tgctgaaact ttgtagttaa 900 ttttttaaaa aggtttcatt ttcctattgg tctgatttca caggaacatt ttacctgttt 960 gtgaggcatt ttttctcctg gaagagaggt gctgattggc cccaagtgac tgacaatctg 1020 gtgtaacgaa aatttccaat gtaaactcat tttccctcgg tttcagcaat tttaaatcta 1080 tatatagaga tatctttgtc agcattgcat cgttagcttc tcctgataaa ctaattgcct 1140 cacattgtca ctgcaaatcg acacctatta atgggtctca cctcccaact gcttccccct 1200 ctgttcttcc tgctagcatg tgccggcaac tttgtccacg gacacaagtg cgatatcacc 1260 ttacaggaga tcatcaaaac tttgaacagc ctcacagagc agaaggtgag tacctatctg 1320 gcaccatctc tccagatgtt ctggtgatgc tctcagtatt tctaggcatg aaaacgttaa 1380 cagctgctag agaagttgga actggtggtt ggtggcagtc cagggcacac agcgaggctt 1440 ctccctgcca ctcttttttc tgagggtttg taggaagttt cctcagttgg agggagtgag 1500 agctgctcat

caaggacttc tctgtccggt tggaggttaa ctctgtctct tgctctctca 1560 tttctgcctg gaccaagact ctgtgcaccg agttgaccgt aacagacatc tttgctgcct 1620 ccaaggtaag aagccgtccc acggtctgtt ttagcaaatg gggagatcca tccccaaatg 1680 tctgaacaag aaacttgtct aatggaaaac gagcgggccc aaattaactc taaggtgtta 1740 gatgttttca aagaacgaga agtctgatct ttactcttaa gcatgttttg gtctttctgg 1800 tttcacttga tttagaagac atgtaataga aagcttacat gctgtagtcc tgactcagat 1860 cctggtcaaa gaaaagccct cttgggtttt acttagcttt ggcatagtgc ctggaacgta 1920 ggaggcactc aataaatgcc tgttgaatga gagaattttt ctggcccata catttctgaa 1980 aaaccaaata ctctcacaga aacagatatt gagatgacag gttgagggag ctttcatttt 2040 gtctaagaga cttcctatgg caacagaaaa ggtatcgcca gagcccctcc tcttccacag 2100 cctggccacc taacagccct ctggttccgg ggctgccgtc cagagctctc agcttgctct 2160 ggccggccga actcccctcc agctcggtct ggaaccatcc tgctgggcag cgtccagcac 2220 atccctgctt cgggctgcct gggcacctcg cctctctgcc tcctgtgctg cctcaccccc 2280 acccctctat ctgtagtggg agggagatag atttgacagc tgatagtgca ttttctctga 2340 caaacacatg actacagccg tatcaatagt tttgtgcatt tcagttcctg ttttcatgga 2400 aacacacggc tgagaatgaa agccccaaag cctcaatttc acagtggtct cctaactacc 2460 tgctttccat gcaaactagg gagatgatat ggccaggagt gaagccctgt gtgttgggca 2520 gggtcacact ccagcaccca gaccatagaa cagggcccat cctgcttcat gagggaaact 2580 gctcttcggg cctttagctg gactatctca tttcattagt tatcccggga gtccgataca 2640 ggatgagatt ctgaagggca aatacacact tttttttttt ttttgagata gggtcttgtt 2700 ctgtcaccca ggctggagtg cagtggtgcg atttcagctc atagcagcct ccacctccca 2760 ggctcaagct atcttcctac ctcagcctcc caagtagccg ggacgacagg tgtgcaccac 2820 cacgcctggc taatttttgt atttttttgt agagatggag tcttgccatt ttgcccaggc 2880 ttgtctcgaa cttctgggct caagcaatcc gtccacctcg gcctctcaaa gtgctgggat 2940 tagccactgc acctgggcaa cagtttatgt gtgtgtgtgt gtgtgtgtgt atatatgtgt 3000 gtgtgtgtat atatatgtgt gtatgtatat atgtgtatgt atatgtgtgt gtgtgtgtgt 3060 gtgtgtgtgt gtgtataaaa tctccaagtc catccaaccg agatggctcc tactagaagc 3120 caagagtcca ccgggttgag cactgggtct ctggaggcct gtcggactgc tgagaaggct 3180 ctaacaaagc caagggaagg gccacctcac tagaagccag gcctggagga agggtgaggg 3240 ctgagggctg gaggtaagac tgcctgtggt tttagaccca ggctctgcca ctgactagct 3300 gtgtggctgg ccttcagaca tcttcacagc tctctgcacc tcagtttcca catgtgaaga 3360 tatgaaagtg attctgaagg tgattgcaag gttgattgga atccagctct tgagttagtg 3420 caaagtgtta ttgtgagatg atataaccac gattaaaagc aagaacaggt gcagagaagc 3480 gatgattcta agaaggaggg gaccgggttg gaaaggatca aaccatccag gatgccgagt 3540 ctggggcaat ccatctgggc tgtttctgga agacccccgg gtgcaggcca ggacactgct 3600 gccctcccgt ccttaactcc cctcttcact cagtcctcac tcacctccct ctcacacaca 3660 caaacatctc ctagaataat ccccactgcc tgccttcact cttacccgtc tcatttgcct 3720 cccctgaact tcatcctcct ggagttcacg atctcactct tcactctttt cttcccctcg 3780 aagattcagc actgcttact tacatgttaa gatatttcag aacagtgaaa tgttgctatt 3840 ttcaaaaacc tacaaaggtg gtatgcagag gaaaaggtac ttctttgtgt tcccaaagaa 3900 aacatctttc caaaatccag cctattgatt ttatttcttc gggggaacaa gaattttagt 3960 atctctaagt tgggtagcat tctactcttg gcagttgctg gaaagaaggc actggtctag 4020 gtcctgggct tcacaggtaa cacctgtcag ggtgtctatg aagtcaaggc tgtctgagga 4080 acagcaaagt gggaagaagc aagctggctg gctgatgaag ggtttcttgg gtggacaagt 4140 agttggagcg atttcctatt taccaaagag agctaaagtt cataattcta cagagagttc 4200 cataatgaac ctcaaatacc tctgtttttt gaaggagttt ctcatataca gcactagctg 4260 actatcctgg gcaggatggg agataatgaa tgcagtgcca atcgggctgg atttatatgg 4320 tcctagtgag gctggtcaag aaccgagtta gaactctcac agagtcactg cccacagaag 4380 aaatctccca agtggctgtt tcctgacatt cccgggaggc aggcctcctt ctgagtcact 4440 ccctaagcag ttctgaactg tgaggtcagc caggctgtcc aagtgcactc cctgagccac 4500 tggcagacac actcagcagc cagagctaga caggcaggtg gtaggagtcc agggccacgg 4560 cagggatgga gtgtcgcccc ctcgctgcga taccagagca agtaaaacgt taaggccttg 4620 cactaaagct gcccttagga tgcattcttt taaagttttt ccatttaatg cagactcttt 4680 tcaattctta ttttatcctt gtttccttta gaaagtcctt tgaaaaatat ctttagaggg 4740 ttttttccta tactatgtgg ccatatacgg gtcaaaatta agtttaattt ccaggctcca 4800 agccagcgtt tcagaaaaat ctcaccaagg tttgtggtaa aagaagcaaa gggctgactt 4860 tttggttttc ttgaatctca ctgttccctc tgcagcagca tgcatgtctg cccacctcca 4920 gacacacagg caccatctgc cgccccccat cagcccgtgt cccttccacc tcgactcgcc 4980 tacaaagccc agagaggtct gtttcttggc ccccagagcc caaagatact gacacactct 5040 tacatttcca actagaatca ggaacgagga gtgactctca gtcagttcat taagtaaatg 5100 tctttctaac cgctctgccc atgggacatc acgccccaca ggggaaaggg gaagcttctg 5160 tagcctggga ttctggtgcc tcagtctggg tctagacttt cctgaaaaaa cgttaaaata 5220 tgaactgcat tcctagaatt tagcctacat aaataagaga tgaacacaaa gatttctata 5280 gtttactcac tgccgcttat ttacagaagc aaaaatctgc cacgataggg gcctgacaaa 5340 tgacagtacc actgtgcaat gcgtttctac gcagctctca atcccatgtt ctctaatacc 5400 accgaaggct taggaaatgc ttatggtata tgtaaagagt aaagaagtta caaacagtat 5460 caacagttga cccctatttt aaaaagtatt tttgaaaagt gtgacgatat ttaccaaaat 5520 attaacgagc aatagttacc tctggctggt gggatgagtg aatgtatttt tgttgaatat 5580 atgttacctt tatagtaaat atatgttatc ttgatcatca gaaaaaaaaa tatgtaagaa 5640 cttgaaagct gcttggacag cgctgctgat agaaacccct gagcatcttg tcactgttct 5700 tctgattcag agggtctggg tggggcaggg gtggtctgag attctgtatt tctaagaagc 5760 tcccagtgat gtccatgctg ctgtccatgg accacacttt gagtatcaag ggaccagagc 5820 atgtcggggg agaggctggg gatagctttc tttatctgaa ctggataaag gaactgggct 5880 caagctaaga accctctcca ggttctgcat ctttgttctt cagtgaaaaa tgagaggaca 5940 caccaggcca ggttcagact gagacacaat ccctctcctg ggttcccaat gacttgtctc 6000 ttgtccattc ccttctctaa ggctaagggc cccccaggaa gagccatgtg gccagaccct 6060 cacagttgct ggcattccaa ggagattctc actccgcatc attggtccaa aaggcccctt 6120 acagaagctc tgcccaaggc tcagatcaat ggcacctgct cccagagctc ctctgatctc 6180 ccaggacacc tttccctgat ctgtgcactt atctcttgct gcctggcaaa atgtcttagc 6240 tcctcacttg ggccatgtgc tgctctcctc tcccatgggg agagccacac ggagagtgct 6300 ggccaaagca gcagagttca ggccaaagga tgtgcactca tttattcaac aggcatgcag 6360 gatttccagg gaaagctgga ttttaaaacc tctgggaaca agagcagaac ctgactgaga 6420 gctcatgtgg gcacttttca tagcagaata gctcatgagg tatagagaca cggacgcaga 6480 acgtgggctg tagcgacaga tggtcctgca ttctagtccc cactgtgcct tttcctcatg 6540 ggatgacttt attcaggtac cctttcggca aaatcctcca agagaaagga aactgggagg 6600 ttctggggag aaggctgctg cgtttgcaat tgggagaggt tgttgacaga ggtttatgtc 6660 tgtggcaagc agccttcctt cagtggaata cttgaagaca ggtctgtagt tgagcaaact 6720 cacctccatt tgtcctcctg gaaagaagaa atcaagagga aaaatctctc tcccatcctc 6780 caaatggagc tggcacattg ctatctgtgg catttgtctt tccagaacac aactgagaag 6840 gaaaccttct gcagggctgc gactgtgctc cggcagttct acagccacca tgagaaggac 6900 actcgctgcc tgggtgcgac tgcacagcag ttccacaggc acaagcagct gatccgattc 6960 ctgaaacggc tcgacaggaa cctctggggc ctggcgggct tggtaagctg cactgtattc 7020 ctggcaagcc ggccgcgtgg ctcctggtgg acagcagcct cacttctaaa cactccttag 7080 gagctgcagc acccttggtc aacccattca ttcattcact cattcaataa gtatttgctg 7140 aagttccaca agtgctgggt gtggttctag gtgctgagga cgtgtcacta aagacagcag 7200 gccgagtccc tgttctcatg gaatgttcta atgggagagt tagaaaaaca aacatgtaaa 7260 atgatggcca gcagtgatac gtgctacaaa gaaaaacata gaaataaaga acataagagt 7320 catgggggag ggggctgact taggagctgg tgacattatc tgagcagata tttgaattga 7380 gggagcaggc cacatgacta actagggaga ccattccagg gagaaggagg aggtatgcaa 7440 aggccttagg atggaaatga actaacttcc tgtatttaaa gaccagtagg aaggccagtg 7500 tggctggatc agagtgagtg aggggtagtt tccaggacag cagatcacac aaggccttta 7560 gattccacca cgagtatgga gggaacacct gcagagcttt gggcaggaca aagactgtac 7620 aatctgattt acgtgattta aaagggtcag tctggctact gtgtggtaaa taggctgaaa 7680 gggggaaagc atagaagcaa gatggcctgt tgggaggcta ccacagtaaa ccaggctaga 7740 gatgatggtg gcgtggacag aatgaagcaa gatggcctgt tgggaggcta ccacagtaaa 7800 ccaggctaga gatgatggtg gcgtggacag aatgaagcaa gatggcctgt tgggaggcta 7860 ccacagtaaa ccaggctaga gatgatggtg gcgtggacaa atggagcagt tgaggtgaac 7920 agatttggga tatgactaaa aataaaacca gaagatttgc tgacagatcg gttgtagggg 7980 gtaagataca ggggaggaaa agatgacctc tttgttcctg cccaaacccc tctggcgatg 8040 gtcagtactg tttacagaga gatgaaagac tggcggcaag gcagggctgg aggttcagca 8100 gaagatcaag agttcaattt tgtacatcgt acatgtaagg tggctcttgg atagccaagt 8160 gaaggtgttg agaagatggt tagaaaagtc tggaacttag gggagaggtc agaacttgca 8220 atacaaaaag gagagtcctt agatagatac tgctgaaaat ctgaatgaca gaaagggaga 8280 gatcaaagga ctgagcctga gatcaacaca tggaggtcag gagaggagga tccagccaag 8340 gggcctgagg aggagtgacc agtgaggcag gagaacatgg agagtgggcg gtaccccagg 8400 aagccggtga ggacactcaa ggagggaggg ttgactgtgt caaatgtact gaaaggacag 8460 gtcaggtgag gaccaagaaa ggcccctggg tttggctgat ggaggccatg ggtgaggctg 8520 atgtaaatgg agaggcagga aggaaagccc agctggagtg ggctcaccga ggatagggtg 8580 gcgagaggag acaaagaagg aacagtgagg gcagacaact ctttgaagat gtttagctat 8640 aaggctgcag agaaactgag cccacagctg cagggtggtt atggagtgag ggaagctctt 8700 ttaaggttgg gggtataccc agcatgttaa tgcacctggg ggaatggtcc agtggagcag 8760 gaagaactga agagagcaga aagaggaaga atcattaggg ggcagaagtc cttgtagccc 8820 agagtggatg ttatctaata tcgagtggag gaattaattg gctttagagg agaacaagga 8880 catgtatccc ctctctgggc ctatcacctt gtagacaatg ggataggtca tgggatagga 8940 acttggcaca acacatgttc tctcttttaa ttctctccat tatcttatga agcaggcaag 9000 taggcaaaca attgtcccaa ctttacaaaa gaaactgaag cttttataaa ttaagtagta 9060 catcctaagc aatacaatta ataaatggta gagctgagat tcaaactgaa gcagtggcct 9120 gggggtagca tctggaatcc ttcccacctt tagggctgct gtgctgcggt gctgctgttt 9180 aatggcacag agggccagat gactgaatct ctctcagcag tccaggcagt catgcagaag 9240 gcccagtaga gcaccgggca ggtctgagcc agcatcttca agttccaccc tgtgagcaag 9300 cacttagctg tgacacactt ctcgagagac tggactcccc cccgcgcaac ccacccaaaa 9360 gcagataggt aatggtatac agtaaccatt tctagaagtg taagtagtat gcacccaaaa 9420 taggcaaaac ctgctggcct agtgatagag acaactccca gtcaggctag actggaggcc 9480 ttggttttat aagtgttcag gtgacaagtg ccacagtagg cttgatcaag tagacaggca 9540 ggcaagacaa atgcttacca atgcaagcta atgaaatgtt tcttttgcag aattcctgtc 9600 ctgtgaagga agccaaccag agtacgttgg aaaacttctt ggaaaggcta aagacgatca 9660 tgagagagaa atattcaaag tgttcgagct gaatatttta atttatgagt ttttgatagc 9720 tttatttttt aagtatttat atatttataa ctcatcataa aataaagtat atatagaatc 9780 taacagcaat ggcatttaat gtattggcta tgtttacttg acaaatgaaa ttatggtttg 9840 caacttttag ggaaatcaat ttagtttacc aagagactat aaatgctatg gagccaaaac 9900 90 4740 DNA Homo sapiens 90 ggatccccgc tgacaatcta gaaacaagca acaggaccct ctgatgtagc catctgtgcc 60 gcgcctctcc gcaccgcccg ccacgccttg gtccctggag accaccctcc agggcagggg 120 ctgccgctcg gccgggcccg cggggtccct cggcctgaca tggccggtgc tggagcggca 180 cgtgcgcgcc tcggcccctc ggccgctccc gcccctcgcc ggtgcgcacc ggcgctcggg 240 gagccgctgg cccgggtgtc cagccggccc ttgccctgcc tggcgctcgg accgccacct 300 ttgccgcccc ctcgccagcc tccgcagctt ccagactggc cggtctgcgc gcccacccct 360 gcctcccgga ccggccaccg ccgaggcgcg gaggagggcc cggccgcgca gatcccgctt 420 atcggcccat ctcccgttac ataaggccac ccccctatct ccgcgggcca tcgccgccgc 480 aaccgccgcg ccagcgcctt ctcccacgcg cgggggcgcc cctgcccacc gctcccggca 540 gggcttttgg tggccatggg ggataagggg cgttgactca cccgggcggg gctccgggag 600 ttgcacagac caaggtagtt ccccgctcct tcccccatca cggagaccct gtgggagatg 660 ccgtgggccc tctactacag attaggaaac aggcccgtag aggggtcaca cggccaagta 720 gcggcactcc aggcactggg ggccctcgag gggaaggggc agacttctgg gagtcagagc 780 cagcagctgg gctgggaagc ttcgagtgtg gacagagagg gtgggaatga cgttccctgt 840 gggaagagag ggtggcaagc ctgggatgcc tctgagcggg aatccagcat gccttgtgag 900 gagggtcaca agcacaccct tgtgaggagg ttgagcccca tcgaggacag gacggaggga 960 gcctgagcag gcagagaggg ggcctgggga ggcgctggtt cggggaggaa gtgggtaggg 1020 gagaaatctt gacatcaaca cccaacaggc aaatgccgtg gcctctgctg tgggggtttc 1080 tggaggactt ctaggaaaac gagggaagag caggaaaagg cgacatgctg cagagactgg 1140 tgagcaaagg ggatcacccc aagccccagt ggcactagga acacttacaa tctctgacct 1200 ggactaaggc tgccagctgg cccagttaag agtttcccag aaggatggcc catacacttt 1260 aaattaaagg ggccagacac gtgcacacta cttccagcca ctctggaagc tgaggtgggg 1320 ggatcgcttg agtctgggag ttggaggcca gcctaggcag gcaacatagt gagaccccat 1380 ctccaaaaaa acaaaacaaa acaaaacaaa aaaacaccaa aaaagctccc agaaagacct 1440 ctgaatcttt ctggatctct cagtggagac cttggaaatc tgaactttga caatccctct 1500 cacagtgggg ccaaggagga attaggcaag ccaaaagaag tgaactttac tcttctattg 1560 cctgtttgaa ttttgtatcc aagcaagtgt tacttaagta atttaagaga ctggttcatc 1620 gaaaaaataa aactccccaa attcccatag ctggtagact gtggtcacag ccacagtgca 1680 ctaagactat ctgctcagca cttctggtga cccaaaaggg tctgaggaca ggagctcaga 1740 gttgggtcag ctgtccaggt actcagggtt gtcacaggca aaactgctgg aactcagggc 1800 agcattgcaa atgccacgcc gctctcaggg ccccttgcct gccgctggaa ttaaacccac 1860 ccagatcttg gaaactctgc cctggaccct tctcaataag tccatgagaa atcaaactct 1920 ttcctttatg cgacactgga ttttccacaa agtaaaatca agatgagtaa agatgtggtt 1980 tctagatagt gcctgaaaaa gcagagacca tggtgtcagg cgtcaccact tgggcctata 2040 aaagctgcca caagacgcca aggccacaag ccacccagcc tatgcatccg ctcctcaatc 2100 ctctcctgtt ggcactgggc ctcatggcgc ttttgttgac cacggtcatt gctctcactt 2160 gccttggcgg ctttgcctcc ccaggccctg tgcctccctc tacagccctc agggagctca 2220 ttgaggagct ggtcaacatc acccagaacc agaaggtgag tgtcggctag ccagggtcct 2280 agctatgagg gctccagggt gggtgattcc caagatgagg tcatgagcag gctgggcctg 2340 gtcctaagat gcctgtaggt caggaaaaat ctccatggac caaggcccgg cccagccatg 2400 agggagagag gagctgggct ggggggctca gcactgtgga tggacctatg gaggtgtctg 2460 gcagactccc cagggactac ctgctctcct ggcctggcct tgtctgccac tgccagctcc 2520 tactcagcca ttcctgaaca gaggacagca gagaagggtc cagcaccctc ccagaaccat 2580 gtggcatttg ccaactggat tttgaccata acaatgcagc cattctcccc agcaccatca 2640 taggcccgcc cttacaggag gattcgttag tagagtcgct ccttgcccca ctagtaacag 2700 ctcacatgtc ttgagcactg cttacaccag gcctggtgca cgtgctttat gtgtcatttc 2760 atcactgcca gccacctcaa gaggcaggta cgatgaaccc attctgctaa ggttcgtgag 2820 gttaagtgac agaggctgga ttcaagccag gcctggccaa caccagagtg tccatgctcc 2880 taactgcagt gttccctcac catcagaagg cagggcattt aatacaccag atccccaccg 2940 cctcccatct gatttgtctt ggtcaacatg gcccaggcca ctcctacttc actcgtcccc 3000 accctgaccc ttcccgcagg cccctgtcct cctgccctga ctatggcaag ccttgcatgc 3060 agcttgtccc ttactagtgg tgtcaatttt tttctctcag ctccaagacc ctaaacagtg 3120 ggacctcacc cctatgcctg ctgttcaaag cagaaaacga agctcaggaa tgctgagggg 3180 ctgccaggcc tgcctctgtg ccacaccagg gatgcttgtg gggcctgtgc tggggcagac 3240 ctggcctggg ctgccagggc aggcccacaa cccctgccag cactctgctc actgtcactt 3300 tgctcccaca gcgtccgctc tgcaatggca gcatggtatg gagcatcaac ctgacagctg 3360 gcatggtaag gacctttggg tgcagggagg atggggcaga ggctccaggc cttgggctta 3420 tcttctctga gcctcccttc catggctggg gttccaagca agcttcaagt gctctcctcc 3480 ctcccgccat aatctggccc cttcccgccc accacccaga ctcacctgcg ccaggcatct 3540 cagccccatc ttcctgcaga ctcacaaaag gcagctgccc aagcagggcc tgacccctcg 3600 gtgtcccctc cccacagtac tgtgcagccc tggaatccct gatcaacgtg tcaggctgca 3660 gtgccatcga gaagacccag aggatgctga gcggattctg cccgcacaag gtctcagctg 3720 gggtaaggca tcccccaccc tctcacaccc accctgcacc ccctcctgcc aaccctgggc 3780 tcgctgaagg gaagctggct gaatatccat ggtgtgtgtc cacccagggg tggggccatt 3840 gtggcagcag ggacgtggcc ttcgggattt acaggatctg ggctcaaggg ctcctaactc 3900 ctacctgggc ctcaatttcc acatctgtac agtagaggta ctaacagtac ccacctcatg 3960 gggacttccg tgaggactga atgagacagt ccctggaaag cccctggttt gtgcgagtcg 4020 tcccggcctc tggcgttcta ctcacgtgct gacctctttg tcctgcagca gttttccagc 4080 ttgcatgtcc gagacaccaa aatcgaggtg gcccagtttg taaaggacct gctcttacat 4140 ttaaagaaac tttttcgcga gggacggttc aactgaaact tcgaaagcat cattatttgc 4200 agagacagga cctgactatt gaagttgcag attcattttt ctttctgatg tcaaaaatgt 4260 cttgggtagg cgggaaggag ggttagggag gggtaaaatt ccttagctta gacctcagcc 4320 tgtgctgccc gtcttcagcc tagccgacct cagccttccc cttgcccagg gctcagcctg 4380 gtgggcctcc tctgtccagg gccctgagct cggtggaccc agggatgaca tgtccctaca 4440 cccctcccct gccctagagc acactgtagc attacagtgg gtgcccccct tgccagacat 4500 gtggtgggac agggacccac ttcacacaca ggcaactgag gcagacagca gctcaggcac 4560 acttcttctt ggtcttattt attattgtgt gttatttaaa tgagtgtgtt tgtcaccgtt 4620 ggggattggg gaagactgtg gctgctggca cttggagcca agggttcaga gactcagggc 4680 cccagcacta aagcagtgga ccccaggagt ccctggtaat aagtactgtg tacagaattc 4740

Claims

1. A method for determining an individual's risk for type 1 diabetes comprising detecting the presence of at least one type 1 diabetes-associated IL4R, IL4, or IL13 polymorphism in a nucleic acid sample of the individual, wherein the presence of said polymorphism indicates the individual's risk for type 1 diabetes.

2. The method of claim 1, further comprising detecting at least one other polymorphism and wherein said polymorphisms comprise a haplotype.

3. The method of claim 1, wherein the nucleic acid sample comprises DNA.

4. The method of claim 1, wherein the nucleic acid sample comprises RNA.

5. The method of claim 1, wherein the at least one polymorphism is detected by amplification.

6. The method of claim 5, wherein the at least one polymorphism is detected by a polymerase chain reaction.

7. The method of claim 1, wherein the at least one polymorphism is detected by sequencing.

8. The method of claim 1, wherein the at least one polymorphism is detected by amplifying a target region containing the at least one polymorphism; hybridization of at least one sequence-specific oligonucleotides that hybridizes under stringent conditions to the at least one polymorphism; and detecting the hybridization.

9. The method of claim 1, wherein the at least one polymorphism is selected from the list of polymorphisms listed in Table 2.

10. The method of claim 1, wherein the at least one polymorphism is selected from the list of polymorphisms listed in Table 21.

11. The method of claim 10, wherein the at least one polymorphism is a combination of two or more polymorphisms selected from the polymorphisms listed in Table 21.

12. The method of claim 1, wherein the at least one polymorphism is an IL4R polymorphism.

13. The method of claim 12, further comprising detecting at least one IL13 polymorphism.

14. The method of claim 12, further comprising detecting at least one IL4 polymorphism.

15. A kit for determining an individual's risk for type 1 diabetes, where the kit comprises at least one sequence-specific oligonucleotide that comprises a sequence that hybridizes under stringent condition to at least one type 1 diabetes-associated IL4R, IL4, or IL13 polymorphism.

16. The kit of claim 15, further comprising reagents for the amplification of a target region, wherein said target region comprises the at least one polymorphism.

17. The kit of claim 16, wherein said reagents for amplification comprise at least one primer.

18. The kit of claim 15, wherein said at least one sequence-specific oligonucleotide is labeled.

19. The kit of claim 15, further comprising a reagent to detect the label.

20. The kit of claim 15, wherein the at least one polymorphism is selected from the list of polymorphisms set forth in Table 21.