JP2001245666A - New polypeptide - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for obtaining a new G protein conjugated receptor polypeptide and searching a ligand, an agonist, an antagonist or a functional modulator of the polypeptide. SOLUTION: This DNA encoding the new G protein conjugated receptor polypeptide can be obtained by randomly sequencing a cDNA clone selected from a cDNA library derived from a human stomach cancer cell. A searching system or a screening kit of a ligand, an agonist, an antagonist or a functional modulator of the polypeptide is obtained by using the polypeptide encoded by the DNA or its partial polypeptide. A compound obtained by using the searching system or the screening kit and an antibody to the polypeptide can be provided. A nonhuman mammal deficient in the DNA can be provided by using DNA.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel G protein-coupled receptor polypeptide derived from the human thalamus, a partial peptide of the polypeptide, a DNA encoding the polypeptide or the partial peptide, and the DNA incorporated therein. A recombinant vector, a transformant carrying the recombinant vector,
A method for producing the polypeptide or the partial peptide, an antibody recognizing the polypeptide, a method for screening a ligand, agonist, antagonist or functional modifier of the polypeptide using the polypeptide or the partial peptide, the polypeptide or A kit for screening a ligand, agonist, antagonist or functional modifier of the polypeptide containing the partial peptide,
The present invention relates to a compound obtained by the screening method or the screening kit or a pharmacologically acceptable salt thereof, an animal deficient or partially modified in a gene encoding the polypeptide, and a method of using the same.

[0002]

2. Description of the Related Art G protein-coupled receptors (hereinafter, GPCRs)
Is sometimes present on the surface of each functional cell in living cells and organs and functions as a receptor for molecules that regulate the function of living cells and organs, such as hormones, neurotransmitters, and biologically active substances. In doing so, it plays a very important role physiologically or pathologically. GPCRs are
It also functions as a receptor for light, taste substances, and odor substances. As specific ligands for GPCRs, a wide variety of proteins, peptides, biogenic amines, lipid mediators, photons, calcium, sugars, nucleic acids and the like are known. Heterotrimeric _{(G α,} G βγ) G protein (guanine nucl
EOtide-binding protein) and transmits a signal into cells through activation of G protein. GPCRs are also called seven transmembrane receptors because they have seven transmembrane domains.

[0003] GPCRs are excellent targets for drug discovery, and natural ligands, agonists or antagonists of GPCRs have been used as drugs so far. Approximately 60% of currently marketed drugs target GPCRs. GPCRs also cause genetic diseases and are important targets in the diagnosis and treatment of genetic diseases [Trends in Pharmacological Science, 18 , 430
(1984)].

Therefore, obtaining a novel GPCR and analyzing its function provides a very useful means for developing a drug closely related to its function. For example, in the central nervous system organs such as the brain, the physiological functions of the brain are regulated by many hormones, hormone-like substances, neurotransmitters or biologically active substances, but unknown in the brain. It is thought that hormones, hormone-like substances, neurotransmitters or bioactive substances are present. It is known that the physiological functions of organs such as the stomach, intestine, and pancreas are also regulated by many hormones, hormone-like substances, neurotransmitters or bioactive substances,
It is thought that unknown hormones, hormone-like substances, neurotransmitters or biologically active substances exist in organs such as the stomach, intestine and pancreas. In organs such as the brain, stomach, intestine, and pancreas, receptors (eg, GPC) corresponding to the above hormones, hormone-like substances, neurotransmitters or bioactive substances
R) is also known to be present, but it is likely that unknown receptors (eg, GPCRs) also exist in these organs. New subtypes may also exist for known GPCRs.

[0005] If a novel GPCR gene expressed in the brain, stomach, intestine, and pancreas can be obtained, the amino acid sequence of the GPCR can be compared with the amino acid sequence of a known GPCR, and the expression distribution of the transcript of the GPCR gene can be determined. By investigating, the function of the GPCR can be estimated and useful information for drug development can be obtained. In addition, if a new GPCR gene can be obtained, a natural ligand, agonist or antagonist for the GPCR can be efficiently screened. The natural ligand, agonist or antagonist is expected as a pharmaceutical.

[0006]

If a G protein-coupled receptor polypeptide and a DNA encoding the polypeptide, which have not been known, can be obtained, a ligand, an agonist, an antagonist or a function modifying substance of the polypeptide may be obtained. Screening becomes possible, and substances obtained by the screening are useful as pharmaceuticals.

The present invention relates to a novel G protein-coupled receptor polypeptide derived from the human thalamus or a partial peptide thereof, a DNA encoding the polypeptide or the partial peptide,
A recombinant vector containing the DNA, a transformant carrying the recombinant vector, a method for producing the polypeptide or the partial peptide, a ligand for the polypeptide,
Screening method and screening kit for agonist, antagonist or function modifying substance, containing the screening method and ligand, agonist, antagonist or function modifying substance obtained by using the screening kit and said ligand, agonist, antagonist or function modifying substance It is intended to provide a drug, an antibody against the polypeptide or a partial peptide thereof, and the like.

[0008]

Means for Solving the Problems The present inventors have conducted intensive studies and found that cDNA derived from human gastric cancer cell line KATOIII was used.
By randomly sequencing the sequence of each cDNA clone in the library, a cDNA encoding a portion of a novel GPCR that was previously unknown was successfully isolated. Since the cDNA lacked the C-terminal portion, the present invention was completed by obtaining a cDNA encoding the full length of the novel GPCR polypeptide from the human thalamus and determining and analyzing the entire nucleotide sequence.

The present invention relates to the following (1) to (35). (1) A G protein-coupled receptor polypeptide having the amino acid sequence of SEQ ID NO: 1. (2) a polypeptide having an amino acid sequence in which one or more amino acids have been deleted, substituted or added in the amino acid sequence of the polypeptide described in SEQ ID NO: 1, and substantially the same as the polypeptide described in (1) A polypeptide having the same activity as (3) A partial peptide which is a partial peptide of the G protein-coupled receptor polypeptide according to (1) or (2), and has a binding ability to a ligand, agonist, antagonist or functional modifier of the polypeptide. (4) A DNA encoding the G protein-coupled receptor polypeptide according to (1) or (2). (5) DN having a base sequence represented by base numbers 175 to 1287 in the DNA set forth in SEQ ID NO: 2.
A. (6) A DNA that hybridizes under stringent conditions with a DNA selected from the DNAs of (4) or (5), and substantially the same as the G protein-coupled receptor polypeptide of (1). DNA encoding a polypeptide having the same activity as the above. (7) A DNA encoding a partial peptide of the G protein-coupled receptor polypeptide according to (3). (8) a DNA encoding a partial peptide of the polypeptide encoded by the DNA according to (6), and the ligand of the polypeptide according to (1) or (2);
DNA encoding a partial peptide capable of binding to an agonist, antagonist, or function modifying substance. (9) The DNA according to any one of (4) to (8)
And a recombinant DNA obtained by incorporating the DNA into a vector. (10) A transformed cell, transformed plant or transformed non-human animal having the recombinant DNA according to (9). (11) Using the transformed cell, transformed plant or transformed non-human animal according to (10), (i) culturing the transformed cell in a medium, and (ii) performing the transformation in the culture. Cultivating a plant, and (iii) breeding the transformed non-human animal to produce the polypeptide according to (1) or (2) or the partial peptide according to (3) in the animal Collecting the polypeptide or the partial peptide from the culture, the plant or the animal, and collecting the polypeptide or the peptide according to (1) or (2). Production method. (12) The polypeptide according to (1) or (2),
Or an antibody that recognizes the partial peptide according to (3). (13) The method for immunologically quantifying the polypeptide according to (1) or (2) or the partial peptide according to (3) using the antibody according to (12). (14) A method for determining cancer or dysfunction of the thalamus or cerebellum using the quantification method according to (13). (15) A method for determining cancer or dysfunction of the thalamus or cerebellum by measuring the amount of mRNA encoding the polypeptide according to (1) or (2). (16) A method for determining cancer, or dysfunction of the thalamus or cerebellum by detecting deletion, substitution or addition of the gene encoding the polypeptide according to (1) or (2). (17) the polypeptide according to (1) or (2),
Alternatively, the partial peptide according to (3) is brought into contact with a test sample, and a ligand, agonist, antagonist or function-modifying substance of the polypeptide according to (1) or (2) is selected from the test sample. A method for screening for a ligand, agonist, antagonist or function modifying substance of the polypeptide according to (1) or (2). (18) the polypeptide according to (1) or (2),
Or a cell expressing the partial peptide according to (3),
A test sample is brought into contact with the test sample, and a ligand, an agonist of the polypeptide according to (1) or (2),
A method for screening for a ligand, agonist, antagonist or function-modifying substance of a polypeptide according to (1) or (2), which comprises selecting an antagonist or a function-modifying substance. (19) (i) The polypeptide according to (1) or (2) or the partial peptide according to (3) is brought into contact with a ligand and (ii) the polypeptide according to (1) or (2). Or the partial peptide described in (3) is compared with the case where the ligand and the test sample are brought into contact with each other, and the agonist, antagonist, or antagonist of the polypeptide described in (1) or (2) is compared with the test sample. A method for screening a polypeptide agonist, antagonist or function-modifying substance according to (1) or (2), which comprises selecting a function-modifying substance. (20) (i) contacting a cell that expresses the polypeptide according to (1) or (2) or the partial peptide according to (3) with a ligand; and (ii) (1) or ( A cell expressing the polypeptide according to 2) or the partial peptide according to (3) is compared with a case where a ligand and a test sample are brought into contact with each other.
Alternatively, the method for screening for an agonist, antagonist or function modifying substance of the polypeptide according to (1) or (2), comprising selecting an agonist, antagonist or function modifying substance for the polypeptide according to (2). (21) the polypeptide according to (1) or (2),
Or a kit for screening a ligand, agonist, antagonist or function-modifying substance of the polypeptide according to (1) or (2), which comprises the partial peptide according to (3). (22) A ligand, agonist, or antagonist of the polypeptide according to (1) or (2), which is obtained by using the screening method according to (17) to (20) or the screening kit according to (21). Or a function modifier, or a pharmacologically acceptable salt thereof. (23) A cancer or a thalamus containing a substance selected from the antibody according to (12), the ligand, agonist, antagonist and function-modifying substance according to (22), or a pharmacologically acceptable salt thereof. Drugs for cerebellar dysfunction. (24) (i) Contacting a cell expressing the polypeptide of (1) or (2) with (ii) a cell expressing the polypeptide of (1) or (2) and a test sample. (1) or (2) from the test sample
A method for screening a compound that changes the expression level of a DNA encoding the polypeptide according to (1) or (2), which comprises selecting a compound that alters the expression of the gene encoding the polypeptide according to (1). . (25) The method according to (24), wherein the variation in the expression level is measured by the method described in (13) or the method for quantifying the amount of mRNA encoding the polypeptide according to (1) or (2). ). (26) A test sample is brought into contact with a transformant containing a reporter gene-linked DNA downstream of a region controlling transcription of the gene encoding the polypeptide according to (1) or (2), and Selecting from the sample a compound that changes the expression level of the DNA encoding the polypeptide according to (1) or (2),
A method for screening a compound that changes the expression level of a gene encoding the polypeptide according to (1) or (2). (27) The region controlling transcription is 20 of SEQ ID NO: 15.
DNA represented by nucleotide sequence from position 202 to position 25202
Having a continuous base sequence of 50 to 5000 bp
The screening method according to (26), which is a region defined by NA. (28) A compound obtained by the screening method according to any one of (24) to (27) or a pharmacologically acceptable salt thereof. (29) A therapeutic agent for cancer or dysfunction of the thalamus or cerebellum, comprising the compound according to (28) or a pharmacologically acceptable salt thereof. (30) an oligonucleotide having the same sequence as 5 to 60 consecutive bases in the base sequence of a DNA selected from the DNA of (4) and (6) and the DNA of SEQ ID NO: 2, complementary to the oligonucleotide; Selected from oligonucleotides having specific sequences and oligonucleotide derivatives of these oligonucleotides. (31) DN according to (4) to (6) and (30)
Transcription of DNA encoding the polypeptide according to (1) or (2) or mR using a DNA selected from A
A method to suppress translation of NA. (32) All or a part of a gene containing a DNA encoding the polypeptide according to (1) or (2) is deleted or replaced, and the expression level of the polypeptide according to (1) or (2) is changed A non-human animal with a deleted or substituted gene. (33) The animal according to (32) is inoculated with a test sample, or the test sample is brought into contact with an organ, tissue or cell of the animal, and the test sample is used to treat cancer or thalamus or cerebellar dysfunction. Characterized by choosing a drug,
A method for screening a therapeutic agent for cancer or dysfunction of the thalamus or cerebellum. (34) A compound obtained by the screening method according to (33) or a pharmacologically acceptable salt thereof. (35) A therapeutic agent for cancer or dysfunction of the thalamus or cerebellum, comprising the compound according to (34) or a pharmacologically acceptable salt thereof.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION (1) G protein-coupled receptor polypeptide of the present invention or a partial peptide thereof The polypeptide of the present invention is a G protein-coupled receptor polypeptide, for example, represented by SEQ ID NO: 1. Or a polypeptide having an amino acid sequence in which one or more amino acids have been deleted, substituted or added in the amino acid sequence of the polypeptide, and having the amino acid sequence represented by SEQ ID NO: 1. A polypeptide having substantially the same activity as the polypeptide can be mentioned.

The origin of the polypeptide of the present invention is not particularly limited. Examples of the origin include humans and mammals (eg, guinea pig, rat, mouse, chicken, rabbit, pig, sheep, cow, monkey, etc.). Cells,
Alternatively, a tissue in which the cell exists can be mentioned.

Specific examples of the cells include spleen cells, nerve cells, glial cells, pancreatic β cells, bone marrow cells, mesangial cells, Langerhans cells, epidermal cells, epithelial cells, endothelial cells, fibroblasts, fiber cells, muscle cells Cells, fat cells, immune cells (eg, macrophages, T cells, B cells, natural killer cells, mast cells, neutrophils, basophils, eosinophils,
Monocytes), megakaryocytes, synovial cells, chondrocytes, osteocytes, osteoblasts, osteoclasts, mammary cells, hepatocytes or stromal cells, or precursors, stem cells or cancer cells of these cells Can be. Specific examples of the tissue include the brain, various parts of the brain (eg, olfactory bulb, nucleus pulposus, basal sphere
Hippocampus, thalamus, hypothalamus, hypothalamus nucleus, cerebral cortex, medulla, cerebellum, occipital lobe, frontal lobe, temporal lobe, putamen, caudate nucleus, brain stain, substantia nigra), spinal cord, pituitary gland, stomach, pancreas, Kidneys, liver, gonads,
Thyroid, gall bladder, bone marrow, adrenal gland, skin, muscle, lung, gastrointestinal tract, blood vessels, heart, thymus, spleen, submandibular gland, peripheral blood, peripheral blood cells, intestinal tract, prostate, testicle, testis, ovary, placenta, uterus,
Examples include bones, joints, small intestine, large intestine, and skeletal muscle. In particular, the brain and each part of the brain are preferable as tissues.

The polypeptide of the present invention may be a polypeptide synthesized by chemical synthesis.

A polypeptide comprising an amino acid sequence in which one or more amino acids are deleted, substituted or added, and having substantially the same activity as the polypeptide having the amino acid sequence represented by SEQ ID NO: 1 , Molecular cl
oning, A laboratory manual, Second Edition. (1989)
(Hereinafter abbreviated as Molecular Cloning 2nd Edition),
Current Protocols in Molecular Biology, John and W
ily & Sons (1987-1997) (hereinafter abbreviated as Current Protocols in Molecular Biology), Nu
cleic Acids Research, 10 , 6487 (1982), Proc. Natl.
Acad. Sci. USA, 79 , 6409 (1982), Gene, 34 , 315 (1
985), Nucleic Acids Research, 13 , 4431 (1985), Pro
c. Natl. Acad. Sci. USA, 82 , 488 (1985), etc., using a site-directed mutagenesis method, for example, to DNA encoding a polypeptide having the amino acid sequence represented by SEQ ID NO: 1. It can be obtained by introducing a site-specific mutation. The number of amino acids to be deleted, substituted or added is not particularly limited, but is a number that can be deleted, substituted or added by a known method such as the above-described site-directed mutagenesis, and is about 1 to several tens, Preferably 1-2
The number is about 0, more preferably 1 to 10, and still more preferably 1 to 5.

In order for a polypeptide having an amino acid sequence in which the amino acid has been deleted, substituted or added to have substantially the same activity as the polypeptide shown in SEQ ID NO: 1, BLAST [J. Mol. Biol. ., 215 , 403 (1990), Nucle
ic acids Research, 25 , 3389 (1997)), FASTA (Metho
d in Enzymology, 183 , 63 (1990), Proc. Natl. Acad.
Sci. USA, 85 , 2444 (1988)], the amino acid sequence represented by SEQ ID NO: 1 is about 70% or more, preferably about 80% or more, more preferably about 90%. It is preferable to have an amino acid sequence having the above homology.

The above-mentioned substantially identical activities include, for example, a ligand binding activity and a signal information transmitting activity of the peptide represented by the amino acid sequence of SEQ ID NO: 1. Substantially identical indicates that their activities are identical in nature. Therefore, the quantitative factors such as the ligand binding activity, the degree of signal transduction, and the molecular weight of the protein may be different.

As the polypeptide of the present invention, further, in the above-mentioned polypeptide, the amino group of the N-terminal methionine residue is protected with a protecting group (for example, a C1-6 acyl group such as a formyl group and an acetyl group). , A glutamyl group formed by cleavage of the N-terminal side in vivo and a pyroglutamate, a substituent on the side chain of an amino acid in the molecule (for example, -OH, -COOH, an amino group, an imidazole group, an indole group, Guanidino group) protected with an appropriate protecting group (eg, a C1-6 acyl group such as a formyl group or an acetyl group), or a complex protein such as a so-called glycoprotein to which a sugar chain is bound. .
Further, the C-terminus of the polypeptide is an amide (-CONH
₂ ) or an ester (-COOR) is the polypeptide of the present invention. Here, R of the ester group is methyl, ethyl, n-propyl, isopropyl or n-
C1-6 alkyl groups such as butyl, for example, C3-8 cycloalkyl groups such as cyclopentyl and cyclohexyl, for example, C6-12 aryl groups such as phenyl, α-naphthyl, etc., for example, phenyl-C1 such as benzyl, phenethyl and the like. In addition to a C7-14 aralkyl group such as an -2 alkyl group or an α-naphthyl-C1-2 alkyl group such as α-naphthylmethyl, a pivaloyloxymethyl ester generally used as an oral ester may be used. When the polypeptide of the present invention has a carboxyl group (or carboxylate) other than at the C-terminus, those in which the carboxyl group is amidated or esterified are also included in the polypeptide of the present invention. Examples of the ester at this time include the above-mentioned C-terminal ester and the like.

As the salt of the polypeptide of the present invention, a physiologically acceptable acid addition salt is particularly preferable. Such salts include, for example, inorganic acids (eg, hydrochloric acid, phosphoric acid,
Salts with hydrobromic acid, sulfuric acid, or organic acids (for example,
Examples thereof include salts with acetic acid, formic acid, propionic acid, fumaric acid, maleic acid, succinic acid, tartaric acid, citric acid, malic acid, oxalic acid, benzoic acid, methanesulfonic acid, and benzenesulfonic acid.

The partial peptide of the present invention is a partial peptide of the polypeptide of the present invention and a peptide capable of binding to the ligand, agonist, antagonist or functional modifier of the polypeptide of the present invention. GPC
It is known that the ligand binding region of R is an extracellular region, a transmembrane region, or both an extracellular region and a transmembrane region [Current Opinion of Cell Biology, 6 , 191
(1994), EMBO J., 18 , 1723 (1999)]. Therefore, examples of the partial peptide of the present invention having the ability to bind to the ligand, agonist, antagonist, or function-modifying substance of the polypeptide of the present invention include, for example, in the cell expressing the polypeptide, the outside of the cell membrane. (Extracellular region) or a partial peptide containing a membrane-bound region. Further, the partial peptide of the present invention may be a peptide individually containing each domain or a partial peptide containing a plurality of domains simultaneously.

[0020] The membrane-binding region of any GPCR may be a known GPCR.
It can be predicted based on homology with PCR [EM
BO J., 12 , 1693 (1993)]. Therefore, the extracellular region and intracellular region of any GPCR can be predicted based on the membrane-bound region predicted by the method. Hydropathy analysis (analysis software MacMolly3.
5) and the membrane-bound region prediction analysis (using the analysis software SOSUI system ver1.0 / 10 purchased from Mitsui Informational Development) can also be used to determine the membrane-bound region, extracellular region, and intracellular region of any GPCR. The region can be predicted.

Therefore, by performing the above analysis on the polypeptide of the present invention, specific extracellular regions (hydrophilic regions), membrane-bound regions (hydrophobic regions), and intracellular regions (hydrophilic regions) can be predicted. can do.

Specifically, for example, in the polypeptide of the present invention having the amino acid sequence represented by SEQ ID NO: 1, the extracellular region includes the first to 49th amino acids of the amino acid sequence represented by SEQ ID NO: 1. , 107th to 1st
21st, 187th to 208th or 298th
And the region represented by the amino acid sequence at positions 50-75 and 81-106 of the amino acid sequence represented by SEQ ID NO: 1. Th, 122nd to 14th
7th, 161st to 186th, 209th to
234th, 272nd to 297th or 3rd
The region represented by the 10th to 335th amino acid sequences can be exemplified.

As the partial peptide of the present invention, further, in the above partial peptide, the amino group of the N-terminal methionine residue is protected by a protecting group (for example, a C1-6 acyl group such as a formyl group or an acetyl group). , A glutamyl group formed by cleavage of the N-terminal side in vivo and a pyroglutamate, a substituent on the side chain of an amino acid in the molecule (for example, -OH, -COOH, an amino group, an imidazole group, an indole group) , A guanidino group or the like) protected with a suitable protecting group (eg, a C1-6 acyl group such as a formyl group or an acetyl group), or a complex peptide such as a so-called glycopeptide having a sugar chain bonded thereto. It is. Further, a carboxyl group (-COOH) or a carboxylate (-
COO ^-) is, as with the polypeptides of the present invention described above,
It may be amide or esterified. When the partial peptide of the present invention has a carboxyl group (or carboxylate) other than the C-terminus, those in which the carboxyl group is amidated or esterified are also included in the partial peptide of the present invention. Examples of the ester at this time include the above-mentioned C-terminal ester and the like. As the salt of the partial peptide of the present invention, a physiologically acceptable acid addition salt is particularly preferable. Such salts include:
For example, salts with inorganic acids (eg, hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid) or organic acids (eg, acetic acid, formic acid, propionic acid, fumaric acid, maleic acid, succinic acid, tartaric acid, citric acid, apple) Acid, oxalic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid) and the like. (2) DNA encoding the G protein-coupled receptor polypeptide of the present invention or a partial peptide thereof The DNA encoding the polypeptide of the present invention may be any DNA as long as it encodes the polypeptide of the present invention.
A, and specific examples thereof include (a) DNA having a base sequence represented by base numbers 175 to 1287 in the base sequence represented by SEQ ID NO: 2, and (b) DN having the base sequence represented by (a).
DNA that hybridizes with A under stringent conditions and encodes a polypeptide having substantially the same activity as the polypeptide having the amino acid sequence of SEQ ID NO: 1 can be mentioned.

The DNA that hybridizes under the above-mentioned stringent conditions and encodes a polypeptide having substantially the same activity as the polypeptide having the amino acid sequence of SEQ ID NO: 1 is the above-mentioned (a). D described in
A DNA obtained by using a colony hybridization method, a plaque hybridization method, a Southern blot hybridization method, or the like with NA as a probe. After hybridization at 42-65 ° C. in the presence of 0.7-1.0 mol / L NaCl, and then 0.1- to 2-fold concentration of SSC (saline-sodium citrat).
e) Solution (The composition of a 1-fold concentration SSC solution is 150 mmol
/ L sodium chloride and 15 mmol / L sodium citrate) and washing the filter under the conditions of 42 to 65 ° C. Hybridization is performed using Molecular Cloning 2nd Edition, Current Protocols in
Molecular biology, DNA Cloning 1: Core Te
chniques, A Practical Approach, Second Edition, Oxf
It can be performed according to the method described in ord University Press (1995) and the like. Hybridizable DN
As A, specifically, a DNA having at least 60% or more homology with the DNA described in (a) above, and preferably 80% or more when calculated using analysis software such as BLAST or FASTA. DNAs having homology, more preferably DNAs having homology of 95% or more can be mentioned.

DNA encoding the partial peptide of the present invention
May be any DNA as long as it encodes the partial peptide of the present invention, and specific examples thereof include (a)
In the nucleotide sequence of SEQ ID NO: 2,
A DNA having a partial nucleotide sequence selected from the nucleotide sequence represented by No. 1287, and a portion having a binding ability to a ligand, agonist, antagonist or function modifying substance of the polypeptide having the amino acid sequence of SEQ ID NO: 1 A DNA encoding a peptide, (b) a DNA having a partial nucleotide sequence selected from a nucleotide sequence representing a DNA that hybridizes under stringent conditions with a DNA encoding the polypeptide of the present invention, and described in SEQ ID NO: 1. And DNA encoding a peptide capable of binding to a ligand, agonist, antagonist, or function-modifying substance of a polypeptide having the following amino acid sequence.

DNA encoding the partial peptide of the present invention
Is a ligand, an agonist of the polypeptide of the present invention,
DNA encoding a partial peptide of the polypeptide of the present invention, which is capable of binding to an antagonist or a function-modifying substance, for example, a portion containing an extracellular region or a membrane-bound region predicted by the method described in (1) above. Examples include DNA encoding a peptide. Specifically, the first to 49th amino acid sequences represented by SEQ ID NO: 1
No. 175 of the nucleotide sequence represented by SEQ ID NO: 2, which is a DNA encoding a partial peptide having an amino acid sequence of the 107th to the 121st, the 187th to the 208th or the 298th to the 309th amino acid Th ~
321nd, 493rd to 537th, 733rd
Th to 798 th or 1066 th to 1101 th
And a DNA having the second base sequence.
In addition, the 50th to 75th, the 81st to the 106th, and the 122nd of the amino acid sequence represented by SEQ ID NO: 1.
Is a DNA encoding a partial peptide having an amino acid sequence at positions 147th, 161st to 186th, 209th to 234th, 272th to 297th, or 310th to 335th. No. 322 to No. 399, No. 415 to No. 492, No. 538 to No. 6 of the nucleotide sequence represented by SEQ ID NO: 2.
The 15th, 655th to 732rd, 799th to 876th, 988th to 1065th or 1102th to 1179th base sequences can be mentioned. Further, the DNA encoding the partial peptide of the present invention may be a DNA encoding each partial peptide individually.
A or a DNA containing a plurality of DNAs encoding individual partial peptides at the same time.

(3) Obtaining the DNA encoding the polypeptide of the present invention, and producing the DNA and the oligonucleotide The DNA encoding the polypeptide of the present invention may be, for example,
All cells of humans and mammals (eg, guinea pig, rat, mouse, chicken, rabbit, pig, sheep, cow, monkey, etc.) (eg, spleen cells, nerve cells, glial cells, pancreatic β cells, bone marrow cells, mesangial cells) , Langerhans cells, epidermal cells, epithelial cells, endothelial cells, fibroblasts, fibrocytes, muscle cells, adipocytes, immune cells (eg, macrophages, T cells, B cells, natural killer cells, mast cells, neutrophils, Basophils, eosinophils, monocytes), megakaryocytes, synovial cells, chondrocytes, osteocytes, osteoblasts, osteoclasts, mammary cells, hepatocytes or stromal cells, or precursor cells of these cells, Stem cells or cancer cells), or any tissue in which these cells are present, for example, the brain, various parts of the brain (eg, olfactory bulb, nucleus pulposus, basal sphere, hippocampus, thalamus, hypothalamus, thalamus Nucleus, cerebral cortex, medulla, cerebellum, occipital lobe, frontal lobe, temporal lobe, putamen, caudate nucleus, brain stain, substantia nigra), spinal cord, pituitary, stomach, pancreas, kidney, liver, gonad, thyroid, gall bladder , Bone marrow, adrenal gland, skin, muscle, lungs,
Gastrointestinal tract, blood vessels, heart, thymus, spleen, submandibular gland, peripheral blood, peripheral blood cells, intestinal tract, prostate, testes, testis, ovaries, placenta, uterus, bones, joints, small intestine, large intestine, skeletal muscle, etc. Genomic DNA, genomic DNA library, cDNA derived from the above cells or tissues, or c
Each genomic DNA selected from a DNA library etc.
Alternatively, it can be obtained by randomly sequencing cDNA.

Preparation of genomic DNA and preparation of a genomic DNA library can be carried out, for example, using the above-mentioned various cells, organs or tissues in accordance with a conventional method. Specific methods include Molecular Cloning 2nd Edition and Current Cloning
Examples include the methods described in Protocols in Molecular Biology.

The preparation of cDNAs and cDNA libraries can be performed, for example, by using m-cells, organs, or tissues derived from
It can be prepared by an ordinary method using RNA. In particular,
Molecular cloning 2nd edition, Current Protocols in Molecular Biology, DNA
Cloning 1: Core Techniques, A Practical Approach,
Second Edition, Oxford University Press (1995), etc., a method for preparing full-length cDNA [Methods in
Enzymology, 303 , 19 (1999), Gene, 138 , 171 (199
4)] or a commercially available kit such as Superscript Plasmid System for cDNA Synthesis and Plasmid Cloning [SuperScrip
t Plasmid System for cDNA Synthesis and Plasmid Cl
oning; Gibco BRL) or Zap-c
DNA Synthesis Kit [ZAP-cDNA Synthesis Ki
t, manufactured by Stratagene).

As a cloning vector for preparing a library, any phage vector, plasmid vector, or the like can be used as long as it can replicate autonomously in E. coli K12 strain. Specifically, ZAP Express
(Strategies, 5 , 58 (1992), manufactured by Stratagene)
pBluescript II SK (+) (Nucleic Acids Research, 17 , 9
494 (1989)], λzap II (manufactured by Stratagene),
λgt10, λgt11 [DNA Cloning, A Practical
Approach, 1 , 49 (1985)], λTriplEx (Clontech), λExCell (Pharmacia), pT7T
318U (Pharmacia), pcD2 [Mol. Cell. Bi
ol., 3 , 280 (1983)], pUC18 [Gene, 33 , 103
(1985)], pAMo [J. Biol. Chem., 268 , 22782-227.
87 (1993), also known as pAMoPRC3Sc (Japanese Patent Laid-Open No. 05-336963), p
AMo-d (see Example 1) and the like.

As the host microorganism used for preparing the library, any microorganism belonging to Escherichia coli can be used. Specifically, Escherichia coli XL1-B
lueMRF '[Stratagies, 5 , 81 (19
92)), Escherichia coli C600 [Genetics, 39 , 440 (1
954)), Escherichia coli Y1088 (Science, 222 , 778
(1983)), Escherichia coli Y1090 (Science, 222 , 7
78 (1983)), Escherichia coli NM522 (J. Mol. Bio
l., 166 , 1 (1983)], Escherichia coli K802 (J. Mo.
l. Biol., 16 , 118 (1966)], Escherichia coli JM10
5 (Gene, 38 , 275 (1985)), Escherichia coli SOLR
^TM Strain (commercially available from Stratagene) and E. coli
LE392 (Molecular Cloning 2nd Edition) or the like can be used.

More specific methods for preparing a cDNA library include the following methods.

MRNA is extracted from human gastric cancer cells KATOIII according to the method described in Molecular Cloning, Second Edition, and cDNA is synthesized by the oligocap method [Gene, 138 , 171 (1994)]. Next, protein nucleic acid enzyme,
41 , 197 (1996) and Gene, 138 , 171 (1994). After synthesizing a first-strand cDNA, using a primer designed on the 5 'end and 3' end of the cDNA,
se chain reaction [Molecular Cloning 2
Plate and PCR Protocols Academic Press (1990), hereinafter abbreviated as PCR). The cDNA was cleaved with restriction enzyme Sfi I, cloning vector pME18SFL3 (GenBank AB00986 cut with Dra I
4, Expression vector, 3392bp)
A DNA library can be prepared.

For the nucleotide sequence of cDNA, each E. coli clone constituting a cDNA library is randomly selected, a plasmid DNA is prepared from the clone, and the nucleotide sequences at both ends of the cDNA contained in the plasmid are usually used. Nucleotide sequence analysis method, for example, the dideoxy method of Sanger et al. [Proc. Natl. Acad. Sci. USA, 74 , 546].
3 (1977)] or a nucleotide sequence analyzer such as a 373A DNA sequencer (manufactured by Perkin Elmer).

The presence of the thus obtained gene having homology to the base sequence or a protein having homology to the amino acid sequence encoded by the base sequence is examined by database search. Blast, FrameSearch method (Co
mpugen) can be used.
As the database, a public database such as GenBank can be used, or a private database can also be used. As a result of the search, the entire nucleotide sequence of a cDNA showing homology with a known GPCR at the base or amino acid level is determined, and the entire amino acid sequence of the polypeptide encoded by the cDNA is determined.

When the cDNA does not encode a full-length polypeptide, a cDNA encoding the full-length polypeptide can be obtained as follows.

Using a single-stranded cDNA library prepared from various organs or cells or a cDNA library prepared by the method described above as a template, PCR is performed using a primer set specific to the cDNA, An organ or cell expressing the gene corresponding to the cDNA is specified, and cDN derived from the specified organ or cell is identified.
The library A was newly subjected to a colony hybridization method (molecular cloning, second edition) using the cDNA as a probe to newly obtain the cDNA.
Can be selected from a cDNA library.

Further, using a single-stranded cDNA library or a cDNA library derived from an organ or a cell expressing the gene corresponding to the cDNA as a template, 5 ′ RAC
E method and 3 'RACE method (Proc. Natl. Acad. Sci. USA, 85 , 89
98 (1988)], a 5′-end fragment and a 3′-end fragment of the cDNA are obtained, and by linking both fragments, a full-length cDNA can be obtained. In addition, commercially available kits (for example, Boehringer's 5 '/ 3' RA
The 5 'RACE method and the 3' RACE method can also be performed using a CE kit.

Single chain cD derived from various organs or cells
The NA library can be prepared by a conventional method or a commercially available kit. For example, the NA library can be prepared by the following method.

From various organs or various cells, a guanidium thiocyanate phenol-chloroform method [Anal.
Biochem., 162 , 156 (1987)]. If necessary, the total RNA is treated with deoxyribonuclease I (manufactured by Life Technologies) to degrade chromosomal DNA that may be contaminated. Total RNA obtained
For each, SUPERSCRIPT ^™ Preamplification using oligo (dT) or random primers
System for First Strand cDNA System (LifeTechnolo
gies) to produce a single-stranded cDNA library. The single-stranded cDNA library prepared as described above includes, for example, a single-stranded cDNA prepared from the human thalamus.
NA libraries can be given.

The entire nucleotide sequence of the cDNA encoding the full-length polypeptide obtained as described above is determined, and the amino acid sequence of the polypeptide is again searched in the database in the same manner as described above, and the known GPCR and Can be examined for homology. Further, a hydrophobicity plot analysis is performed using the amino acid sequence of the polypeptide, and it is determined whether the polypeptide has a seven-transmembrane region common to GPCRs. The polypeptide has a seven-transmembrane region, If the polypeptide shows homology with a known GPCR, the polypeptide can be considered to be a GPCR. If the polypeptide is different from the known GPCR, the polypeptide can be considered a novel GPCR.

Further, based on the determined amino acid sequence of the novel polypeptide, DN encoding the polypeptide is
The target DNA can also be prepared by chemically synthesizing A. Chemical synthesis of DNA can be performed using a DNA synthesizer manufactured by Shimadzu Corporation using a thiophosphite method, a DNA synthesizer model 392 manufactured by Perkin-Elmer Inc. using a phosphoramidite method, or the like.

Using the oligonucleotides described below as sense primers and antisense primers, the mRN of cells expressing mRNA complementary to these DNAs was used.
The target DNA can also be prepared by performing PCR using the cDNA prepared from A as a template.

As the DNA encoding the novel G protein-coupled receptor polypeptide obtained by the above method,
For example, a DNA encoding the polypeptide represented by SEQ ID NO: 1 and the like can be mentioned. Specifically, in the nucleotide sequence of SEQ ID NO: 2, base numbers 175 to 1287
And the like having a base sequence represented by No. DNA having a nucleotide sequence represented by nucleotide numbers 175 to 1287 in the nucleotide sequence of SEQ ID NO: 2
Is encoded by hydropathy analysis, membrane-bound region prediction analysis method, and ENBO J., 12 , 1693
According to the method described in (1993), the following structure is expected (see FIGS. 1 and 2 to 10). N-terminal extracellular domain (49 amino acids), first transmembrane domain (26 amino acids), first intracellular loop (5 amino acids), second transmembrane domain (26 amino acids), first extracellular loop (15 amino acids) , Third transmembrane domain (26 amino acids), second intracellular loop (13 amino acids), fourth transmembrane domain (26 amino acids), second extracellular loop (22 amino acids), fifth transmembrane domain (26 amino acids) , Third intracellular loop (37 amino acids), sixth transmembrane region (26 amino acids), third extracellular loop (12 amino acids), seventh transmembrane region (26 amino acids), C-terminal intracellular region (36 amino acids) ).

From the hydropathic analysis of the amino acid sequence, it is considered that the polypeptide does not have a signal peptide (see FIG. 1).

DN having the nucleotide sequence of SEQ ID NO: 2
Examples of the plasmid containing A include pBS-KAT
O6734L. pBS-KA
Escherichia coli harboring TO0673L
DH5α / pBS-KATO0673L is FERM B
P-6967 has been deposited with the Institute of Biotechnology and Industrial Technology, Institute of Industrial Science and Technology, 1-3-1 Higashi, Tsukuba, Ibaraki, Japan (Postal Code 305-8566) on December 8, 1999.

Using the DNA and DNA fragment of the present invention obtained by the above-mentioned method, the DNA is synthesized by a conventional method described in Molecular Cloning, 2nd edition, etc., or by a DNA synthesizer using the nucleotide sequence information of the DNA. Oligonucleotides such as antisense oligonucleotides and sense oligonucleotides having a partial sequence of the DNA of the present invention can be prepared.

Examples of the oligonucleotide include a DNA having the same base sequence as 5 to 60 consecutive bases in the base sequence of the DNA of the present invention or a DNA having a sequence complementary to the DNA. Examples of the DNA include a DNA having the same sequence as 5 to 60 consecutive nucleotides in the base sequence of the DNA described in SEQ ID NO: 2 or a DNA having a sequence complementary to the DNA. When used as a sense primer and an antisense primer, it is preferable to select an oligonucleotide whose melting temperature (Tm) and the number of bases of both do not extremely change. Specifically, for example, a primer set of an oligonucleotide having the nucleotide sequence represented by SEQ ID NO: 11 and SEQ ID NO: 12, a primer set of an oligonucleotide having the nucleotide sequence represented by SEQ ID NO: 11 and SEQ ID NO: 17, or And primer sets of oligonucleotides having the nucleotide sequences represented by SEQ ID NOs: 13 and 14.

Further, derivatives of these oligonucleotides (hereinafter referred to as oligonucleotide derivatives) can also be used as the oligonucleotide of the present invention. Examples of the oligonucleotide derivative include an oligonucleotide derivative in which a phosphodiester bond in an oligonucleotide is converted to a phosphorothioate bond, and a phosphodiester bond in an oligonucleotide converted to an N3′-P5 ′ phosphoramidate bond. Oligonucleotide derivative, the ribose and the phosphodiester bond in the oligonucleotide are converted to a peptide nucleic acid bond, the uracil in the oligonucleotide is C-
Oligonucleotide derivatives substituted with 5-propynyluracil, oligonucleotide derivatives in which uracil in the oligonucleotide is substituted with C-5 thiazoleuracil, oligonucleotide derivatives in which cytosine in the oligonucleotide is substituted with C-5 propynylcytosine, oligo Oligonucleotide derivative in which cytosine in nucleotide is replaced by phenoxazine-modified cytosine, oligonucleotide derivative in which ribose in oligonucleotide is replaced by 2′-O-propyl ribose, or ribose in oligonucleotide Can be exemplified by oligonucleotide derivatives substituted with 2'-methoxyethoxyribose [Cell Engineering, 16 , 1463 (1997)]. (4) Method for Producing Polypeptide and Partial Peptide of the Present Invention The polypeptide of the present invention or a salt thereof can be produced from the above-mentioned method for purifying a protein from human or mammalian cells or tissues, and will be described later. It can also be produced using a transformant containing a DNA encoding the polypeptide of the present invention. Further, it can also be produced according to the peptide synthesis method described later. When producing from human or mammalian tissues or cells, for example, after homogenizing human or mammalian tissues or cells, extraction is performed with an acid or the like, and the extract is subjected to chromatography such as reverse phase chromatography or ion exchange chromatography. Isolation and purification can be performed by combining the chromatography.

The partial peptide of the present invention or a salt thereof can be synthesized according to a known peptide synthesis method, or can be produced by cleaving the G protein-coupled receptor polypeptide of the present invention with an appropriate peptidase. can do. As a method for synthesizing a peptide, for example, any of a solid phase synthesis method and a liquid phase synthesis method may be used.
That is, the target peptide can be produced by condensing a partial peptide or amino acid capable of constituting the partial peptide of the present invention with the remaining portion, and removing the protective group when the product has a protective group. Known condensation methods and elimination of protecting groups include, for example, the following (a) to
The method described in (e) is mentioned. (A) M. Bodanszky and MA Ondetti, peptide
Synthesis (Peptide Synthesis), Interscience Publi
shers, New York (1966) (b) Schroeder and Luebke, The Pepti
de), Academic Press, New York (1965) (c) Nobuo Izumiya et al., Fundamentals and experiments of peptide synthesis, Maruzen
(1975) (d) Haruaki Yajima and Shunpei Sakakibara, Laboratory of Biochemical Experiments 1,
Protein Chemistry IV, 205, (1977) (e) Supervised by Haruaki Yajima, Development of Continuing Drugs Volume 14 Peptide Synthesis Hirokawa Shoten Also, after the reaction, conventional purification methods such as solvent extraction, distillation, and column chromatography -The partial peptide of the present invention can be purified and isolated by a combination of liquid chromatography, recrystallization and the like. When the partial peptide obtained by the above method is a free form, it can be converted to an appropriate salt by a known method, and conversely, when it is obtained by a salt, it can be converted to a free form by a known method. Can be.

In addition to the above method, the polypeptide or partial peptide of the present invention can be produced by expressing the DNA of the present invention obtained in the above (3) in a host cell.

That is, a recombinant DNA in which the DNA of the present invention is inserted downstream of a promoter of an appropriate expression vector is constructed, and the recombinant DNA is introduced into a host cell, whereby the polypeptide or partial peptide of the present invention is obtained. A polypeptide or partial peptide of the present invention can be produced by obtaining a transformant that expresses and then culturing the transformant.

As the host cell, any prokaryotic cell, yeast, animal cell, insect cell, plant cell, or the like can be used as long as it can express the gene of interest. Moreover, an animal individual or a plant individual can also be used.

As the expression vector, those which can replicate autonomously in the above-mentioned host cells or can be integrated into a chromosome and which contain a promoter at a position suitable for transcription of the DNA of the present invention can be used. .

When a prokaryote such as a bacterium is used as a host cell, the DNA expression vector of the present invention is capable of autonomous replication in a prokaryote, and has a promoter, a ribosome binding sequence, a novel receptor gene, a transcription termination sequence. , It is preferable to be constituted. A gene that controls the promoter may be included.

As an expression vector, for example, pBTr
p2, pBTac1, pBTac2 (all commercially available from Boehringer Mannheim), pSE280 (Invitrogen), pGEMEX-1 (Promega),
pQE-8 (manufactured by QIAGEN), pKYP10 (JP-A-58
-110600), pKYP200 [Agric. Biol. Che
m., 48 , 669 (1984)], pLSA1 [Agric. Biol. Che.
m., 53 , 277 (1989)], pGEL1 [Proc. Natl. Aca
d. Sci., USA, 82 , 4306 (1985)], pBluescript II SK
(-) (STRATAGENE), pTrs30 (FERM BP)
-5407), pTrs32 (FERM BP-540)
8), pGHA2 (FERM BP-400), pGK
A2 (FERM B-6798), pTerm2 (JP-A-3-22979, US4686191, US4939)
094, US5160735), pKK233-2 (Ph
armacia), pGEX (Pharmacia), pET system (Novagen), pSuex, pUB11
0, pTP5, pC194, pTrxFus (Invitrog
en company), pMAL-c2 (New England Biolabs) and the like.

The promoter may be any promoter as long as it can be expressed in host cells such as E. coli.
For example, trp promoter (P trp), lac promoter (P lac), P _L promoter, such as P _R promoter,
Promoters derived from E. coli, phage, etc., SPO1
Promoter, SPO2 promoter, penP promoter and the like can be mentioned. In addition, artificially designed and modified promoters such as a promoter in which two Ptrps are connected in series (P trp x2), a tac promoter, a lacT7 promoter, and a let I promoter can also be used.

As the ribosome binding sequence, Shine-
It is preferable to use a plasmid in which the distance between the Shine-Dalgarno sequence and the initiation codon is adjusted to an appropriate distance (for example, 6 to 18 bases).

Although a transcription termination sequence is not always necessary for expression of the DNA of the present invention, it is preferable to arrange a transcription termination sequence immediately below a structural gene.

As the host cell, microorganisms belonging to the genus Escherichia, Serratia, Bacillus, Brevibacterium, Corynebacterium, Microbacterium, Pseudomonas, etc., for example, Escherichia coli XL1-B
lue, Escherichia coli XL2-Blue, Escherichia coli D
H1, Escherichia coli MC1000, Escherichia coli KY32
76, Escherichia coli W1485, Escherichia coli JM10
9, Escherichia coli HB101, Escherichia coli No.4
9, Escherichia coli W3110, Escherichia coli NY49,
Escherichia coli BL21 (DE3), Escherichia coli BL21
(DE3) pLysS, Escherichia coli HMS174 (DE3), Escheric
hia coli HMS174 (DE3) pLysS, Serratia ficaria , Serra
tia fonticola , Serratia liquefaciens , Serratia mar
cescens , Bacillus subtilis , Bacillus amyloliquefac
iens , Corynebacterium ammoniagenes , Brevibacterium
immariophilum ATCC14068, Brevibacterium saccharol
yticum ATCC14066, Corynebacterium glutamicum ATCC13
032, Corynebacterium glutamicum ATCC14067, Coryneb
acterium glutamicum ATCC13869, Corynebacterium ace
toacidophilum ATCC13870, Microbacterium ammoniaphi
lum ATCC15354, Pseudomonas sp. D-0110 and the like.

As a method for introducing a recombinant vector, any method for introducing DNA into the above host cells can be used. For example, electroporation [Nu
cleic Acids Res., 16 , 6127 (1988)], a method using calcium ions [Proc. Natl. Acad. Sci. USA, 69 ,
2110 (1972)], the protoplast method (JP-A-63-24839).
4), Gene, 17 , 107 (1982) and Molecular & General Gen
etics, 168 , 111 (1979).

When a yeast strain is used as a host cell, as an expression vector, for example, YEp13 (ATCC37)
115), YEp24 (ATCC37051), YCp50 (ATCC37
419), pHS19, pHS15 and the like. Any promoter can be used as long as it can be expressed in yeast strains.
Promoter, PGK promoter, GAP promoter, ADH promoter, gal 1 promoter, g
Examples of the promoter include an al10 promoter, a heat shock protein promoter, a MFα1 promoter, and a CUP1 promoter.

As the host cell, Saccharomyces sp.
Yeast strains belonging to the genus Schizosaccharomyces, Kluyveromyces, Trichosporone, Siwaniomyces and the like can be mentioned, and specifically, Saccharomyces cerevisi
ae , Schizosaccharomyces pombe , Kluyveromyces lacti
s , Trichosporon pullulans , Schwanniomyces alluvius
Etc. can be given. Alternatively, a mutant suitable for GPCR expression can be used [Trends in Biotechnolog
y, 15 , 487 (1997), Mol.Cell.Biol., 15 , 6188 (199
5), Mol. Cell. Biol., 16 , 4700 (1996)].

As a method for introducing a recombinant vector, any method for introducing DNA into yeast can be used. For example, an electroporation method [Methods.
n Enzymol., 194 , 182 (1990)), spheroplast method (Proc. Natl. Acad. Sci. USA, 84 , 1929 (1978)),
Lithium acetate method (J. Bacteriol., 153 , 163 (1983)),
Natl. Acad. Sci. USA, 75 , 1929 (1978).

When an animal cell is used as a host cell, pcDNAI / Am may be used as an expression vector.
p, pcDNAI, pCDM8, pAGE107, pR
EP4, pAGE103, pAMo, pAMoA, pA
Mod-d (see Example 1), pAS3-3 and the like can be exemplified.

Any promoter can be used as long as it can be expressed in animal cells. For example, the IE (imme) of cytomegalovirus (human CMV) can be used.
diateearly) promoter, early promoter of SV40, long terminal repeat promoter of Moloney Murine Leukemia Virus
at promoter, a retrovirus promoter, a heat shock promoter, an SRα promoter, a metallothionein promoter, and the like. Further, an enhancer of the IE gene of human CMV may be used together with the promoter.

As host cells, mouse myeloma cells, rat myeloma cells, mouse hybridoma cells, CHO cells, BHK cells, African green monkey kidney cells, Namalwa cells, Namalwa KJM-1 cells, human fetal kidney cells, human leukemia Cell, HBT5637 (JP-A-63-299), human colon cancer cell line, frog oocytes, frog melanocytes and the like.

Further, as mouse / myeloma cells,
YB2 / 0 for rat myeloma cells such as SP2 / 0 and NSO
HEK293 etc. as human fetal kidney cells, BALL-1 etc. as human leukemia cells, COS-1 and COS-7 as African green monkey kidney cells, HCT-15 etc. as human colon cancer cell line. it can.

As a method for introducing a recombinant vector, any method for introducing DNA into animal cells can be used. For example, an electroporation method [Cytote
chnology, 3 , 133 (1990)], calcium phosphate method (Japanese Patent Laid-Open No. 2-227075), lipofection method [Proc. Natl. Ac
ad. Sci. USA, 84 , 7413 (1987)], Virology, 52 , 456.
(1973). Obtaining and culturing the transformant can be performed according to the method described in JP-A-2-227075 or JP-A-2-25791.

When an insect cell is used as a host, for example, baculovirus expression vectors, a laboratory manual [Baculovirus Exp.
ression Vectors, A Laboratory Manual, WH Freema
n and Company, New York (1992)], Molecular Biology A Laboratory Manual (Molecular
Biology, A Laboratory Manual), Current Protocols in Molecular Biology, Bio / T
The polypeptide can be expressed by the method described in Echinology, 6 , 47 (1988) and the like.

That is, after a recombinant gene transfer vector and a baculovirus are co-transfected into insect cells to obtain a recombinant virus in the culture supernatant of insect cells, the recombinant virus is further infected into insect cells, and the polypeptide is transferred to the insect cells. Can be expressed.

The gene transfer vector used in the method includes, for example, pVL1392, pVL1393, pBlueBacII
I (all manufactured by Invitrogen) and the like.

Examples of the baculovirus include, for example, Autographa californica nuclea polyhedrosis virus, which is a virus that infects night moth insects
(Autographa californica nuclear polyhedrosis viru
s) etc. can be used.

As insect cells, Spodoptera frugiperd
a ovarian cells, ovarian cells of Trichoplusia ni, can be used cultured cells derived from silkworm ovary. Spodoptera f
The ovarian cells of rugiperda include Sf9 and Sf21 (Baculovirus Expression Vectors A Laboratory Manual) and the like. The ovarian cells of Trichoplusia ni are High 5, BTI-TN-5B1-4 (manufactured by Invitrogen)
Bombyx mori N4 is a cultured cell derived from silkworm ovary.
Etc. can be given.

As a method for co-transferring the above-mentioned recombinant gene transfer vector and the above baculovirus into insect cells for preparing a recombinant virus, for example, a calcium phosphate method (Japanese Patent Laid-Open No. 2-227075), a lipofection method [Proc.
Natl. Acad. Sci. USA, 84 , 7413 (1987)].

In addition, DNA can be introduced into insect cells using the same method as that for introducing DNA into animal cells. For example, electroporation [Cytotech.
nology, 3 , 133 (1990)], calcium phosphate method (Japanese Patent Laid-Open No. 2-227075), lipofection method [Proc. Natl. Aca
d. Sci. USA, 84 , 7413 (1987)].

When a plant cell or a plant individual is used as a host, a known method [tissue culture, 20 (1994), tissue culture, 21 (1995), Trends in Biotechnology, 15 , 45 (1)
997)].

As the promoter used for gene expression, any promoter can be used as long as it can be expressed in plant cells. Examples thereof include the cauliflower mosaic virus (CaMV) 35S promoter and the rice actin 1 promoter. Further, by inserting intron 1 and the like of a maize alcohol dehydrogenase gene between the promoter and the gene to be expressed,
Gene expression efficiency can also be increased.

The host cells include potato, tobacco,
Corn, rice, rape, soy, tomato, wheat,
Plant cells such as barley, rye, alfalfa, flax and the like can be mentioned.

[0080] Introduction of the recombinant vector can be carried by any of the methods for introducing DNA into plant cells, for example, Agrobacterium (Agrobacterium
m ) (JP-A-59-140885, JP-A-60-70080, WO94 / 0097)
7), electroporation [Cytotechnology, 3 ,
133 (1990), JP-A-60-251887], a method using a particle gun (gene gun) (Patent Nos. 2606856, 251
7813).

The cells or organs of the plant into which the gene has been introduced can be cultured in large quantities using a jar fermenter. Further, a plant individual (transgenic plant) into which the gene has been introduced can also be created by redifferentiating the plant cell into which the gene has been introduced.

The polypeptide of the present invention can be produced using an animal individual. For example, known methods [American
Journal of Clinical Nutrition, 63 , 639S (1996), A
American Journal of Clinical Nutrition, 63 , 627S (1
996), Bio / Technology, 9 , 830 (1991)], and the polypeptide of the present invention can be produced in an animal into which a gene has been introduced.

As the promoter, any promoter that can be expressed in animals can be used.
Mammary cell-specific promoters such as α-casein promoter, β-casein promoter, β-lactoglobulin promoter, whey acidic protein promoter and the like are preferably used.

A transformant derived from a microorganism, animal cell, or plant cell having a recombinant DNA into which DNA encoding the polypeptide or partial peptide of the present invention has been incorporated is cultured according to a conventional culture method. The polypeptide can be produced by producing and accumulating the peptide and collecting the polypeptide from the culture.

When the transformant is an individual animal or plant, the animal is bred or cultivated according to a usual method to produce and accumulate the polypeptide, and the polypeptide is collected from the individual animal or plant. And the polypeptide can be produced.

That is, in the case of an animal individual, for example, a non-human transgenic animal having the DNA of the present invention is bred and the polypeptide or partial peptide of the present invention encoded by the recombinant DNA is produced in the animal. -The polypeptide or partial peptide of the present invention can be produced by accumulating and collecting the polypeptide or partial peptide from the animal. Examples of the production / accumulation site in the animal include cell membrane fractions of various cells, milk, eggs, and the like of the animal.

In the case of a plant individual, for example, the DNA of the present invention
Cultivating a transgenic plant carrying the recombinant DNA, producing and accumulating the polypeptide or partial peptide of the present invention encoded by the recombinant DNA in the plant, and collecting the polypeptide or partial peptide from the plant. Thus, the polypeptide or partial peptide of the present invention can be produced.

When the transformant for producing the polypeptide or partial peptide of the present invention is a prokaryote such as Escherichia coli, or a eukaryote such as yeast, the medium for culturing these organisms may be a carbon-containing medium capable of assimilating the organism. Any of a natural medium and a synthetic medium may be used as long as the medium contains a source, a nitrogen source, inorganic salts, and the like, and can efficiently culture the transformant.

The carbon source may be any as long as the transformant can assimilate, such as glucose, fructose, sucrose, molasses containing these, carbohydrates such as starch or starch hydrolysate, acetic acid, propionic acid and the like. Alcohols such as organic acids, ethanol, and propanol are used.

Examples of the nitrogen source include ammonium salts of various inorganic and organic acids such as ammonia, ammonium chloride, ammonium sulfate, ammonium acetate, and ammonium phosphate, other nitrogen-containing compounds, peptone, meat extract, yeast extract, and corn steep. Liquor, casein hydrolyzate, soybean meal and soybean meal hydrolyzate, various fermented cells and digested products thereof can be used.

As the inorganic salt, potassium (II) phosphate, potassium (II) phosphate, magnesium phosphate, magnesium sulfate, sodium chloride, ferrous sulfate, manganese sulfate, copper sulfate, calcium carbonate and the like can be used.

The cultivation is performed under aerobic conditions such as shaking culture or deep aeration stirring culture. The culturing temperature is preferably 15 to 40 ° C, and the culturing time is usually 16 to 96 hours. During culture
H is kept between 3.0 and 9.0. The pH is adjusted using an inorganic or organic acid, an alkali solution, urea, calcium carbonate, ammonia, or the like.

[0093] If necessary, an antibiotic such as ampicillin or tetracycline may be added to the medium during the culture.

When a microorganism transformed with an expression vector using an inducible promoter is cultured, an inducer may be added to the medium, if necessary. For example, when culturing a microorganism transformed with an expression vector using a lac promoter, culturing a microorganism transformed with an expression vector using a trp promoter such as isopropyl-β-D-thiogalactopyranoside (IPTG). At times, indole acrylic acid (IAA) or the like may be added to the medium.

When the transformant for producing the polypeptide or the partial peptide of the present invention is an animal cell, the culture medium for culturing the cell may be a commonly used RPMI1640 medium.
Medium (The Journal of the American Medical Associat
ion, 199 , 519 (1967)], Eagle's MEM medium [Sc
ience, 122 , 501 (1952)), DMEM medium (Virology,
8 , 396 (1959)), 199 medium (Proceeding of the Soc
Society for the Biological Medicine, 73 , 1 (1950)], or a medium obtained by adding fetal bovine serum or the like to such a medium.

The cultivation is usually carried out at pH 6-8, 30-40 ° C.
This is performed for 1 to 7 days under conditions such as the presence of 5% CO ₂ .

During the culturing, kanamycin,
An antibiotic such as penicillin may be added to the medium.

As a medium for culturing a transformant obtained by using an insect cell as a host cell, a commonly used medium is used.
NM-FH medium (Pharmingen), Sf-900 II SFM medium (Gibco BRL), ExCell400, Ex
Cell 405 (manufactured by JRH Biosciences), Grace's Ins
ect Medium [Nature, 195 , 788 (1962)] and the like can be used.

The culture conditions are pH 6-7, and the culture temperature is 25-
Preferably, the cultivation time is usually 1 to 5 days at 30 ° C. Also,
If necessary, an antibiotic such as gentamicin may be added to the medium during the culture.

The polypeptide or partial peptide of the present invention can be expressed as a secretory protein or a fusion protein in accordance with the method described in Molecular Cloning, 2nd edition, in addition to direct expression. Examples of proteins to be fused include β-galactosidase, protein A, an IgG binding region of protein A,
Chloramphenicol acetyltransferase,
Poly (Arg), poly (Glu), protein G, maltose binding protein, glutathione S-transferase, polyhistidine chain (His-tag), S peptide,
DNA binding protein domain, Tac antigen, thioredoxin, green fluorescent protein, F
LAG peptide and epitopes of any antibodies [Akio Yamakawa, Experimental Medicine, 13 , 469-474 (199)
Five)〕.

The polypeptide or partial peptide of the present invention can be produced in a host cell, secreted out of the host cell, or produced on the host cell membrane. The method can be selected by changing the structure of the polypeptide to be produced.

In the case of secretory expression outside the host cell or expression on the host cell membrane, a signal sequence suitable for the host is added to the N-terminal side of the polypeptide or partial peptide of the present invention, if necessary. In some cases, it is better to add the above-mentioned secretion signal after partially deleting the N-terminus of the polypeptide or partial peptide. When the host is a bacterium belonging to the genus Escherichia, an alkaline phosphatase signal sequence, an OmpA signal sequence, or the like is used. When the host is a bacterium belonging to the genus Bacillus, an α-amylase signal sequence, a subtilisin signal sequence, or the like is used. If the host is an animal cell, an insulin signal sequence, an α signal sequence, an invertase signal sequence, etc.
-Interferon signal sequence, antibody molecule / signal sequence, etc. can be used.

The polypeptide or partial peptide of the present invention can be expressed by adding a tag for purification or detection. Tags can be added anywhere,
When expressed in mammalian cells, it is preferable to add the gene to the extracellular region or intracellular region described above.

Examples of tags for purification and detection include β-galactosidase, protein A, an IgG binding region of protein A, chloramphenicol acetyltransferase, poly (Arg), poly (Glu), protein G, and maltose binding protein. , Glutathione S-transferase, polyhistidine chain (His-tag), S peptide, DNA binding protein domain, Tac antigen,
Examples include thioredoxin, green fluorescent protein, FLAG peptide, and an epitope of any antibody [Akio Yamakawa, Experimental Medicine, 13 , 469-47.
4 (1995)].

Further, according to the method described in Japanese Patent Application Laid-Open No. 2-227075, the production amount can be increased by utilizing a gene amplification system using a dihydrofolate reductase gene or the like.

In order to isolate and purify the polypeptide of the present invention from the culture of the transformant for producing the polypeptide or the partial peptide of the present invention, a usual method for isolating and purifying the polypeptide is used.
Purification methods can be used.

For example, when the polypeptide or partial peptide of the present invention accumulates in a lysed state in the cells of the transformant for producing the polypeptide or partial peptide of the present invention, the culture is centrifuged. After collecting the cells in the culture and washing the cells, the cells are crushed by an ultrasonic crusher, a French press, a Manton-Gaurin homogenizer, a Dynomill or the like to obtain a cell-free extract.

The supernatant obtained by centrifuging the cell-free extract was subjected to a solvent extraction method, a salting-out method using ammonium sulfate, a desalting method, a precipitation method using an organic solvent, diethylaminoethyl (DEAE) -Sepharose, DIAION. Anion exchange chromatography using a resin such as HPA-75 (Mitsubishi Kasei), cation exchange chromatography using a resin such as S-Sepharose FF (Pharmacia), butyl sepharose, phenyl sepharose, etc. Purified samples can be obtained using techniques such as hydrophobic chromatography using resins, gel filtration using molecular sieves, affinity chromatography, chromatofocusing, and electrophoresis such as isoelectric focusing. it can.

When the polypeptide or partial peptide of the present invention accumulates on the cell membrane of the transformant for producing the polypeptide or partial peptide of the present invention, the cells are similarly recovered, crushed, centrifuged or filtered. , And the polypeptide or the partial peptide is solubilized from the membrane using a surfactant such as Triton X-100, and then the purified sample is isolated and purified in the same manner as described above. Obtainable.

When the polypeptide or the partial peptide is expressed by forming an insoluble substance in the cells, the cells are similarly collected, crushed, and centrifuged to obtain a precipitate fraction. After recovering the polypeptide or the peptide by an ordinary method, the insoluble form of the polypeptide or the partial peptide is solubilized with a denaturant. The lysate is
After the denaturant is not contained or the concentration of the denaturant is diluted to a dilute solution such that the polypeptide or the partial peptide is not denatured, or dialyzed to form the polypeptide or the partial peptide into a normal three-dimensional structure. A purified sample can be obtained by the same isolation and purification method as described above.

When the polypeptide or the partial peptide is secreted extracellularly, the culture is treated by a technique such as centrifugation to obtain a soluble fraction. From the soluble fraction, a purified sample of the polypeptide or the partial peptide can be obtained by the same method as the above-mentioned isolation and purification method from the cell-free extract supernatant.

Further, the polypeptide of the present invention or the partial peptide can be produced as a fusion protein with another protein, and purified by affinity chromatography using a substance having an affinity for the fused protein. [Akio Yamakawa, Experimental Medicine, 13 , 469-474 (199
Five)〕. For example, the method of Lowe et al. [Proc. Natl. Acad. Sci.
USA, 86 , 8227 (1989), Genes Develop., 4 , 1288 (1
990)], the polypeptide or partial peptide of the present invention is produced as a fusion protein with protein A, and purified by affinity chromatography using immunoglobulin G according to the method described in JP-A-05-336963 and WO94 / 23201. can do. Further, the polypeptide or partial peptide of the present invention can be produced as a fusion protein with a FLAG peptide and purified by affinity chromatography using an anti-FLAG antibody [Proc. Natl. Acad. Sci. USA, 86 , 8227 ( 1989), G
enes Develop., 4 , 1288 (1990)].

Further, it can be purified by affinity chromatography using an antibody against the polypeptide of the present invention or the partial peptide itself.

Further, a known method [J. Biomolecular NMR,
6 , 129, Science, 242 , 1162 (1988), J. Biochem., 1
10 , 166 (1991)], the polypeptide or partial peptide of the present invention can be produced using an in vitro transcription / translation system.

Further, the polypeptide or the partial peptide of the present invention can also be produced by a chemical synthesis method such as the Fmoc method (fluorenylmethyloxycarbonyl method) and the tBoc method (t-butyloxycarbonyl method).
Advanced ChemTech, Perkin-Elmer,
Pharmacia Biotech, Protein Technology Instr
Chemical synthesis can also be carried out using a peptide synthesizer such as ument, Synthecell-Vega, PerSeptive, Shimadzu Corporation.

The structural analysis of the purified polypeptide of the present invention is carried out by a method usually used in protein chemistry, for example, a method described in Protein Structural Analysis for Gene Cloning (Hisashi Hirano, Tokyo Chemical Dojin, 1993). It is feasible.

When the polypeptide or partial peptide of the present invention obtained by the above method is obtained in a free form, it can be converted into a salt by a known method or a method analogous thereto, and conversely, can be obtained by a salt. In this case, it can be converted to a free form or another salt by a known method or a method analogous thereto. The polypeptide or partial peptide of the present invention can be arbitrarily modified or partially removed by the action of an appropriate protein modifying enzyme. As protein modifying enzymes,
For example, trypsin, chymotrypsin, arginyl endopeptidase, protein kinase, glycosidase and the like are used.

The activity of the polypeptide or partial peptide of the present invention or the salt thereof which can be produced by the above method is as follows.
It can be measured by a binding experiment with a labeled ligand, an enzyme immunoassay using a specific antibody, or the like. (5) Antibody recognizing the polypeptide of the present invention (5-1) Production of antibody recognizing the polypeptide of the present invention (I) Preparation of polyclonal antibody Polypeptide or portion obtained by the method described in (4) above A polyclonal antibody can be prepared by using a peptide as an antigen and administering it to an animal.

The animals to which the antigen is administered include rabbits,
Goats, rats, mice, hamsters, etc., 3 to 20 weeks old can be mentioned. The dose of the antigen is preferably 50 to 100 μg per animal. When a peptide is used as an antigen, the peptide is used as keyhole limpet hemocyanin
(ole limpet haemocyanin) or bovine thyroglobulin, which is covalently bonded to a carrier protein. A peptide serving as an antigen can also be synthesized by a peptide synthesizer.

The administration of the antigen is performed 1-2 times after the first administration.
Perform 3 to 10 times every week. Blood is collected from the fundus venous plexus 3 to 7 days after each administration, and the reaction of the serum with the antigen used for immunization is determined by enzyme immunoassay [enzyme immunoassay (ELIS)].
A method): 1976, published by Medical Shoin, Antibodies-A Laborat
ory Manual, Cold Spring Harbor Laboratory (1988))
Confirm with etc.

[0121] A polyclonal antibody can be obtained by obtaining serum from the above animal whose serum shows a sufficient antibody titer against the antigen used for immunization, and separating and purifying the serum.

Methods for separation and purification include centrifugation,
Salting out with 40-50% saturated ammonium sulfate, caprylic acid precipitation [Antibodies, A Laboratory manual, Cold Spri
ng Harbor Laboratory, (1988)] or DEAE-Sepharose column, anion exchange column, protein A
Alternatively, there is a method in which chromatography using a G-column or a gel filtration column or the like is performed alone or in combination. (II) Preparation of Monoclonal Antibody (a) Preparation of Antibody-Producing Cells The above animal whose serum showed a sufficient antibody titer against the partial peptide of the polypeptide of the present invention used for immunization was used as a source of antibody-producing cells. Can be served as

The spleen is excised 3 to 7 days after the last administration of the antigenic substance to the above animal showing the antibody titer. The spleen is shredded in a MEM medium (manufactured by Nissui Pharmaceutical Co., Ltd.), loosened with forceps, centrifuged at 1,200 rpm for 5 minutes, and the supernatant is discarded. The spleen cells in the obtained precipitate fraction are treated with a Tris-ammonium chloride buffer (pH 7.65) for 1 to 2 minutes to remove erythrocytes, and then washed three times with a MEM medium. Use as cells. (b) Preparation of myeloma cells As myeloma cells, cell lines obtained from mice or rats are used.

For example, 8-azaguanine-resistant mice (BA
LB / c-derived) myeloma cell line P3-X63Ag8-U1 (hereinafter P3-U
1) [Curr. Topics. Microbiol. Immunol., 81 ,
1 (1978), Europ. J. Immunol., 6 , 511 (1976)), SP2
/ 0-Ag14 (SP-2 ) [Nature, 276, 269 (1978)], P3-X63-Ag
8653 (653) [J. Immunol., 123 , 1548 (1979)], P3-X63
-Ag8 (X63) [Nature, 256 , 495 (1975)] or the like can be used.

These cell lines were prepared in an 8-azaguanine medium [glutamine (1.5 mmol) in an RPMI-1640 medium.
/ L), 2-mercaptoethanol (5 × 10 ⁻⁵ mmol /
L), gentamicin (10 μg / ml) and fetal calf serum (FCS) (manufactured by CSL, 10%) (8% azaguanine (15 μg / ml) were further added to a medium (hereinafter referred to as a normal medium). Medium), and cultured in a normal medium 3 to 4 days before cell fusion, and use 2 × 10 ⁷ or more of the cells for fusion. (c) Preparation of hybridoma The antibody-producing cells obtained in (a) and the myeloma cells obtained in (b) were cultured in MEM medium or PBS (disodium phosphate 1.
83 g, monopotassium phosphate 0.21 g, salt 7.65
g, 1 liter of distilled water, pH 7.2)
The cells are mixed so that the number of cells becomes antibody-producing cells: myeloma cells = 5 to 10: 1, centrifuged at 1,200 rpm for 5 minutes, and the supernatant is discarded.

[0126] The cells of the obtained precipitate fraction were loosened well,
To the cell group, while stirring, at 37 ° C., 10 ⁸ antibody-producing cells per polyethylene of glycol -1000 (P
EG-1000), 0.2 to 1 ml of a mixed solution of 2 g of MEM, 2 ml of MEM and 0.7 ml of dimethylsulfoxide (DMSO) is added, and 1-2 ml of MEM medium is added several times every 1 to 2 minutes. After the addition, a MEM medium is added to adjust the total volume to 50 ml.

[0127] After centrifuging the prepared solution at 900 rpm for 5 minutes, the supernatant is discarded. The cells of the obtained precipitate fraction are loosened loosely, and then slowly sucked and blown out with a mesipette into a HAT medium [hypoxanthine (1 in normal medium).
0 ^-4 mmol / L), thymidine (1.5 × 10 ^-5 mmol / L)
Medium supplemented with aminopterin (4 × 10 ⁻⁷ M)
Suspend in 100 ml.

The suspension was added to a 96-well culture plate in an amount of 100
aliquots per well, in a 5% CO ₂ incubator,
Incubate at 37 ° C for 7-14 days. After culturing, a part of the culture supernatant is removed and used for antibodies (Antibodies, A Laboratory
manual, Cold Spring Harbor Laboratory, Chapter 14
(1988)).
A hybridoma that specifically reacts with a partial peptide of the polypeptide of the present invention is selected.

Specific examples of the enzyme immunoassay include the following methods. In the case of immunization, a partial peptide of the polypeptide of the present invention used as an antigen is coated on an appropriate plate, and the hybridoma culture supernatant or After reacting the purified antibody obtained in (d) below as a first antibody, and further reacting an anti-rat or anti-mouse immunoglobulin antibody labeled with biotin, an enzyme, a chemiluminescent substance or a radioactive compound as a second antibody A reaction according to the labeling substance is performed, and a substance that specifically reacts with the polypeptide of the present invention is selected as a hybridoma that produces a monoclonal antibody against the polypeptide of the present invention.

Using the hybridoma, cloning was repeated twice by the limiting dilution method (the first time, an HT medium (a medium obtained by removing aminopterin from the HAT medium), and the second time, a normal medium was used). Those having a strong antibody titer are selected as hybridoma strains producing anti-polypeptide antibodies of the polypeptide of the present invention. (d) Preparation of Monoclonal Antibody To a mouse or a nude mouse of 8 to 10 weeks old, which was treated with pristane (0.5 ml of 2,6,10,14-tetramethylpentadecane (Pristane) was intraperitoneally administered and bred for 2 weeks). , 5-20 hybridoma cells producing the polypeptide monoclonal antibody of the present invention obtained in (c)
× 10 ⁶ cells / mouse are injected intraperitoneally. In 10 to 21 days, the hybridoma becomes ascites cancer.

[0131] Ascites was collected from the mouse with ascites cancer,
Centrifuge at 3,000 rpm for 5 minutes to remove solids. From the obtained supernatant, a monoclonal antibody can be purified and obtained by a method similar to the method used for polyclonal.

The subclass of the antibody is determined using a mouse monoclonal antibody typing kit or a rat monoclonal antibody typing kit. The protein content is
It is calculated from the Lowry method or the absorbance at 280 nm. (5-2) Use of the Antibody of the Present Invention (a) Immunological Detection and Quantification of the Polypeptide of the Present Invention Using the Antibody of the Present Invention As an immunological detection method of the polypeptide of the present invention, a microtiter plate is used. E used
LISA method, fluorescent antibody method, Western blot method, immunohistological staining method [Separate volume Experimental Medicine, The Protocol Series, Immunostaining / in situ Hybridization, Yodosha (1997), Journal of Immunological Methods, 150 ,
5, (1992)].

As an immunoassay, a sandwich E using two kinds of monoclonal antibodies having different epitopes among antibodies reacting with the polypeptide of the present invention in the liquid phase is used.
LISA, radioimmunoassay using the polypeptide of the present invention labeled with a radioisotope such as ¹²⁶ I, and an antibody recognizing the polypeptide of the present invention, and the like.

The above detection or quantification method can be used for diagnosis of colorectal cancer, gastric cancer and the like. In addition, cells or cell lines that express the polypeptide of the present invention can be identified using the above detection or quantification methods. Cells or cell lines that express the polypeptide of the present invention are useful for searching for a ligand, agonist or antagonist of the polypeptide, and for analyzing the function of the polypeptide. (B) Medicine containing the antibody of the present invention The antibody of the present invention can be used as a medicine, for example, a therapeutic agent for diseases such as abnormalities of the thalamus or cerebellum, colon cancer, and gastric cancer.

Although the drug containing the antibody of the present invention can be administered alone as a therapeutic agent, it is usually mixed with one or more pharmacologically acceptable carriers. It is desirable to provide it as a pharmaceutical preparation manufactured by any method well known in the field of pharmaceutical science.

It is desirable to use the most effective route for treatment.
Parenteral administration, such as in the respiratory tract, rectally, subcutaneously, intramuscularly and intravenously, can be mentioned. Administration forms include sprays, capsules, tablets, granules, syrups, emulsions, suppositories, injections, ointments, tapes and the like.

Formulations suitable for oral administration include emulsions, syrups, capsules, tablets, powders, granules and the like. Liquid preparations such as emulsions and syrups include water, sugars such as sucrose, sorbitol, fructose, glycols such as polyethylene glycol, propylene glycol, oils such as sesame oil, olive oil, soybean oil, p-hydroxybenzoate. It can be produced using preservatives such as acid esters and flavors such as strawberry flavor and peppermint as additives. Capsules, tablets, powders, granules, etc. are excipients such as lactose, glucose, sucrose, mannitol, disintegrants such as starch, sodium alginate, lubricants such as magnesium stearate, talc, polyvinyl alcohol, hydroxy It can be produced using a binder such as propylcellulose and gelatin, a surfactant such as fatty acid ester, and a plasticizer such as glycerin as additives.

Preparations suitable for parenteral administration include injections, suppositories, sprays and the like. For example, injections
It is prepared using a carrier comprising a salt solution, a glucose solution, or a mixture of both. Suppositories are prepared using carriers such as cocoa butter, hydrogenated fats or carboxylic acids. Also,
Sprays are prepared using the compound itself or a carrier which does not irritate the oral and respiratory tract mucosa of the recipient and which disperses the compound as fine particles to facilitate absorption. Specific examples of the carrier include lactose and glycerin. Formulations such as aerosols and dry powders are possible depending on the properties of the compound and the carrier used. Also, in these parenteral preparations, the components exemplified as additives in the oral preparation can be added.

The dose or the number of administrations varies depending on the intended therapeutic effect, administration method, treatment period, age, body weight, etc., but is usually 10 μg / kg to 8 mg / kg per day for an adult.
It is. (6) Use of polypeptide of the present invention or partial peptide of the present invention (6-1) Method for screening ligand for G protein-coupled receptor polypeptide of the present invention Using polypeptide of the present invention or partial peptide of the present invention Searching for a ligand for the polypeptide of the present invention,
Can decide.

As a method of searching for or determining the ligand, for example, a method of contacting a polypeptide or a partial peptide of the present invention or a salt thereof with a test substance and selecting a ligand for the polypeptide of the present invention from the test substance. Or a method of contacting a cell expressing the polypeptide or partial peptide of the present invention or a membrane fraction of the cell with a test substance, and selecting a ligand for the polypeptide of the present invention from the test substance. .

As test substances, known ligands [for example, angiotensin, bombesin, cannabinoid, cholecystokinin, glutamine, serotonin, melatonin, neuropeptide Y, opioid, purine, vasopressin, oxytocin, PACAP (Pichuitari)
Adenylate cyclase activating protein), secretin, glucagon, calcitonin, adrenomedullin, somatostatin, GHRH (growth hormone-releasing hormone), CRF (corticotropin-releasing factor), ACTH (adrenocorticotropic hormone), melanin stimulant Ration hormone, GRP (gastrin-releasing peptide), PTH (parathyroid hormone), VIP (vasoactive intestinal and reduced polypeptide), dopamine, motilin, amylin, bradykinin, CGRP (calcitonin gene-related peptide), leukotriene,
Pancreatastatin, prostaglandin, thromboxane, adenosine, adrenaline, α and β-chemokines [eg, IL-8 (interleukin-8), G
ROα (Growth Relayed Gene α), GRO
β, GROγ, NAP-2 (neural calmodulin binding protein-2), ENA-78
(Epserial Cell-Derived Neutrofil-Activating Protein-78), PF4 (Platelet Factor 4), IP10 (Interferon-γ Inducible Protein of 10k)
d), GCP-2 (granulosite chemotactic protein-2), MCP-1 (monosite chemoretractant protein-1), HC14, MCP-
3, I-309, MIP1α (macrophage inflammatri protein 1α), MIP-1β, RANT
ES (Regulated on Activation, Normal T Cell Expressed and Secreted), endothelin, enterogastrin, histamine, neurotensin, TRH, pancreatic polypeptide, galanin, urotensin I and I
I, neuropeptide FF, orexin and melanin concentrating hormone, etc.], as well as, for example, tissue extracts of humans or mammals (eg, mice, rats, pigs, cows, sheep, monkeys, etc.) and those derived from the extracts. Biological samples such as purified products, cell culture supernatants and purified products derived from the supernatants, known proteins, recombinant proteins produced using recombinant techniques, microbial cell extracts and purifications derived from the extracts Products, microbial culture supernatants, purified products derived from the supernatants, known compounds, compounds synthesized using combinatorial chemistry, and the like.

As a specific method for searching for or determining a ligand for the polypeptide of the present invention, a method for searching for a polypeptide or a partial peptide of the present invention or a salt thereof, or a cell expressing the polypeptide or partial peptide of the present invention may be used. Examples of the method include a method of measuring the amount of a test substance bound to a polypeptide or a partial peptide of the present invention or a salt thereof, or detecting a response of a cell expressing the polypeptide of the present invention.

More specifically, (a) when the labeled test substance is brought into contact with the polypeptide or partial peptide of the present invention or a salt thereof, the polypeptide or partial peptide of the labeled test substance or their salts (B) a method for determining a ligand for the polypeptide of the present invention, which comprises measuring the amount of binding to a salt of the present invention and selecting a ligand for the polypeptide of the present invention or a salt thereof from the test substance; Is contacted with a cell containing the polypeptide or partial peptide of the present invention or a membrane fraction of the cell, the amount of the labeled test substance bound to the cell or the membrane fraction is measured, and the test substance Selecting a ligand for the polypeptide of the present invention from a ligand for the polypeptide of the present invention. Determination method, (c) a test substance, when in contact with the membrane fraction of the cell or the cell containing the polypeptide of the present invention, the labeled GTPyS G
A method for determining a ligand for the polypeptide of the present invention, which comprises measuring the amount of binding to the α protein and selecting a ligand for the polypeptide of the present invention from the test substance. A ligand for the polypeptide of the present invention, which comprises measuring the GTPase activity when the cell is contacted with a cell containing the polypeptide or a membrane fraction of the cell, and selecting a ligand for the peptide of the present invention from a test substance. How to determine
(E) polypeptide-mediated cell stimulating activity (eg, arachidonic acid release, acetylcholine release, intracellular Ca ²⁺ release, intracellular) when a test substance is brought into contact with a cell containing the polypeptide of the present invention. cAMP production, intracellular cGMP production, inositol phosphate production, cell membrane potential fluctuation, intracellular protein phosphorylation, c-fos activation, p
H decrease, cell proliferation activity, aggregation or diffusion of melanin pigment, or activity to promote or suppress reporter gene expression, etc.) and select a ligand for the polypeptide of the present invention from the test substance. And a method for determining a ligand for the polypeptide of the present invention.

Hereinafter, the method for searching for or determining the ligand of the present invention will be described in more detail. (I) Method for Measuring the Amount of Test Substance to be Bound As shown in (a) and (b) above, the polypeptide or partial peptide of the present invention or a salt thereof, or the polypeptide or partial peptide of the present invention is expressed. The labeled test substance is allowed to act on the cells, the amount of the test substance bound to the polypeptide of the present invention is measured, and the ligand for the polypeptide of the present invention is selected from the test substance, whereby the ligand for the polypeptide of the present invention is obtained. Can be searched or determined.

As test substances, for example,^ThreeH,¹⁴C,
¹²⁵I,³⁵S,³²Known labeling with radioactive isotopes such as P
GPCR ligands [angiotensin, Bombesi
, Cannabinoids, cholecystokinin, glutamine,
Rotonin, melatonin, neuropeptide Y, opioid
Do, pudding, vasopressin, oxytocin, PACA
P, secretin, glucagon, calcitonin, adreno
Medulin, somatostatin, GHRH, CRF, AC
TH, melanin stimulation hormone, GR
P, PTH, VIP, dopamine, motilin, amiri
, Bradykinin, CGRP, leukotriene, punk
Reastatin, prostaglandin, thromboxane,
Adenosine, adrenaline, α and β-chemokines
(For example, IL-8, GROα, GROβ, GROγ,
NAP-2, ENA-78, PF4, IP10, GCP
-2, MCP-1, HC14, MCP-3, I-30
9, MIP1α, MIP-1β, RANTES, etc.),
Endothelin, enterogastrine, histamine,
-Rotensin, TRH, pancreatic polypeptide
Id, galanin, urotensin I and II, neuro
Peptide FF, orexin and melanin concentrate
And other hormones). Ma
Was ^ThreeH,¹⁴C,¹²⁵I,³⁵S,³²Radioactive isotopes such as P
Using any protein, peptide or compound labeled with
You can also.

In the above method, the polypeptide or the partial peptide of the present invention can be used directly as the polypeptide or the partial peptide, but the cell itself containing the polypeptide or the partial peptide or the cell membrane fraction thereof is used. You can also. As the partial peptide when the cell membrane fraction is used, a partial peptide having both a hydrophilic site capable of binding ligand and a hydrophobic site that is a cell transmembrane region is preferable. When cells containing the polypeptide or partial peptide are used, the cells may be immobilized with glutaraldehyde, formalin, or the like.

As the polypeptide or partial peptide of the present invention, any of a naturally occurring polypeptide or partial peptide, and a recombinant polypeptide or a recombinant partial peptide prepared by using a gene recombination technique may be used. Can be. As the polypeptide of the present invention, among the polypeptides encoded by the mutant DNA obtained by introducing a mutation into the DNA having the base sequence represented by SEQ ID NO: 2 by the method described in (3), The activated mutant polypeptide is particularly useful.

Among G protein-coupled receptors (GPCRs), there are those that, when a GPCR polypeptide is overexpressed in a cell, pass a signal even in the absence of a ligand. It is called a type GPCR. Also,
It is known that even GPCRs that are not originally constitutively active become constitutively active by introducing mutations such as amino acid substitutions and deletions. Since it is known that a mutant GPCR changed to a constitutively active form may have an increased affinity for an agonist, a constitutively active mutant GPCR is considered to be useful in the search for a ligand [Journal of Pharmacology and Experimental Ther
apeutics, 275 , 1274 (1995), Endocrinology, 137 , 39
36 (1996), J. Biol. Chem., 272 , 1822 (1997)].

The constitutively active GPCR can be obtained according to a known method [J. Biol. Chem., 271 , 1857].
(1996), Science, 268 , 98 (1995), Journal of Pharm
acology and Experimental Therapeutics, 275 , 1274
(1995), J. Biol. Chem., 272 , 1822 (1997), Journal
of Receptor and Signal Transduction Research, 17 , 5
7 (1997), WO98 / 46995].

The above-mentioned cell membrane fraction refers to a fraction containing a large amount of cell membranes obtained by disrupting cells and then obtaining the cell membrane by a method known per se. Potter
-A method of crushing cells with an Elvehjem-type homogenizer, crushing with a Waring blender or a polytron (manufactured by Kinematica), crushing with ultrasonic waves, crushing by ejecting cells from a thin nozzle while applying pressure with a French press, and the like. For the fractionation of cell membranes, fractionation by centrifugal force such as fractionation centrifugation or density gradient centrifugation is mainly used. For example, the cell lysate is slowed (50
(3000 rpm) for a short period of time (usually about 1-10 minutes), and the supernatant is further centrifuged (15000-30)
000 rpm) for 30 minutes to 2 hours, and the resulting precipitate is used as a membrane fraction. The membrane fraction is rich in the expressed receptor protein and membrane components such as cell-derived phospholipids and membrane proteins.

As the cells expressing the polypeptide of the present invention, as described in the above (4), a plasmid obtained by introducing a recombinant DNA containing a DNA encoding the polypeptide into an appropriate host cell can be used. Cells expressing the polypeptide in large amounts, such as transformed cells, can also be used.
In addition to microorganisms such as Escherichia coli, Bacillus subtilis, and yeast, insect cells, frog oocytes, frog melanocytes, animal cells, plant cells, and the like can be mentioned, and the polypeptide of the present invention produced by the transformant can be used. In order to maintain the higher-order structure and maintain the binding property to the ligand, it is preferable to express in yeast, insect cells, frog oocytes, frog melanocytes, animal cells, plant cells and the like.

The amount of the polypeptide or the partial peptide in the cell containing the polypeptide or the partial peptide of the present invention or the cell membrane fraction thereof is preferably, for example, 10 ³ to 10 ⁸ molecules per cell, and preferably 10 ⁵ to 10 ⁵ molecules. Preferably it is 10 ⁷ molecules. The higher the expression level, the higher the ligand binding activity (specific activity) per membrane fraction, which makes it possible not only to construct a highly sensitive screening system, but also to measure a large number of samples in the same lot. .

Hereinafter, specific examples will be described.

A sample of the polypeptide of the present invention is prepared by suspending a cell containing the polypeptide or the partial peptide of the present invention or a cell membrane fraction of the cell in an appropriate buffer. The buffer may be any buffer as long as it does not inhibit the binding between the ligand and the polypeptide of the present invention, for example, pH 4 to 10 (preferably pH 6 to 8).
Phosphate buffer or Tris-HCl buffer. In addition, CHA was used for the purpose of reducing non-specific binding.
Surfactants such as PS, Tween-80, digitonin, deoxycholic acid and various proteins such as bovine serum albumin and gelatin can also be added to the buffer. In addition, PMSF, leupeptin, E
Protease inhibitors such as -64 and pepstatin can also be added. 10 μl to 10 ml of the polypeptide preparation is allowed to coexist with a radioactive test substance labeled with a radioisotope ( ³ H, ¹²⁵ I, ¹⁴ C, ³⁵ S, ³² P, etc.). A reaction tube containing a large excess of unlabeled test substance is also prepared to determine the amount of non-specific binding (NSB). The reaction is carried out at about 0 to 50 ° C, preferably about 4 to 37 ° C, for about 20 minutes to 24 hours, preferably for about 30 minutes to 3 hours. After the reaction, the mixture is filtered with a glass fiber filter or the like, washed with an appropriate amount of the same buffer, and the radioactivity remaining on the glass fiber filter is measured with a liquid scintillation counter or a γ-counter. From the total binding amount (B) to the non-specific binding amount (NS
A test substance having a count (B-NSB) less than 0 cpm minus B) can be selected as a substance that binds to the polypeptide of the present invention. Among them, the binding activity to the cell containing the polypeptide of the present invention or the cell membrane fraction of the cell is strong, and the binding activity to the cell not containing the polypeptide of the present invention or the cell membrane fraction of the cell is weak. Substances can be selected as ligands for the polypeptide of the invention.

In the above method, as a cell containing the polypeptide or the partial peptide of the present invention, a host cell not expressing the polypeptide of the present invention is transformed into a vector DNA by encoding a DNA encoding the polypeptide or the partial peptide.
Using the polypeptide or the partial peptide-expressing cells obtained by introducing the recombinant DNA incorporated into the host cell as a cell not containing the polypeptide of the present invention, by introducing only a vector into the host cell. By using control cells that do not express the polypeptide, the ligand can be more accurately determined. (II) Method for measuring the amount of GTPγS bound to Gα protein As shown in (c) above, a test substance is brought into contact with a cell containing the polypeptide of the present invention or a membrane fraction of the cell,
By measuring the amount of labeled GTPγS bound to Gα protein (membrane fraction), the ligand of the polypeptide can be searched or determined [Molecular Pharmacolo
gy, 47 , 848 (1995), WO98 / 46995].

As the test substance, any substance can be used. For example, a known peptide, a known GPCR ligand,
Biological samples and biological samples such as known proteins, recombinant proteins produced using recombinant techniques, cell extracts and purified products derived from the extracted solutions, cell culture supernatants and purified products derived from the supernatants, serum and the like Use of a purified product derived from a microorganism, a cell extract of a microorganism, a purified product derived from the extract, a microorganism culture supernatant or a purified product derived from the supernatant, a known compound, a compound synthesized using combinatorial chemistry, or the like. Can be. GTP labeled
As γS, for example, GTPγS labeled with ³⁵ S can be used.

As the cells containing the polypeptide of the present invention or the membrane fraction of the cells, those described in the above (I) can be used.

Hereinafter, specific examples will be described.

A cell or cell membrane fraction preparation containing the polypeptide of the present invention is prepared by the method described in the above (I). A test compound, a radioactive amount of GTPγS labeled with a radioisotope (such as ³⁵ S), and GDP are allowed to coexist with 10 μl to 10 ml of the polypeptide preparation.
If it is necessary to know the amount of non-specific binding (NSB), prepare a reaction tube to which a large excess of unlabeled GTPγS has been added. The count (B-NSB) obtained by subtracting the non-specific binding amount (NSB) from the total binding amount (B) is the specific binding amount.
The reaction is carried out at about 0 to 50 ° C, preferably about 4 to 37 ° C, for about 2 hours.
The reaction is performed for 0 minute to 24 hours, preferably for about 30 minutes to 3 hours.
After the reaction, the mixture is filtered with a glass fiber filter or the like, washed with an appropriate amount of the same buffer, and the radioactivity remaining on the glass fiber filter is measured with a liquid scintillation counter. The same operation is performed using cells or cell membrane fractions that do not express the polypeptide of the present invention, and the radioactivity is measured.

When a cell containing the polypeptide of the present invention or a membrane fraction of the cell is used, GTPγS binds to the cell or the membrane fraction of the cell, and does not express the polypeptide of the present invention. The binding activity of GTPγS to a cell or a cell membrane fraction when using a cell or a membrane fraction of the cell is compared, and a cell containing the polypeptide of the present invention or a membrane fraction of the cell is determined from a test sample. A substance having a strong activity of enhancing the binding of GTPγS to the membrane fraction when used can be selected as a ligand of the polypeptide of the present invention.

In the above method, as a cell containing the polypeptide of the present invention, a host cell that does not express the polypeptide of the present invention is obtained by introducing a recombinant DNA obtained by incorporating a DNA encoding the polypeptide into a vector. A control cell that does not express the polypeptide of the present invention prepared by introducing only a vector into the host cell as a cell that does not contain the polypeptide of the present invention, using a cell that expresses the polypeptide of the present invention as a cell that does not contain the polypeptide of the present invention. By using this, the ligand can be determined more accurately. (III) Method for measuring GTPase activity As shown in the above (d), a test substance is brought into contact with a cell containing the polypeptide of the present invention or a membrane fraction of the cell,
By measuring GTPase activity, the ligand of the polypeptide of the present invention can be searched or determined [J. Biol. Chem., 271 , 1857 (1996), J. Biol. Che.
m., 271 , 1857 (1996), WO98 / 46995].

As the test substance, the substances described in the above (II) can be used.

As the cells containing the polypeptide of the present invention or the membrane fraction of the cells, those described in the above (I) can be used.

The following is a specific example.

A cell or cell membrane fraction preparation containing the polypeptide of the present invention is prepared by the method described in the above (I). To the polypeptide preparation of 10Myueru～10ml, test compounds, coexisting GTP radioactive isotopes (such as ³² P) constant amount of radioactivity labeled with (e.g. [γ ³² P] GTP). The reaction is carried out at about 0-50 ° C, preferably about 4-37 ° C.
For about 20 minutes to 24 hours, preferably for about 30 minutes to 3 hours. After the reaction, the supernatant of the reaction solution was recovered and released [γ
The radioactivity of < ³² > P] Pi is measured with a liquid scintillation counter. After filtering the reaction solution with a glass fiber filter or the like and washing with an appropriate amount of the same buffer, the radioactivity in the filtrate may be measured with a liquid scintillation counter. Similarly, the radioactivity in the filtrate is measured for cells or cell membrane fractions that do not contain the polypeptide of the present invention.

GTPase activity when cells containing the polypeptide of the present invention or a membrane fraction of the cells were used, and GTPase activity when cells not expressing the polypeptide of the present invention or a membrane fraction of the cells were used. The GTPase activity of the polypeptide of the present invention is compared with a compound having a strong activity of enhancing GTPase activity when a cell containing the polypeptide of the present invention or a membrane fraction of the cell is used. Can be selected as

In the above method, the cells containing the polypeptide of the present invention may be obtained by introducing a recombinant DNA having a DNA encoding the polypeptide into a vector into a host cell that does not express the polypeptide of the present invention. A control cell that does not express the polypeptide of the present invention prepared by introducing only a vector into the host cell as a cell that does not contain the polypeptide of the present invention, using a cell that expresses the polypeptide of the present invention as a cell that does not contain the polypeptide of the present invention. By using this, the ligand can be determined more accurately. (IV) Method for Detecting Cell Response As shown in (e) above, a test substance is brought into contact with a cell expressing the polypeptide of the present invention, and the activation of the polypeptide is detected using the cell response as an index. By doing so, the ligand of the polypeptide can be searched or determined. Cell responses include, for example, arachidonic acid release, acetylcholine release, intracellular Ca ²⁺ release, intracellular c
AMP production, intracellular cAMP reduction, intracellular cGMP production, inositol phosphate production, cell membrane potential fluctuation, intracellular protein phosphorylation, c-fos activation, pH decrease, cell growth activity, melanin pigment aggregation or diffusion, Or the expression level of a reporter gene can be raised (J.
Biol. Chem., 271 , 1857 (1996), Science, 268 , 98.
(1995), Journal of Pharmacology and Experimental
Therapeutics, 275 , 1274 (1995), J. Biol. Chem., 27
2 , 1822 (1997), Journal of Receptor and Signal Tr
ansduction Research, 17 , 57 (1997), Endocrinology
138 , 1400 (1997), Endocrinology 138 , 1471 (1997),
Nat. Biotechnol., 16 , 1334 (1998), Biochem. Bioph.
ys. Res. Commun., 251 , 471 (1998); Brit. J. Pharma.
col., 125 , 1387 (1998), Trends Biotechnol., 15 , 48
7 (1997), Anal. Biochem., 252 , 115 (1997), Nature,
358 , 325 (1992), Nature, 393 , 272 (1998), Cell, 9
2 , 573 (1998), J. Biol. Chem., 272 , 27497 (1997),
Trends Pharmacol. Sci., 18 , 430 (1997), Trends Pha
rmacol. Sci., 20 , 370 (1999), WO98 / 46995].

When the response of a cell is monitored using a reporter system, for example, a cell expressing the polypeptide of the present invention may be appropriately placed downstream of a promoter sequence of a gene whose expression is induced by activation of the polypeptide. By introducing a DNA linked to any reporter gene, the activation of the polypeptide can be measured by the expression of the reporter gene. As the promoter, for example, IC
AM-1 gene promoter, c-fos promoter, Krox-24 promoter [Biochem. J., 3
20 , 145 (1996)]. Further, the promoter may be an artificial promoter consisting of a binding sequence of an appropriate transcription factor and a basic promoter. Examples of transcription factor binding sequences include CRE (CREB binding element),
TRE (TPA responsive element), SRE (serum re
sponsive element) can be used. Reporter genes include chloramphenicol acetyltransferase gene, β-glucuronidase gene,
The galactosidase gene, β-lactamase gene, aequorin gene, luciferase gene, green fluorescent protein gene and the like can be used.

As the cells containing the polypeptide of the present invention used in the above method, those described in the above (I) can be used.

As the cells expressing the polypeptide of the present invention, as described in the above (4), a plasmid obtained by introducing a recombinant DNA containing a DNA encoding the polypeptide into an appropriate host cell can be used. Cells expressing the polypeptide in large amounts, such as transformed cells, can also be used.
Examples of the host cell include microorganisms such as Escherichia coli, Bacillus subtilis, and yeast, as well as insect cells, frog oocytes, frog melanocytes, animal cells, plant cells, and the like. In order for the polypeptide to retain its higher-order structure and retain its binding property and functionality with a ligand, it is expressed in yeast, insect cells, frog oocytes, frog melanocytes, animal cells, plant cells, etc. It is preferred that Further, a mutant strain of yeast, a yeast expressing the modified Gα protein, or the like can also be used as a host [Tren
ds in Biotechnology, 15 , 487 (1997), Mol. Cell. Bi
ol., 15 , 6188 (1995), Mol. Cell. Biol., 16 , 4700.
(1996)].

As the test substance, the substances described in the above (II) can be used.

Specific examples are shown below.

The cells expressing the polypeptide of the present invention are cultured in a multiwell plate or the like. After the culture, the medium is replaced with a fresh medium or an appropriate buffer that is not toxic to cells, if necessary, and the test compound is added thereto and incubated for a certain period of time. Thereafter, cellular responses (eg, arachidonic acid release, acetylcholine release, intracellular Ca ²⁺ release, intracellular cAMP production, intracellular cGMP production, inositol phosphate production, cell membrane potential fluctuation, intracellular protein phosphorylation, c- fos activation, pH lowering, cell growth activity, aggregation or diffusion of melanin pigment, activity to promote or suppress reporter gene expression, etc.)
Is measured. For example, using the cell extract or supernatant, the product produced by the response of the cells is quantified according to a conventional method. When production of a substance (for example, arachidonic acid or the like) as an indicator of cell stimulating activity is difficult to be assayed by a degrading enzyme contained in a cell, an inhibitor for the degrading enzyme may be added for quantification. In addition, for activities such as cAMP production inhibition, cells are cAMP-expressed with forskolin or the like.
It can be detected as a production inhibitory effect on cells whose production amount has been increased.

The same operation is performed for cells that do not express the peptide of the present invention, and the response of the cells is measured.

The response of the cell when using the cell containing the polypeptide of the present invention was compared with the response of the cell when using the cell not expressing the polypeptide of the present invention. Substances that strongly detect cellular responses when cells containing the polypeptide of the present invention are used can be selected as ligands for the polypeptide of the present invention.

In the above method, the cells containing the polypeptide of the present invention may be obtained by introducing a recombinant DNA having a DNA encoding the polypeptide into a vector into a host cell that does not express the polypeptide of the present invention. A control cell that does not express the polypeptide of the present invention prepared by introducing only a vector into the host cell as a cell that does not contain the polypeptide of the present invention, using a cell that expresses the polypeptide of the present invention as a cell that does not contain the polypeptide of the present invention. By using this, the ligand can be determined more accurately. (6-2) Kit for screening ligand for G protein-coupled receptor polypeptide of the present invention Kit for screening ligand binding to polypeptide of the present invention or a salt thereof comprises polypeptide of the present invention or a part of the present invention It contains a peptide or a salt thereof, a cell containing the polypeptide of the present invention or the partial peptide of the present invention, or a membrane fraction of the cell. Examples of the kit for screening a ligand of the present invention include the following. (A) Ligand screening reagent Measurement buffer and washing buffer Hanks' Balanced Salt Solution (Gibco)
One containing 05% bovine serum albumin (manufactured by Sigma). Sterilized by filtration through a 0.45 μm filter, 4 ° C
Or may be prepared at the time of use. Preparation of polypeptide or partial peptide of the present invention CHO cells expressing the polypeptide or partial peptide of the present invention were subcultured on a 12-well plate at 5 × 10 ⁵ cells / well, and 5% CO ₂ , 95% air Cultured at 37 ° C. for 2 days in an incubator. Labeled test compound A compound labeled with commercially available [ ³ H], [ ¹²⁵ I], [ ¹⁴ C], [ ³⁵ S], or the like, or a compound labeled by an appropriate method. An aqueous solution of the compound at 4 ° C or -20
Store at ℃ and dilute to 1 μmol / L with a measuring buffer before use. Test compounds that are poorly soluble in water are dissolved in dimethylformamide, DMSO, methanol and the like. Unlabeled test compound An unlabeled compound prepared at a concentration of 100 to 1000 times that of the labeled compound. (B) Measuring method CHO cells expressing the polypeptide of the present invention, which were cultured using a 12-well tissue culture plate, were placed in a measuring buffer 1
After washing twice with ml, 490 μl of measurement buffer is added to each well. 5 μl of the labeled test compound is added and reacted at room temperature for 1 hour. In order to know the amount of non-specific binding, 5
What has been added may be prepared. Remove the reaction solution and wash 3 times with 1 ml of washing buffer. The labeled test compound bound to the cells is dissolved in a lysis solution (0.2 m
ol / L NaOH, 1% SDS) and mixed with 4 ml of liquid scintillator A (manufactured by Wako Pure Chemical Industries). Liquid scintillation counter (Beckman)
The radioactivity is measured using.

The ligand of the polypeptide of the present invention includes, for example, substances present in the brain, hypothalamus, pituitary gland, pancreas and the like, but the ligand of the present invention does not include known ligands. Known ligands include:
Angiotensin, bombesin, cannabinoid, cholecystokinin, glutamine, serotonin, melatonin, neuropeptide Y, opioid, purine, vasopressin, oxytocin, PACAP, secretin, glucagon, calcitonin, adrenomedullin, somatostatin, GHRH, CRF, ACTH, ACTH Simulation hormone, GRP, PTH, VIP, dopamine, motilin, amylin, bradykinin, CGRP,
Leukotriene, pancreastatin, prostaglandin, thromboxane, adenosine, adrenaline, α
And β-chemokines (eg, IL-8, GROα,
GROβ, GROγ, NAP-2, ENA-78, PF
4, IP10, GCP-2, MCP-1, HC14, M
CP-3, I-309, MIP1α, MIP-1β, R
ANTES, etc.), endothelin, enterogastrin, histamine, neurotensin, TRH, pancreatic polypeptide, galanin, urotensin I
And II, neuropeptide FF, orexin and melanin concentrating hormone. (6-3) Method for Quantifying Ligands for G Protein-Coupled Receptor Polypeptides of the Present Invention Since the polypeptides of the present invention or partial peptides thereof or salts thereof have binding properties to ligands, Ligand concentration in the body can be quantified with high sensitivity. The quantification method of the present invention can be used, for example, by combining it with a competition method. That is,
The ligand concentration in the subject can be measured by contacting the subject with the polypeptide of the present invention or a partial peptide thereof or a salt thereof. Specifically, for example, it is carried out according to a known method ("Radio Immunoassay" edited by Hiroshi Irie (Kodansha, published in 1974), "Radio Immunoassay" (edited by Hiroshi Irie) (published in Kodansha, 1974) or a method according thereto. be able to. (6-4) Method of Screening for Agonist, Antagonist or Functional Modifier of G Protein-Coupled Receptor Polypeptide of the Present Invention Polypeptide of the present invention or a partial peptide thereof or a salt thereof, or polypeptide or partial peptide of the present invention The cells that express E. coli and the membrane fraction of these cells are useful as reagents for selecting agonists, antagonists or functional modifiers for the polypeptide of the present invention.

As a method for screening for agonists, antagonists or functional modifiers of the polypeptide of the present invention, for example, (A) a method in which the polypeptide of the present invention or its partial peptide or a salt thereof is contacted with a ligand; Comparison between the case where the polypeptide of the present invention or a partial peptide thereof or a salt thereof is brought into contact with a ligand and a test substance is carried out, and an agonist, an antagonist or a function modifying substance of the polypeptide of the present invention is selected from the test substance. (B) a cell expressing the polypeptide of the present invention or a partial peptide thereof, or a cell membrane fraction thereof, and a ligand, and the polypeptide of the present invention or a partial peptide thereof. Expressing cells or the cell membrane fraction, ligand and test substance (C) a method characterized by comparing with the case of contacting and selecting an agonist, antagonist or function modifying substance of the polypeptide of the present invention from the test substance; A method characterized by using the same method as the method for searching for the ligand of the polypeptide of the present invention described in (d).

In the method for searching for a ligand of the polypeptide of the present invention described in (a) to (d) of (6-1), the constitutively active mutant polypeptide described in (6-1) is used. A screening method for an agonist, antagonist or functional modifier of the polypeptide of the present invention, which is characterized in that it is used, can also be mentioned.

More specifically, (a) the case where the labeled ligand is brought into contact with the polypeptide of the present invention or the partial peptide or a salt thereof, and the case where the labeled ligand and the test substance are brought into contact with the polypeptide of the present invention or the same. When contacted with a partial peptide or a salt thereof, the amount of the labeled ligand bound to the polypeptide or the partial peptide or a salt thereof is measured and compared, and an agonist of the polypeptide of the present invention is obtained from the labeled ligand; A method for screening for an agonist, antagonist or function-modifying substance of the polypeptide of the present invention, which comprises selecting an antagonist or a function-modifying substance; and (b) a labeled ligand containing the polypeptide or partial peptide of the present invention. Contacted with cells or membrane fraction of the cells When the labeled ligand and the test substance are brought into contact with the cell or the membrane fraction of the cell containing the polypeptide or partial peptide of the present invention, the amount of the labeled ligand bound to the cell or the membrane fraction Measuring and comparing the agonist, antagonist or function modifier of the polypeptide of the present invention from the labeled ligand, screening method of agonist, antagonist or function modifier of the polypeptide of the present invention, (C) When the ligand is brought into contact with the cell containing the polypeptide of the present invention or the membrane fraction of the cell, and when the ligand and the test substance are brought into contact with the cell containing the polypeptide of the present invention or the membrane fraction of the cell. Labeled GTPγS when contacted
Is measured and compared to the Gα protein (membrane fraction),
A method of screening for an agonist, antagonist or function-modifying substance of the polypeptide of the present invention, which comprises selecting an agonist, antagonist or function-modifying substance of the polypeptide of the present invention from a test substance; GTPase activity was measured in the case of contacting with a cell containing the or a membrane fraction of the cell, and in the case of contacting a ligand and a test substance with a cell containing the receptor protein of the present invention or a membrane fraction of the cell. (E) a method of screening for an agonist, antagonist or function-modifying substance of the polypeptide of the present invention, which comprises selecting an agonist, antagonist or function-modifying substance of the polypeptide of the present invention from the test substance. Ligand is added to a cell containing the polypeptide of the present invention or the cell. A cell containing the polypeptide or partial peptide of the present invention or a cell mediated by the polypeptide of the present invention in a case where the ligand and the test substance are brought into contact with the membrane fraction of the cell. Stimulation activity (eg, arachidonic acid release, acetylcholine release, intracellular Ca ²⁺ release, intracellular cAMP production, intracellular cGMP production, inositol phosphate production, cell membrane potential fluctuation, intracellular protein phosphorylation, c-fos activation ,
pH decrease, cell proliferation activity, aggregation or diffusion of melanin pigment, or activity to promote or suppress the expression level of a reporter gene, etc.). A method for screening for an agonist, antagonist or function-modifying substance of the receptor protein of the present invention, which comprises selecting an antagonist or a function-modifying substance; (f) a cell containing the polypeptide of the present invention as a test substance or the cell; GT in contact with the membrane fraction of
Measuring the amount of binding of PγS to Gα protein (membrane fraction) and selecting an agonist or functional modifier of the polypeptide of the present invention from the test substance; A method for screening a substance, (g) measuring the GTPase activity when a test substance is brought into contact with a cell containing the polypeptide of the present invention or a membrane fraction of the cell, and determining whether an agonist of the polypeptide of the present invention or A method for screening for an agonist or a function-modifying substance of the polypeptide of the present invention, which comprises selecting a function-modifying substance; and (h) a method for contacting a test substance with a cell containing the polypeptide of the present invention. Cell stimulating activity via the polypeptides or partial peptides of the invention (eg, arachidonic acid release, acetylcholine Release, intracellular Ca ²⁺ release, intracellular c
AMP production, intracellular cGMP production, inositol phosphate production, cell membrane potential fluctuation, intracellular protein phosphorylation, c-
fos activation, pH reduction, melanin pigment aggregation or diffusion, or activity to promote or suppress reporter gene expression, etc.) is measured, and an agonist or functional modifier of the polypeptide of the present invention is determined from the test substance. A method of screening for an agonist or a function modifier of the polypeptide of the present invention, characterized by selecting
(I) In the above (a) to (h), an agonist, antagonist or function-modifying substance of the polypeptide of the present invention, characterized in that a polypeptide mutated to a constitutively active form is used as the polypeptide of the present invention. Screening methods.

The substances selected in the above (a) and (b) can be obtained by using the above methods (c) to (i)
It is possible to distinguish between antagonists or functional modifiers. On the other hand, among the substances selected by the above methods (a) to (i), substances which are not agonists or antagonists of the polypeptide of the present invention but which can modify the function of the polypeptide of the present invention (function modifying substances) ) Is also included. For example, among the substances selected by the methods (a) to (i),
It is considered that a substance that inhibits or enhances coupling between the polypeptide of the present invention and a G protein is also included. Also,
It is considered that the substances selected by the methods (c) to (i) include substances that inhibit or enhance signals downstream of the polypeptide of the present invention. These functional modifiers are also useful as drug candidates.

A detailed description of the method for screening for an agonist, antagonist or functional modifier of the polypeptide of the present invention by the above method will be given below. (I) Method for Measuring the Amount of Ligand Binding As shown in (a) and (b) above, the polypeptide or partial peptide of the present invention or a salt thereof, or the polypeptide or partial peptide of the present invention is expressed. A test substance and a labeled ligand are allowed to act on cells, and the amount of the ligand bound to the polypeptide or partial peptide of the present invention or a salt thereof is measured. Screening can be performed.

As the labeled ligand, a labeled ligand, a labeled ligand analog compound and the like can be used. For example, [ ³ H], [ ¹²⁵ I], [ ¹⁴ C], [
³⁵ S] or the like.

As a test substance, any substance can be used. For example, a known peptide, a known GPCR ligand,
Biological samples and biological samples such as known proteins, recombinant proteins produced using recombinant techniques, cell extracts and purified products derived from the extracted solutions, cell culture supernatants and purified products derived from the supernatants, serum and the like Use of a purified product derived from a microorganism, a cell extract of a microorganism, a purified product derived from the extract, a microorganism culture supernatant or a purified product derived from the supernatant, a known compound, a compound synthesized using combinatorial chemistry, or the like. Can be.

The polypeptide or partial peptide of the present invention used in the present method may be any as long as it contains the polypeptide or partial peptide, and may be a cell containing the polypeptide or partial peptide. The polypeptide or the partial peptide purified from the above may be used, or a cell containing the polypeptide or the partial peptide itself or a cell membrane fraction thereof may be used. When cells containing the polypeptide or partial peptide are used, the cells may be immobilized with glutaraldehyde, formalin, or the like.

[0186] The polypeptide or partial peptide of the present invention may be a naturally occurring polypeptide or partial peptide, or a recombinant polypeptide or a recombinant partial peptide prepared using a gene recombination technique. Was constitutively active in the polypeptide encoded by the mutated DNA obtained by introducing a mutation into the DNA having the base sequence represented by SEQ ID NO: 2 by the method described in (3). Variant polypeptides are particularly useful.

As described in (6-1), regarding the GPCR mutated to the constitutively active form, it is known that the mutant GPCR may have an increased affinity for agonists. Mutant GPCRs of the type are useful in the search for agonists. Searching for an antagonist usually requires the use of a ligand, but in the case of a GPCR whose ligand is unknown (called an orphan GPCR), the ligand cannot be used. However, if a natural or mutant constitutively active GPCR is used, it is possible to search for an antagonist without a ligand.
For example, an antagonist can be selected by searching for a signal that flows when a constitutively active GPCR polypeptide is overexpressed in a cell or a substance that suppresses activation of a G protein. At this time, a GPCR function modifier may be selected together with the antagonist. Also,
By searching for a signal that flows when the constitutively active GPCR polypeptide is overexpressed in cells and a substance that enhances the activation of G protein, an agonist or a function modifying substance can also be selected.

A number of diseases caused by mutations in GPCRs and changes to constitutively active forms have been known [Japanese clinical practice, 56 , 1658 (1998), Japanese clinical practice, 56 , 1843 (1998),
Japanese clinical, 56 , 1856 (1998), Japanese clinical, 56 , 1931 (199)
8), Trends in Endocrinology and Metabolism, 9 , 27
(1998), Endocrinology, 137 , 3936 (1996)]. Antagonists capable of suppressing the activity of these constitutively active mutant GPCRs (called inverse agonists) are useful for treating diseases caused by constitutively active mutant GPCRs. Antagonists are classified into neutral antagonists and inverse agonists. Inverse agonists can suppress the activity of constitutively active GPCRs, whereas neutral antagonists cannot suppress constitutive activity. The constitutively active mutant GPCR is useful for searching for an inverse agonist.

The cell membrane fraction used in the above method can be prepared by the method described in the above (6-1).

As the cells expressing the polypeptide or partial peptide of the present invention, as described in the above (4), a recombinant DNA containing the DNA encoding the polypeptide is introduced into an appropriate host cell. Cells expressing the polypeptide in large amounts, such as the resulting transformed cells, can also be used. Examples of the host cell include microorganisms such as Escherichia coli, Bacillus subtilis, and yeast, as well as insect cells, frog oocytes, frog melanocytes, animal cells, and plant cells. In order that the polypeptide of the present invention expressed by the transformed cell retains a higher-order structure and retains binding with a ligand, yeast, insect cells, frog oocytes, frog melanocytes, animal cells, It is preferably expressed in plant cells and the like. A mutant strain of yeast or a yeast expressing the modified Gα protein can also be used as a host [Tr
ends in Biotechnology, 15 , 487 (1997), Mol. Cell.
Biol., 15 , 6188 (1995), Mol. Cell. Biol., 16 , 4700.
(1996)].

The amount of GPCR in cells containing the polypeptide or partial peptide of the present invention or in the cell membrane fraction thereof is as follows:
It is preferably 10 ³ to 10 ⁸ molecules per cell, preferably 10 ⁵ to 1 molecule.
0 ⁷ a molecule of is preferable. The higher the expression level, the higher the ligand binding activity (specific activity) per membrane fraction, which makes it possible not only to construct a highly sensitive screening system, but also to measure a large number of samples in the same lot. .

Hereinafter, specific examples will be described.

A preparation of the polypeptide or the partial peptide of the present invention is prepared by the method described in the above (6-1). To the polypeptide or partial peptide preparations of 10Myueru～10ml, the test substance and radioactive isotopes ^{(3 H, 125 I, 14} C,
³⁵ S, ³² P, etc.) and coexist with a certain amount of radioactive ligand. A reaction tube containing a large excess of unlabeled ligand is also prepared to determine the amount of non-specific binding (NSB). The reaction is carried out at about 0 to 50 ° C, preferably about 4 to 37 ° C,
The reaction is performed for about 20 minutes to 24 hours, preferably for about 30 minutes to 3 hours. After the reaction, the mixture is filtered with a glass fiber filter or the like, washed with an appropriate amount of the same buffer, and the radioactivity remaining on the glass fiber filter is measured with a liquid scintillation counter or a γ-counter. The count (B-NSB) obtained by subtracting the non-specific binding amount (NSB) from the total binding amount (B) is the specific binding amount. A specific binding amount of the ligand in the absence of the test substance, and comparing the specific binding amount of the ligand in the presence of the test substance, a substance that decreases the specific binding amount of the ligand, an agonist of the receptor protein of the present invention, It can be selected as an antagonist or a function modifier.

In the above method, as a cell containing the polypeptide or the partial peptide of the present invention, a host cell that does not express the polypeptide of the present invention is transformed into a vector DNA by encoding the DNA encoding the polypeptide or the partial peptide.
By using cells expressing a large amount of the polypeptide or the partial peptide obtained by introducing the recombinant DNA incorporated into the receptor, the agonist, antagonist or functional modifier of the receptor protein of the present invention can be selected with higher sensitivity. Can be. In addition, cells that do not express the polypeptide obtained by introducing only the vector into the same host cell, and other cells that are obtained by introducing an expression plasmid of another G protein-coupled receptor polypeptide into the same host cell. By performing similar experiments using cells expressing the G protein-coupled receptor polypeptide, it is possible to examine the specificity of the obtained agonist, antagonist or functional modifier for the G protein-coupled receptor polypeptide of the present invention. it can. (II) Method for measuring the amount of GTPγS bound to Gα protein As shown in (c) above, a test substance and a ligand are brought into contact with the membrane fraction of a cell containing the polypeptide of the present invention,
By measuring the amount of labeled GTPγS bound to the Gα protein (membrane fraction), it is possible to screen for agonists, antagonists or functional modifiers of the GPCR [Molecular Pharmacology, 47 , 848-854 (19)
95), WO98 / 46995].

Further, as shown in (f) above, the test substance was brought into contact with the membrane fraction of the cells containing the polypeptide of the present invention, and the amount of labeled GTPγS bound to the Gα protein (membrane fraction). By measuring the molecular weight, an agonist or a functional modifier of the polypeptide can be screened [Molecular Pharmacology, 47 , 848-854 (1992).
5), WO98 / 46995].

As a test substance, any substance can be used. For example, a known peptide, a known GPCR ligand,
Biological samples and biological samples such as known proteins, recombinant proteins produced using recombinant techniques, cell extracts and purified products derived from the extracted solutions, cell culture supernatants and purified products derived from the supernatants, serum and the like Use of a purified product derived from a microorganism, a cell extract of a microorganism, a purified product derived from the extract, a microorganism culture supernatant or a purified product derived from the supernatant, a known compound, a compound synthesized using combinatorial chemistry, or the like. Can be. GTP labeled
As γS, for example, GTPγS labeled with ³⁵ S can be used.

As the cells containing the polypeptide of the present invention or the membrane fraction of the cells, those described in the above (I) can be used.

Hereinafter, specific examples will be described.

The polypeptide preparation of the present invention was prepared using the above (6-
Prepared by the method described in 1).

When screening for agonists, 10
A test substance, a radioactive isotope (such as ³⁵ S) -labeled GT with a certain amount of radioactivity
PγS and GDP coexist. If it is necessary to know the non-specific binding amount (NSB), prepare a reaction tube to which a large excess of unlabeled GTPγS has been added. The count (B-NSB) obtained by subtracting the non-specific binding amount (NSB) from the total binding amount (B) is the specific binding amount. The reaction is about 0
The reaction is carried out at 50 ° C., preferably about 4-37 ° C., for about 20 minutes to 24 hours, preferably for about 30 minutes to 3 hours. After the reaction, the mixture is filtered with a glass fiber filter or the like, washed with an appropriate amount of the same buffer, and the radioactivity remaining on the glass fiber filter is measured with a liquid scintillation counter. A substance having an activity of enhancing the binding of GTPγS to the membrane fraction can be selected as an agonist or a function modifier of the polypeptide of the present invention.

When screening for antagonists or functional modifiers, a ligand, a test substance, a radioactive isotope (such as ³⁵ S) -labeled GTPγS, And G
A similar experiment is performed in the presence of DP. Comparing the amount of GTPγS binding in the absence of the test substance with the amount of ligand binding in the presence of the test substance, and selecting a substance that reduces the amount of GTPγS binding as an antagonist or a functional modifier of the polypeptide of the present invention. Can be. On the other hand, a substance that increases the amount of GTPγS bound can be selected as a function-modifying substance or an agonist of the polypeptide of the present invention.

As a cell containing the polypeptide of the present invention, a polypeptide obtained by introducing a recombinant DNA obtained by incorporating a DNA encoding the polypeptide into a vector into a host cell that does not express the polypeptide of the present invention. By using cells expressing a large amount of the above, an agonist, an antagonist or a functional modifier of the receptor protein of the present invention can be selected with higher sensitivity. In addition, cells that do not express the polypeptide obtained by introducing only the vector into the same host cell, and other cells that are produced by introducing an expression plasmid for another G protein-coupled receptor polypeptide into the same host cell. By performing the same experiment using cells expressing the G protein-coupled receptor polypeptide, it is possible to examine the specificity of the obtained agonist, antagonist or functional modifier for the G protein-coupled receptor polypeptide of the present invention. it can. (III) Method for measuring GTPase activity As shown in (d) above, a ligand and a test substance are brought into contact with a membrane fraction of a cell containing the polypeptide of the present invention,
By measuring the GTPase activity, an agonist, antagonist or functional modifier of the polypeptide can be screened [J. Biol. Che., 27
1 , 1857-1860 (1996), WO98 / 46995].

Further, as shown in the above (e), the test substance is brought into contact with the membrane fraction of the cell containing the polypeptide of the present invention, and the GTPase activity is measured to determine the agonist or function of the polypeptide. Modifiers can be screened [J. Biol. Che., 271 , 1857-18.
60 (1996), WO 98/46995].

As the test substance, any substance can be used. For example, a known peptide, a known GPCR ligand,
Biological samples and biological samples such as known proteins, recombinant proteins produced using recombinant techniques, cell extracts and purified products derived from the extracted solutions, cell culture supernatants and purified products derived from the supernatants, serum and the like Use of a purified product derived from a microorganism, a cell extract of a microorganism, a purified product derived from the extract, a microorganism culture supernatant or a purified product derived from the supernatant, a known compound, a compound synthesized using combinatorial chemistry, or the like. Can be.

As the cells containing the polypeptide of the present invention or the membrane fraction of the cells, those described in the above (I) can be used.

Hereinafter, specific examples will be described.

The polypeptide preparation of the present invention was prepared using the above (6-)
Prepared by the method described in 1).

When screening for agonists, 10
the Myueru～10 ml of the polypeptide preparation, test compound coexist GTP radioactive isotopes (such as ³² P) constant amount of radioactivity labeled with (e.g. [γ ³² P] GTP). The reaction is carried out at about 0 to 50 ° C, preferably about 4 to 37 ° C, for about 20 minutes to 24 hours, preferably for about 30 minutes to 3 hours. After the reaction, the supernatant of the reaction solution was recovered, and the released [γ ³² P] Pi
Is measured with a liquid scintillation counter. After filtering the reaction solution with a glass fiber filter or the like and washing with an appropriate amount of the same buffer, the radioactivity in the filtrate may be measured with a liquid scintillation counter. GTPase
A substance having an activity of enhancing the activity can be selected as an agonist or a function modifier of the polypeptide of the present invention.

When screening for antagonists or functional modifiers, 10 μl to 10 ml of the polypeptide preparation is added to a ligand, a test compound, a radioactive isotope (such as ³² P), and a certain amount of radioactive GTP ( For example, [γ ³²
P] GTP) is used in the same experiment. By comparing the GTPase activity in the absence of the test substance with the GTPase activity in the presence of the test substance, a substance that decreases the GTPase activity can be selected as an antagonist or a function modifier of the polypeptide of the present invention. On the other hand, a substance that increases GTPase activity can be selected as a function modifier or an agonist of the polypeptide of the present invention.

As a cell containing the polypeptide of the present invention, a recombinant DNA obtained by introducing a DNA encoding the polypeptide into a vector is introduced into a host cell that does not express the polypeptide of the present invention. By using cells expressing a large amount of the peptide, the agonist, antagonist or functional modifier of the receptor protein of the present invention can be selected with higher sensitivity. Also,
A cell that does not express the polypeptide obtained by introducing only a vector into the host cell, or another G protein that is obtained by introducing an expression plasmid for another G protein-coupled receptor polypeptide into the host cell By performing a similar experiment using cells expressing the conjugated receptor polypeptide, the specificity of the obtained agonist, antagonist, or function modifying substance for the G protein-coupled receptor polypeptide of the present invention can be examined. (IV) Method of Detecting Cell Response As shown in (e) above, a cell expressing the polypeptide of the present invention is brought into contact with a ligand and a test substance, and the activation of the polypeptide is used as an indicator of the cell response. As a result, an agonist, antagonist or functional modifier of the polypeptide can be screened.

[0211] As shown in the above (h), a test substance is brought into contact with cells expressing the polypeptide of the present invention.
By detecting the activation of the polypeptide using the response of the cell as an index, an agonist or a functional modifier of the polypeptide can be screened.

The cell response includes, for example, arachidonic acid release, acetylcholine release, intracellular Ca ²⁺ release, intracellular cAMP generation, intracellular cAMP reduction, intracellular cGMP
Production, inositol phosphate production, cell membrane potential fluctuations, phosphorylation of intracellular proteins, c-fos activation, pH reduction,
Cell proliferation activity, melanin pigment aggregation or diffusion, etc. are measured. The response of cells can also be monitored using a reporter system. For example, by introducing into a cell that expresses a functional polypeptide of the present invention, a DNA linked with an appropriate reporter gene downstream of the promoter sequence of a gene whose expression is induced by activation of the polypeptide, Activation of the polypeptide can be measured by expression of the reporter gene. Examples of the promoter include a promoter of ICAM-1 gene, c-
A fos promoter, a Krox-24 promoter and the like can be used. Further, the promoter may be an artificial promoter consisting of a binding sequence of an appropriate transcription factor and a basic promoter. As the binding sequence of the transcription factor, for example, CRE, TRE, SRE and the like can be used. Examples of the reporter gene include a chloramphenicol acetyltransferase gene, a β-glucuronidase gene, a β-galactosidase gene, a β-lactamase gene, a luciferase gene, an aequorin gene, and a green fluorescent protein gene.

As the cells expressing the polypeptide of the present invention, as described in (4) above, a trait obtained by introducing a recombinant DNA containing a DNA encoding the polypeptide into an appropriate host cell. Cells expressing the polypeptide in large amounts, such as transformed cells, can also be used. Examples of the host cell include microorganisms such as Escherichia coli, Bacillus subtilis, and yeast, as well as insect cells, frog oocytes, frog melanocytes, animal cells, and plant cells. In order that the polypeptide of the present invention expressed by the transformed cell retains a higher-order structure and retains binding and functionality with a ligand, yeast, insect cells, frog oocytes, and frog melanocytes It is preferably expressed in animal cells, plant cells and the like. A yeast mutant or a yeast expressing the modified Gα protein can also be used as a host.

As the test substance, any substance can be used. For example, a known peptide, a known GPCR ligand,
Biological samples and biological samples such as known proteins, recombinant proteins produced using recombinant techniques, cell extracts and purified products derived from the extracted solutions, cell culture supernatants and purified products derived from the supernatants, serum and the like Use of a purified product derived from a microorganism, a cell extract of a microorganism, a purified product derived from the extract, a microorganism culture supernatant or a purified product derived from the supernatant, a known compound, a compound synthesized using combinatorial chemistry, or the like. Can be.

Specific examples will be described below. Agonist screening method Cells expressing the polypeptide of the present invention are cultured in a multiwell plate or the like. If necessary, replace with a fresh medium or an appropriate buffer that is not toxic to cells, add the test substance, and incubate for a certain period of time. afterwards,
Cell response (eg, arachidonic acid release, acetylcholine release, intracellular Ca ²⁺ release, intracellular cAMP production, intracellular cGMP production, inositol phosphate production, cell membrane potential fluctuation, intracellular protein phosphorylation, c-fos activity , P
H reduction, melanin pigment aggregation or diffusion, or the activity of promoting or suppressing the expression level of a reporter gene, etc.). For example, using a cell extract or supernatant, a product produced by the cell response is quantified according to a conventional method. Substance as an indicator of cell stimulating activity (for example,
When the production of arachidonic acid or the like is difficult to be assayed by a degrading enzyme contained in cells, an inhibitor for the degrading enzyme may be added for quantification. As for activities such as cAMP production inhibition, cells were cAA-expressed with forskolin or the like.
It can be detected as a production inhibitory effect on cells whose MP production has been increased. A substance having an activity of enhancing a cellular response can be selected as an agonist or a function modifier of the polypeptide of the present invention. Screening for antagonists or functional modifiers Cells expressing the polypeptide of the present invention are cultured in a multiwell plate or the like. If necessary, replace the medium with a fresh medium or an appropriate buffer that is not toxic to cells, add the ligand and the test substance, and incubate for a certain period of time. Thereafter, the response of the cells is measured by the same method as described above. By comparing the cellular response in the absence of the test substance with the cellular response in the presence of the test substance, a substance that reduces the cellular response can be selected as an antagonist or a functional modifier of the polypeptide of the present invention. On the other hand, substances that increase the cellular response can be selected as functional modifiers or agonists of the polypeptide of the present invention.

The cells containing the polypeptide of the present invention
Host cells that do not express the polypeptide of the present invention.
DNA encoding the repeptide was incorporated into the vector
The size of the polypeptide obtained by introducing the recombinant DNA
The receptor protein of the present invention is obtained by using
Agonists, antagonists or functional modifiers
Can be selected with high sensitivity. In addition, the host cell
The polypeptide obtained by introducing only the vector
Cells that do not express the peptide or host cells that contain other G proteins
Introduce an expression plasmid for a coupled receptor polypeptide
G protein-coupled receptor polypeptide obtained by the method
Similar experiments were performed using peptide-expressing cells.
Acquired agonists, antagonists or functional
G protein-coupled receptor polypeptide of the present invention as a decorating substance
The specificity can be determined. (6-5) G protein-coupled receptor polypeptide of the present invention
Of agonists, antagonists or functional modifiers
Cleaning Kit Agonists, antagonists and the like of the polypeptide of the present invention.
Or a kit for screening for a functional modifier
Or a partial peptide thereof or
Or a polypeptide of the present invention or a partial peptide thereof.
A cell containing a tide-containing cell or a membrane fraction of the cell.
And so on. The screening kit of the present invention
For example, the following reagents and measurement methods can be used.
You. (A) Screening reagent Measurement buffer and washing buffer Hanks' Balanced Salt Solution (Gibco)
05% bovine serum albumin (Sigma) was added.
of. Sterilized by filtration through a 0.45 μm filter, 4 ° C
Or may be prepared at the time of use. Polypeptide preparation of the present invention CHO cells expressing the polypeptide of the present invention
5 × 10 on hole plate ^FivePassage by individual / hole, 5% CO_Two, 9
Cultured at 37 ° C for 2 days in a 5% air incubator
What you did. Labeled ligand^ThreeH], [¹²⁵I], [¹⁴C], [³⁵S]
Labeled ligand. Aqueous solution at 4 ° C or
Store at −20 ° C., and use 1 μmol /
Dilute to L. Ligand standard solution 0.1% bovine serum albumin (Sigma)
And dissolved at 1 mmol / L in PBS containing -20 ° C.
To save. (B) Measuring method The poly of the present invention cultured on a 12-well tissue culture plate
Peptide-expressing CHO cells were washed twice with 1 ml of measurement buffer.
After washing, 490 μl of measurement buffer was added to each well.
You. 10^-3-10^-Ten Add 5 μl of mol / L test substance solution
After adding 5 μl of labeled ligand, react for 1 hour at room temperature
Let it. To know the amount of non-specific binding
10^-3 Add 5 μl of mol / L ligand. Remove the reaction solution and wash 3 times with 1 ml of washing buffer
You. The labeled ligand bound to the cells is lysed (0.2 mol / L)
 NaOH, 1% SDS).
Mix with Chilator A (manufactured by Wako Pure Chemical Industries). Liquid scintillation counter (Beckman)
Radioactivity was measured using Percent Maximum Binding
(PMB) is calculated by the following equation. PBM = (B−NSB) ÷ B0 × 100 PMB: Percent Maximum Binding B: Value when a sample is added NSB: Non-specific Binding (Non-specific binding amount) B0: Maximum binding amount The above (6-4) Screening method or this screen
The substance or its salt obtained using the kit for
Agonists, antagonists and the like of the polypeptide of the present invention.
Or a functional modifier. Such substances include peptides,
Protein, non-peptide compound, synthetic compound, fermentation production
Substances, and even if these substances are new substances,
Or a known substance, but the known substance is
Agonist, antagonist or functionally modified product of the present invention
Not included in quality. Jaw for the polypeptide of the present invention
Nists have ligands for the polypeptides of the invention.
Has the same effect as the physiological activity of
It is useful as a safe and low toxic pharmaceutical composition
is there. Conversely, antagonies to the polypeptides of the invention
The strike has a ligand for the polypeptide of the present invention.
Biological activity can be suppressed,
It is useful as a safe and low toxic pharmaceutical composition
You. In addition, the functional modifier of the polypeptide of the present invention
Physiological activity of the ligand for the polypeptide of the invention
Can be enhanced or suppressed.
Safe and low toxic pharmaceutical composition to enhance or suppress
And useful.

When the compound or its salt obtained by using the screening method of (6-4) or this screening kit is used as the above pharmaceutical composition, it can be formulated according to a conventional method. For example, sugar-coated tablets as needed, capsules, elixirs, orally as microcapsules, or a sterile solution with water or other pharmaceutically acceptable liquid,
Alternatively, it can be used parenterally in the form of injections such as suspensions. For example, the compound or a salt thereof is mixed with physiologically acceptable carriers, flavors, excipients, vehicles, preservatives, stabilizers, binders and the like in a unit dosage form required for generally accepted pharmaceutical practice. Can be manufactured. The amount of the active ingredient in these preparations is such that an appropriate dose in the specified range can be obtained. Examples of additives that can be mixed with tablets, capsules, and the like include binders such as gelatin, corn starch, tragacanth, and acacia, excipients such as crystalline cellulose, corn starch, gelatin, and alginic acid. Useful bulking agents, lubricants such as magnesium stearate, sweeteners such as sucrose, lactose or saccharin, flavoring agents such as peppermint, reddish oil or cherry and the like are used. When the preparation unit form is a capsule, a liquid carrier such as oil and fat can be further contained in the above-mentioned type of material. Sterile compositions for injection can be formulated according to normal pharmaceutical practice such as dissolving or suspending the active substance in vehicles such as water for injection, naturally occurring vegetable oils such as sesame oil, coconut oil and the like. .

As the aqueous solution for injection, for example, physiological saline, isotonic solution containing glucose and other adjuvants (eg, D-sorbitol, D-mannitol, sodium chloride, etc.) and the like are used. It may be used in combination with a solubilizing agent, for example, an alcohol (eg, ethanol), a polyalcohol (eg, propylene glycol, polyethylene glycol), a nonionic surfactant (eg, polysorbate 80 (TM), HCO-50). Good. As the oily liquid, for example, sesame oil, soybean oil and the like are used, and may be used in combination with solubilizers such as benzyl benzoate and benzyl alcohol. In addition, buffers (eg, phosphate buffer, sodium acetate buffer), soothing agents (eg, benzalkonium chloride, procaine hydrochloride, etc.), stabilizers (eg, human serum albumin, polyethylene glycol, etc.), storage Agents (eg, benzyl alcohol, phenol, etc.), antioxidants and the like. The prepared injection is usually filled in a suitable ampoule. Since the thus obtained preparation is safe and has low toxicity, it can be administered to, for example, humans and mammals (eg, rats, rabbits, sheep, pigs, cows, cats, dogs, monkeys, etc.). . The dose of the compound or a salt thereof varies depending on the administration subject, the target organ, the condition, the administration method, and the like. However, in the case of oral administration, generally, in an adult (60 kg), about 0.1 per day is used.
-100 mg, preferably about 1.0-50 mg, more preferably about 1.0-20 mg. In the case of parenteral administration, the single dose varies depending on the administration subject, target organ, symptoms, administration method and the like. About 0.01 to 30 mg, preferably about 0.1
Conveniently, about 20 mg, more preferably about 0.1 to 10 mg is administered by intravenous injection. In the case of other animals, the dose can be administered in terms of 60 kg. (7) Use of DNA or oligonucleotide of the present invention (7-1) Obtaining chromosomal gene encoding polypeptide of the present invention and use of the gene (I) Obtaining chromosomal gene encoding polypeptide of the present invention Using the DNA or oligonucleotide of the invention as a probe, a known method [New Cell Engineering Experimental Protocol, edited by Cancer Research Division, Institute of Medical Science, The University of Tokyo, Shujunsha (1993)
Year)] can be used to obtain a chromosomal gene encoding the polypeptide of the present invention and an expression control region of the gene.

By comparing the sequence of the human cDNA encoding the polypeptide of the present invention with the sequence of the human chromosomal gene registered in a public database, the human chromosome encoding the polypeptide of the present invention can be compared. It may be possible to identify genes and elucidate their structure. If a chromosomal gene sequence that matches the cDNA sequence is registered, the exon and intron structures of the chromosomal gene encoding the polypeptide of the present invention are compared by comparing the sequence of the chromosomal gene with the sequence of the DNA of the present invention, and The expression control region (eg, promoter region, etc.) of the chromosomal gene can be determined.

Examples of the promoter region include all promoter regions involved in transcription of a gene encoding the polypeptide of the present invention in mammalian cells.
Specifically, for example, the human thalamus, cerebellum, whole brain, hippocampus,
Substantia nigra, fetal brain, fetal kidney, fetal liver, heart, liver, mammary gland,
Placenta, prostate, salivary glands, skeletal muscle, thymus, thyroid, uterus,
In human colorectal cancer cells or human gastric cancer cells, a promoter region involved in transcription of a gene encoding the polypeptide of the present invention can be mentioned.

As a specific example of the promoter region, the base sequence shown in SEQ ID NO: 15
D having the nucleotide sequence represented by Nos. 0202 to 25202
A DNA having a continuous base sequence of 50 to 5000 bp in NA can be mentioned. (II) Chromosomal gene encoding polypeptide of the present invention and method of using the gene As shown in (8) below, the promoter region is used for screening for a substance that controls transcription of the gene encoding the polypeptide of the present invention. Useful. Further, a decoy DNA for suppressing transcription of a gene encoding the polypeptide of the present invention using the sequence information of the promoter region.
(Nippon Rinsho-Japanese Journal of Clinical Medi
cine, 56 , 563 (1998), Circulation Research, 82 , 10
23 (1998), Experimental Nephrology, 5 , 429 (199
7), Nippon Rinsho-Japanese Journal of Clinical M
edicine, 54 , 2583 (1996)]. (7-2) Measurement of Transcript Amount of Gene Encoding G Protein-Coupled Receptor Polypeptide of the Present Invention Northern Hybridization Method (Molecular Cloning 2nd Edition), PCR Method [PCR Protocols, Acade
mic Press (1990)], quantitative PCR (Proc. Natl. Aca
d. Sci. USA, 87 , 2725 (1990)], Real Time PCR (J
Unko Stevens, Experimental Medicine (extra edition), 15 , 46 (1997)] and the like can measure the amount of the transcript of the gene encoding the polypeptide of the present invention. In particular, the quantitative P
The CR method and the Real Time PCR method are preferred methods in that they have excellent quantitative properties. By quantifying the transcript,
Diagnosis of diseases based on abnormal expression of the polypeptide of the present invention is possible. Therefore, the DNA or oligonucleotide of the present invention is useful as a genetic diagnostic agent for quantifying a transcript of a gene encoding the polypeptide of the present invention. (7-3) Detection of Mutations and Polymorphisms in the Gene Encoding the G Protein-Coupled Receptor Polypeptide of the Present Invention It is known that mutations and polymorphisms exist in the GPCR gene or the expression control region of the GPCR gene. I have. For example,
It is known that the mutation of the GPCR gene inactivates or activates the function of the GPCR and causes various diseases [Japanese clinical, 56 , 1658 (1998), Japanese clinical, 56 , 1836.
(1998), Japanese clinical practice, 56 , 1843 (1998), Japanese clinical practice, 56 , 1
848 (1998), Japanese clinical study, 56 , 1856 (1998), Japanese clinical study, 5
6 , 1871 (1998), Japanese clinical practice, 56 , 1876 (1998), Japanese clinical practice, 56 , 1931 (1998), Trends in Endocrinology and Me
tabolism, 9 , 27 (1998)]. In addition, GPCR gene or GPCR gene expression control region mutation
Expression level increases or decreases, and various diseases may occur. Therefore, by examining the mutation of the gene encoding the polypeptide of the present invention or the expression control region of the gene, inactivation or activation of the function of the receptor protein of the present invention, or expression of the polypeptide of the present invention, Diagnosis of a disease based on an increase or decrease can be made.

Further, the properties of the GPCRs and the GPCR
It is known that the expression level of the disease changes, and the disease incidence rate and progression rate differ (Cancer. Res., 59 , 3561 (1999), A
nnu. Rev. Immunol., 17 , 657 (1999), Japanese clinical practice, 56 ,
1871 (1998)]. For example, the beta ₃ adrenergic receptor 64
It is known that a person having the mutant receptor protein in which the second tryptophan is substituted with arginine has a high incidence of obesity and diabetes. Therefore, by examining the mutation of the gene encoding the G protein-coupled receptor polypeptide of the present invention or the expression control region of the gene, the disease incidence rate based on changes in the properties and expression level of the polypeptide of the present invention can be improved. A prediction of the speed of progress can be made.

Most of the drugs currently used are G
Although it is targeted for PCR, the GPR gene and the polymorphism of the expression control region of the GPCR gene may cause
It is known that the properties of CR and the expression level of GPCR change, and the sensitivity to drugs changes [Journal of Pharmac.
olgy and Experimental Therapeutics, 275 , 1274 (199
5), J. Bilo. Chem., 272 , 1822 (1997), Pharmacogene
tics, 5 , 318 (1995), J. Bilo. Chem., 274 , 12670 (1
Natl. Acad. Sci. USA, 95 , 9608 (199).
8), Science, 286 , 487 (1999)]. Therefore, by examining the mutation of the gene encoding the polypeptide of the present invention or the expression control region of the gene, it is possible to predict drug sensitivity based on changes in the properties and expression level of the polypeptide of the present invention.

Detection of a mutation or polymorphism in the gene encoding the polypeptide of the present invention or the expression control region of the gene can be performed using the nucleotide sequence information of the gene or the control region. Specifically, gene mutations and polymorphisms can be detected by Southern blotting, direct sequencing, PCR, DNA chip method, etc. [Clinical test, 42 , 1507-1517 (1998), clinical test , 42 , 15
65-1570 (1998)].

By using a DNA encoding the polypeptide of the present invention or a partial peptide thereof and a DNA having a nucleotide sequence in the expression control region of a gene encoding the polypeptide of the present invention as a probe or a primer, Alternatively, it is possible to detect a gene encoding the polypeptide of the present invention in a mammal (eg, rat, rabbit, sheep, pig, cow, cat, dog, monkey, etc.) and a mutation or polymorphism in the expression control region of the gene. it can. Therefore, a DNA encoding the polypeptide of the present invention or a partial peptide thereof, and a DNA having a nucleotide sequence in the expression control region of the gene encoding the polypeptide of the present invention may be a gene for detecting the above mutation or polymorphism. Useful as a diagnostic agent. (7-4) A therapeutic agent for a disease in which the expression level or function of the G protein-coupled receptor polypeptide of the present invention is enhanced. Antisense RNA / DNA technology [Bioscience and Industry, 50 , 322 (1992), Chemistry, 46 , 681 (199
1), Biotechnology, 9 , 358 (1992), Trends in Biotec
hnology, 10 , 87 (1992), Trends in Biotechnology,
10 , 152 (1992), Cell Engineering, 16 , 1463 (1997)), Triple Helix Technology (Trends in Biotechnology, 10 ,
132 (1992)), ribozyme technology (Current Opinion in
ChemicalBiology, 3 , 274 (1999), FEMS Microbiology
Reviews, 23 , 257 (1999), Frontiers in Bioscience,
4 , D497 (1999), Chemistry & Biology, 6 , R33 (199
9), Nucleic Acids Research, 26 , 5237 (1998), Trend
s In Biotechnology, 16 , 438 (1998)) or decoy DNA method [Nippon Rinsho-Japanese Journal of Cli
nical Medicine, 56 , 563 (1998), Circulation Resear
ch, 82 , 1023 (1998), Experimental Nephrology, 5 , 4
29 (1997), Nippon Rinsho-Japanese Journal of Cli
nical Medicine, 54 , 2583 (1996)].

For example, the DN of the present invention described in the above (2)
A, using an oligonucleotide or a derivative thereof, suppresses transcription of a DNA encoding the polypeptide of the present invention;
Alternatively, translation of a transcript encoding the polypeptide of the present invention can be suppressed. That is, by administering the DNA, oligonucleotide or derivative thereof of the present invention, production of the polypeptide of the present invention can be suppressed.

For example, when there is a patient whose physiological activity via a receptor is enhanced due to increased expression of the polypeptide of the present invention or increased expression of a ligand of the polypeptide,
The physiological action can be suppressed by administering the DNA, oligonucleotide or derivative thereof of the present invention to a patient. That is, the DNA, oligonucleotide, or derivative thereof of the present invention is useful as a therapeutic agent for a disease in which the expression level or function of the polypeptide of the present invention is enhanced.

When the DNA, oligonucleotide or derivative thereof of the present invention is used as the above therapeutic agent, the DNA, oligonucleotide or derivative thereof of the present invention may be used alone or in a retrovirus vector, adenovirus vector, adenovirus associated virus vector. After insertion into an appropriate vector such as
It can be formulated, formulated and administered according to the usual method described in 5). (7-5) Gene preventive / therapeutic agent for a disease in which the expression level or function of the G protein-coupled receptor polypeptide of the present invention is reduced, DNA encoding the polypeptide of the present invention, the expression level of the polypeptide of the present invention Alternatively, it can be used as a medicament such as an agent for preventing or treating a disease with reduced function. For example, when there is a patient who cannot expect the physiological action of the ligand due to a decrease in the expression or mutation of the polypeptide of the present invention, (i) DNA encoding the polypeptide of the present invention
Is administered to the patient and expressed, or (ii) after inserting and expressing the DNA encoding the polypeptide of the present invention in cells of interest and transplanting the cells into the patient, etc. Can increase the amount of the polypeptide of the present invention in the body of the subject and sufficiently exert the action of the ligand. Therefore, the DNA encoding the polypeptide of the present invention is useful as a safe and low-toxic gene preventive / therapeutic agent for diseases in which the expression level or function of the polypeptide of the present invention is reduced.

DNA encoding the polypeptide of the present invention
When used as the above prophylactic / therapeutic agent, the D of the present invention
After inserting NA alone or into an appropriate vector such as a retrovirus vector, an adenovirus vector, an adenovirus associated virus vector, etc., it can be formulated, formulated and administered according to the conventional method described in the above (7-4). . (7-6) Screening method for substance controlling transcription or translation of gene encoding G protein-coupled receptor polypeptide of the present invention Transcription process of gene encoding polypeptide of the present invention
Alternatively, a compound that promotes or suppresses the process of translating a transcript into a protein can control cell functions exerted through the polypeptide by controlling the expression of the polypeptide. Therefore, a compound that promotes or suppresses the transcription process of a gene encoding the polypeptide of the present invention or the translation process from a transcript to a protein can enhance or suppress the physiological activity of the ligand for the polypeptide of the present invention. Therefore, it is useful as a safe and low toxic pharmaceutical composition that enhances or suppresses the activity of the ligand.

The compound can be obtained by the following methods (a) to (c). (A) [i] a cell expressing the polypeptide of the present invention;
[Ii] In the presence of a test substance, cells expressing the polypeptide of the present invention are cultured for 2 hours to 1 hour by the culture method described in (4) above.
After culturing for a week, the amount of the polypeptide in the cells is determined according to the above (5).
A method for measuring and comparing using the antibody of the present invention described in the above, and selecting and obtaining a compound having an activity of increasing or decreasing the amount of the polypeptide.

As a measurement method using the antibody of the present invention, for example, an ELISA using a microtiter plate is used.
Method, fluorescent antibody method, Western blot method, immunohistochemical staining and the like. (B) [i] cells expressing the polypeptide of the present invention;
[Ii] In the presence of a test substance, cells expressing the polypeptide of the present invention are cultured for 2 hours to 1 hour by the culture method described in (4) above.
After weekly culture, DN encoding the polypeptide in the cells
The transcript amount of A is measured and compared using a method such as the Northern hybridization method or the PCR method described in the above (7-2), and a compound having an activity of increasing or decreasing the transcript amount is selected. How to get. (C) A plasmid in which a reporter gene-linked DNA is incorporated downstream of the promoter obtained in the above (7-2) is prepared by a known method, and introduced into animal cells according to the method described in the above (5). , To obtain a transformant.
[I] In the presence of the transformant and [ii] a test substance, the transformant is cultured for 2 hours to 1 hour by the culture method described in the above (5).
After culturing for a week, the expression level of the reporter gene in the cells can be determined by a known method (New Cancer Engineering Protocol, Shujunsha (1993), Biotechniques, Ed.
20 , 914 (1996), J. Antibiotics, 49 , 453 (1996),
A method for selecting and obtaining a compound having an activity of increasing or decreasing the expression level by measuring and comparing the results using Trends in Biochemical Sciences, 20 , 448 (1995), Cell Engineering, 16 , 581 (1997)].

Examples of the reporter gene include chloramphenicol acetyltransferase gene, β-glucuronidase gene, β-galactosidase gene, β-lactamase gene, luciferase gene, aequorin gene, and green fluorescent protein (GFP). Genes and the like can be mentioned. (7-7) Preparation of animal deficient or substituted for DNA of the present invention An animal deficient or substituted for the DNA of the present invention has all or a part of the gene containing the DNA of the present invention deleted or substituted. An animal in which the expression level of the polypeptide of the present invention has changed. The animal, or an organ, a tissue or a cell of the animal, can be used for evaluation of a medicament obtained by the above method, for example, a therapeutic drug for a disease such as colorectal cancer, gastric cancer or thalamus, cerebellar dysfunction, etc. It is useful as a drug evaluation model animal, organ, tissue or cell.

As the animal, a vector containing the DNA of the present invention is used, and the desired animal, for example, cow, sheep, goat,
Pigs, horses, chickens, embryonic stem cells of non-human mammals such as mice (embryonic stem cells), a DNA encoding the polypeptide of the present invention on the chromosome in a known homologous recombination technique (for example, Nature, 326 , 295 (1987), Cell,
51 , 3, 503 (1987), etc.] to produce a mutant embryonic stem cell clone inactivated or substituted with an arbitrary sequence [Nature, 350 , 243 (1991)].

Using the embryonic stem cell clone thus prepared, a chimera comprising an embryonic stem cell clone and normal cells by a technique such as injection chimera method or assembly chimera method of injecting a fertilized egg of an animal into a blastcyst. Individuals can be created. By crossing the chimeric individual with a normal individual, it is possible to obtain an individual having an arbitrary mutation in the DNA encoding the polypeptide of the present invention on the chromosome of whole body cells. A homozygous individual (knockout animal) having both mutations can be obtained.

Thus, in an animal individual, a mutation can be introduced into the chromosome at any position of the DNA encoding the polypeptide of the present invention. For example, by introducing a mutation such as deletion, substitution or addition of a base into the translation region of the DNA encoding the polypeptide of the present invention on the chromosome, the activity of the product can be changed.

By introducing similar mutations into the expression control region, it is also possible to modify the degree, timing, tissue specificity, etc. of the expression. Furthermore, by combining with the Cre-loxP system, the expression timing, expression site, expression amount, and the like can be more positively controlled.

[0237] Examples of such a case include the use of a promoter expressed in a specific region of the brain and deletion of the target gene only in that region [Cell, 87 , 1317 (1996)] and C
Using adenovirus expressing re, at the desired time,
An organ-specific deletion of a target gene [Science, 27
8 , 5335 (1997)].

Accordingly, the expression of the DNA encoding the polypeptide of the present invention on the chromosome can be controlled at any time and in any tissue, or the deletion, substitution or addition of any base can be controlled by its translation region or expression. It is possible to create an animal individual having the control region.

Such an animal may induce, at any time, at any degree, or at any site, various diseases caused by the polypeptide of the present invention, for example, cancer, dysfunction of the thalamus or cerebellum. Can be.

Therefore, the knockout animal of the present invention becomes an extremely useful animal model in the treatment and prevention of various diseases caused by the polypeptide of the present invention, for example, cancer and dysfunction of the thalamus or cerebellum. In particular, it is very useful as a therapeutic drug, a preventive drug, and a model for evaluating functional foods, health foods and the like.

[0241]

EXAMPLES The methods described in the following examples were carried out according to the method described in Molecular Cloning, Second Edition, unless otherwise specified. Example 1 Novel G protein-coupled receptor (KAT0673)
Cloning of cDNA Encoding 4L Polypeptide) (1) Preparation of KATOIII Cell-Derived cDNA Library mRNA was extracted and purified from human gastric cancer tissue cell line KATOIII. The oligocap method (Gen
e, 138 , 171 (1994)]. Oligo-cap linker (DNA having the base sequence represented by SEQ ID NO: 3) and Oligo dT primer (SEQ ID NO: 4
Using a DNA having a base sequence represented by the following formula: protein nucleic acid enzyme, 41 , 197 (1996), Gene, 200 , 149 (1997)
BAP (Bacterial Alkaline Phosphat)
ase) treatment, TAP (Tobacco Acid Pyrophosphatase) treatment, RNA ligation, first-strand cDNA synthesis and RN
A was removed. Next, double-stranded DNA was obtained by PCR using the DNA having the nucleotide sequence represented by SEQ ID NO: 5 (5 'end) and the DNA having the nucleotide sequence represented by SEQ ID NO: 6 (3' end) as a primer set. After converting into cDNA, it was cut with restriction enzyme SfiI . The cDNA
The vector pME18SFL3 (GenBank acc.
esion no. AB009864, Expression vector, 3392 bp) to prepare a cDNA library. By the above method, the cDNA is unidirectionally integrated so that expression is possible. (2) Random sequence Plasmid DNA was obtained from each E. coli clone of the cDNA library prepared in (1) according to a conventional method,
The nucleotide sequence at the 5 'end of the cDNA contained in each plasmid was determined. The nucleotide sequence is determined by the Dye Terminator Cycle
Sequencing FSReady Reaction Kit, dRhodamine Termi
nator Cycle Sequencing FS Ready Reaction Kit or
BigDye Terminator Cycle Sequencing FS Ready Reacti
on Kit (PE Biosystems) and DNA sequencer
This was performed using ABI PRISM 377 (manufactured by PE Biosystems).
DNAs having the nucleotide sequences represented by SEQ ID NOS: 7 and 8 were used as primers for nucleotide sequence determination.

[0242] The obtained base sequence was determined by Blast and Fast.
a) Analysis of homologous genes and proteins was performed using programs such as FrameSearch. As a result, pME-
CD contained in a plasmid named KAT06734
NA (referred to as KAT06734 cDNA) was found to encode a protein having homology to human vasopressin receptor and human luteinizing hormone releasing hormone receptor. (3) Determination of the entire nucleotide sequence of KAT06734 cDNA The nucleotide sequence at the 3 ′ end of the cDNA (KAT06734 cDNA) contained in pME-KAT06734 obtained in (2) above was also determined, and the entire nucleotide sequence of the cDNA was determined. did. To determine the nucleotide sequence, use PerkinElmer's DN
A Sequencer 377 and Applied Biosystems' reaction kit (ABI Prism ^™ BigDye ^™ Terminator Cycle Se
quencing Ready Reaction kit) was used.

The KAT06734 cDNA is 880 bp.
Encoded a polypeptide consisting of 143 amino acids (SEQ ID NO: 19). From the homology analysis and the hydropathy analysis, the protein encoded by the present cDNA was considered to be a novel GPCR lacking the C-terminus. (4) Full-length cDNA from human thalamus (KAT0673)
Acquisition of 4L cDNA) The 3'-RACE method was performed using total RNA from human gastric cancer cell line KATOIII and colon cancer cell line Colo205 as a template to obtain a 3'-terminal cDNA fragment deleted in KAT06734 cDNA. did. The 3'-RACE method includes 5 '/
Using a 3 'RACE kit (Boehringer), a DNA having the nucleotide sequence of SEQ ID NO: 9 (06734-5-
1) was used as a KAT06734 cDNA specific primer. The sequence of the cDNA fragment obtained by the 3′-RACE method was determined, and a sequence (KAT06733-1 ′) linked to the sequence of the KAT06734 cDNA was prepared (FIG. 11).
To 13). In this array, Open Reading
Since the frame was shifted, it was decided to try to obtain the full-length cDNA again by PCR. At that time, it was thought that there was a possibility of splicing abnormality in the cancer cell line.
Was used.

Using the single-stranded cDNA derived from the human thalamus prepared by the method of Example 4 below as a template, a DNA having the nucleotide sequence of SEQ ID NO: 11 (06734SP5 in FIG. 7) and a DNA represented by SEQ ID NO: 12 DNA (see FIG. 10)
PCR using 06734-3-3) as a primer set
Was performed to obtain a new cDNA fragment. In the PCR reaction, 10 pmol of a forward primer (DNA of SEQ ID NO: 11) was added to the single-stranded cDNA (6.25 μl) derived from the human thalamus synthesized in (a) of Example 4 and
10 pmol of the reverse primer (DNA of SEQ ID NO: 12) and 2.5 mmol / L dNTP mixed solution
μl, 5 units / μl of TaKaRa Ex Taq ^™ (Takar
a) (0.1 μl, 10 × reaction buffer (Takar)
a) was added and sterilized water was added to make a total amount of 2 μl.
It was adjusted to 5 μl. Thermal cycler DNA eng
ine (manufactured by MJ Research) at 95 ° C
After denaturing the DNA by heat treatment for 5 minutes, 35 cycles of a reaction consisting of 96 ° C. for 15 seconds and 68 ° C. for 1 minute and 48 seconds were performed. Thereafter, the reaction was further performed at 72 ° C. for 10 minutes.

The sequence of the cDNA fragment amplified by the above PCR was compared with ABI Prism ^™ BigDye ^™ Terminator Cycle Sequenci.
ng Ready Reaction Kit (PE Applied Biosystems)
Was determined by direct sequencing using As the primer, a DNA having the nucleotide sequence represented by SEQ ID NO: 9, 11, or 12, and a DNA having the nucleotide sequence represented by SEQ ID NO: 18 (06734-3-2 in FIG. 9) were used. Determined nucleotide sequence and KAT06734 cDNA
5'-terminal sequence and the 3'-terminal sequence of KAT0673-3-3 'were prepared (see FIGS. 7 to 10).
The cDNA having the synthetic sequence was designated as KAT0673Lc
It is called DNA, and its base sequence is shown in SEQ ID NO: 2. Book c
The DNA encoded a protein of 371 amino acids (see FIGS. 7-10). The polypeptide having the amino acid sequence is called KAT0673L polypeptide, and the amino acid sequence is shown in SEQ ID NO: 1.

From the hydropathy profile, the polypeptide was considered to have seven membrane-bound regions (FIG. 1). EMBO J., 12 , 1
693 (1993).

The polypeptide had homology at the amino acid level to known G protein-coupled receptors. It showed high homology with the receptor, human luteinizing hormone-releasing hormone receptor, and vasotocin receptor in white soccer. FIG. 14 shows a dendrogram created using the KAT0673L polypeptide and the amino acid sequence of the known G protein-coupled receptor. Dendrogram CL
USTAL X Multiple Sequence Alignment Program ( ftp: /
/ftp-igbmc.u-strasbg.fr/pub/ClustalX/ ). FIGS. 2 to 6 show the results of aligning the KAT0673L polypeptide and the amino acid sequence of the known GPCR using the same program. KAT06734
The seven predicted transmembrane regions in the L polypeptide (TM
1 to TM7) are underlined.

At the amino acid level, KAT0673L polypeptide interacts with human vasopressin 1A receptor by 24.8.
%, Mouse vasopressin 1A receptor 26.4%, human vasopressin 1B receptor 27.8%, human vasopressin 2 receptor 22.9%, human oxytocin receptor 25.6%, human luteinizing hormone release It showed 19.0% homology with the hormone receptor and 26.7% with white soccer vasotocin receptor.

From the above results, it was revealed that the polypeptide is a novel G protein-coupled receptor. Human and mouse vasopressin 1A receptor, human vasopressin 1B receptor, human vasopressin 2 receptor,
Since natural ligands of human oxytocin receptor, human luteinizing hormone-releasing hormone receptor, and white soccer vasotocin receptor are all peptides, it is estimated that the ligand of the novel G protein-coupled receptor is also a peptide. . In addition, since the novel G protein-coupled receptor shows 19.0 to 27.8% homology with the known G protein-coupled receptor, the ligand of the novel G protein-coupled receptor is It was presumed to be different from the ligand of the G protein-coupled receptor. Example 2 Expression of KAT0673L polypeptide in a human cultured cell line (1) Construction of pBS-KAT0673L L Using the single-stranded cDNA derived from the human thalamus prepared by the method of Example 4 below as a template, represented by SEQ ID NO: 13 PCR was performed using a DNA having a base sequence (06734-F in FIG. 7) and a DNA having a base sequence represented by SEQ ID NO: 14 (06734-R in FIG. 9) as a primer set, and KAT0 was obtained.
A cDNA fragment encoding the 6734L polypeptide was obtained. The PCR reaction was performed using the human thalamus-derived single-stranded cDNA (6.25 μl) synthesized in Example 4 (a) described below.
10 pmol of a Forward primer (DNA having the base sequence represented by SEQ ID NO: 13) in Rev
rse primer (DNA having the base sequence represented by SEQ ID NO: 14) at 10 pmol, 2.5 mmol / L dNTP
2.5 μl of the mixture, 5 units / μl of TaKaRa Ex Taq ^{T M}
0.1 μl (manufactured by Takara) and 2.5 μl of 10 × reaction buffer (manufactured by Takara) were added, and sterilized water was added to adjust the total volume to 25 μl. Thermal cycler D
The DNA was denatured by heat treatment at 95 ° C. for 5 minutes using NA engine (manufactured by MJ Research), and then denatured at 94 ° C. for 30 seconds, 65 ° C. for 1 minute, and 72 ° C. for 2 minutes.
One cycle of the reaction was performed for 35 minutes, and 35 cycles were performed. Thereafter, the reaction was further performed at 72 ° C. for 10 minutes.

[0250] limit the cDNA fragment enzyme Hin dIII
After the cut with Xba I, Hin of 1.1kb dIII- X
The ba I fragment was obtained. Also, pBluescript II SK
(+) After cleavage with restriction enzymes Hin dIII and Xba I,
It has acquired the Hin dIII- Xba I fragment of 3.0kb. By combining the above two fragments, pBS-KAT
0673L was produced. The absence of errors in the PCR amplified fragment was confirmed by sequencing. pBS-KAT
Escherichia coli DH5, which is Escherichia coli containing 06734L
α / pBS-KAT06734L was submitted to FERM BP-696 on December 8, 1999 by
No. 7 has been deposited. (2) KAT06734L was cleaved with polypeptide limit pBS-KAT06734L which was constructed with the expression plasmid pAMo-KAT06734L of Construction (1) enzyme Hin dIII and Not I, 1.1 kb
It has acquired the Hin dIII- Not I fragment. Also, p
AMo [J. Biol. Chem., 268 , 22782 (1993), aka p
AMoPRC3Sc (Japanese Unexamined Patent Publication No. 05-336963)]
After the cut with Hin dIII and Not I, of 8.7kb Hi
n dIII- Not acquired the I fragment. By combining the above two fragments, pAMo-KAT0673L was constructed. (3) Expression of KAT0673L polypeptide in human cultured cell lines Control plasmid (pAMo) and KAT06
734L polypeptide expression plasmid (pAMo-K
AT0673L) was dissolved in TE at a concentration of 1 μg / μl, and then electroporation [Cy
totechnology, 3 , 133 (1990)] to transform the cells into Namalwa KJM-1 cells [Cytotechnology, 1 , 151 (1988)].

After introducing 4 μg of plasmid per 1.6 × 10 ⁶ cells, 8 ml of RPMI1640 · ITPSG was introduced.
Medium [7.5% NaHCO ₃ 1/40 volume, 200 mmol / L L-glutamine solution (GIBCO) 3%, penicillin-streptomycin solution (GIBCO, 5000 units / ml penicillin,
(5000μg / ml streptomycin) 0.5%, 10mmol / LN
-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic acid (N-2-hydroxyethylpiperazi
ne-N'-2-hydroxypropane-3-sulfonic acid; HEPES),
3 μg / ml insulin, 5 μg / ml transferrin,
RPMI supplemented with 5 mmol / L sodium pyruvate, 125 nmol / L sodium selenite, and 1 mg / ml galactose
1640 medium (manufactured by Nissui Pharmaceutical Co., Ltd.)] and cultured at 37 ° C. for 24 hours in a CO ₂ incubator. Then G418
(Manufactured by Gibco) was added to a concentration of 0.5 mg / ml, and the mixture was further cultured for 14 days to obtain a stable transformant. The transformant was RPMI1640.IT containing 0.5 mg / ml G418.
Passaged in PSG medium.

The stable transformant or the membrane fraction thereof was obtained according to the method described in the above (6-1), using KAT0673.
A ligand for a 4L polypeptide (a novel G protein receptor),
It can also be used to search for agonists, antagonists or functional modifiers. Example 3 Structural Analysis of KAT06734L Chromosome Gene By comparing the sequence of the KAT06734L cDNA of the present invention with the sequence of the human chromosome gene registered in the database, the human chromosomal gene encoding the KAT06734L polypeptide of the present invention (KAT0673L) was compared.
4L chromosome gene) and its promoter region, exon and intron structure were determined by the following method.

The base sequence of KAT0673L cDNA (SEQ ID NO: 2) and GenBank [Nationa on the Internet
l Accessible from the Center for Biotechnology Information (NCBI) homepage (http://www.ncbi.nlm.nih.gov/).
Registration numbers AC005493, AC005680, AC005174, AC005862, AC
Part of the human chromosomal DNA sequence of 005853 is KAT067
It turned out that it matched a part of base sequence of 34L cDNA. As a result of the analysis, it was revealed that the KAT0673L chromosome gene is a very large gene composed of 9 exons and 8 introns. AC0054
The sequence of 93 and AC005680 were linked, and contained exon 1 and exon 2 (DNA having the nucleotide sequence represented by SEQ ID NO: 15). The sequences of AC005174, AC005862, and AC005853 were linked and included exons 3 to 9 (DNA having the nucleotide sequence represented by SEQ ID NO: 16). The exon 1 upstream sequence (1 to 5 kb) corresponds to the promoter region of the KAT0673L chromosome gene (including the transcription control region).
It was considered. The sequence with accession number AC005680 is a human chromosome.
Since it is located at 7p14-15, it was found that the KAT0673L chromosome gene was located at human chromosome 7p14-15. The location and structure of the KAT0673L chromosome gene on the chromosome (promoter region, exon region, intron region) were determined by the present study to be KAT06734L cDN.
This was the first time that the structure of A was identified.

About the upstream sequence (5 kb) of exon 1, Transcription of sequence analysis software GENETYX-MAC 10.1
Factor Database (Nucleic Acids Research 18 , 1749
(1990), Trends in Biochemical Sciences 16 , 455 (199
1), Nucleic Acids Research 20S , 2091 (1992), Nucleic Acids Research
ic Acids Research 21S , 3117 (1993)], the Motif Search Program was used to analyze the presence of a consensus sequence of transcription factor binding sequences, and found that the sequence had a promoter region. It was judged.

KAT06734L cDNA, KAT0
As a result of comparing the base sequences of KAT06734L-3 ′ and KAT06734L chromosomal gene, KAT07634 cDNA and KAT0III
The DNA fragment obtained from the cells by the 3'-RACE method was considered to be a variant generated by splicing abnormality. Example 4 Examination of Expression Level of Transcripts from KAT06734L Chromosomal Gene in Various Human Cells Transcripts from KAT06734L chromosomal gene (KA
Quantification of T0673L transcript) is performed by a conventional method [PCR
Protocols, Academic Press (1990)].

Also, the transcript of β-actin, which is considered to be expressed to the same extent in any cell, was simultaneously determined, and the amount of mRNA between cells and the difference in mRNA between samples by reverse transcriptase were determined. It was confirmed that there was not much difference in the conversion efficiency to the main-strand cDNA.

The quantification of the β-actin transcript was carried out by a conventional method [Pr
oc. Natl. Acad. Sci. USA, 87 , 2725 (1990), J. Bio
l. Chem., 269 , 14730 (1994), JP-A-06-181759], and a quantitative PCR method. (A) Synthesis of single-stranded cDNA derived from various cells and cell lines As human cell lines, T cell lines (Jurkat, Molt-3, Mol
t-4, HUT78), B cell line (Namalwa KJM-1, Daudi, Ra
ji), granulocyte / monocyte cell lines (HL-60, U-937, THP-
1), vascular endothelial cell lines (IVEC, HUVEC), melanoma cell lines (WM266-4, WM115), neuroblastoma cell line SK-N-MC,
Lung cancer cell line (PC-9, HLC-1, QG90), prostate cancer cell line PC
-3, gastric cancer cell line KATOIII, pancreatic cancer cell line (Capan-1, Capa
n-2), colon cancer cell lines (Colo205, SW1116, LS180). Jurkat, QG90 and SW1116 were obtained from Aichi Cancer Center. HLC-1 was obtained from Osaka University Cancer Institute. K
ATOIII and PC-9 were obtained from the Institute for Immunobiology. HUVE
C (human umbelical vascular endothelial cell) was obtained from Kurabo. IVEC [J. Cell. Physiol., 157 ,
41 (1993)]. T. L. Obtained from FRANCE. Molt-4 and Daudi are Japanese Collection of Research
Bioresources (JCRB) cell bank [Internet address http://cellbank.nihs.go.jp/] Other cells are from the American Type Culture Collection (American Type Culture Collection).
n) Obtained from

In addition, polymorphp from peripheral blood of healthy adults
Polymorphonuclear leukocytes and mononuclear cells were separated and obtained using rep ^TM (manufactured by Nycomed Pharma). Next, Ny was obtained from the obtained mononuclear cells.
T cells were obtained using ronFiber (manufactured by Wako Pure Chemical Industries, Ltd.).
The method followed the instructions of Nyron Fiber. J. Immunol., 15
0 , 1122 (1993), the obtained T cells were cultured under the following three conditions to obtain activated T cells. Using RPMI 1640 medium containing 10% FCS,
50 ng / T cells seeded at × 10 ⁶ cells / ml
ml of interleukin 2 (IL-2), 1 μg / ml ph
ytohemagglutinin-P (PHA-P) and 5 ng / ml transforming growth factor β (TGF-P
β) was added, and the cells were collected after culturing for 2, 4, 6, or 8 days. Using RPMI 1640 medium containing 10% FCS,
50 ng / T cells seeded at × 10 ⁶ cells / ml
ml of IL-2 and 1 μg / ml of PHA-P were added and
After culturing for 6 days or 8 days, the cells were collected. 10% FCS in culture vessel coated with anti-CD3 antibody
1 × 10 ⁶ cells / RPMI1640 medium containing
T cells were seeded in ml and cultured for 2, 4, 6, or 8 days in the presence of 50 ng / ml IL-2, and the cells were collected.

The total RNA of each cell was determined by a conventional method [Biochemistry,
18 , 5294 (1977)]. Synthesis of single-stranded cDNA from total RNA is performed using a kit (SUPERSCRIPT ^™ Prea
mplification System; manufactured by BRL).
For cell lines, 5 μg of total RNA to single-stranded cDNA
Was synthesized, diluted 50-fold with water, and used as a template for PCR. As a primer, an oligo (dT) primer was used.

In addition, mRNA derived from various human organs (Clon
tech) was synthesized in the same manner.
Single-stranded cDNA was synthesized from 1 μg of mRNA, and
A 40-fold dilution was used as a template for PCR. As a primer, an oligo (dT) primer was used. mR
As NA, mRNA derived from the following 35 kinds of organs was used. 1 adrenal gland, 2 brains, 3 caudate nuclei, 4 hippocampus, 5 substantia nigra,
6 thalamus, 7 kidneys, 8 pancreas, 9 pituitary gland, 10 small intestine, 11 bone marrow, 12 amygdala, 13 cerebellum, 14 corpus callosum, 15 fetal brain, 1
6 fetal kidney, 17 fetal liver, 18 fetal lung, 19 heart, 20
Liver, 21 lungs, 22 lymph nodes, 23 mammary glands, 24 placentas, 2
5 prostate, 26 salivary glands, 27 skeletal muscle, 28 spinal cord, 29 spleen, 30 stomach, 31 testis, 32 thymus, 33 thyroid, 34 trachea, 35 uterus. (B) Quantitative PCR for Standard and preparation pBS-KAT06734L internal control cutting out the cDNA partial restriction enzyme Hin dIII and Not was cut with I linear DNA of
After converting to 1 μg /
It was serially diluted with ml of water and used as a standard for quantification.

Also, cDNA encoding β-actin
Containing pUC119-ACT, and β- actin containing cDNA were missing part of the cDNA encoding pUC119-ACTD respective cDNA portion was cut with restriction enzymes Hin dIII and Asp 718 linear DNA of After conversion to yeast transfer RNA
Was serially diluted with water containing 1 μg / ml and used as a standard and an internal control, respectively, for quantifying β-actin transcripts [J. Biol. Chem., 269 , 1].
4730 (1994), JP-A-06-181759]. (C) Quantification of KAT0673L transcript using PCR method PCR was performed using single-stranded cDNAs derived from various tissues and cell lines prepared in (a) as templates. In the PCR, SEQ ID NO: 9 (0634-5 ′ in FIG. 7) and SEQ ID NO: 17
DNA having the nucleotide sequence represented by (06734-3 ′ in FIG. 8)
Was used as a primer set. In addition, a calibration curve was prepared by similarly performing PCR using the standard prepared in (b) above as a template.

The PCR reaction was carried out using the single-stranded c synthesized in (a).
10 pmol of the forward primer (DNA of SEQ ID NO: 9) was added to DNA (5 μl),
e primer (DNA of SEQ ID NO: 17) at 10 pm
ol, 2.5 mmol / L dNTP mixture 1.6 μl, dimethyl sulfoxide 1 μl, 5 units / μl Recombin
ant Taq DNA Polymerase (Takara) 0.1
2 μl of 10 μl reaction buffer (Takara)
Then, sterile water was added to adjust the total volume to 20 μl. Thermal cycler DNA engine (MJ Res
DNA, denatured by heat treatment at 94 ° C for 3 minutes, and 2 cycles of a reaction consisting of 94 ° C for 1 minute, 60 ° C for 1 minute, and 72 ° C for 1 minute.
Five to 35 cycles were performed. After subjecting a part (8 μl) of the reaction solution to agarose gel electrophoresis, the gel was subjected to SYBR Green
The cells were stained with I nucleic acid stain (Molecular Probes). The pattern of the amplified DNA fragment is converted to a fluoro imager (FluorImager SI; manufactured by Molecular Dynamics).
The amount of the amplified DNA fragment was measured by the analysis described in (1). For more accurate transcript quantification, PC
The same PCR was performed by changing the number of cycles of R. The amount of the standard was changed according to the number of PCR cycles.

For quantification of the transcript of β-actin,
J. Biol. Chem., 269 , 14730 (1994) and JP-A-06-1
It carried out similarly to the method of 81759.

The KAT0673L transcript was relatively frequently expressed in the thalamus and cerebellum in normal human tissues. The whole brain,
Hippocampus, substantia nigra, fetal brain, fetal kidney, fetal liver, heart, liver,
Mammary gland, placenta, prostate, salivary gland, skeletal muscle, thymus, thyroid,
Expression was also seen in the uterus (FIG. 15). KAT06734
The L polypeptide is presumed to perform important physiological functions in the expression tissues.

In the human cultured cell line, a colon cancer cell line (LS-1
80, Colo205) and KATO in gastric cancer cell line (KATOIII).
Expression of the 6734L transcript was observed. No expression was observed in other cell lines (FIG. 16). KAT06734
The cell line expressing the L transcript is (6-
According to the method described in 1), it can be used for searching for ligands, agonists, antagonists or functional modifiers of the KAT0673L polypeptide.

[0266]

[Sequence List Free Text]

 SEQ ID NO: 3-Description of Artificial Sequence: Synthetic DNA SEQ ID NO: 4-Description of Artificial Sequence: Synthetic DNA SEQ ID NO: 5-Description of Artificial Sequence: Synthetic DNA SEQ ID NO: 6-Description of Artificial Sequence: Synthetic DNA SEQ ID NO: 7-Artificial Sequence Description: Synthetic DNA SEQ ID NO: 8-Description of Artificial Sequence: Synthetic DNA SEQ ID NO: 9-Description of Artificial Sequence: Synthetic DNA SEQ ID NO: 10-Description of Artificial Sequence: Synthetic DNA SEQ ID NO: 11-Description of Artificial Sequence: Synthetic DNA Sequence No. 12—Description of Artificial Sequence: Synthetic DNA SEQ ID No. 13—Description of Artificial Sequence: Synthetic DNA SEQ ID No. 14—Description of Artificial Sequence: Synthetic DNA SEQ ID No. 17—Description of Artificial Sequence: Synthetic DNA SEQ ID No. 18—Description of Artificial Sequence Description: Synthetic DNA

[0267]

[Sequence List] SEQUENCE LISTING <110> KYOWA HAKKO KOGYO CO., LTD <120> Novel Polypeptides <130> H11-1931A4 <140> <141> <160> 18 <170> PatentIn Ver. 2.0 <210> 1 <211> 371 <212> PRT <213> Homo sapiens <400> 1 Met Pro Ala Asn Phe Thr Glu Gly Ser Phe Asp Ser Ser Gly Thr Gly 1 5 10 15 Gln Thr Leu Asp Ser Ser Pro Val Ala Cys Thr Glu Thr Val Val Thr Phe 20 25 30 Thr Glu Val Val Glu Gly Lys Glu Trp Gly Ser Phe Tyr Tyr Ser Phe 35 40 45 Lys Thr Glu Gln Leu Ile Thr Leu Trp Val Leu Phe Val Phe Thr Ile 50 55 60 Val Gly Asn Ser Val Val Leu Phe Ser Thr Trp Arg Arg Lys Lys Lys 65 70 75 80 Ser Arg Met Thr Phe Phe Val Thr Gln Leu Ala Ile Thr Asp Ser Phe 85 90 95 Thr Gly Leu Val Asn Ile Leu Thr Asp Ile Asn Trp Arg Phe Thr Gly 100 105 110 Asp Phe Thr Ala Pro Asp Leu Val Cys Arg Val Val Arg Tyr Leu Gln 115 120 125 Val Val Leu Leu Tyr Ala Ser Thr Tyr Val Leu Val Ser Leu Ser Ile 130 135 140 Asp Arg Tyr His Ala Ile Val Tyr Pro Met Lys Phe Leu Gln Gly Glu 145 150 155 160 Lys Gln Ala Arg Val Leu Ile Val Ile Ala Trp Ser Leu Ser Phe Leu 165 170 175 Phe Ser Ile Pro Thr Leu Ile Ile Phe Gly Lys Arg Thr Leu Ser Asn 180 185 190 Gly Glu Val Gln Cys Trp Ala Leu Trp Pro Asp Asp Ser Tyr Trp Thr 195 200 205 Pro Tyr Met T hr Ile Val Ala Phe Leu Val Tyr Phe Ile Pro Leu Thr 210 215 220 Ile Ile Ser Ile Met Tyr Gly Ile Val Ile Arg Thr Ile Trp Ile Lys 225 230 235 240 Ser Lys Thr Tyr Glu Thr Val Ile Ser Asn Cys Ser Asp Gly Lys Leu 245 250 255 Cys Ser Ser Tyr Asn Arg Gly Leu Ile Ser Lys Ala Lys Ile Lys Ala 260 265 270 Ile Lys Tyr Ser Ile Ile Ile Ile Leu Ala Phe Ile Cys Cys Trp Ser 275 280 285 285 Pro Tyr Phe Leu Phe Asp Ile Leu Asp Asn Phe Asn Leu Leu Pro Asp 290 295 300 Thr Gln Glu Arg Phe Tyr Ala Ser Val Ile Ile Gln Asn Leu Pro Ala 305 310 315 320 Leu Asn Ser Ala Ile Asn Pro Leu Ile Tyr Cys Val Phe Ser Ser Ser 325 330 335 Ile Ser Phe Pro Cys Arg Glu Gln Arg Ser Gln Asp Ser Arg Met Thr 340 345 350 Phe Arg Glu Arg Thr Glu Arg His Glu Met Gln Ile Leu Ser Lys Pro 355 360 365 Glu Phe Ile 370 <210> 2 <211> 1714 <212> DNA <213> Homo sapiens <400> 2 agcacgtaga tcctccctgt catcaggcag agctcttcag tgaggtgggc tcagggaggg 60 ctctgtgcct ccgttcagca gagctgcagc tgctgcccag ctctcaggag gcaagctgga 120 ctccctcact cggctgcagg agcaaggaca gtgaggctca accccgcctg agcc atg 177 Met 1 cca gcc aac ttc aca gag ggc agc ttc gat tcc agt ggg acc ggg cag 225 Pro Ala Asn Phe Thr Glu Gly Ser Phe Asp Ser Ser Gly Thr Gly Gln 5 10 15 acg ctg gat tct tcc cca gtg gct tgc act gaa aca gtg act ttt act 273 Thr Leu Asp Ser Ser Pro Val Ala Cys Thr Glu Thr Val Thr Phe Thr 20 25 30 gaa gtg gtg gaa gga aag gaa tgg ggt tcc ttc tac tac tcc ttt aag 321 Glu Val Val Glu Gly Lys Glu Trp Gly Ser Phe Tyr Tyr Ser Phe Lys 35 40 45 act gag caa ttg ata act ctg ctg tgg gtc ttc ttt gtt att gtt 369 Thr Glu Gln Leu Ile Thr Leu Trp Val Leu Phe Val Phe Thr Ile Val 50 55 60 65 gga aac tcc gtt gtg ctt ttt tcc aca tgg agg aga aag aag aag tca 417 Gly Asn Ser Val Val Leu Phe Ser Thr Trp Arg Arg Lys Lys Lys Ser 70 75 80 aga atg acc ttc ttt gtg act cag ctg gcc atc aca gat tct ttc aca 465 Arg Met Thr Phe Phe Val Thr Gln Leu Ala Ile Thr Asp Ser Phe Thr 85 90 95 gga ctg gtc aac atc ttg aca gat att aat tgg cga ttc act gga gac 513 Gly Leu Val Asn Ile Leu Thr Asp Ile Asn Trp Arg Phe Thr Gly Asp 100 105 110 ttc acg gca cct gac ctg gtt tgc cga gtg gtc cgc tat ttg cag gtt 561 Phe Thr Ala Pro Asp Leu Val Cys Arg Val Val Arg Tyr Leu Gln Val 115 120 125 gtg ctg ctc tac gcc tct acc tac gtc gtg tcc ctc agc ata gac 609 Val Leu Leu Tyr Ala Ser Thr Tyr Val Leu Val Ser Leu Ser Ile Asp 130 135 140 145 aga tac cat gcc atc gtc tac ccc atg aag ttc ctt caa gga gaa aag 657 Arg Tyr His Ala Ile Val Tyr Pro Met Lys Phe Leu Gln Gly Glu Lys 150 155 160 caa gcc agg gtc ctc att gtg atc gcc tgg agc ctg tct ttt ctg ttc 705 Gln Ala Arg Val Leu Ile Val Ile Ala Trp Ser Leu Ser Phe Leu Pcc 165 170 175 att ccc acc ctg atc ata ttt ggg aag agg aca ctg tcc aac ggt 753 Ser Ile Pro Thr Leu Ile Ile Phe Gly Lys Arg Thr Leu Ser Asn Gly 180 185 190 gaa gtg cag tgc tgg gcc ctg tgg cct gac gac tcc tac tgg acc cca 801 Glu Val Gln Cys Trp Ala Leu Trp Pro Asp Asp Ser Tyr Trp Thr Pro 195 200 205 tac atg acc atc gtg gcc ttc ctg gtg tac ttc atc cct ctg aca atc 849 Tyr Met Thr Ile Val Ala Phe Leu Val Tyr Phe Ile Pro Leu Thr Ile 210 215 220 225 atc agc atc atg tat ggc att gtg atc cga act att tgg att aaa agc 897 Ile Ser Ile Met Tyr Gly Ile Val Ile Arg Thr Ile Trp Ile Lys Ser 230 235 240 aaa acc tac gaa aca gtg att tcc aac tgc tca gat ggg aaa ctg tgc 945 Lys Thr Tyr Glu Thr Val Ile Ser Asn Cys Ser Asp Gly Lys Leu Cys 245 250 255 agc agc tat aac cga gga ctc atc tca aag gca aaa atc aag gct atc 993 Ser Ser Tyr Asn Arg Gly Leu Ile Ser Lys Ala Lys Ile Lys Ala Ile 260 265 270 aag tat agc atc atc atc att ctt gcc ttc atc tgc tgt tgg agt cca 1041 Lys Tyr Ser Ile Ile Ile Ile Leu Ala Phe Ile Cys Cys Trp Ser Pro 280 285 tac ttc ctg ttt gac att ttg gac aat ttc aac ctc ctt cca gac acc 1089 Tyr Phe Leu Phe Asp Ile Leu Asp Asn Phe Asn Leu Leu Pro Asp Thr 290 295 300 305 cag gag cgt ttc tat gcc att tgt g atc ac ctg cca gca ttg 1137 Gln Glu Arg Phe Tyr Ala Ser Val Ile Ile Gln Asn Leu Pro Ala Leu 310 315 320 aat agt gcc atc aac ccc ctc atc tac tgt gtc ttc agc agc tcc atc 1185 Asn Ser Ala Ile Asle Pro Leu Tyr Cys Val Phe Ser Ser Ser Ile 325 330 335 tct ttc ccc tgc agg gag caa aga tca cag gat tcc aga atg acg ttc 1233 Ser Phe Pro Cys Arg Glu Gln Arg Ser Gln Asp Ser Arg Met Thr Phe 340 345 350 cgg gag aga act gag agg cat gag atg cag att ctg tcc aag cca gaa 1281 Arg Glu Arg Thr Glu Arg His Glu Met Gln Ile Leu Ser Lys Pro Glu 355 360 365 ttc atc tagaccctag ggcagtgcca gtgctaggcacct gtgccct gagcaccatcgc gctccctcgc gtgctaggc 1397 tcattacctg ggagatgcac aagacaaatg ttctaatgac tgcatgcact gcttaagtat 1457 tggccaacac gaactcccca gttattcatg ccagccagga aggaaacgcc ttccttcccc 1517 accattccca gccctccttc ccactggcca gcacctgaac ccagtgaaca caggcatcag 1577 tggtccaggg tcctggcttg gagccagtga gtagacaggc aagcagaggg gacaaaggta 1637 gctgggttat acatgaatat tctcatta ca ataggagaaa ataaaagact taattaagcc 1697 caaaaaaaaa aaaaaaa 1714 <210> 3 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 3 agcaucgagu cggccuuguu ggccuacugg 30 <210> 4 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 4 gcggctgaag acggcctatg tggccttttt tttttttttt tt 42 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 5 agcatcgagt cggccttgtt g 21 <210> 6 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 6 gcggctgaag acggcctatg t 21 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 7 cttctgctct aaaagctgcg 20 <210> 8 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 8 tgtgggaggt tttttctcta 20 <210> 9 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 9 agagggcagc ttcgattcca gtg 23 <210> 10 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 10 gaatggggtt ccttctacta ctcc 24 <210> 11 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 11 ggctgcagga gcaaggacag tg 22 <210> 12 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 12 aacccagcta cctttgtccc ctc 23 <210> 13 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 13 tgacaagctt accccgcctg agccatgcca gcc 33 <210> 14 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 14 tagtgcggcc gctggcactg ccctagggtc tag 33 <210> 15 <211> 80578 <212> DNA <213> Homo sapiens <220> <221> exon <222> (25203) .. (25523) <220> <221> exon <222> (51516) .. (51648) <400>   Fifteen   gatctttgac   aaacctgaca   aaaacaagaa   atggggaaag   gattccctat   ttaataaatg   60   gtgctgggaa   aactggctag   ccatatgtag   aaagctgaaa   ctggatccct   tccttacacc   120   ttatataaaa   attaattcaa   gatggattaa   agacttaaac   gttagaccta   aaaccataaa   180   aaccctagaa   gaaaatctag   gcaataccat   tcaggacata   ggcatgggca   aggacttcat   240   gtctaaaaca   ccaaaagcaa   tggcaacaaa   agccaaaatt   gacaaatggg   atctaatcaa   300   actaaagagc   ttctgcacag   caaaagaaac   taccatcaga   gtgaacaggc   aacctacaga   360   atgggagaaa   agttttgcaa   tctactcatc   tgacaaaggg   ctaatatcca   gaatctacaa   420   tgaactccaa   caaatttaca   agaaaaaaac   aaacaacccc   atcaacaagt   gggcaaagga   480   catgaacaga   cacttctcaa   aagaagacat   ttatgcagcc   aaaagacaca   tgaaaaaatg   540   ctcatcatca   atggccatca   aagaaatgca   aatcaaaact   acaatgagat   accatctcac   600   accagttaga   atggcaatca   ttaaaaagtc   aggaaacaac   aggtgctgga   gaggatgtgg   660   agaaatagga   acacttttac   actgttggtg   ggactgtaaa   ctagttcaac   cattgtggaa   720   gtcagtgtgg   cgattcctca   gggatctaga   actagaaata   ccatttgacc   cagcaatccc   780   attactgggt   atacacccaa   aggattataa   atcaagctgc   tataaagaca   catgcacatg   840   tatgtttatt   gtggcactat   tcacgatagc   aaagacttgg   aaccaaccca   aatctccaac   900   aatggtagac   tggattaaga   aaatgtggta   catatacacc   atggaatact   atgcagccat   960   aaaaaatgat   gagttcatgc   cctttgtagg   gacacggatg   aagatggaaa   ccatcactct   1020   cagcaaacta   tcgcaaggac   aaaaatccaa   acactgtatg   ttctcactca   taggtgggaa   1080   ttgaacaatg   agaacacatg   gacacaggaa   ggggaacatc   acacacaggg   gcctgttgtg   1140   gggtgtgggg   aggtggggag   ggatagcatt   tggagatata   cctaatgtta   aatgacgagt   1200   tactgggtgc   agcacaccaa   catggcatat   gtatacatat   gtaactaacc   tgcacattgt   1260   gcacatgtac   cctaaaactt   aaagtataat   aaataacaaa   aaaaaaaaaa   gaaaaggaac   1320   aaacaaatcc   tctgagaaat   atagaactat   gtgaaaagac   caaatctacg   tttgatgggt   1380   gtacctgaaa   gtgatgggga   gaatggaacc   aagttggaaa   acattcttca   ggatatgatc   1440   caggagaact   tccccaacct   actaagacag   gccaatattc   aaattcagga   agtacatgga   1500   acaccacaaa   gatactcctc   gagaagagca   accccaaaac   acataatcat   cagattcacc   1560   aagtttgaaa   tgaaggaaaa   aatattaaca   gcagctagag   agaaaggtcg   ggttacccac   1620   aaaagggagc   ctatcagact   aacagtggat   ctctctgcag   aaaccctaca   agccagaaga   1680   gagtaggggc   caatattcaa   cattcttaaa   gaaaataatt   ttcaacccag   aatttcatat   1740   ccagtcatac   taagcttcat   aagcaaagga   gaaacaaaat   cctttacaga   caagcaaatg   1800   ctgagagatt   ttgtcaccac   cagacctgac   ttaaaagagc   tcctgaagga   agcactaaat   1860   atgaaaagga   aaaacccata   ctagccactg   caaaaacata   ccaaattgta   aagaccatca   1920   acactatgaa   gaaactgcat   caactaatga   gcaaaataac   cagctagcat   cacaatgaca   1980   ggatcaaatt   cacactaaca   atattaacct   caaatgtaaa   caaactaaat   gccccaatta   2040   aaagacacag   actggcaaat   tggataagca   gtgaagaacc   actggtgtgc   tgtattcagg   2100   agacccatct   cacagacaaa   gacatacaca   ggctcaaaat   gaagggatgg   aggagaattt   2160   accaagaaaa   tggaaagcaa   aaaaaagcag   gggttgcaat   cctagtctct   gataaaacag   2220   actttaaacc   aacaaagatc   aaaaaagaca   aagaagagca   ttacataatg   gtaaagggat   2280   caaagcaaca   agaagagcta   actatcctaa   atatatatgc   acccaataca   ggagcaccca   2340   gattcataaa   gcaagttctt   agagaactac   aaagagactt   agaatctcac   acaataatag   2400   tgggagactt   taacaaccca   ctgtcaatat   tagacagatc   aacaagacag   aaaattaaca   2460   aggatattca   ggacttgaac   tcagctctgg   attgagcaga   cctaatagac   atctccaccc   2520   caaattaaca   gaatatacct   tcttctcagc   acctcatggc   acttattcta   aaattgacca   2580   cataattgga   agtgaaacac   tcctcagcaa   atgcaaaaga   acggaaatca   taacagtctc   2640   tcagaccaca   gtgcaatcaa   attagaaccc   aagattaaga   aactcattca   aaactgcaca   2700   accacaagga   aactgaacaa   cctgcccctg   aatgactact   gggtacataa   tgaaatcaag   2760   gcagaaataa   ataagttctt   tgaaaccaat   gagaacaaag   acacagtgta   ccagaatctc   2820   tgggacacag   ctaaagcagt   gtttagagga   aaatttatag   cactaaatgc   ccacaggaga   2880   aagtagaaaa   gatctaaaat   caacacccta   acatcagaat   taaaagaact   agagaagcaa   2940   gggcaaagaa   attcaaaagc   tagcagaagg   caagaaataa   ctaagatcag   agcagaactg   3000   aaggagatag   agacatgaaa   aacccttaaa   aatatcaacg   aacccaggag   ctggtgtttt   3060   gaaaagatta   acaaaataga   tagaccgcta   gccagactaa   taaagaagaa   aatagagaaa   3120   aatcaaatag   acacaataaa   aaatgataaa   gggataccat   cactgatccc   acagaaagac   3180   aaaccaccat   cagagaatac   tataaacatc   tctatgcaaa   taaactagaa   catctagaag   3240   aaatggataa   attcctggac   acatacacac   tcccaagacc   aaaccaggaa   gaagttgatc   3300   cctgaataga   ccagtaacaa   gttctgaaat   tgaggcagta   attaatagcc   taccaaccaa   3360   aaaaaccaca   ggaccagatg   gattcactgc   caaattctac   cagaggtaca   aagaggagct   3420   ggtactattc   cttctgaaat   ttttccaaac   aacaaaaaaa   gagggactcc   tccctaactc   3480   attttaagag   accagcatca   tcctgatact   aaaacctggc   agagacacaa   caaaaaaaga   3540   aattttaggc   caatatccct   gatgaacatc   aatgtgaaaa   tcctcaataa   aatactggca   3600   aaccaaatcc   agcagcacat   caaatagctt   atccaccaca   atcaagttgg   cttcatacct   3660   gggctgcaag   gctagtttaa   cttatggaaa   tcaataaacg   taatccatca   cataaacaga   3720   accaatgaca   gaaaccacat   gattatctca   atagatgcag   aaaaggcctt   tgacaaaatt   3780   caacacccct   tcatgctaaa   aactctcaat   aaactaggta   ttgatggaac   atatctcaaa   3840   ataataagag   ctatttatga   caaacccaca   gccagtatca   cactgaacgg   gcaaaagctg   3900   gaagcattcc   ctttgaaaac   cagcacaaga   caaggatgcc   ctctctctcc   actcctattc   3960   aacatagaat   tggaagttct   ggccagggca   atcaggcaag   agaaagaaat   aaagcgtatt   4020   caaaaagaaa   ggaaatccaa   ctgtctctgt   ttgcagatga   catgactgta   tacctagaaa   4080   accccattgt   ctcagcccaa   aatctccttc   agctgataag   caacttcagc   caagtctcag   4140   gatacaaaat   caatgtgcaa   aaatcagaag   aattcctatg   caccaatcat   acacaaaaag   4200   agagaaagat   tatgagtgaa   ctcccattca   caatggctac   aagagaataa   atacctagga   4260   atccaactta   cttacaaggg   atgtgaagga   cctcttcaag   aactacaaac   cactgctcaa   4320   ggaaataaaa   gaggacacaa   acacatggaa   aaacattcca   tgctcatgga   taggaagaat   4380   caatatcgtg   aaaatggcca   tactgcccaa   agtaatttag   agattcaatg   ctatccccat   4440   caagctacca   ttaactttct   tcacagaatt   agaaaaaaac   tactttaaat   ttgatacgga   4500   accaaaaatg   aggctgtata   gccaagaaaa   tcctaagcaa   aaataacaaa   gctggaggca   4560   tcacactacc   tgacttcaaa   ctatactaca   aacagcatgg   tactggtacc   aagacagata   4620   tatagaccaa   tggaacagaa   cagaggcctc   agaaataatg   acatatatct   acaatcatct   4680   gatctttgac   aaacctgaca   aaacaagcaa   tgaggaaagg   attccctgtt   taataaatgg   4740   tgctgggcaa   actggctagc   catatgcaaa   aaacacagaa   actggacccc   ttccttacac   4800   cttatacaaa   aattaagcca   agatggatta   aagacttaag   tgtaagacct   aaagccataa   4860   aaaccctaga   agaaaacctg   ggcaatacca   ttcaggacat   aggcatgggc   aaagacttca   4920   tgactaaaac   accaaaagca   aaggcaacaa   aagccaaaat   tgacaaatgg   gatctaatta   4980   aactaaagag   cttctgcaca   gcaaaataaa   ctatcatcag   agtaaacagg   caacctacag   5040   aatgggaaaa   aaattttgca   aactattcat   ctgaaaaagg   gctaatatcc   agaatctaca   5100   aagaacttaa   agaaatttac   aagaaaaaaa   atcccatcaa   aaagtgggtg   aaggatatga   5160   atagacactt   ctcaaaagaa   gatatttatg   tggtcaaaaa   acatatgaaa   aaaagctcat   5220   catcactggt   cattagagaa   atgcagatca   aaaccacaat   gagataccgt   ctcatgccag   5280   ttagaatggc   gatcattaca   aagtcaggaa   acaacagatg   ctggagagga   tgtggagaaa   5340   taggaaagct   tttacactac   tggtgggagt   gtaaattagt   tcaaccattg   tggaagacag   5400   tgtggcgatt   cctcaaggat   ctagaaccag   aaataccatt   tgacccagcc   atcccattac   5460   tgggtatata   cccaaaggat   tataaatcat   tctactataa   agatgcatgc   atacgtaggt   5520   ttattgcagc   actagtcaca   atagtaaaca   cttggaacca   acccaaatgc   ccatcagtaa   5580   tagactggat   aaagcaaatg   tggcacatat   acaccatggg   atactatgca   cctataaaaa   5640   aggatgggtt   catgtccttt   gcagggatgt   gaatgaagct   ggaaaccatc   attctcagca   5700   aactaacaca   gaaacagaaa   accaaacact   gcatgttctc   actcataagt   gggagttgaa   5760   caatgagaac   agatggacac   agggaggaca   tcacacacca   tggcctgcca   gggggttaga   5820   ggctagggga   gagatagcat   tcaagaaata   cctaatgtag   atgacaggtt   gatgggtgca   5880   gcaaaccaac   atggcatgtc   catacctatg   taacaaacct   gcacattctg   cacatgtatc   5940   ccagaactta   aagtataatt   ttaaaaaagt   aggttggaga   ttcatattta   aagtgaaact   6000   aagccaaaat   ttgtattatt   tttcttaata   atttccaaaa   tgcaatctaa   gttttacctt   6060   ttaagccaat   ctgatcaact   tctcatggca   ctaccttgag   aaaaactcta   aggtatacta   6120   agacacagtg   gttctcagtg   ccataatcca   tcagtgaatg   tggggacagg   cagcaggcac   6180   agagagtggg   gagtgtgccc   agctctgaca   taaagaacaa   caatatgttg   atgcaacatc   6240   aagaagttgc   ttctctcctg   ggcttccttg   tagccgttat   cagctattac   ccaactcagc   6300   tactctacag   tcattttcaa   cacttttctt   cctttctgtg   tcccattgcc   aagtctttaa   6360   aagccaggca   ctcatttttc   aagacccctt   tcataaggcc   aggggttgct   gtataaccta   6420   gttattgcca   gtaggatgta   aagagataat   tgtcatcatc   ttcataagca   tcatgaaaat   6480   attttcctct   ctgaacagag   aaacagatga   agaagaatgt   cttcctctac   caccagcatc   6540   ctcccccacc   ctccgaccta   ttttgttgtt   gttgattttt   gtttcttgtt   tgagacggag   6600   tctcgctctg   tcgcccaggc   tggagtgcag   tggcgcaatc   ttggctcact   gcaacctctg   6660   cctcccaggt   tcaagcgatt   ctcctgcctc   agcctcccga   gtagttggga   ttacaggcac   6720   cctgccccca   cgcctggcta   atttttgtat   ttttagtaga   gatgaggttt   caccatgttg   6780   gccaggctgg   tctcctaacc   tcaagtgatc   ctcttaacct   caagtgatct   gcctgtcttg   6840   gcctcccaaa   gtgctgggat   tatatgcatg   agccaccacg   cctagcctcc   tccccgcctt   6900   tttttaaacc   cattaccatc   cttgctttct   gatttggact   ggcattgaag   atatgattat   6960   taggaatctg   actgccaact   tgtgatcgtg   aaggaagaca   ttggcaacat   gataaaaata   7020   gcatagagga   ggatgaaagg   agtctggatg   tcctggggac   ttgttgagct   accaaaccca   7080   ccctgatact   gcccagcttc   tgttctttgt   ggtataaaca   atacatggct   tggcggtttc   7140   atctactgtg   agttaagtct   tctgctcctt   gctatcgagt   gtatcctaaa   accaatacac   7200   cagcaaatga   gatgcaagaa   taactgcctg   tttgctctgt   catttttgca   atgtagacac   7260   cacctagaac   aagaaaatta   ctgaatctct   gaatgacatt   tttgtaaagc   catagaagca   7320   aaaacacaag   tttaaaagac   ctagatttta   attattagag   tcagattctt   gagaaaattt   7380   ctgatggcct   tgaagccagc   agtttataca   gtgaggcagg   acaaggagaa   atctatttga   7440   gtgttgtttc   ctccccgcac   acagataaaa   tcagaacatt   ttctagagag   gattgggaat   7500   ggagaagaca   gcagatgcta   atgttgggct   ctgcttcctg   gctttccctt   cctacagggt   7560   tgacccattg   atggggtgga   taaaataggc   ttggagatct   gaatgtagaa   aggacttgcc   7620   ccacacttcc   tgcctcgaac   tcagagagga   gatggccttc   aagaatgcct   tgcagcaacg   7680   tgggtagaca   gcattgggct   cagaataggg   gcacagcata   gcaaaaccaa   ctcatctcag   7740   gtcttggaag   agacttcaaa   aagctcccaa   gcttccccca   aaaaggagct   caaaggacag   7800   cagagaatga   agagctggaa   tgcatcagga   attcacatcc   gcatatcaga   gactcagaaa   7860   gtgattatgt   cagataaatg   gaagaaaaat   aagtagaaag   agaatcattc   aggcagacta   7920   aattttaaaa   taaattactc   tgaaatgaga   agttaagagt   cttttgggtc   ttgtcccata   7980   tagaacaaaa   gagctatgtt   aatttatctt   gtaaagaaag   ttttgtttca   tgcatccgag   8040   ttgataatgt   aaaatttaaa   gttgcttcag   ataatgtgac   aaccctggag   tataatacat   8100   taggactcat   ctctctgaac   gaacagcaac   tttgcaaaag   aggaaacggg   gctgcctgtt   8160   ctgttactgt   cacatctgat   ttttatttgt   atatgacttg   accagccttt   atagctctca   8220   ccacacagca   cttccatgtt   ccataatgga   cctgttcaaa   ggaagccata   tcactccact   8280   tgatttgctt   ctgataccag   ttccaggggt   attgatctgt   cttgagtaat   gtgttattgt   8340   tgacagagag   gtttcttgac   tcactgtagg   tgacttttta   tgccctttaa   agttctgaaa   8400   agcccataaa   gataatgttt   aaattacaag   tcatacacaa   cagaagagag   tctggagcag   8460   ccacacaggc   aactgcatag   taccagaagg   actgtggaca   tggaatcaga   ctacacagtt   8520   ccatagctga   ctgggtatat   tactctttcc   tcaatttcct   catatgtgaa   atggtaaggc   8580   taatatctac   ctagcatagt   tttctgaaga   ttttataaga   gatcacatat   ataaagcacc   8640   taaccttgtt   aatccataga   agaaatgtaa   taaatatcag   aaaaagaaga   gatttttttt   8700   ctcccttctt   ttgttatttc   atgctataaa   ttttaccatt   ttctgggttt   aagtacaatg   8760   cgtttatgct   aaggtttaac   acggtggctt   catcttttag   tcattctaat   taatttcagc   8820   attgactgaa   cccagtggcc   cctgtagata   aaaacaggac   caaccactaa   acattaaact   8880   ctcaagcacc   cacctgtaat   cacttaaaca   gctagaggtg   gctccactga   tacttattaa   8940   acagatgttc   actctgtaac   aaaagtgtac   caatctcatc   ctgaagccca   gaaaatagaa   9000   aagaagatag   gacagcctct   gttaccaatg   gcagaaatat   gacagggcta   tatgaggcta   9060   agataaagga   tatcaaactc   aaaaccaaga   gtgataagca   aggagactgt   gttagactgt   9120   ttcaataatt   aaggtgagaa   atgatggcag   cctgaagtgt   gattttcatg   atggagatgg   9180   ggagaaatag   atagtttaaa   aactgttaaa   aaggagcatt   ggcagaactc   attaattgaa   9240   cagatgcaga   tagcaggtag   gctgggggag   ggaggtgtta   catatgacaa   catgacatat   9300   tctgttaata   ctaattgaag   gcctctatgg   tggaatcact   tacttaggtg   acttgggaat   9360   acagggaggt   aaagtcacat   tcctggctcc   caagaaatgt   atagtacatt   taaagggata   9420   agacataaca   acatcttttt   agtatctgct   ctagtttatt   attttatttt   ttagggccct   9480   attctgttct   caaaaaaaaa   aaaaaaaaaa   gtagtaagga   tgaaaagggg   tagggcatct   9540   ggaaaaaact   ttggagaagc   ctcaggaaca   tcccgcttga   aacctccagc   caattatttc   9600   agtcttatag   agagagtgga   ttaaatgaat   aggttgtgtc   actatttttg   agaagcctga   9660   gtgcctcttc   taagagctat   tctcatttcc   acaaaaagga   agggaaattg   cggtatctaa   9720   aagtagtgtt   tgaaagtcac   ttcttaagaa   ttatatcgag   tcaaaacact   ttcatttaaa   9780   gcccttaata   tttgaacatt   gttatttatt   tttgctgttt   ggatttttat   gtgtgtgttt   9840   gtgtacctat   gaaaaactat   tcatgtctca   aatgaaataa   aagaaggtgg   ctgtgatgta   9900   ttagtatagg   ttctcaattt   ctgtcctagg   taagatgaaa   agcatgaaga   tgcctccttc   9960   aatcccgtca   tggccacagt   ctccttgtca   tacccatata   ggctctacct   caagttgtgg   10020   gcaaacttct   ttacccacct   gtcatgaaac   tgtgcttttt   ctatgctatc   tctagggccc   10080   caagactgta   tggatagctg   cagaccacca   cattcaggaa   gaggggccag   atccattagg   10140   gagtatattt   ggcattgagc   tgatagagaa   taaaggagaa   aaacttcctg   agtagctcca   10200   gtcttaccaa   cccagcttct   gtttcaaagt   gagttcatga   tcaacggtgg   tctcgcctgg   10260   agtcattagt   ccagagaaca   tcccaacaaa   gaaggtgtag   gaagttttcc   taaacctaat   10320   ttttaggtca   aaaaaacaag   acagagagag   aacaagatta   ggaaataaat   gggaagttgg   10380   aggagcccgg   atcaagcaga   agtaagccca   agaatgcaag   ggaaaggtct   gaggtataat   10440   ggtataggtc   tgagacagga   agagctacgg   aataagcaga   gcaagaaagt   gcagagggca   10500   ataaggaatc   ctttcagatg   cttttcttct   actccaaccc   caattagata   cacatgggca   10560   ttggtaattt   tgccttctga   atctctggga   taatacatca   accacgtcat   ttgccctggg   10620   gagagggtct   ttgaaattag   cacaatagct   ctgatttcct   gccaaatggt   taatcagaag   10680   gggacctaat   acataaagag   caggaatcta   ttgttttcat   agactcctta   tgccttgggt   10740   aagacataat   tctaacttgc   tattaccccc   ttctctgatt   ttacactaac   tagggatata   10800   acagccacct   caaatgtccc   cagatagtcc   acatcccagc   aacagccgac   ttcgccaaat   10860   tggggcagcg   atttcacctt   catctcacag   agttcaatga   gtatgttgcc   aagggacatt   10920   ttgagtgcca   attcactaat   tttacagaaa   ctctgtaatt   ggttggaatt   gctaaggtca   10980   gggattgtac   atttcttgtc   accttccttc   agagggctaa   tttctgtttt   ctctctcgga   11040   tgcattctat   agctcaaaca   gcagatgcat   tctcttgcag   tggtgaataa   aggccattct   11100   tggacaggcc   caattgctct   actaagttag   aaggaacctc   ctttttagct   gacatataat   11160   caatcagcat   gcattcggtt   tagagcaatg   aatcttctgt   aagtgacaag   agactggatt   11220   tgttaatttt   gctttcagca   aatgtacctt   aatatggata   tttctaagat   tcttggcagc   11280   aatataattt   aaaacatggt   ttagtgtgac   aatacaagag   catgatgttc   tgattctgat   11340   aaaattttgt   cacatatgag   agaaggaaga   aaaaaaaggt   tagtagttca   gataagagaa   11400   aataattggc   tgaaaagctt   ttatatttga   gaatgatata   aaagtggaaa   gtgaaatcat   11460   tgacatgaac   tgagaacttt   gacggacctt   tactctggaa   ccgcaattgc   ccagctgtgc   11520   ctgacatgat   cccatttgaa   gtcaggggtc   atgatataaa   cctctcatct   ctgattttcc   11580   cctaaccagg   gctccaatgg   acatctcaga   attcctcaga   acccccacac   tctccaacag   11640   ctgactttga   tagtttgcct   gttacgaatc   agggcagcaa   tttgatcact   tctcatatgg   11700   tccaatacat   atgttgtcaa   gtaaagtaaa   aagatagcac   aagaaatttt   tactagatat   11760   aattgcattt   gaatcacatc   caactatgca   tgaaaacagc   atagatatca   cctaagactc   11820   ttactaacga   aagttcccca   caaaatatct   tagtgcagtg   attctcaaag   ttgtggtata   11880   tatcggaatc   accaggggga   acttgttaaa   atgcagaagc   ccaggctttt   gtctacttga   11940   tgaatctgag   cccctattct   ttattaactc   ccccaggtaa   ttctaaaaca   cttctgagtt   12000   tgagaaccac   tgatgtagtg   ttatagaaag   taagtagaga   aaaggatgag   ctatagataa   12060   gagggcaaca   ctttctgccc   cttttcccac   tgcatgtagg   acatgttgag   gagtggttta   12120   gctggatggg   aaagcaagtg   tcttttttca   tcacttttca   ctgagcaggc   agtttctcaa   12180   ttgcagcgcc   ttctccctct   gactccatgc   ccatacacac   attatgaatc   cgagggccgg   12240   tgcccatgca   gccatacaat   agagacaagg   accaatttac   ttgccaacta   aacatgtagg   12300   gacatgctgt   ctatcctcaa   cacagataaa   acctgcaggt   gctcttcctt   tgttgagccc   12360   agtgcttctc   aaccttggtt   gcacattaca   attgaagaat   gtttcaaact   ccccattgcc   12420   caagctgtaa   cacagtctag   ttacagctaa   attgggagga   gagcacaggt   tttggtggtt   12480   tataaggctt   cccaggtgat   tcaacatcta   accaaaattg   aaaacaactg   tctctaattc   12540   agattctcac   gaatgaggca   gattcatcct   gcatttaaga   gagcaacctg   taactctaag   12600   gttacttctt   ctattgtata   gtctcatgat   ttggtatcag   attaagtagt   ttaaaatatg   12660   agaaaatgtg   cacgaagaaa   agtcccagag   atctgattgc   ccattccttc   tcctttccta   12720   cagtcttcag   tctaggtgac   aatgcaaata   taaacctgct   cttttcagga   agaaaatacc   12780   caccacacca   acatatggaa   atagaaagac   tgtgtaaacc   tgattctttt   ataaggcgtg   12840   taaagagaga   aatgagaaca   gcacatttga   aattcagatt   taatcaagaa   gtcttaaact   12900   caataaaaag   acaactcaat   ttttaaatgg   acagaagatt   ttagtagata   tttttatcaa   12960   agaagatata   caaataacta   ataagcacct   gaaaaaaagc   tcaacattat   tggctattag   13020   gaaatgcaaa   tcaaaaccac   atgaaataac   acttactcat   cactgaaatg   gctataatca   13080   aaaaaacaga   caataaaagt   gatgtcaatg   atgtggaaaa   actaaaacgc   tcattaatac   13140   attgctggtg   ggaatgtaaa   atggtgtggc   cactttggaa   agcagtatgc   caatttctta   13200   aaaattaaac   acaaagttac   catgcaaatc   agcaattcta   ctcctagtag   tctctccaag   13260   agagatgaaa   atgaaagatg   gccacacaaa   gacttgtatg   tgaatgttta   ttatggataa   13320   tacccccaaa   gtggaaacaa   cgcaaatgtc   tttcaactgg   taaatggata   aacaaaatgt   13380   gatatagcca   caaaatggaa   taatttacaa   caataaaaag   taatacagta   ctgatatagg   13440   ccacactatg   catgaacttc   agaaaattat   gctgcacaaa   agaaatccaa   cacaaagtcc   13500   acatatttta   tgatttaatt   tctacaaaat   gtctagaaaa   gataaatctg   cagagacaaa   13560   aagctgatta   gtggttgccg   gggactggag   tggatataaa   gaatgactac   aaatgggcct   13620   gagagatact   atttgcgtga   gagaaatgtt   ctaaaattgg   attgtggcga   tggttgcaca   13680   actctgtaaa   tttactagaa   gttaattaag   ttgtgtactt   aaaatgggtg   aattttatga   13740   tatatatatt   ataataaagc   tatttagaaa   aacatgtaga   attgacttta   caattctgca   13800   aaaataatgt   aaaaatttaa   gtagatataa   catgcaaaaa   taaagcttat   tactttggaa   13860   aaaattactt   caaaaatatg   aaaatagttt   aagctgcagg   aattttttta   aaaaaacaaa   13920   ctctaaaatg   ataatattga   gcattgacta   tattccaggc   attattctaa   gtgctttata   13980   tcattttgtc   ctataaatga   gcctctaagt   ttgatgttat   gttccattgt   actagtattg   14040   aaactgaggt   ctaactagag   ataaaagact   gatgctagaa   taagaagaca   gattaaaagg   14100   aagatggtag   tactaaagaa   ataatttgag   taatttatag   tgcgtttgca   taattgtaac   14160   tggtattaaa   aagaatgaaa   aagagaatag   agacccaaat   ctagtgttga   tcagatgaag   14220   attaaagaca   tcacagacaa   tgcagagaaa   aaggagaaag   agattaaggt   gatcagagaa   14280   tagatgatag   atagatacag   atgacataaa   atgaggaaca   gagtagggat   aattagtgtt   14340   cttggggtgc   agaggagaat   caacataaaa   atattaaaga   tattatttaa   aatgtctttt   14400   tagctgaaga   gaattctaaa   cataaataat   actatcatta   gcaagaataa   atatcaggca   14460   ataatgtacc   acaactgcac   tgagcacaca   tcagctcctt   gaaccatcac   aacaacccta   14520   ggaagcatga   actataattt   ccattttaca   aaggatccct   agagctaaga   tgactagata   14580   tcttgtttgc   ttaggacaat   tctggtttac   tgcatgtaaa   ctacctaaaa   tacccatttt   14640   attctcaaat   ttatgagtct   ggacaataaa   ttacatgatc   accctatttt   gagtctaaga   14700   actacaggtg   gtaaactgca   gaaccaaaac   ttattggaga   agtctccaaa   gcctttgttc   14760   ttaaccattt   tgttatacta   aatctgcaga   gctaaagtgt   tcatcacaaa   gcgacacaac   14820   ttgtaaaaaa   taagtgatta   taccaagaaa   ttctgcattt   caaagataaa   taaacaactc   14880   tatatgcatt   cagtagaagt   aagtcattta   aaaaggaaaa   agatattaat   taatttcata   14940   agtaaatgaa   aaatagtacc   ctggatttct   tctttacaac   attaaatgcc   agaaaacaat   15000   gtctacaaag   tcaagttaga   attaatggaa   gctctcaaat   atacaagtag   ctaagaatca   15060   agaagtcaag   caactactta   aagacattgt   ccagctaatt   taggtagaaa   tcaaaattta   15120   gacttcacaa   acacataagt   aagatacaga   agtacttata   gtgggcaatg   aatccacttt   15180   aatatcaaat   tatgtaacag   ttgatctggt   aaatatcatg   tctagacaaa   gtatacatgt   15240   gatatttctt   ggaagacaat   atgcaccata   tctaacttcg   atcctgcatc   aaaaattcca   15300   ctggtttgag   aagaggccga   gggagcagtg   tgggaagaaa   tgcatatgag   ctgcttttcc   15360   cattttcaca   aggggaatcc   aataaataat   gtcttattcc   tgttgttgat   aaactagcag   15420   tctgaaattt   agtcaaaatt   tttaagttgt   aaaaatatac   atcagtagag   caaaaaaaat   15480   tgtattatct   ttcatgttgc   agaagaaaat   agaattaaag   aaaaccttag   aaagataaca   15540   taaagaaaaa   atgcaaaggt   aatactaaaa   ctaaagaaaa   tattttaaat   agcagtaaaa   15600   gacagaaaac   aaaggtaaaa   atcaggaagt   atatagagag   accaaacaca   ggatgtttaa   15660   tactaaatac   aaatggttaa   atcactctct   tattaaaaga   aagatagcct   catattaaac   15720   aaaacaataa   gtaaaatata   tttacaagag   acaccaaatg   ccaatgatta   cagaatgtag   15780   agaataaaag   aatcagtaaa   gatatgcaaa   agaaactaaa   atataataca   aatttaaaaa   15840   tacagtataa   cagctattta   cctagcattt   acattgtatt   taattaatct   agagatgatt   15900   ttaagtatat   gtaaggatat   gcataggtta   tatgcaaata   ctacaccgtt   ttatataagc   15960   gacttgagca   tcttggattt   ggcatccacg   ggagtcttac   aacaaatcct   ccatgcatac   16020   caagggattt   gtctgcttat   ctgacttact   taaagaatcc   tgattttata   gagcgtgacc   16080   atattctcag   cctcccttgc   aaataggaga   agctatgtga   ctaagttctg   accagtgaaa   16140   tataagtgga   agttactggg   agggcttcca   ggaaaattcc   tttaaaaaga   gctgagctcc   16200   cttttgtctt   tcccctttcc   ttcttccatt   ggctggaatg   caaacacgat   gaagctgaaa   16260   tgtatatctt   acaaccatat   ggtgacctta   aagatggaag   cacttcatca   aggaaaacag   16320   agcaagaaga   tacgatgagc   ctgggtcact   aatgatatcc   tggaaccact   tcaccacccc   16380   ttgattcatg   ttaccactaa   ttccccattg   ctttcctaca   ttaattcaaa   cttgactttg   16440     taaaaattat   tttctggcat   ataatgttgt   aaagaaaaaa   ttcaagatac   tgttttttta   16500   tttacttact   aaattaatat   agtatttctt   tattagtgac   tcacttttac   tgaatgcatg   16560   taaaacttgt   ttatccttga   aatacaggat   tccttggtac   aaccactttt   ttttctaaaa   16620   ctagtttgtt   tctttatgat   gaacactata   atagctctgc   agatttagtg   taataaaagg   16680   gttaagaaca   atagctttgg   agacttccat   gactgaaagt   tttggttctg   caacttacca   16740   actctagttc   ttaggcccaa   agtaaggtga   tcatgttatt   taccttgtgt   cttcaccaca   16800   ataccttgac   tctatctttc   ttgagggaaa   gacaaacaag   cattctgttt   tgttcaaatt   16860   gctctttttt   taagatctct   gtacttggac   taaaaagcaa   ttattaactg   atatcagcac   16920   ataaaattag   tttataacaa   atcaaaatcc   aaggtgaaaa   aattatatga   gagtcttttt   16980   acttgatggt   tgacaaattc   aacataatca   cgtgtgtcat   gaatcttttt   atgacaacaa   17040   tacagcatca   aaatgtacag   tgtagaccta   gaggaaatac   aaagagaaac   acagggcccc   17100   aataataatg   gaaaacataa   acaaaccttt   ctgtgccttt   gacaaattag   gtagacaaaa   17160   gctaaaaggt   aaagaaatga   gattctagag   acttcagtag   aactaagagt   acctaataaa   17220   atgcttctaa   aaatatgtgt   gtgtgcgtgc   gcacatgtgc   acacacacag   gcatttgtga   17280   aattatctta   taaatagaat   tccaggtatt   ttttcaaaaa   ctgaacatat   atatataatg   17340   ttttaaaata   tttcaaaaga   tagaacttgc   cttcaaaata   taattcaaga   aataaacaaa   17400   acttgaaatt   aaactgaaca   aaaatataaa   catttagaag   tcacttccaa   gtaattcttt   17460   ggtctaagaa   gaaatcagac   acacattgta   tttattgtag   aaaataatga   aaatgaaaat   17520   accacatatc   agactctcca   ggtcacagct   aaaagcagat   tcagaggaaa   attcgttgcc   17580   ttaaatgctt   caattataaa   tatgaatgaa   tgaaaataaa   cgttaagtat   ttaatccaag   17640   cattcagatt   ttttaaagac   agcaaaatta   agaaagaaaa   agaaggaatt   gatgaagatg   17700   tagtaatagc   aattgtactt   caataaaata   ttttgaggaa   ataatcagac   aagagctcaa   17760   gaatgtttgt   tcggtgctgt   ttctggaaac   tcagaaaaaa   ctttaatctt   tattataggg   17820   atgattgaat   aaatcataac   acatttcatg   ttagcagaac   aaagagacac   taaaatatat   17880   accttaatac   aaaaatctac   aacaagtgga   attaaaaagc   agaatgcaga   gctgaatata   17940   agaatcacaa   ttgggcactg   aggaatacat   tagcagttaa   tataagttgt   gtctttacat   18000   cttcaggtat   ttatttaaag   gaactacagt   ttggtgttat   aattagaaaa   atacaaccct   18060   aaaagtatgg   actgtgaaat   catatttatt   ctagtgtcct   cctttaagta   gaacattgca   18120   ttccatttct   ttagggactg   tatttcttaa   tcattgtaac   aggtataacc   tcttgtaccc   18180   attacagcaa   gcaattacta   aatataatat   accacatttg   tcttttctgc   aggatcccaa   18240   gaatttctct   gccctcaaag   caattttctg   tttaatgcta   tgctgaggat   actcttattt   18300   catactaatg   atttagaacc   ctcaccacca   tatcagaatt   ccaagccaaa   actaaggtac   18360   tatgaaaagt   taactggtgt   ggaaagtcaa   tggtcatttc   ttatcttatt   tctctacttg   18420   aagagttatt   ccttaccaag   agctctccgc   accttcacta   tggggaaata   tgcacctact   18480   ttaaggaaat   attatgggaa   ttaaatgttg   tgacatcttt   caggcaccca   gcacagtggt   18540   gagatggagt   tcccccttta   ccttctccca   tattctttcc   tgatcctatc   tgagtaatta   18600   caactcttcc   aaccaatcca   cttttacaca   gtttttttta   tagaataaac   atccaaatgc   18660   agcatctgtg   atatatctga   attaaatagt   gggtgcaaca   acatggctat   ttcaagctta   18720   aatgtcagct   aatttttttg   aaagcaaagt   catgggagtg   agtgtctgtt   tacacctaat   18780   gctgagaaat   ttaatctctg   caggctgtga   cacattctga   tatcagagga   tcaaaatccc   18840   actggtggaa   gttaatttcc   tttgctctac   aatccacact   taccattctg   cactcaggtg   18900   ggaaaataaa   acttgtgttc   ttcatccaaa   gttactaaaa   acacccaagc   ccaaatttaa   18960   acctcaaaac   tatgagctct   gcttccttgg   aactcaaaag   aataattctc   tttgctactc   19020   agagattaat   cacactgcct   ttatcaccta   ctatgcttgt   gttattttac   tcttcttttg   19080   ttatttaact   gagagcattt   gtgtctattc   tacccaacca   aaccatgaat   ttgcatgtgt   19140   cagggaagct   ctggcaaaat   ctgatacaac   ttattcctca   tatattaatt   gcttaataca   19200   gtttttgtga   ttgaattttg   agctaataac   atattttaat   tcaatacagc   atgcattttt   19260   aaacactctg   agaatgtcca   aaaacaattt   ggcacttagg   taagaagaga   ctttattcag   19320   gctgggtgcg   gtgggtggct   cacgcctgta   atcccagccc   ttcgggaggc   cgaggcgggc   19380   ggatcacaag   gtcaagaaat   cgagaccatc   ctggacaaaa   tggtgaaacc   cagtctctac   19440   taaaaataca   aaaattagct   gggcgtggtg   ccatgggcct   gtagtcccag   ctactgggga   19500   ggctgagaca   ggagaattcc   ttgaacccgg   gaggcggaga   ttgcaataag   ctgagatcac   19560   gccagtgcac   tccagcctgg   caacagagca   agactccatc   ttaaaataga   ataaaataaa   19620   ataaagagac   tttattcgaa   aggattagtg   caagagggag   actggaccat   tacagtaagg   19680   agaaaggggc   tgttgtagtt   tggagaacat   tcttatcatg   agatctggaa   gtatctcaag   19740   ggttaggaaa   aaagagattc   tctttcatag   gaaagaatag   acaaggctag   aaagaaccag   19800   gtgtggagaa   gtaccatgac   aggagtgaca   cgatctgcca   gtagatgagg   gaatatttta   19860   ccctgaactc   tgcagcgtac   tccagggaga   ggctgtggag   gaggagctgt   gtgctggctg   19920   aggctgaggg   cgggacaaag   ttcagcctag   gagaagcaaa   gaatcttaac   caaagttggt   19980   ttagcctgca   tttttttctg   attgatcagt   ggggacaaaa   cagcttagct   aatatcattt   20040   atgagacaaa   caaaaggaat   ttagagggtc   tgcgtctggc   cttgttataa   gtgttgatgg   20100   ctatttactc   tgtgttgcta   caaaaatata   tgtgaaattg   ggtaatttaa   aaagaacaga   20160   ggttcatttg   gttcacagtt   ctgcaggctg   tacaagaagc   atagtgtcag   catttgcttc   20220   tggtgagggc   cttgggaagc   ttacaatcat   ggcagaaggc   aaagtaggaa   caggcacgcc   20280   acatggcaag   agagggagca   agagagaaag   gaggaggtgc   caggctcttt   gaacaaccag   20340   atctcatgag   aactaaaatc   aagaactcac   tcattactgc   aagaacagca   ccaagccctt   20400   catgagggat   ccacccccat   gatcgaaaca   cctcccacca   ggccccacct   ccaacattag   20460   aggtcacatt   tcaacatgag   atttgaaggg   gacaaaacat   ttgaaccata   tcaatgccca   20520   caaggcagca   ttggtgaatc   tgatttaagt   cacatgggga   ataatggtta   gctgcacaaa   20580   gccctttctg   gaacacaaag   cagtgggggc   atttattaac   cttttctgtt   ttccaggatc   20640   acagggttct   ggtaaagttc   aaacttgtca   gaggatacaa   ggagaaaagg   tgcagtgtct   20700   ccattcagga   atttggagac   taaagaaaga   gtcaaggaaa   aattgttcac   atttttaaaa   20760   atagtaacga   ttaagcagaa   gacaagaagg   aagacaatta   gatctcattt   gatttccaat   20820   accttcatga   tgcaggcacc   acagtctctc   agttaacaga   tgagaagaaa   gaggcttaga   20880   actgttaagt   aaataatcca   gggactcaca   gacaggacat   agcagagcca   agtttgaatc   20940   cagatcattt   tgacagccca   gggcacagtt   catatatgaa   aaggccaact   aacaaaatgt   21000   tagctaacaa   atgcatcact   gtggacagtc   aggaaatacc   tgcaagctcc   agaactttct   21060   catttacata   gcactaacag   aaacttttgg   actttccctg   caatgaggca   gagtgtcaat   21120   ccctactgag   atggacccca   ataaccatta   aaaatacata   tataaaaaac   aagtttaaat   21180   attttgccta   taacttttat   tccaaagctt   atgaactctt   ctcttttagc   attacaggaa   21240   aatgatgtat   ttgttcaagg   atataaaaat   tgtggccagg   gctcccatag   tgaatgtatg   21300   aagcaggaag   ctggggggat   ccatggaacc   ttgagcattc   tcttccagaa   ctctccatcc   21360   ctgctaaagc   taaaatgcct   gtatcatggg   caaagcttgt   cacactagaa   aggtcaaaac   21420   ttaaaaatgt   tcttaccaag   tatctcccag   tagtatgtgg   ctgatatgag   actgaggcaa   21480   agcttctcct   agcagaaaag   aagggactca   gtaacctggg   aaaggtgacc   ttcatctaga   21540   aaaggaagag   acagaatttc   tacaaatgag   ttcaggtaag   aatgtgtatt   agagttctcc   21600   agagaaacag   aaatagaact   aataggaggg   gtgtgtgtgt   gtgtgtgtgt   gtgtgtgtgt   21660   gtgtgtgtgt   gtagggaggg   gggagagaga   gaaacagaga   gagaaattta   ttatagggaa   21720   tacaatatga   aataatcatt   cacacaatta   atagagacta   agaagtccca   agatctgcaa   21780   gcagcaagtg   tcaggcctct   gagcccaagc   caagccatcg   catcccctgt   gacttgcacg   21840   tatacgccca   gatggcctga   agtaactgaa   gaatcacaag   agaagtgaaa   aggccctgcc   21900   ccgccttaac   tgatgacatt   ccaccattgt   gatttgttcc   tgccccaccc   taactgatca   21960   aggtactttg   taatctcccc   cacccttaag   aaggttcttt   gtaattctcc   ccacccttga   22020   gaatgtactt   tgtgagatcc   acccctgccc   gcaaaacatt   gctcttaact   tcaccgccta   22080   tcccaaaacg   tataagaact   aatgataatc   caccaccctt   tgctgactct   cttttcagac   22140   tcagcccacc   tgcacccagg   tgaaataaac   agccatgttg   ctcacacaaa   gcctgtttgg   22200   tgctctcttc   acacggacgc   gcatgaaagc   aagctggaga   cccaggagag   ctgatgtgta   22260   gttccagtct   gagtatgaag   gcctgagaac   caggagagcc   aatggtgtag   ttccagtctg   22320   aaagctggca   ggcttgagac   ccaggaagaa   ctgatgtttc   agttcaagcc   caaaagccag   22380   aaaaaaaaca   atgtcccagc   tcaaagaagt   caggcaagag   gagttccccg   cttttcacag   22440   aagaattagc   ctatttattc   tattcaggcc   tttaattgaa   tggatgagag   ccattcacat   22500   tagggagggc   aatctgcttt   actcagtctg   ccaattcaaa   tgctaatctc   atccagagcc   22560   acctcacaga   cacacccaga   ataatgttta   actatttggg   cactccatgg   cccagtcaaa   22620   ctgacacata   cagttaatca   ttataccatg   ttagtttgtc   ttcttgtcca   ctcaactttc   22680   ttctaactac   tgtatagtat   tccattgtgt   tatatacatg   cacagttata   ctttataaag   22740   atgaaagggc   tgggcacagt   ggctcacacc   tgtaatccca   gcactttggg   aggccgaggc   22800   aggtggatca   cgaggtcagg   agatcaagac   catcctggct   aacatggtga   aaccctgtct   22860   ctactaaaaa   tacaaaaaat   tagccaggcg   tggtggcagg   cgcctgtagt   cccagctact   22920   tgggaggttg   aggcaggaga   atggcgtgaa   cccaggaggc   agaacttgca   gtaagccgag   22980   atcgtgccat   tgcactccag   cctgggcaac   agaaaaagac   tccatctcaa   aaaaaaaaaa   23040   aaaaaaaaag   gaaacaacca   tatttattat   tttgcttaat   attatgtttg   taagttccat   23100   ttatgttatt   tttggctaca   gatcattttc   ttattgctat   atagtatttc   attgtgtgac   23160   tacatgaaaa   tttattcatc   cagttgatgg   acatttgggt   ggtttccagt   ttgggatgat   23220   tcattgattt   tgacatcatg   aatatttagg   ttgcttctga   tacgtttctt   taatactttg   23280   cgaaggcatc   attttgcatg   actgcttgta   tagatacata   aatgtttctc   ttgggtggcc   23340   acaggttaaa   caatggtcta   tctggatcat   gaatggagag   aaaaggaaac   aaccagaaag   23400   ctctctctcc   ccacctattc   tctgtccatg   gcatctgcca   ctttctctca   ctctgcccct   23460   accacactgt   gacctaacca   gcttctacct   tctcccagga   gttcctcaca   gattttgtct   23520   tataaacatt   ttcttgaagt   aaaacatacc   tacagaaaag   tgcatatgtt   ttaagtatgt   23580   agtgcaaaga   atttatgtga   attgagcata   tgtaagtaac   ccagatagag   atgaaaaatt   23640   attagcatct   agaaatggaa   caatgcatta   gaggatatgt   aattccagct   ttactaaata   23700   ctgctgaatt   gcttttcaaa   aattaccaca   tcaactaact   ctcaccagaa   gtaaagatgt   23760   acacccgttt   cccttcaatc   ttgtcagcac   ttggggatag   ccacacgcta   gctttcacta   23820   taataatagg   taaacattct   tactggtttt   agttattttc   cttaattttt   aatatggttg   23880   agcctatctt   catactcatt   tgttagttgg   aatatgctgc   acttgttttg   ctattgagtt   23940   gttgtcctgt   tttgaacctt   tccttaagag   tccacagctt   atccagtttt   ctacacaact   24000   cacttctgtt   tgctgccctt   cattatagac   tcaatgcctt   ctcattgcta   gagcactgtg   24060   ccctccttgg   ggctcgatgg   tttataataa   agcttcccca   ggagatctca   tgggggccag   24120   gttctttagt   ggtggccact   aaagctacta   cctcttattc   caggtgccat   aaaccagtgg   24180   ctggacctgg   ttggttcaca   gagtgttctc   aatgtgagga   atcaggtatt   gtgcttggtt   24240   gtcatataag   cagcttagtt   cccattgttt   tatacctggc   ctgctaccct   cattcatgtt   24300   accttcatgc   cccatagaca   tttgagattt   gatgcaaacc   agtctgttac   agtaaaagca   24360   ccactgtcta   ggaagttctg   gagatgggtt   atcagctcat   ttacatcagc   agctgcggga   24420   tcttgggcaa   gtcatttcac   ctattggagc   cactctctgc   tagcataaag   gagaacataa   24480   agataccttc   cctgcctatc   tcacaaggtt   gtttgggaaa   ctgaatgaaa   gaacgtgtaa   24540   atgtgctttg   caggtttttt   gtgaaacagg   aatagctgtt   agtattctcg   cctagccctg   24600   tctgtggtgc   ccaggatgct   gcattacctc   ctttctaccc   tcgccctgag   tattggtcct   24660   aggtagaagc   tgaggtgaat   atactgaaag   cagtaagaat   cttcaccagg   cccctcccag   24720   tcatatcaca   aggtgttgta   gcttattgac   aaacacagac   ctcatgtttc   tctgagaacc   24780   gccctgtgaa   cacctagata   gaactaaggg   tgtgaaatac   aaacgtacta   tttgcctaca   24840   cttacggaaa   taagagcaca   tgacacagta   gatggcaagg   tgaggcttta   agtcagaaag   24900   caacctcagc   tgaaacaaat   gcataattat   aatttgattt   cagaagtgga   aacatgaccc   24960   gaattcatgc   acctactgat   tagtccaaat   tcctttattg   ttgaagctgg   agcagaagtg   25020   ggggtccacc   gtggaagtaa   ggaaaagcaa   aacagaacag   gggtctcatc   cagggagcca   25080   tgtcgatggc   tatggggagc   tccaccttgc   tcttggggtg   gggccctggc   cctgggtggg   25140   actgtggggt   gatgagggtt   gtgctatgct   ggctacctgg   tggtaattgc   caaggagaga   25200   gcagcacgta   gatcctccct   gtcatcaggc   agagctcttc   agtgaggtgg   gctcagggag   25260   ggctctgtgc   ctccgttcag   cagagctgca   gctgctgccc   agctctcagg   aggcaagctg   25320   gactccctca   ctcagctgca   ggagcaagga   cagtgaggct   caaccccgcc   tgagccatgc   25380   cagccaactt   cacagagggc   agcttcgatt   ccagtgggac   cgggcagacg   ctggattctt   25440   ccccagtggc   ttgcactgaa   acagtgactt   ttactgaagt   ggtggaagga   aaggaatggg   25500   gttccttcta   ctactccttt   aaggtaagtt   tcttgcctgc   gactctgaac   actgacttat   25560   aacaatgaga   ctgctggaac   ttaagagtgt   caattgaagt   atcatagcca   gtattgtgaa   25620   tgagtgttat   tttctttact   aaaaaggatt   tttaaaagtc   tgaagtgcta   aaacaacaag   25680   ctgtagtgtg   cagagaccta   gattgtagtt   tcttaggaat   acaggtgacg   tttttctttc   25740   tgatgctgct   ggaaatgtgt   gaatcgtgag   taacatttat   gagtggatga   tttttacttt   25800   ccccctttcc   ttaggtgaaa   gcacttttaa   ttaatcctat   agcagtactt   gaaatgtctg   25860   tttctagtag   gggttagagc   tatgagtttt   ttgctaggaa   caccagagat   gcggaaagct   25920   cacaatcccc   aggacagtga   ctgatatgat   gagaaacaac   attgaataag   gcaggaagga   25980   cgacactttc   tttgatgatc   cttttcatta   aacatcagtg   aggaatatat   ttctttgagg   26040   gtagttgatg   atgccaccat   ttgcaacaaa   ttacctccta   gaacaaatga   aggacagaga   26100   gtttggtgag   tttcaccttc   aaagtatcca   agggccacac   tgaggaagga   tcttcattaa   26160   gctaatactg   tattgtttct   tattccgcaa   caagatcttg   tggacagaaa   tgaaccgcac   26220   atttgggaaa   agaacagttt   cctgcttctt   gcctcagaat   gctgggttgt   caggttattt   26280   cacacctgtc   taacgatatg   tttagtcatt   gcaataattc   acaggtctca   aactggtcct   26340   ttccactgaa   gaaaaaatac   ttcaaaaaga   ttctagttgt   tgctttgttg   ggttttgttg   26400   ttgttgttgt   tgttaatcct   ctgtatgttc   ttgaaatgat   ttggttttta   cacaatgaga   26460   tagcattact   gtaacatcca   cctccctact   ggaagttcac   ttcttttgca   aatattcaga   26520   ataaaacttt   cagcagggtg   ctgtcccagg   agtcagactt   aaacttccag   ggccctaatg   26580   atcctgcagc   aaagcagaac   ctgagcaaac   tactcctcca   ctggcttcag   tgaaaccctg   26640   tcccttggaa   tcacctccta   atgcatatgc   acatacgcag   cccagctgca   cagctccacg   26700   atgtagctta   gaaagtaaaa   ttagagtttg   ttgccactta   atgtcgactg   aataatcccc   26760   tttatgaaat   cagcctgcag   acccagagag   tctgtcttgg   caggctgtgt   gcacccagga   26820   agcacaggac   tccactcagt   attgctgcct   gtgtagcctc   tttgatattc   ttagaacatg   26880   atagcaagca   gaatgaagag   aggccctcca   ttgcaaaggg   aataaaatca   gcaataaggt   26940   ggttattctg   agactctggc   aagaatcaat   ctcagctgaa   atctcatttg   ccattttttg   27000   aatatagcac   ctacagggat   atttggaata   tgtgaagaaa   caaatcccac   cttgctctct   27060   atgagtgttt   gtcctctcta   tcctagcaac   atcatcgcca   gacactaaag   agaatcacag   27120   catctctagc   tgattcctat   tgatatgttt   gcagcaaaac   ggctggacca   ctggacaagg   27180   agaaaagcag   aaagacagga   attgtgtttg   gccaggagct   tagaatgctc   agattcatta   27240   tttgagagat   acgtgtattg   actaaaaccc   tgtgattttt   agttacacag   tgattactgg   27300   tggtatttgt   gagaacttag   cttatatttc   ttagtgtaac   tttttttcta   tgatcccctt   27360   tgggcacaag   caattctaat   gcattgttga   tacagaaata   gttccaatgc   aagcccttgt   27420   tcactgcaat   tcattttatc   tgacatactt   ctagagaaga   aattcgaaac   catgctccct   27480   ttactgtctc   ccagatgtat   cccctttaat   ccccattgcc   ttcatcaaaa   gcagtaaaaa   27540   aaatagcatg   aaaataaaaa   tagcagcagc   agtaaaagtc   acagaaataa   cagcaacaac   27600   catacgtctc   atttactttg   ggcattcttg   gtgccaagag   tcatagcaag   actcacttag   27660   caagtgactt   ctgtgaagta   tctcatttaa   ttctcacaat   ccatggtgta   tctttccttc   27720   cggtcctgcc   ttccatcttc   atctgtttat   ctcaatttta   cccagtctat   aaagctcagc   27780   ttgggttgct   tcctccatga   agccctcctt   gactcatccc   tccctgactc   atccctccct   27840   gagcttcatt   agtgtcacgg   actgtacctc   tcttcttgcc   acttaatgca   gttctgtctt   27900   aaactgctat   ttccatatgt   gttggctgaa   tgtgttacag   tcctcaagct   ccttgagtaa   27960   aatgaatgcc   ttatatttgt   ttcatatttt   ctcccaggac   ctaccactat   gttagtcata   28020   ctcaataaac   attgactgca   taaacaaatt   gtgatgtgtt   ttttcttcca   attaccaaca   28080   ttttgtcaaa   taagcaacag   aaagcatgta   ttccctgtga   cagaggtggg   tgggggctct   28140   cattaaccaa   gtttccaccc   atcagcccct   accctgcctg   gtctctatac   tcacccaggt   28200   ccctttctgc   aagcatgttc   agcttgtccc   tattccattt   ctaagccaat   gagaccaaag   28260   tcggttctag   attccagtgg   gtggggtatg   gaattagctg   aggcaccatg   agctggtacc   28320   aagtacctgg   tccagccctg   gggaatccat   ctctattgca   tggcagcctt   acctagtctc   28380   agctgtcttg   ctccgtccac   ttacctaggc   tcctgacttc   gtttcccatc   tcacattcta   28440   gttcttatcc   tctgaactcc   aaagctcttt   caggaggaga   caaagcaccc   cgtctagatc   28500   ttactggcac   ttccttcaca   ggcttagagt   ctgttccttc   ctcccacctc   cacccttgaa   28560   ttctggcttc   tgcttccacc   cctctaacct   actctacctt   cctaagcatg   tgaggctctt   28620   ggaaatgagc   agggaggtct   agggtgggtt   cacccatccc   atcttccacc   atgggccttg   28680   tgtaactaac   cctacgtggt   cttgggttca   agattctcag   ctcagaccct   ggctcccttc   28740   acagtttcca   gttttgtgtg   cttttaagat   cacatccagc   tcacctcttc   aagacctagg   28800   tcttggcact   aaggaattat   atgcgtttca   agaatctacc   agaagttttc   aaggggaaat   28860   gggaaccagc   tcatcagaaa   ataaatgagg   tatctgaata   taaggctgtc   caaatgcaaa   28920   acaaatacaa   aactcttcac   acagaaagcc   cttaattttg   ctggtgaatt   caccagcaat   28980   gtttcctgaa   tctctgatac   ttaatgagaa   acctaattag   aagtaactgg   gagggagacc   29040   tgtcttgatt   agtgaattat   ctaaattata   aaataatgta   aatgaaactt   agttttatta   29100   ctcataggtg   agcacagaaa   catgaactaa   gactgctaaa   tatcacctaa   taacaatctt   29160   ttattgcgcc   tccttcggtg   gatagttaca   agtgatacac   aattagtcta   ttgtttatat   29220   gtaaatggtt   gcttctaagt   gcttgagggt   gatttgacac   agtttgttcc   ctttgtttgt   29280   acaccctccc   tgccacgctg   ccttactgtt   aattttctta   acacactctt   cctccattaa   29340   tataacaaaa   gcaaatttca   actcaacgca   tgtcccaatt   atctcacgct   atgcttttgc   29400   aagggagctc   cttcctgccc   tggccccagg   acagtgtcag   ggatgtcctt   cctggaatcc   29460   agaatccctc   atgtcccaac   ccccaaatgt   atctcctgca   atttgtaacg   agaggtcttg   29520   catttacatc   atgcattttt   tattcttttc   taattagtct   cgctcagtat   gttcaatttg   29580   agtttctaca   gttgaatact   gtggctgctc   ctaaggaaat   tactaccagt   tttgctcggt   29640   taaatcatta   ttaaacatat   tattccttcc   tgccagttct   gctacccaga   cctagtgctc   29700   tgctttccaa   tccctgaccc   tgggtaaact   ctaccgtgca   gtgtgtccaa   tttcacttca   29760   cacaaatgag   aaaggctgag   gatggcacac   aatttccaaa   ctatagacgc   tagtatttga   29820   aaaggaaatc   ttcctcagct   aaatttctct   ttgcattact   aatacatttg   atggcaggaa   29880   gcataatctt   agctaattca   gaataaacct   agtcattgca   gagcttttca   gcccaggaag   29940   agttttcaca   ttttctggct   tcgggaaggc   atctaactcc   aggggcagtg   cttctgtagt   30000   gcatttctct   ctctctctct   ctctctctct   ctgtgtgtgt   gtgtgtatgt   gtgtgtggcg   30060   ggggggagtg   ggggtagagg   atggatggtg   aagagagtct   gatctactga   gggatctgag   30120   caagttggca   tcagctgaaa   caagacctgt   ctgtccctca   gatgtggtca   tcctcgttct   30180   ggactttcat   ctgggttccc   atccgacctg   tgtcaggccc   acattgcttt   ctggggtaat   30240   gtcccttggc   cacattctct   cctggtacag   ggtgcttgaa   tccctgtctc   tcaatcccag   30300   cgataatttc   tgcctccttc   ctaagaatag   aaagtttgac   tgctaccaac   tttgggctgc   30360   agaaaacaaa   atggagcatc   tctggccctc   tctcatccca   catagccacc   tacgctatac   30420   aacttccacc   tcatgataga   tgggaaaatc   tgggagactg   tacttccctg   gtccctacac   30480   agtaagtgga   cctcagcgcc   cttccaatcc   tgtgctcccc   aaatgtagca   aatgttctct   30540   cttcctccgc   tttttcctgc   taacccttgg   aggggggcaa   tccaataccc   tcttgtcatt   30600   gtctcagttg   cctcgcatct   ccccagcatc   tgacaccact   gctgcgccat   caaactcttc   30660   catcgccctc   agcgaccccc   ctgtcatggg   gaaattgatg   acacagtgtt   cttccgaatt   30720   tattattcca   ggctctctgt   tctgaatgct   caaaagtcac   accctcttgc   aacagaaaaa   30780   acccatttct   cttccagctg   tttaagatat   cctcactgag   gtattttgaa   agcccagtgc   30840   tgacttgttt   ctgtccatct   gcattgataa   agagtgcagt   ctgaggacag   tggggctgcc   30900   tgggctggtg   gacagtgtat   caaagaagtg   gaaagaagca   catccattca   tgcttgacaa   30960   tgtcattctg   ccatccttca   aataatttcc   tgttgatgaa   gtaagaatgt   ttcaagccat   31020   gcactcaggt   agataagaaa   tctaacacta   tttttcttaa   ataaactgag   cttctctctg   31080   gacctctcag   tcccccaagt   aaactcaggc   ttttgagctt   tctccatcat   ttcaatgagg   31140   cagtgtggct   cagagagcag   gctttagagc   cagacttctg   gggtcaaatc   ccagtgccac   31200   accatctagc   agagtgactt   tgggcacttt   acttccaccg   gtgtctcagt   ttcctcatcc   31260   aacaaacaaa   gctaataatt   gcatgtcctg   tagagagttg   ttgtgaatag   ttaagtgaga   31320   gaatgcttgt   atagccttaa   agaacagcaa   gcggccccca   ggagtgttca   ttattcataa   31380   ttagagtccc   aggaattcgt   cattccactc   ttttctcaag   acaactactg   agaggtgagg   31440   aggcagcatt   agctgtaagg   aagaagtttc   ggaagatgct   gagagagacc   actgcaggtg   31500   gaagcacttc   aggacagctt   cgatatgcag   gtggtcctca   aacaatggtc   atgaattaag   31560   ctgtttcttt   tctttttttt   aaaaaaaaaa   aatgagacca   tgaaaatgta   gaataagata   31620   tagatataga   gggaaaattt   ccatttctta   ctttgtcatc   ctgatattat   gtctagctat   31680   tatctaagca   aacttaatgg   cccactacat   gaaaggctat   ttgcagaaag   agcaaattat   31740   gccctatctg   gagaaagaaa   gaaatggctt   cttctctgct   gtgaagattt   tccaggccac   31800   agatggcaaa   atcacccaag   gtcatgagga   agttgtaagg   aataatcaag   tttcttgttt   31860   tcccacttag   gcaatattga   tggctgccta   aaaatgatag   atctattact   cttgtcagca   31920   ctttacagtt   tgcaatatga   tttcacaata   tcacctttga   acctcccatc   agctctaggg   31980   ggatttaaaa   tagcattctt   tttctttgag   cagatatgaa   actaaagtct   gagggaagtt   32040   aaaggtgtaa   acttcctaaa   agttaagata   aggtaaaaat   aaaataagaa   agaaaagaaa   32100   gagaattctg   agctgatttt   ggtctggata   ggttaagctc   tccaacagta   aaaaacaaac   32160   cctaaatctc   cgtgatttaa   cacaccagag   gtttattgtt   ccctcatatc   acagtagcaa   32220   acaggtcaac   ttgtagctac   accttctaga   acaagtggct   tccaggggtg   atggtaaatt   32280   ttctcttgct   gtctttggcc   tgaaatgaca   catgactttg   actgaattcc   attggcaaga   32340   attaattacc   tcaataggca   acagaaacgc   ccccagggtc   gagggtataa   tgcatggctt   32400   taaagatttg   atccttacct   tctggcagat   aaaacccaaa   tccttgggcc   tggcctactg   32460   gcctttcaca   ttctgaccca   atcaccttct   ctactcctct   cttcacatcc   tggaccacgt   32520   tgtactgact   gtcttaccat   agtctgagga   tacagggtcc   tgtttacctt   ttctggatat   32580   gcctttctca   cacctttttc   ttctgatatc   acgctaacct   caattgcctt   ctcttctagg   32640   aaggcttcag   aattgttcag   tagtccagca   tcatcacatc   actacccttc   tgctcctaaa   32700   gcacttagaa   tctctgtgca   atatggttcc   tctcttgaca   tgcatgttca   cctacataca   32760   cacacacaca   cgcatacaca   cacacccatg   agcacataca   catggacaca   tacaaataca   32820   cagtgacaga   cacacatgcc   cacacacaca   cagactagat   gtcagttatt   tagggacacc   32880   tttagatcct   actattccta   gtacagtgac   tgacacacag   cagataaaag   gtcagtaaat   32940   attttgttga   gtgaaagagt   atatcaagaa   ataatctgta   aaacaagttt   actgctaata   33000   tctcaataag   ctcatgtgaa   tgaatttggg   tagataaagt   cttaatcctt   attcaccaac   33060   catttggctt   gaatttatag   gtgtcataat   ttgggaggga   ttcgtgacat   cacagtaagc   33120   catgttctat   attatttagt   aatgatgagg   aataatgatg   atgctatcgg   attttgggaa   33180   aaatccgtag   gattggtgag   tagaggcact   acacaagcct   ggaaaaggga   aaggaaaaga   33240   gaacaggttt   ctgtaaccca   ggatgacatt   tggttgaaac   aatacttagt   ttcaacctcc   33300   tcctcacttt   cccagctaag   tactcactct   cctagtctcc   cttgcagtta   aaggtagcac   33360   agtttgggaa   gcttaattgg   aagtctctga   gatttctaat   aaagattttt   ccttttctga   33420   tgaaagcgga   aagatattgc   tggcattctg   atgtccccct   gacttgaatt   cagacataat   33480   gcatggagcc   gcagcagcca   ttttgtgacc   acaggaaaaa   aaagccaagg   gaatcttagc   33540   aaattgagtt   cagcagcaac   atatcttcag   atttcttatt   atatgaaaaa   aaataaatct   33600   ttgtttaagc   cactgctaag   ctgaatactt   tcctaattga   tataaatact   aagtgtaata   33660   cgacattgac   agtattgaag   ccaagatctt   aaaacatggc   ccccagagag   tcaagcagac   33720   attgcattat   tacacatttg   gagggtacag   aggctggatc   tgcccatggt   ttttgtcttg   33780   ttattatcta   cattacaaag   agattgaata   gattcttata   aaaactgatt   cagactagat   33840   tgtatggttt   tgagtaacca   agaaagtaaa   agtttctccc   cctcctcaaa   gaaaagggcc   33900   ttacattttt   caaatgattc   tctacattcc   agataataac   aatgccattc   acttatgtaa   33960   ttctatgaaa   aaatactaaa   accattctga   aaacatcagc   tcaaaaaact   acccaaaaca   34020   tttgctcatt   ttttcatgat   ggctcaataa   gaagattttt   ttattctcct   ccaaaaagca   34080   taaggaatca   catcctttcc   tgacagctta   aaacatccaa   gcaaaatgtg   agctggagcc   34140   atgggctctg   gacctggcag   gcaggtgtgg   accttcagca   caccacaggg   ctgctctagc   34200   cccaggcatt   tctgccctgg   gtacagaaca   ataaccacca   tccctgcctg   gatggtgaag   34260   taggaatgga   gccagttctc   atgtgggagg   aagagttttg   agagacagga   gaaagggggg   34320   tagtcatgga   ggagttctgc   ttactaatct   gtgacaatcc   tctccaagcc   cagagaaggt   34380   gggaggtagg   ggaggaccga   tgctcttccc   catcaaaggc   cagcctccta   gactctccat   34440   tcatccagct   tcttcaccct   cctggcctct   ccactttccc   cgcctttcca   cttactcaga   34500   ttctccacct   ttccagacac   tccatcctca   aagactctcc   acccacccag   gctagtcatg   34560   cattgctaaa   gataaataaa   aaagaaactt   cctgggactc   tttcctcctt   ctaaagacag   34620   gagtaggtgg   ttggaaagga   ataagatgca   atcataactt   tgacaagact   cacaagaacc   34680   ctcatgaaga   ttccctgccc   tctcacaagc   ctctcttttc   ccagccttaa   ctgcttgccc   34740   tcatccttac   cagggcattc   agcttggact   ttgcacgctc   agtttttaaa   aaacagttct   34800   gtctccccct   ttctcaccat   ccctctttta   tctcatcaga   acttctttct   tcaataaact   34860   gattccaagt   attagcaaaa   aaaaccattc   taagtaaaag   agttctgagt   tccatccctc   34920   aagtgtctct   tcttataacc   cccctaacat   atgttagggg   ggttaatatt   aatgtaaaat   34980   aaactcttac   tcctcatgcc   ctctggacag   agtctaaagg   attcttggag   ttccttaaag   35040   aagccaaaac   tcttttgctg   ggctacagcc   acgattcacc   caggcctctg   aatcagaccc   35100   aagatatccc   cacagcactc   agaattccat   tcacataaag   atattgaacg   aaggtgttca   35160   cacaaggagc   attattcaca   agtcataatt   taataaaaaa   tgtcagggaa   tattgtgagt   35220   gctcaaatgc   aggtttctca   tctttcacgt   attattgtag   actctaaccc   ttagcaacac   35280   agagcaaccc   atgcagatag   aagatattct   tttggctaag   aatacattta   ttccttttcc   35340   aatgtattta   gtcccttttc   aggatttttg   gtctaggtta   agattaaaat   gcaaagagta   35400   ttcaagaaca   cagaaactat   cagcctaaaa   taaaacttag   aataatgtaa   tgaatgcaat   35460   tctatcaggc   cactcagtct   tttctattga   tcatttcatt   catcaatcag   gttaagcaag   35520   cttatgctga   ggaaacaaat   aactggatat   tctcagtggg   ttaaaatcat   aatattttaa   35580   tgtttgtttc   ctactcatgt   cccatgtcca   tcacaggtta   acagggtgac   tccgtgtatt   35640   gtaatctctc   agggaaccag   ttgatggaag   ccccatgtca   acaaaggctt   ccataatact   35700   gaagtcagaa   gagcaagagc   tagagagata   cagagtaacg   aaccttgccc   tggctctcat   35760   aagcttctgc   ccagaaataa   cacatatgac   ttccactcac   tattcattgg   ccaaagtaag   35820   ttcatggtca   cgtctggtat   cagtggagca   gagaaatgta   atcctcctgc   tcggaaaggc   35880   agggaatctt   ggtgctcggt   aatgtctaat   gcaactccta   aagctctgag   acaggcatta   35940   tcctctttga   tttagagaga   agaaaagtga   ggctctgaga   agataagttc   acataatagt   36000   taataatcca   aaaactttta   cccagatatt   ggttaaaatt   ataacccaga   taaggcgcat   36060   gctctttcca   ccatatcaga   acttcccaaa   agttgttcca   cagaactcta   attatgagag   36120   ttcttaagca   gttgtcaggg   gaatatgaaa   taaagaattt   tggaagcacc   acatattata   36180   caccattata   tacttagagg   atgcattcac   catttaactc   tctgagaagt   cttacagtaa   36240   ggaaacttgt   ttatggtggc   tcacgcctgt   aatcccagca   ctttgggaga   tcaaagcggg   36300   tggatcattt   gaggtcagga   gtttgagacc   agcctggcca   acatgataaa   accccatctc   36360   tactaaaaat   acaaaaatta   gccaggcgtg   gtggcaggtg   cctgcagtcc   cagccactca   36420   ggaggctgag   gctgaggcag   gagaattgct   tgatcccggg   aggcagagga   tgcagtgagc   36480   caagatcaca   ccattacact   ccagcctggg   cgacagagcg   agactctgtt   tcaaaaaaaa   36540   aaaaagaaag   aaagaaaaga   aacttgttta   attctttaat   atgcattttc   caagtctatt   36600   tgaccacaga   gcattacaat   attacctcta   ataatgacca   ctggcacaca   cttcagaaaa   36660   cactctgtta   tgtgttttcc   tgaaagagcc   taccagacaa   ccaaacagaa   ctagctcttt   36720   gagcagatga   ggcccagaga   agtaaagtga   ccttcccaag   gttacatagc   tggtagatgg   36780   tagagtcagg   acaagaactg   caatctctaa   ctatccaagt   tggagcttct   gctcagctgt   36840   cgagaggatc   gtgaaaaaac   gagagtgttt   tataaactgg   aaagcgctat   tcgaatttaa   36900   gatagcatta   attctctagg   tgaagcatta   ctgaaatata   tagaatacat   ggaaggacat   36960   gaggtgcttc   aagataggat   gattaaattt   ggtgaagatg   tcaattctcc   ccaaattagc   37020   atataatgtt   tgaaataatt   ttaatcaaaa   cttcaatgtg   attgttttaa   attgattaaa   37080   taatttgaaa   tttttattta   aaaaataggt   gagaaaagct   cagaaatttg   aacacaaaca   37140   actatgggga   agaaagatgt   ttccctatta   gttattaata   aatgtatatc   cacagtaagc   37200   aaaacattac   agaactagca   aaataattga   cagatggatc   aataataaga   aatattttgc   37260   actgaaaata   aagtcctggt   acatatatga   acttaatgtt   acaaagaaac   taccacagtt   37320   agtaaaaaag   gaactagtat   ttttgcaaca   aaagtagtgg   gtgaataatg   gttcttatat   37380   aaagttgtaa   atctcagcta   gattgcggaa   ctaaattttt   acaaaatagc   tagaaaaatt   37440   atataatgat   gacactaatt   tcaggactag   caagcattat   tcaagcctaa   aaacaaaaat   37500   ttaaagggaa   acaacaaact   aagaaaatta   tttgcaataa   aaataaatat   ataatattat   37560   agctaatagt   atatacatta   attatataaa   aatataacat   ggttaaagtc   cttaatatgt   37620   aaagagccct   tgagggaaaa   taaataatat   ttctaatgga   aaagtgggca   tgggaaataa   37680   aaacaatttt   agattaaata   cagagttaaa   agaatgtgtg   aaaaaatatt   gtttattagt   37740   ggtctgaaca   atacaaactc   aataattcac   cataatatga   gatatatcaa   ataaagatca   37800   aagcaaaata   atatataaag   cacagaacag   ttctgtaaaa   ctgtttgtgg   gagagtaaat   37860   cagaacaact   taacccaatt   tttctggaga   acaatttagt   aatatacagc   aaacttaaat   37920   gatgtattca   ggcactgctt   agtaattcta   cttgtaggat   tataccataa   gtatactaaa   37980   gaaacattca   aagacagaaa   caaagactgc   attaagttgt   tttattaaaa   agtaagtttc   38040   tagcaatgga   ataataaaat   tgtgatataa   ccatagaatg   gaatattaag   gaatattatg   38100   caactataat   aaccatttct   ctttgtgcaa   tgttaaaggt   ttttaaaagc   acaataaata   38160   taatatattt   caagttttat   atacactcat   aaaatataga   agaaacaaat   accaagatgg   38220   aaaagtaaga   aattctctct   gaaagttact   ttgcagatgg   tttttatttt   atctttgtgt   38280   aaaagcttac   tgtattttct   agagtgagca   catattatat   ttattattct   aaacgacaaa   38340   ataaccatgt   aagatacacc   atgccacctc   tcaaataaat   tgtgtgcatt   gtatttccat   38400   gacagtaaat   aggaagaatc   tatttactct   gcagaagccc   agaaatgaac   ccaagcaagc   38460   gaggcttgca   tgacccagag   agcacattaa   aggatccaga   aatgcaggca   agggaacaag   38520   acagccaatg   aaaaccaagg   cagatgactg   gctgtatgaa   ttgagtgcac   accaactcat   38580   tagcctttta   cattttgcta   cccaaccacc   caacttagat   gcacagaaaa   caaagcttgc   38640   aagagtagca   attccagact   ctgtagcagg   aatccttctg   ctgagcctct   ctcatgcccc   38700   ctcccctact   gttctcatct   cccaggatgt   tcaaatccat   tccctggtcc   ttacatctgc   38760   ttctgcccat   gcatgtgttc   gctcctcctg   gtttcttttg   gctacttgat   tttgagtctc   38820   ccggatggat   tttaaatacc   aggtccaagc   caaagtgggc   cctccagggg   aaagtttctc   38880   agacactttc   cctttttatt   agcacctcca   gaacagaaaa   caaattcctg   aaggtttatt   38940   attttaatta   cgtggatcat   caggaataat   acattctggg   tatttttatt   atatagtaat   39000   tactttggtt   cctacgacat   tgtagagatt   tggacttaat   ttgctcaaat   gttagttttc   39060   catatttcaa   gagtctagtg   cttaaatagc   aatttaaatg   ttaatctcag   ttcttgttat   39120   gtagctgaca   tttcattcgt   ttattcagca   aatattaatt   actacataga   cttgaagccc   39180   agtgggtttc   caaatgattt   ctaacatctg   tggtttgcta   tattatggga   attcagatgg   39240   aatctcagaa   tttaattgtt   tcaaagcttt   caatacatgc   tattagtggt   taatttggaa   39300   gtcctcttgc   gatccaagaa   caaattgcaa   gaagagattc   acgttaaaat   gattgcagaa   39360   cattggcatt   tctgtttctg   gtgaaattca   aggattcacc   agtaaccaga   aaactgaatt   39420   cgaaagtaac   gcaagtgcag   tcccacatgc   caagccctga   tcgaaatgag   aacgtttagg   39480   atgtgggaga   ttaacattgt   taataaagca   tgatctacgg   cccatggtac   tgatgataaa   39540   aactccaaac   caatgactat   gtctttctgg   tcagagtaga   aatgactaat   gcagttattc   39600   aacatgtcct   ggagactaaa   ctaagaaatc   aatgtcaggc   agtaatccag   agtccactga   39660   atcactttcc   tgtacagtcc   cagtctgaag   gtgctagtat   tacattctgt   cacttgtaat   39720   catggtgttg   agaggtaagg   caaaggcaat   tcaggtgaag   acaactgagg   aagatcgcag   39780   atggacaaga   atctctcctc   ctgacagcag   attcttctaa   ctccggacaa   tgacacaggc   39840   cctggacctt   gtcccattat   tgaaataatt   ggctttcaga   ccgagctcca   gcttgctcat   39900   tgtaaacctt   tctctgctgt   tactacctca   aaagtgcctg   aatggccact   atctgtcctc   39960   actaacctgc   agttctcact   ctctgccttc   acttacgctg   ttgccaggca   ccctacttct   40020   tagcttggct   acttctactc   attgtttcag   actcagctca   cataacagtc   tctctaggct   40080   atacctgctg   accaaccatg   accagctcct   ccccaggttc   tattacatgt   tccttatttg   40140   ggtttctaca   acatctttat   catagaagtt   cacattgaaa   attctggttg   acgtacctgc   40200   ctccaccatc   agactataag   ctccttgtct   tcgcctccct   tttatcccca   aagtatgttg   40260   aactttgata   ctagattctc   agaaatgttt   atggaatttg   attgtacaga   ttacaaacca   40320   gtataatcat   gtctccagct   ccatgatttt   caggagagtt   tcccacttcc   ggcctgagtg   40380   tcatttctac   accaacatgt   aagccaaggc   tctctcctct   ggataaatgc   ctttccctct   40440   gccatactac   atgggagttc   acagaatgac   tctcagttca   caactacttg   gaacaatcgt   40500   ggtccacagc   cccactctcc   aattcaacag   gcctaggctt   caactgcacc   tcccatccca   40560   ttcacattcc   ctattcttca   tgaagaaggc   ctctgtcagc   ttgtcttgcc   agtctccaga   40620   caagtctaga   ggcattgcag   agggtgtaat   ctctctccat   ccctgttcta   gacccttgtt   40680   aaagtgatat   gagaagcctt   acaaaggtat   taaaacttga   gctaacaacc   acatttatat   40740   gtagctgtat   acagatttct   tacttttttg   tgaatattta   aaaaatttta   tcatacatat   40800   tttcataatt   gagtgttgta   actgatgaca   atcctgggta   atccaggata   gaatccccat   40860   agttcatggt   aaagcagtta   gaaaacgtaa   atgccaacag   gaggtcgtga   tgacaataat   40920   tcaactgtat   ttgcaagcag   gcctcccagt   tgggttactc   aggggttgat   agcaggggag   40980   tggaggagag   acagatttta   aaaaggggca   tcatcggaag   gagatggctg   tcagggagaa   41040   aaatggcagg   ccctcctgaa   ctagcccagt   cctagcttca   ggcctccttg   ttgaccttga   41100   ctggttattg   accacctggc   attgcttctg   tagcagcagg   tagggtgaga   tgtgggaaga   41160   agttgggatg   acctcctgaa   tgacagatta   ccaactgtcc   agctactgcc   caggggtcac   41220   cacatcctca   cattgtgtct   cattctccaa   gtctaattaa   gcagcttcag   gaacacagaa   41280   ctttgctatc   tgctcctacc   tgtttcactt   aaaatcaggt   gcccatgtat   gtctctgtgt   41340   tacactgtga   ctgatcccta   ataaaattag   agattttcaa   tctttgcact   tgatattgcc   41400   aactcaccta   agcaaattaa   tgaggctttg   agtaggcctc   agagcctggt   gctgtgaatg   41460   atggaagcca   cacaagtctc   agggacaggg   cagtagaatt   tcattgtctt   ctcactgaga   41520   agtgacacca   aatcattgaa   accttttttc   ttatacttta   tttcctgtat   tggaaaatgg   41580   cagatatttg   ctccttcctg   tgttagagat   attcatcttc   aacatcacat   acataacaac   41640   cttcctaaaa   caaaccctga   aaacatgagg   ggcacggaaa   gctttgtgac   tcccaggaca   41700     gctgatgtag   ctgcatcttg   tgtgacattt   atcactactg   ttgtgtggag   gccacatgtt   41760   gccatagagt   ccctccattc   ctccaatgca   tggaccttat   gctcagtcct   gagggagaca   41820   gtcatgaatc   caacccagcc   ccagaaaccc   agcttcaggc   aagttttggc   tcagaaagaa   41880   aatgaagatt   tgtctccccc   ttgaaaattg   aaagtaaatg   aggcaacttt   cactatctta   41940   aaccaaaatt   ggggcagaaa   actactagag   taagcagggc   atggtctttc   cagttaagca   42000   aaactacatg   caaatcccag   ctgcatcacc   taatgagcca   gtaacccagg   taagtaactt   42060   ctctgagctt   catttcctta   cctataagat   tgaggacagt   gataactcgt   tggtcaggtt   42120   gttatggtta   agtgagcaat   atgtaagcct   tcttaaatga   tgcttggcac   atggcaatca   42180   atcaaaaata   tttattgtgc   acctactaca   tacattacaa   actcagtaag   ccatgtactg   42240   gattaatgtg   attcactcaa   attatttcat   tccctttcac   cctataattg   ttaaagatac   42300   ttaggctttt   tgtcttccag   ttttcagcac   ctaactgaaa   aaggagatca   ctattccttg   42360   gatctggata   aatccagact   agcctgacag   atccagaatc   aactagagta   actaatgtct   42420   tgtatttcct   gagataattc   ccatttcaaa   tgaccttttc   tgtgattctt   gtaagtagct   42480   ttgcacttgt   catcaaccaa   gtttccatct   gacagagatt   tattaagcac   cttctacggg   42540   ccagacagat   gccaggggtg   gagttgtgac   ccaggcttag   tgcctgccag   catgggcatc   42600   cttttggctc   tggaagcatg   gctcatggag   ccacatccca   gctatcagcc   ctgtggcagc   42660   atcaccatca   tcaaaggaac   cctgcaattc   atctttctaa   atcccaattt   gcaattctct   42720   aagtctgcag   aaaactgatg   tcactgtgta   gcctgactaa   cttgaggtga   aaagccaatc   42780   agtcatgtgc   cagcaaccat   gtaattgagc   tgagaacagg   ccaatcttta   gttattgaaa   42840   cattttaaat   acatttctta   tttccactca   gtattggacc   atctggctgg   aattttgcct   42900   cacatcacca   ctagggggca   ttttatgccc   aactatattc   tgcaatagtt   tgggggttgt   42960   gttgggacac   catcagcaca   gtgtggtggg   gggtgggtag   ggagaatgct   tcttaggctt   43020   agtgaaattc   cgatagactg   gtaccgaaaa   tagggtgaac   agggtgaatt   caagcatcac   43080   tctgaaaagg   cagtgagtta   gttatcaccc   ctcaaatgta   catatgtagt   ccacaaacct   43140   aagggaggag   ataaaccaat   ttttattgca   tatctactat   gtaacaggca   ctgagctaaa   43200   aatctccatc   ttcatcatct   tatctaccca   tcacaactat   cctgagatag   gttttgtctt   43260   catcctcatt   ttacaaataa   gaggtcagag   aggttatcac   ttacaaccta   tcacacatcc   43320   agcaagtagc   aaaaactgag   tccaggtcag   gttgctttca   aagcctatca   acttttcgtc   43380   cacgaggctc   acctagttgt   gatattctca   ctcctctgaa   attgtttcct   cagtccccat   43440   taccatgact   ctccagatta   tcctgggatt   tgtgtgtgtc   tctgtgtgcc   tgtgggtggg   43500   gagggggaca   tgcacgcata   atacggctgc   gaataaagga   gcaagcagct   gagtgttttc   43560   atacttaatg   acccttgtac   tagtcagtga   aacaggattt   aaatttgact   gagtataact   43620   tacacactct   acctgtttta   ttctcttgta   gcaccaacgc   cttacactgt   gtctggcata   43680   gagtaggaaa   ttaatagatg   ttagatgaat   tcataaacag   ataaatcaac   atggctctta   43740   ggaagacccc   aaaagcattt   gcccggggag   gcttcctatg   tctggaagtg   ttctgaagcc   43800   tatgaagtaa   ggtgagatta   tcttccaaca   gaaaactcca   aaagatgtgg   aggtggggag   43860   ggtttgattt   ataggatttg   ctcagcctgc   cctagagaca   tcagccaatc   aaaggaaaat   43920   agactccaac   ctgtttacag   agctcctcgg   acttaccatc   ccaaagcata   gtccaagaac   43980   aaaagcccag   tccaagggat   cactgactca   ccagcccctg   ggaatgattt   ccaattcact   44040   gatacatcaa   accagggtga   tttgatatgc   agtgtttgtc   actctgaact   tccaacaatg   44100   cagatctgag   ccccgggact   actttgaaaa   gactggatcc   cttggctcgg   attgtactct   44160   acaaagatga   agaaggcagc   cattttctgc   ccactgggga   ccagaagcct   cttccattag   44220   gcagaaactt   catttctgga   aatgtgaact   ttgctgccag   agtcatcaat   gccgacgctc   44280   tatgtgaaaa   gcagaccatt   tactcacttc   agcattgatt   ttctggaaac   atggcctgcc   44340   tgtatttata   tgaggaaagg   agaacatgaa   agagagatat   agtctgaaaa   tgtaacgttg   44400   acttccaatg   caaaactacc   ctgagttcac   tgattttatt   ctcttcctct   ttccaattct   44460   catccaaaag   gctatcagaa   tgtaatttca   gttgaaaagg   cttccatatc   agaagtagaa   44520   aatagggaag   gtcgatgact   tccctattcc   cttccacctg   ctggaaaact   ggaggaatgc   44580   ccacaaggtg   tagagtaggt   aagacattct   gagggagggg   ccaacagatt   cgtctgctct   44640   ctgacaggaa   gtaaaggcct   tcctggggag   gtcagtagag   aagagaccag   ccccagggga   44700   ttccactcct   cagagtccat   ccactgaggc   tggggccatc   tccagtgagc   aaatttctgt   44760   gggaggttta   ctgaatgctg   cagacctcac   aaaaaaggat   tttggccagt   gccttctccc   44820   agctaaagga   gtgtggacag   ctggctaagt   aaaacttggg   cgaggccatg   gatggtgcca   44880   gtgaagacag   agcctcagta   aggaaaacag   tcctgtatct   actatcttcc   tcacattcca   44940   cccttgcctc   cttctttggc   tgctcccaga   cattgttctc   acctctccca   ccatggagac   45000   gtggatttct   gatctggcaa   agaaacatca   caagtgcttt   aacttttctc   ttccttcttc   45060   atccaagaat   gaaagttcaa   cagagaccaa   tacagttctg   gacaagtgca   agaaataaac   45120   catcccccag   caccatccag   gctcagaaaa   aatgctggat   atgatacaag   gctttgcaat   45180   tctttttttt   tttttttttt   ttttttgaca   gagtctcatt   ctatctccag   gctggagtgc   45240   agtggcgcca   tctcagctca   ctgcaacctc   cgcctcccgg   gttcaagcga   ttctcctggc   45300   tcagcctccc   gagtagctgg   gactacaagc   acgcaccacc   acgaccagct   aattgcaatt   45360   tttaaagaag   aaattgcttt   aaggcattaa   catgcttctc   tttatgaaac   agatccccta   45420   aaagaaacag   gagtaaatct   tggaaaatct   cttacttgta   ttctcagcag   agcttgaaag   45480   gacattgcat   taagaaaagg   gacacagtgg   ccgggcgcgg   tggctcacat   ctgtaatctc   45540   agcactttgg   gaggcccagg   agggtggatc   atgaggtcag   gagatcgaga   ccatcttggc   45600   taacacagtg   aaaccccgtc   tctactaaaa   atacaaaaaa   ttagctgggc   atggtggtgg   45660   ttgcctgcag   tcccatctac   tcgggaggct   gaggcaggag   aatggcgtga   acccggcagg   45720   cggagcttgc   agtgagccga   gattgcgcca   ctgcactcca   gcctgggtga   cagaactaga   45780   ctctgtctca   aaaaaaagaa   aaaagaaaaa   aaaaagaaat   tcttataaac   ttcactgtaa   45840   acaatggaaa   gcagaatatg   atctcccgaa   aagagaagta   tgttatacta   gaatatgcta   45900   ataatctaaa   ttttaaaatc   acagattatt   ttaacagaac   ctggggttgg   ggaaataaaa   45960   tagttttaaa   gcctcttaat   taattttatt   tctcagtgtg   aagtcaaatt   tgaatgattt   46020   ttaatggtta   cttaaaaatt   aagcattcaa   tgtcttaaat   tttttaaagt   taacatgcga   46080   taaaagagaa   taactgtttc   aaattactag   tgaacaaatt   aaaataaaaa   cacaaatgac   46140   aaagaaaata   tttttaaagt   acaaaaacaa   actttaaaaa   ccttgtaaga   tcttacatat   46200   cagttacaca   ataaattaca   atgaatcagt   gtcattatta   aaaatagact   gtcagtttaa   46260   gtcaaaaaca   aaaattatat   tctacttaca   ggagaaatac   ttggtagcaa   atgaaacaac   46320   atagttagaa   atatatagat   aaacaaagat   ataccattaa   aatagaaata   tgattcaatc   46380   aagtatggca   aaattaatat   taggaaatag   ttcaaggcaa   aaagagttat   gaaagaaaag   46440   aacagtaagc   cctcacttat   catccgagtg   ttttgataag   ttctcggaaa   ctgtgacttt   46500   aagcgaaact   acatataata   aaaccagtga   tttttctttt   tcttatcaat   ggcataatgt   46560   tgaaggaaat   gatgttattc   tgggcctgct   agttcgtttc   acttaaagtc   acagtttcca   46620   agaacctatt   gatgatgata   agtgaggact   tactgtaagt   tatgtaattg   aagatgaaat   46680   agacataaaa   tcacaagcca   aataaaaata   tttcattaaa   atgtgtaaac   aaaaattatt   46740   agtaataaaa   ggtaaacttt   aaaattagta   tagaaatcat   ttaaataata   atatgcaaca   46800   gttttattta   taagtataaa   ctgaaatggt   gctactacta   gttagaaata   aactaatcct   46860   aatactaaat   acaaaatgtt   aaaattgatc   acaaatgaga   taaaaaatat   caaaaattga   46920   tcttatctag   ataattcatc   aacagtataa   aaataattgc   agaaatgtta   tgtaaagaat   46980   tgcatagcaa   atatttacaa   agctcacatg   atatgaccat   aatagataaa   ccaccagaga   47040   aaaccttatt   gattttatgc   ataacaagaa   tgccttctat   catattataa   cctcatcatt   47100   gttctgaaac   ttctaaccaa   tgcaaaaaaa   tgggaaatat   aaataatctt   tgaaagtcgg   47160   aaagaaaatt   atttgccatt   gatatgataa   tctccactaa   aaatacaaaa   gacttaatga   47220   aatgacaatt   aagattagtg   aaacaattta   tatcttggct   ggcttcaaag   tacatatatg   47280   aaagtaagtt   atttttatat   aaaccaacat   tacccagtta   gagaaataca   atggtgtaaa   47340   tgaaatatgt   catttaaaat   agaaatttta   aaatataaaa   cacttaggaa   taaaaaatgt   47400   gaagaaaact   gtaaagctac   ccagaaccac   taaagagagc   atcatggcat   atcctagtca   47460   agtttaagat   ttgtttctcc   catgtcccag   catttccata   cctagaatat   gtactagaga   47520   aaaaaagtac   ggtaccctat   agcattgttt   ctgatagctt   caaactggaa   accacccaac   47580   tgtctatcaa   atacatgaga   tcaataaatc   gcagcatagt   cacaccatgg   agtactatat   47640   aggaataaaa   atgaaaaacc   tgcagctctt   tgcaatcaca   agaacgaata   ttgcaatcat   47700   aaaccaaaga   gtcaagacaa   aagaaacaca   tagcataaaa   ctactctata   aagctggaaa   47760   acaggcagca   ctctttttgg   atgcatgtat   aagtggtaac   agtaaatatt   aaataagtag   47820   aaatcaggtt   agaggtaatt   tctagaagag   acagaataga   gttttacatg   gggcacagga   47880   agggctttct   ggtgttgata   aaggtttttt   ttttttttaa   tgatgttggt   aataattata   47940   tgggctttta   ctctatgatt   actaagtaat   gtgtatgtat   atatattgtg   cactttttat   48000   gttgtatatt   ttgtgtcagg   cagatcgtct   gcatagtcct   aaactcttga   tttcttccac   48060   ttcttgacac   aatcctttgg   gaccctttca   gctcttcatt   ctgtaggctc   aagttctgcc   48120   ctgcctccat   ccccagacca   agtcccaacc   tgagggagag   ctgggtctca   atcttccctt   48180   tactaggatg   ggaaatgcca   acattctttt   tggttaccca   ttcctggagg   atggttccag   48240   ctggcagggg   ccatagaaga   ttgatgcaca   gctgtatttt   cctgtcattt   tgggcacagg   48300   ggtgaggatg   gaactttgcc   cctttgacag   ttttagctcc   agtcaactgc   tgttaacttt   48360   gacctcagga   cacccttcac   cctacatcca   tgaacatgtt   gcaacatcag   gcaactacaa   48420   ggtaagtaga   ggtatgcagg   gccttgtgga   ttcagtttct   tctcccacgt   ggatgaacag   48480   cagcagcgga   agagatagat   agcacctctt   ttccagctgg   atagtgtgct   gaaacacttc   48540   ctcagcctcc   tagcagtgcc   aaagacatgt   agggtggcct   tagccagaaa   ggtgaggcca   48600   tcccagcctc   cagcatgggg   ttgcggaagt   agatataggt   gacccactag   agctgtgaac   48660   tcctttaact   ctagcgttga   agtgccatat   tcaatgagca   cagttctgga   aaggtctcca   48720   gggaccccat   atcacatcat   gaactaaccc   aaacctctaa   atcccctaaa   tctttcaaag   48780   ctgagagaga   gagagagaga   gtaattcacc   tttagggatt   gatatagact   gggctatgat   48840   attttgagcc   cactcactgg   gaaccatgga   tagggtgatt   atataattga   ctatccaacc   48900   aaagacactt   tttgagagag   aaagggcaat   ttaaataatt   atcttgggac   agcagttgac   48960   aactaggact   atcctaggca   aaccaggtta   cataattacc   ctagccatgg   accatacttt   49020   gagaagcact   ttccaaaggc   ctctgccatg   tgcagctgct   catagccaag   ctcatgctac   49080   acaggtggaa   aagcttctgg   aaaattgggg   tgcttcccta   gaagactcag   aatgttaaca   49140   aggaataggg   aagaaagaga   actactgaag   agcttggcac   aggagggctg   gcccagaacc   49200   tgtgtcacag   cactgccctg   ggccattcac   accaaagctc   attccagaga   ctgaaagtat   49260   ttgatggcct   tctgtcatac   cttacctgcc   aggccacctg   cttgatgggc   tctttcctga   49320   acctagacac   ccccagggca   ccatcgtcct   ccctgataga   atattatatt   agaacaattt   49380   gaggaacata   gacacattat   ccccaaccct   gtccctcaac   tgtttcatct   gtcagcacca   49440   tggtgtcagt   gagaccttct   gatggagact   tccaagccac   ctggctgctt   aggttagtct   49500   gcaagttgtt   ccagtgtttt   ttaagagagg   tcctccaaat   tcaaatcact   tgtttatgtc   49560   ccactgctgg   cacaccacct   gaatgggaag   tgtcagtcta   cgacatcaaa   ggcctctcct   49620   gctgtgtctg   ctactgacag   cactgttgtc   catctatctc   aaatgggaga   attctcttac   49680   aaaaactcac   tccctccact   cagtacggtc   ctttgcataa   caattagttt   ttattcttga   49740   tgctttatca   cccatttcaa   gactgctaga   gaaatgagtt   atgcaggata   tgacccctct   49800   agtcatcaaa   gaaccagaga   aaaagtgtgt   ggcttgaagc   tgagcaaaga   tgctgcctcc   49860   aagtgggaag   tgacatatgt   atgctggaca   cgatatatct   gcagagctct   ccaaccctgt   49920   catcctccca   ctctagttgt   acagccctca   tcctcccccc   acatcaccac   aatgcctatc   49980   ctcatttcca   cagttctgca   gttccaaggc   cctgacctcc   acctaggatc   tcagaaactg   50040   atctgtaaat   ccaggatgga   gagcctaaat   atcagattga   taaagtgtag   gactcatgaa   50100   gatgctcata   aaaaatttct   tagaaggcca   ggtgcggtgg   ctcacaccta   taatcccagc   50160   actttgggag   gctgaggcag   gtggatcact   aggtcaggag   attgagacca   tcctggctaa   50220   cacggtgaaa   ccctgtgtct   actaaaaaat   acaaaaaaaa   attcgccggg   cgtggtggtg   50280   ggtgcctgta   gtcccagcta   cttgggaggc   tgaggcagga   gaatggcatg   aacctgggag   50340   gcagagcttg   cagtgagcta   aggtcatgcc   actgcactcc   agcctgggtg   acagagtgag   50400   actctgtcac   aaaaaaaaaa   aaaatatttc   ttagaaggac   tgtcagattc   atagcattct   50460   cgcttttttc   ttttttacta   attaatcatt   tatcttaatc   tgtttcatgc   tgctatgaca   50520   gaatacccaa   gactgagtaa   tttataaaga   aaagtaattt   atttctacag   tgccagggtc   50580   tgggaaggtg   ctggtatctg   gtgagggctt   tcttgctgca   tcattccatg   gcagaaagtg   50640   agagggtgag   agagggacaa   gggaggggaa   ctgaactcat   tcctttatca   gtaacccact   50700   cctgcaataa   ctaatccact   cccacaataa   caacattaat   ctattcatga   gggcacagcc   50760   ttcatgacct   agtcacttct   taaaggttct   accttaactc   cattgctttg   gggattaaat   50820   ttcaacatat   aaacccttgg   aggacacatt   caaaccacag   aaacattctt   cagtagaact   50880   ttaatattac   tgtcttataa   aattctgtca   aatgaacaaa   agataaccca   taattacacc   50940   ctaatatgac   tgcttttaac   attttactgt   atttcagcct   ttttgctatg   tatataattt   51000   tacagagttg   taatcatacc   cagtatatga   ttttatcatg   ttttcccact   taccattata   51060   ggtattttta   atattgctac   atagtcttca   tggttgtcat   tgttaatagc   tatgctgtaa   51120   tagttcactg   aattgaagtg   ctttatttac   ttagctaccc   tattatcttt   aaacaatttc   51180   taatttcttt   ttataataaa   catggacata   tttctgacag   gggtgttctt   tttcacatct   51240   tgacctactt   ttcacatagt   gttacaatta   cctgaccaaa   gaatacaaac   tttttgtctc   51300   ttgacgtata   tttccaaaag   atttttaaaa   ggtgcattaa   tttactctgc   agctggtgta   51360   aatgaagacc   attttgtcat   tgttttcttg   agagtagagc   ttccaaaagt   agggatatgt   51420   ggctaggagg   aagaaatcca   gcctggggca   ggcattctgt   aaagaactcc   agttctcact   51480   ggtacactgg   ttttattttt   ctctgtttct   tgcagactga   gcaattgata   actctgtggg   51540   tcctctttgt   ttttaccatt   gttggaaact   ccgttgtgct   tttttccaca   tggaggagaa   51600   agaagaagtc   aagaatgacc   ttctttgtga   ctcagctggc   catcacaggt   aagtaactat   51660   gcaagtgaga   ggcaggaagc   tatatgtgaa   gtccctatgg   cttcctgctt   ttaatgaatt   51720   ttatcaaaaa   aaaaaaaatg   taacgcatcg   gtcaatttgg   gaataatttc   tgaaagaata   51780   taaaacctat   atttgaatat   ttcctctggc   atacttaaca   catatgaatg   cctctaagat   51840   ttcattataa   aagtaactca   ctacactaat   ggaaatatca   ttatcagtat   caccaaatgg   51900   ttctataggg   tttccctgtt   tttatacaca   gtagcctcca   tagttcacta   aggtgatatt   51960   accatatcat   cctgcattat   aagcttaagg   aatacatccc   agaaaatgta   ccagcactat   52020   ctctattaaa   ttattaatgt   taagtcttga   gtactttgca   gattaaagcc   tgtggacatt   52080   gcttttgaaa   tgcaaatgat   gtgtaacacc   actactgggg   gtagtataat   gaagtaatat   52140   aatctcaagg   atcgggatgc   ttaagtgtag   actttggaaa   tgaccattca   gtagaagagt   52200   gagcacagca   cacctagtag   aggaatggtg   aaatattctc   aaggaagaaa   atgatctgtg   52260   aaagttaggg   ccccaaagac   atgagtagag   ctcatgttca   aaaggaaaag   aaaaacacaa   52320   ccagggaaag   agtcctgttc   tacaggccag   aaagaaagtg   agaaaatgag   attcacagaa   52380   ggtccagcat   aagccacact   tggagtccta   agctctggga   gcaagatggc   tgttatatta   52440   gcttaatttg   gatcagacta   tagaaaaaaa   aatgcaaaag   gaaaaattta   aatattaaat   52500   gcttacaagg   agtgacggca   aatgactttc   agaaagggct   atttggtaca   tttcaacctg   52560   tactgttatc   tcagctgtag   taatggtagt   ggagctcttt   agaatttagt   cttgagccat   52620   aatatttcaa   aaatttatga   cactgtaact   tacaagagaa   tcttagccat   acattgttag   52680   taacctgaaa   ttcccctttt   tctttctaac   aaagtataac   aagcccacct   gcttgagctg   52740   ggctgcagtg   gccagggtaa   acatccaagg   caccagtgaa   aaatacagag   aaggtaaaag   52800   gagcaagagt   tctgaagatg   gaacctggga   tgggggaaag   tttcttcaat   ctttcctacc   52860   aacaagaact   ccaatttttc   actcctataa   ccgtagaagt   agaggtaatt   aggatcatcc   52920   agcaaatgct   tagaggcaaa   tatccctgga   tgaggatgcc   acagcttatt   ttcattatat   52980   ttcttcgatt   acagtgtggt   aatgcatgtt   gtatggaact   acatattctt   tcagaatgaa   53040   aggatttaga   ggtggcaaga   atatcagctt   gaaatttaaa   gttttttcat   aaacaataaa   53100   caaatgataa   ttgaaaattc   actacatatt   atcaaagaca   aaagttgtat   gttctaattc   53160   atcatcgttg   tattaatctg   ggcttaattt   catgtacatc   tcctggacag   tgtttttatt   53220   tgttaattgt   ttctagaaaa   actctagggg   tcaggtcaga   agcattttaa   atgaagaatt   53280   cctgaaatga   aaaacattgc   caaaggtcta   agactgaagc   tcaatggtct   gagattgaat   53340   ttatatatta   ttgaaatttc   cttcatacac   attttagcat   gatcacagat   cctttggtaa   53400   aggaggtgag   aatacatacc   tgagtcttga   gtatgcatat   agaagttacc   caggcatcaa   53460   ctgaggggcc   ctgtgctttg   agacatatat   aaaaatgtgg   tttgagtccc   ccatcatatg   53520   tcaatgtcct   ccttaagatt   tcttcaaatt   ctgtggaatt   tatattttta   cttctctttt   53580   gcttcttgct   tcctccttcc   actaaattct   tgctctctct   caccagtatc   tcatctttct   53640   gagctcctct   ttttttgcgg   ttgtttcatg   ctgtccctaa   cgaggcactc   aaagcaccct   53700   cctttccttt   attttcttta   aaatcatcca   caaaagtatg   actttctttg   tttaaaggac   53760   ttttctgaat   atttcaggta   atataatgat   attgtatcac   agaaaagtaa   atatcttaag   53820   agcaaatatt   tcattttctc   agaacattca   gctgacatca   gaaatgtttc   attttgatca   53880   tgccactgca   ctccagcctg   ggcgacagag   tgagactccg   tctcaaaaaa   aaaaaaaaaa   53940   aaaaagaaat   gtctcagttt   taccttgctt   cagcctcact   attctgctct   ctcttctaaa   54000   tgtctagtat   cttgtgccaa   aaaattttaa   tactttcctc   tgtttttgtt   ttgctttgtt   54060   cttaccacta   ccaaaccgtg   atctcagaga   ccccacaact   tccaaaaaac   aagtatcagg   54120   cttttttttt   ttttaacaaa   ggaagttgct   ccctaaaaaa   gaaccctgga   actagttaca   54180   atgataaaca   gacaaagaaa   aatacacatt   catatttata   gttacatatg   tcatggctaa   54240   aaataaatta   ttctaaattg   tataaaacca   taagatatac   agtgataact   caatagtatg   54300   ttgtattagt   ctgttctcac   actgctatga   agaaattctc   gagactgggt   aatttataaa   54360   ggaaagaggt   ttagttgact   cagagttctg   cattgctagg   gagacctgag   gaaacttatt   54420   aatacaatca   tggcagaagg   caaaggagaa   gcaggtacct   tcttcacaag   gcagcaaggt   54480   ggagtgaggg   caagcagggg   gaatgccaga   tgcttataaa   acatcagatc   ttgtgagact   54540   cactcattat   cacaacaaca   gcatagggaa   accgccctca   tgatccaatt   acctccacct   54600   ggtcctgccc   ttgacacatg   ggggttatta   caattcaagg   tgagttttgg   gtggggacat   54660   agagtcaaac   catatcatat   attatgtgct   gtgcttgcaa   aaattttgaa   gtggttttag   54720   gccaatacgc   ctacctacaa   taattagaaa   agctggacaa   aatttaaaca   tttgttagaa   54780   ggtatcacat   agctgccaaa   gacaattaag   acttgagggg   ccaagatttc   atagagcaga   54840   aaattttatg   agataaatac   attatttgac   accactatcc   ccttaaagta   ttagctgatt   54900   taaaagcaga   aactggaagg   ctgaaaacct   gagtagagct   ttgggcagtt   tcactggact   54960   gatggggcaa   aaattaattt   gccaaagcat   actggctttg   ataaacattc   cagggtttcc   55020   attggtactg   ctgaaaagat   acaacctaca   agtaagagtg   aactaaaaat   ggatcagccc   55080   tcaccaaaaa   attaaataca   acccaaaacc   tgaatgaaat   cagatgatct   acctctgcat   55140   gcacacaaac   catctgccag   tgcaaaaata   aatcttcttt   ggaggaagaa   ggcattatcc   55200   catcatcaaa   ttaactataa   ttttccatgc   acaacgttca   gcacttgatt   aaaaaataac   55260   caggaaggca   acaatacaat   aattgatgaa   aaacgaggag   aaaaatagat   aacgaagagt   55320   tgcacaagag   atccatctat   tggcattatc   aagatacaga   atttataact   gttgctcatc   55380   tgtttagaag   ctacaaaata   agattgaagg   ttgaatgtcc   tatctccagg   cttttggtgc   55440   acctgccccc   ttaacagtgg   agcttgagca   cacaaagaac   tccctttcca   cacaaagaac   55500   ttggtgtagt   ggcagttcct   ccactccaca   cctggatgta   attccagcca   tttaacacac   55560   ctgccccaat   ggactaggag   tttgagccac   ctcttcctcc   catgcaaaga   ccttgtggct   55620   gcagtttctc   tgctctttcc   ccagacatat   cacccaactc   gaggtgcacc   tgctcctcca   55680   gatcaggagc   ttgagctact   cctacattcc   ccaggaaaac   tttgtggcag   tagtggtctc   55740   tctccacctt   gctggggcat   atttccaggc   atttgacaca   cctgattgtc   taaattagaa   55800   gcatgacctg   cccctccgtg   cagagatctt   ggtagagcaa   agctctctgc   actcgatgcc   55860   cagacatatt   tccaggcatt   tgaagcacca   actcttctag   attaggaatt   taagcttccc   55920   ccacttcctg   tgcagagaac   ttgggacagt   ggaggtttcc   agactctatg   cctaggcaca   55980   tctccaggca   cttggcagct   gcccaccaga   ccctgcctca   gagctaatgc   ttctgcctgc   56040   cattgggaga   actgtaggag   agcttgctta   gtctagccca   cccatcttgg   cccccactcc   56100   agaactaagc   agggagctca   gaccactgtg   catttcacag   atcagcccat   tgcccaagtc   56160   agcagaactt   ctcccagtaa   acaagaatca   agtatatatc   catttgagtc   agctgcagct   56220   gaactttatc   cataagcttc   acctactggc   ctggaggttg   aactgcacaa   tacaataaga   56280   aatctggggc   caggcacagt   ggctcacgcc   tgtaatccca   gcactttggg   aggccaaggc   56340   gggcagatca   caaggtcagg   agatcgagac   catcctggct   aatacggtga   tacccggttt   56400   ctactaaaaa   tgcaaaaaat   tagctgggtg   tggtggtgga   cgcctgtagt   cacagctact   56460   cgggaggctg   aggcaggaga   atggcatgac   cccaggaggt   ggagcttgca   gtgagccgag   56520   attgtgccac   tgcactccag   cctggatgac   agagccagac   tctgtctcaa   aaaaaaaaaa   56580   aaaaaaaaaa   aaaaaaaaga   aaagaaaaga   aagaaagaaa   tctaggcagg   gcatggtggt   56640   ttatgcctgt   agtcccagca   ctttgggatg   ctaaggccgg   caaatcactt   gagcttagga   56700   attcgtaacc   aacctgggca   acatagtgag   acccccatct   ctgcaaaaaa   tttttaaaat   56760   tagccaggca   tggtgatgca   cacctgtagt   cccagttgcc   agggaggctg   aggtgggaga   56820   attattggtg   cccagaaggt   caaggcagca   gtgagccaag   atcatgccac   tgcactccca   56880   tctgggtaac   aaagtgggac   cctgtctcta   aaaaaaatta   aaaaaaaaaa   aaggaaaaga   56940   aaagaaatct   gatgacataa   ctgcacagca   ctggaaaatg   taataagcct   cctgagacct   57000   ctgccaccca   gccttatagg   aggcagtgag   cctcctcaca   tacccagtac   acggctacta   57060   caaccagcat   ttgggaaagc   caccatacaa   agactaccca   ttaccaagaa   acgcttatac   57120   agactcgttg   ccactgaaag   cacccagaac   caaatccaaa   ggaccttaca   caacataaac   57180   tatagacatc   tcctcagatg   aggaaaaaag   tcacatccaa   ataaaagcaa   atttaaaaaa   57240   aaataagaag   agatagctta   tctggatgag   aaggaaccag   agaaataact   cggaatgtat   57300   gaataaaata   gagtgctaca   acacccccaa   aggatcacac   taactctcca   gtaatggatt   57360   ctaaccaaga   tgaaatattt   gaaatatcag   gtaaagagct   caaaatattg   atttttaaaa   57420   agctcaatga   gatccaagaa   aaaattgaaa   accaacacaa   agaaaacaat   tcaggacatg   57480   caagaagaga   tagataccac   taaaataaaa   gaaacggaca   tttaaaaatt   attgaaggaa   57540   aaacaaaata   aaagtgaaag   cttcaacaat   aggctagacc   aagtggaaga   aagactttca   57600   gaactagaag   acatcttttg   aattaaacca   gtaagaaaaa   tatacagaaa   aatgaatttt   57660   aagaaatgcc   tagtgccttt   gataaatacg   ggattatgta   aagcacccaa   acctacaatt   57720   tataggtatt   tcaaagggtg   aagaagaaaa   agtatggaaa   acctatttga   ggaaataatt   57780   cagaaaagct   tctttggttt   tgggagagat   ttagaaaccc   agatacaaga   aactcaaaaa   57840   actcctagaa   gacacatcgt   gagaagaaca   tcaccaagac   gtatagtcat   cagactatcc   57900   aaagtcaaca   tgaaggaaaa   aatcctaaga   gtgcccagag   agaagtgtct   aatcacctat   57960   aaaggaaatt   tcataagact   aatagtggac   ttctcagcag   aaaccctaca   tgccagaaga   58020   gatcaggggc   ctatttttag   cctccttaaa   gaacaaatgt   gccagccaag   attttcacat   58080   cctgccatgc   taagctttat   aaatgaagaa   gaaatcaagt   cttaaccaga   caaataaatg   58140   ctaagggaat   ttgtcaatac   tagaccagac   ttacaagata   tgctcaaagg   agttctaaac   58200   atggaaacaa   aagaacaata   ctcatcatca   taaaaggaca   cacagataca   aagctcacag   58260   atcctgtaaa   ccaacaacac   aattgaaact   ccaagtaaca   acactgtgac   aggaacacaa   58320   cctcacatat   taatattaat   cttgaatgca   aatggcctaa   atattccact   taaaatatag   58380   agtggaaaat   tggattaaaa   agagactcaa   ctatctgcta   cctacaagat   atccacttac   58440   tgactaaaga   cagctataga   ctcaaagtaa   aggggtggga   aaatatatat   catgcaaatg   58500   ggaaaaaaag   caagcaggag   tagccattct   tatattagat   aaaacagaca   tcaaaccacc   58560   aacaataaaa   aaacaagaca   aagaatggca   tcatataatg   ataaaaggtt   caaacgacaa   58620   gaagattgaa   ctattgtaaa   gtatacacga   ccaacactgg   agagcccaga   tttacaaaaa   58680   ttacgactac   atataagaac   aaagatgaat   agaaaaaaaa   tagtggtctt   caatactcca   58740   ctgacatcag   tagacagatc   atcaaggcag   gaaatcaaca   aaaaatctct   ggacttattc   58800   tgggtacagt   aactgactct   tgtaatccca   gcactttgag   gggctgaggt   ggtaggattg   58860   tttgaggcca   gttgttcaag   accagcttgg   gcaatatact   gagacccaat   tccacaaaat   58920   aaataaatga   ataaataaat   aaaaattaat   taagcatagt   agcacatgcc   tgtagtctaa   58980   gcaacttgca   aggctgagct   ggaaggattg   tgtgagtgca   ggagtttgag   gttacagtaa   59040   gcaccactgc   aatccagcct   gggcaaaaga   gtgagaccct   gtctcctaaa   aaaacaaaac   59100   aaaacgaaac   aaaataaaaa   gaaagaaaga   aactctggac   ttaaactata   tatagaccaa   59160   atggacctaa   tagacattta   tagaacattt   catttaataa   ctgtaaaata   tacattcttc   59220   tcatctgtac   atggaacatt   ctcccaaatt   gactttattc   ttagccataa   agcaattctc   59280   aataaattaa   aaaaaactga   aatcatatca   agtgttttct   taaaccacag   tggaataaaa   59340   ttataaatca   aaccaacggg   caccttcaaa   aatacacaag   aacatgaaaa   ctaagcaatt   59400   tgctcctgaa   tggccttttg   gtaaacaata   aaattaagac   aaaaataaaa   aacatttcaa   59460   aatgaatgaa   agtagagata   caacatacca   aaacctctgg   gatatagtga   aaacagttct   59520   aagagaaaag   tttatagaat   tcaatgctta   cactaaaaga   tagaaagatt   tcaaattaac   59580   aacctaacat   tacacctcta   aggaacaagg   aaagcgagaa   caagccaaat   ccaaagtaag   59640   cagaaaaaaa   agaaatataa   taacaaagat   cagagcacaa   tgacatgaga   ttgagattta   59700   aaaaaagata   caaaaagtca   acaaaacaaa   atgttggatc   tttgaaaaga   taaatgaaat   59760   tggcagcctt   ctagctagat   gaaacaagaa   acaggagaga   tgattcaatt   aagtacaatc   59820   agaaatgata   aagatgattg   acatagtttg   gatgtgtgtc   ctagaccaaa   tcgcatgttg   59880   aaatacaatc   cccccgtgtt   ggaggtaggg   cctggtggaa   agtgattata   tcatggaggc   59940   agatttctca   ttaatggttt   aggaccatcc   cccttgggca   ccagcctact   gccctcatga   60000   tagtgaatga   gcttttgtga   gatctggtgt   tgcacccccc   tgctctctct   ctcttgattc   60060   ttctctggcc   atgtgatgtg   cctgctccct   cttcgctttg   ccttccatta   tgattgtaag   60120   cttcctgagc   ccctccaaga   agccaagcag   atgccagcat   catacttcct   gcacagcctg   60180   cagaactatg   agccaattaa   acctcttttc   tttataaatt   acccagtctc   agttacttct   60240   ttatagcctt   gtgagaatgg   actaatgcag   tgaaattaca   actgatacta   tagaaataaa   60300   aaaaaatcat   cagagatgac   tatgaacacc   tctgtgcaca   caaactagaa   aacctggagg   60360   aaatggataa   atcccagaaa   catacaaccc   cctaagattg   aaccaggaaa   gagtagaaat   60420   cctgaataga   ccaatagcaa   gtaataaaat   cacatcagta   ataaaaaatc   ttccaacaaa   60480   aaaaatctca   agaccagaca   gattcacagc   tgaattttat   cagatgtaca   aagaaaagct   60540   ggtaccaatc   ttactgaatc   tattccaaaa   catgaatgtg   aagcaattcc   taactgttcc   60600   tataaaacca   gtatcaccct   gataccaaaa   tcatacaagg   acacaacaac   aacaatacaa   60660   aaaaacccac   tacaggccaa   taaccctgat   ggacatagtt   gcaaaatttc   tcatcaagat   60720   accagcaaac   caaatccaac   agcatgttaa   aaagataata   catcacaatc   aagtgagttt   60780   tattccatgg   atgcaaggat   gattcaacat   atgcaaatca   ataaaatcaa   tagacatgat   60840   tcaccacatt   aacaaaacta   aaaacaaaaa   ccatatgatc   atctcaatac   aataaggcgt   60900   caataaaatc   caacatcttt   tgataataaa   aaccctcaac   aaattaggca   tctaaggaac   60960   atacctcaaa   ataagagcca   tctattacaa   acctacagcc   aacattatac   tgaatgagga   61020   aaaattgaaa   gcattatccc   taaaaactgg   aacacaatga   ggatgtccac   attcaccact   61080   cctattcaac   atagcactga   aagtcctagc   tagagcaatc   aggcaagaaa   aagaaatgaa   61140   aggaatccac   actagaaaaa   aaagtcaaat   taccaaatta   tctctgttca   gtaatgacat   61200   gactgtatac   ttagaaaaca   ctctacagac   tccaaaagac   tcctagactt   aacaaacaac   61260   ttcaataaat   tttcaggata   caaaatcaac   atataaaaat   cagtagcatg   tctatacacc   61320   agtagtcttc   aggctgagaa   ccaaatcaag   aacttgatct   taacaatagc   cacaaaaaat   61380   aaaataaaat   aaaataataa   aacacctagg   aatactttta   accaaggaga   tgaaatatct   61440   ctacaaggac   aactacaaaa   agtgatgaaa   gaaattgtag   ataacacaca   tgactggaaa   61500   aacatcccat   gctcatggat   tggaaaattt   aatatcatta   aaatgaccat   acttcccaaa   61560   gcaatctaca   gattcaacac   aatccctatc   atattacaaa   tgccattttt   cagagaatta   61620   gtaaaacaag   attaaagttt   atatagaatt   ttaaaagacc   ctgaatagcc   aaagtaattc   61680   ttttcttttt   ttgaaatgga   gtctcgctct   gtcaccaggc   tggagtgcat   tggcacgatc   61740   ttggctcact   gcaacctaca   cctcctgggt   tcaagagatt   ctcctgcctc   agcctcctga   61800   gtagctggga   ctacaggtgc   gcatcaccat   gcccagctaa   tttttgtatt   tttagtagag   61860   acggggtttc   accatgttgg   ccaggatggt   cttgatctct   tgaccttgtg   atctgccagc   61920   ctcagcctct   caaagtgctg   ggattacagg   tgtgaaccac   cgtgcctagc   atagccaaag   61980   taattctaag   caaaagaaaa   aatccagagg   caccacattg   cctgacttta   tacaacaagg   62040   ctgtagtaac   taaaacagca   tgatactggt   acaaaaatag   acacacagat   caatgaaaca   62100   gaatagagaa   agcagttaaa   atgccacata   cctgcaacca   acccatcttt   aacaaagctg   62160   acaaaaataa   acaatatgga   aaggacacct   tattcaataa   atggtgctgg   gaaaattggc   62220   tagccatatg   cagaagaatg   aaaccctacc   tatgacaata   tacaaacatt   aactcaagat   62280   agatcaaaga   cttaaatgta   agatctgaaa   ctaaaaatct   tagggaaaaa   aacctgggaa   62340   aactctactg   gacattagac   taggcaaaga   atttatgaca   aagaccccca   aaacaaatgg   62400   aacagaaaca   aaaataggca   agtggaactt   atttaaatta   aaaagcttct   gtacaacaaa   62460   agaaataatc   aacaaataga   cagcctattt   ggagaggaag   tatttgcaga   ttatttctcc   62520   aacaaatgac   taatatccaa   aatatacaag   gaactaagac   aaatcaacaa   gaaagaacaa   62580   acaaccccat   taaaaactgg   gcaaaagaca   tgaacagaca   tttctcaaaa   ttagaaatac   62640   tagcagccaa   caaacacatg   aaaaaaggct   caatatcact   aatcatcaga   gaaattaaaa   62700   gtaataccac   aatgagatat   catcttacac   cagtcagaat   ggctattatt   aaaaaatcaa   62760   aaaacaatag   atgctggcaa   ggctgtgaag   aaaagggtag   acttacacac   ttttgaggga   62820   atgtaaatta   gttcaaccac   tatagaaaac   agtatagaga   tttctcagaa   gacttgaaac   62880   agatcttcca   tttcaccaag   caatctcact   actgagtatc   tacccaaagg   aaatgaaatc   62940   actatataaa   aaagacattt   gccctcatat   gttcattgca   gcactattca   cactagcaaa   63000   gtcaggcaat   gaagctcagt   ctccagcaat   ggttgatttg   ataaaaaaaa   aaaaaaaaaa   63060   aactgcggta   tatatacacc   atagaatact   atgcagctat   aaaaaagaat   gaaatcatgt   63120   cctttacaga   aacatagata   aagctagaag   ccattatact   aagtaaaata   actcagaagc   63180   aaataaaata   acatatgttc   tccttcataa   gggggagcta   aatatccatc   catagataaa   63240   aagatggaaa   taatagacac   tagggactcc   aaaagagaga   aaggtgggag   gtgaataaga   63300   gttgaaaagt   tacctattgg   ttacagtgtt   cactatttag   gtaatgggta   cactggaatc   63360   acaatctcca   ctagttacaa   tatgttacaa   tacacctgtg   tattacaata   tgcctgtgta   63420   aaaaactgca   cacatatgcc   tggaatctaa   aattttaaaa   taatgaagat   tgaatgtcct   63480   agaaaggaat   aaaacaaaac   atttaggaac   tgacaattac   aactgaagct   tacaatacaa   63540   tagttaaact   tagcggcaaa   ttaaatgtag   ctaaagaaga   aatcattgaa   ctggaatata   63600   ggtccaaaga   aaataattag   attgaagcat   gaaaaaagaa   aggacaaaaa   atacaaaaag   63660   aaaataaaaa   gcatatggaa   gtagtgaaaa   ggactcccag   aaaaagataa   agcccagact   63720   ggatagaaac   aacacttata   gtggccaata   attttcccaa   actctttaaa   gacatcaaaa   63780   cacaaataca   taaagcacca   taaaggacaa   cagcaagcaa   acaaacaaat   ataaagagaa   63840   ttaaaacaca   tgaaatgata   agactaagta   aaagccccat   aatcttgaat   ttgaaatgaa   63900   aatatttatg   ctagctcatg   atgtatttta   tgcttcagga   gtttttaagg   atacatacca   63960   aaatgtttat   aggtaaactg   ataacatgtc   tgggatttac   ttcaaaataa   tttgaggctg   64020   ggggtgggtt   gagatgtata   gatgtacaaa   attgatattg   ataactgttg   gaggttggtg   64080   gtgagtacat   tggattcttt   atttttttta   tactattctc   tctactttta   tgtaagttta   64140   agattttttt   cataataaaa   ttcaaaacaa   aattatgcat   gattataatg   aaacaaaata   64200   aagagatctg   ggtagttaaa   gatgttaaag   tccttggatt   gcctggatgt   agtaaaagta   64260   ctaatttcta   ttagatttta   atgtcaagga   ggtgtgttaa   aatctctaaa   gtaatcctcc   64320   caaattatga   aaactgacat   tactagcaag   ctaaagaagg   tataaaatgc   agtatttgaa   64380   tatgcaaata   tatatgttta   tatatagagt   tattccccag   tatccatgga   ggattagttc   64440   cagaatcccc   tgccaataca   aaaattcgag   gatgcttaag   tcccttacat   aaaatagata   64500   gtatttgcat   ataacctaca   tacaccctcc   tgtatacttt   aagtcatctc   taaattactt   64560   ataatttcta   atgcaatgta   aattctatat   aaaaattttt   atactatatt   cttaaaattt   64620   gtatcatttt   attattgtat   tgttatttta   tactttctta   atatttttaa   tctgcagttg   64680   atagaatcca   tggatgtgga   acccacagat   aaaaagggcc   aattataata   catacatatt   64740   atataaaatc   aatctaagtt   aagacaggag   agaaaaaata   ttagaattat   tagaacaagt   64800   aagaagaata   ggccagtaca   cttaacctca   aatgtatcag   taattgcatt   aattgtaaat   64860   ggactaaata   ttcccattaa   aataacaaga   ttgtcagagt   agatttttaa   attacctatt   64920   ttaatttgtt   tataaaaggt   gtacaccata   aataaaatga   tgcaggttaa   aattaaaagg   64980   aaaaggatgg   taaagtaaaa   tgataaaaaa   taatgaaaca   aaacaaaaca   caaagtaaaa   65040   gtaagccaat   gcagctacat   atgtgtatca   aaagtttata   aatttgctgt   gattcttgga   65100   atcacttgta   gctttatctg   tgtcttagaa   tttgctttga   aaggaataca   aagacaggag   65160   ggcttctaaa   gctagtaatg   ttctatttct   tgatcttgct   tgtagttacg   tgggcgtgtt   65220   cctcttgtaa   taattcacca   aattatatac   ttatgatttt   gtatactttt   ctttatgtgt   65280   tttatttcaa   tacaaagttt   atttacaaaa   tatatcacat   taatccaggg   caggggttgc   65340   aatctcaaat   ggctttagga   atcaagcagg   caatgaatgt   aaacatgtaa   aagagccaag   65400   gggaagagac   agaatgaaaa   cagagataaa   aaccagagaa   tgaatgccct   gcctaaagac   65460   attaaaatca   atttttgtca   aacactgtgc   tgatcaaaca   aatcacacat   gcaggccaga   65520   tttgacccaa   gggtgacttt   cttacaacaa   ctgatatata   aaatttaaac   atcagaattt   65580   gaaaagtgtg   ccactataaa   attctagccc   acctctgaag   tatgaaccca   ccttcaattt   65640   gtgaggtaga   atattaatct   ttactggtgg   attgtgagaa   gtgaggccaa   gaagttgggg   65700   aggttaacat   ttacaatggt   ttagacaaat   catatgtttg   acaaggttta   aaatcccttc   65760   tgttcaaaga   atatcttata   ttaggatttt   cttctcctaa   tgtgtttatt   attgttttta   65820   ttagtataat   ttttaaagaa   atctgaacta   ttgcacgcct   cagagaccat   aacagtgaga   65880   acaaaaatat   aaaggaaaat   tttcaaatga   tggaaagtgt   aacccatata   acccgaaatc   65940   ttcatgattt   tcagatgacc   atctgattaa   ggagaaaagt   caagataagc   acatattcat   66000   atctgcttat   ttctagaatt   gttctatcag   aagaccatga   attggtcata   tgaggggaag   66060   aaacactgac   atgatactgg   ctctattagg   atgatattaa   aagaggctgc   tggctcaagc   66120   ttagcatgga   tgagttgttc   tcacataaag   atgctgttat   tgaccaggtg   tggtggctca   66180   cgcctataat   cccagcactt   tgggaggctt   tggtgagtag   atcccctgtg   tccaggagtt   66240   caagaccagc   ctgagcaaca   tagtgagacc   ctgtctctac   taaaaaattt   tttaaattaa   66300   aagttaaaaa   attaaatgat   gttgctattg   ttaatgcatt   cctgcaatgc   taacatgctt   66360   tcataaatat   acttatgatt   ttataggtgt   tttcctctta   gcctcaagaa   gggagatgtt   66420   ttagtctagg   gactgtcata   acaaaataat   ataaactgtg   tggctcaaac   aacagaaatt   66480   tatttcctcg   cagtcctgta   ggctgaaagt   ttaacattag   ggtgccagca   tggtgtggct   66540   ctcttcctgg   gttgcagatg   tcaccgtctc   actgcatcct   catgtggcct   ttgtgcaatg   66600   cctctgcaca   aagagagaga   aacagcaagt   tatgtgccgt   ctcttcttgt   aggctcacta   66660   atcctatgga   atcagggccc   cacccttgtt   aaccttaatt   atttccacag   aggtcatatg   66720   ttcaaataca   gtcacattgg   gagttagaga   ttcaacatat   aaattttagg   ggaatacaaa   66780   ccttcagtct   gtaacaggga   acagaggtgc   tagaagaaat   gtagtagtag   caaaaggtgg   66840   ggtggggaga   taataggagc   ttcatatgtc   ttgctaagaa   gttcaggctt   tgcttttagc   66900   acagcccctc   ttctgctagc   tttgttcagg   ccaccatcac   ctttgatgca   gattactaca   66960   aaagccaccc   aaggtgtatg   aaatgagaaa   acacatactt   aaattaaccc   atggccaggt   67020   tttgctggta   acttgggaac   acagtaaggg   aaggaaaata   actttgttgt   ggagagggca   67080   aaccaaggaa   ggtcagaaag   acatagagac   agtggaagct   caaggaaccg   gaggttgtcc   67140   caaagtcata   aggcggatgt   aacgagagca   ccagaacaac   agagctggaa   ggaaggaagg   67200   aaggaaggat   atgtttagaa   agcaagatgt   tcccagatga   atactgcaga   ggtagtataa   67260   ctcagtgcaa   cgattgtact   gggtaggcaa   ggtagggtgg   cagggaaagt   cactagcaaa   67320   catggccaca   ataatggaaa   ctaaactttg   tgtttaccat   gcaccaggct   ctgtgctaca   67380   tgctttctgt   agatcatctc   atttaatcct   cacacaggga   tttaagcctg   aagttttttt   67440   cagagaagac   atacaaatcc   caagtaagag   catgaaaaga   tgttcattag   cattagtcat   67500   tagaggaaat   gcaaatccaa   agatcaggga   gatacgactt   cacaccttca   agaatggcta   67560   ttcccctata   aaataatata   ctaaatcact   ctttgaaaca   taacattttc   cacaaactca   67620   cctctttgaa   ttcacatttt   gctttgccag   tgttttatgt   tattcagaca   tgagtttgaa   67680   tctcagcatt   tctacttctg   agccctatga   cctatgaaaa   gtgacaattt   ctctgaatat   67740   caatttccca   atccaaaaaa   tgggaatacc   tcttggagtt   gtggtgagga   ttaaataaga   67800   taatgaatat   aaagtgccta   gtatagtaag   aggcatatta   ttgttcaata   aatgctagct   67860   gtgaatgcta   ctgctatagt   tatttcattc   actgtagaaa   tgattaattt   aaactccttc   67920   aagtaactta   ctaaagagtg   aaagttctcc   aaaaggcttg   aatttcactg   caaacctctt   67980   tccttgtttt   gctcttttat   tcaggacaat   acactttgct   caggtgtgtc   tttgactctc   68040   aagcattgct   ttaaagaatg   accagacttt   gtcatatacc   attcttaaag   gagttcagga   68100   catgccaccc   caaaatatgc   tgctttcata   tattgattat   tttgagctga   tagcacttca   68160   acagcaaatg   caagagagac   tttttcggaa   ctcagctgcc   taaaaacaga   tcctccaaaa   68220   gaaattcaat   tgccatatat   cctctccctg   ggagttttat   taatcagcaa   agatacactt   68280   ttatcacaaa   acaggaaact   ggaagtcaac   accacctaca   gaaaaacttt   ttcataaact   68340   gttatatctc   ccattgattc   ttctaaaaac   ccattcatct   ttccccagaa   tcatttactc   68400   caccctaagt   gacttacatc   acccctcccc   atcccctgtt   aacacagtat   gtaagctctc   68460   aaatctcttt   taggagtatt   cattattttc   ctgtggcacc   ccatgaacat   aatattaaaa   68520   tgaatatgcc   ttctctctta   ttaatctgcc   tgttgtcagt   ttatttcata   gacacagcta   68580   tcgaacctag   gagagtacag   ggaaattctt   tcctccccta   tatgcttaaa   tcataagatt   68640   caactgacat   cagcaatgaa   acaaacattc   agaaaactaa   attcagagac   attatatctg   68700   gcttttctga   gggagaataa   taagaactaa   taggcattgt   caggtaaggc   aatacagatg   68760   tttttaatta   aatgatgttt   tttcaccatg   acccacatct   tgaaaacacc   ttcctaatta   68820   aagaccagta   gtcttgagtg   attctttgaa   gaacattttt   acttccattt   ggtcttttct   68880   ttattgagtt   gatgaatctg   ctggaattag   attcagcgca   cataacataa   aatccaaata   68940   atgctggctt   aagttcataa   ggatttattt   ttatcttatg   taaagaagtc   cggacgtagg   69000   taatccaggg   ctcgtgctgc   attttcttac   cattgggaac   ccaggctcct   ctgtatttct   69060   actgtattaa   aggactgtca   ccctcaagtt   cagctcatgg   tacagatggc   ggttggttca   69120   agatgactgc   tagaacatct   gtcatcatgc   ccaaattcta   gccagcagga   aggaagaagg   69180   gaggaaggac   agaggggctc   atatcagctc   tttgtcctcc   tcttaattgg   cattgccaga   69240   attcccaccc   attagttttt   ccttgcatct   caatggtaac   gtctaagtga   aaaaaagcct   69300   gggaaatatt   ttcatttatt   attatttttt   taacccaggc   ccattggcca   gaataaaacc   69360   agcttccttt   tcctaaggaa   gaaggagaaa   tagaattaag   gaacagtctc   tggcatagct   69420   tgttctccat   acttttactt   ccatccagaa   ctgagaagtt   tagacaacaa   atctcttttt   69480   gtccacccat   tgcctgcttt   gagaatgctt   cttccagagt   gctacgctct   cttctaatga   69540   cttattatct   tggcagactg   cacttttcac   accctccata   ggctgcagtg   atccattgga   69600   tgattgtccc   attatgtaga   taagttccta   ttgctggctg   tttatgcttt   tctcccattt   69660   ttaggctatt   agcaatcctg   caataaacat   tctcataatt   aaatttgtat   gagaattctg   69720   tattatttat   ttagaaaaag   tttatagaag   tggacttact   gcatcaaaga   aaggcacttt   69780   tgtggctttt   gaattggcta   atgtcttttt   tggttaccca   agttaaaaga   tgatggaaaa   69840   tgtcggggga   caaatgctct   agaacccgct   agcatagacc   tacactaagt   cttattcaaa   69900   aaggtccctg   taaacaaaac   cctagttgac   ttgcagtagc   tacagaatca   taactggttt   69960   tttgagagca   gagattctct   ccgttcctcc   cttagtgata   cttagccagc   tactggtaat   70020   ttgcttatta   tcaaaaggag   cggccaggcg   cggtggctca   tgcctgtaat   cccaccactt   70080   tgggaggcca   aggcgggcgg   atcacgaggt   caggagatcg   agaccatcct   ggctaacacg   70140   gtgaaacccc   gtctctacta   aaaatacaaa   aaatcagcca   ggcgtggtgg   cgggcgcgat   70200   ggcaggcgcc   tgtagtccca   gctactccag   aggctgaggc   gggagaatgg   cgtgaacccg   70260   ggaagcggag   ctcgcagtga   gccgagatcg   cgccactgcc   ctccagcctg   ggtgacagag   70320   caagactccg   tctcaaaata   aataaataaa   taaataaata   aataaataaa   gcattttttt   70380   tttcacttta   gccagcgcat   cagagaagaa   cccacacttg   attgtcttat   ttttccccct   70440   acaatgccca   gagcagtatt   cacacattca   tgaaaagaaa   acacatcact   tagaacattc   70500   gaatagagca   cgtgttttta   ttttgctctt   ctatgtttta   cctccacttc   tccacaattt   70560   tgatcccttc   aaaaaaaaat   cttaactaat   tattctctat   atattctatt   aggaatcttg   70620   gctgtagact   taatcttgag   gttagaaaga   aaatagatct   tgaagccact   attttggcac   70680   attctgtaat   tcttagaatt   tccattctaa   caattttctg   gatgaattta   ataaccgttt   70740   atattttatg   agctaaagtg   ccaattaaaa   ttattcattt   accatcttta   agacttactt   70800   gtctgtttta   ttcaaatgca   ataaaaatgc   ggaactaata   agagtaactc   ccgatgagtg   70860   ttcaacaaag   aaaagaaaga   aaacatattt   tttggcgcac   attcaaatca   tttccttttt   70920   ctggtagggc   caaatcttct   catattggga   aaggaaatta   gctttgcttc   aaagcacttt   70980   ctgaagagca   atttactaat   gagcttagga   attctctgcc   ctcataaccc   tcctatacat   71040   tacctgaacg   cagagaaact   tgagacgttc   agcaagggag   gtaaggccag   gagtgttgag   71100   gcgtccaggt   ccgtctgtgg   agttcactgc   caccttcctg   tgtggatttc   agtccttgtt   71160   ttcctggatt   ttgagttgca   ttctcaatac   attattttta   tttttattca   tagtgaaaat   71220   attgaagact   gaattttttc   tgaattcata   caatttatat   gacatgttat   tttttatttt   71280   ctaaatattt   tactgttgaa   gtagttcctc   tttcacccaa   aaaaactaat   agtttttgct   71340   gtcttttttt   aatttctacg   tgaccatttc   tttaatattt   tgtaattcac   atttaatttt   71400   attgccttgt   agtcaaggaa   tatttctgaa   aggacttctg   ctctttggaa   cttattaaga   71460   gtgtctttga   ggtttaatat   gtaattatta   tcttaaaaat   atttaatgta   tactaaaatg   71520   taagacacaa   tttattgctg   tggaattaaa   caaagaccat   ccaaatgaga   acaagcaatg   71580   gctatttact   cagagtttgg   gagtcagccc   acagtttctt   gtgttgacag   agactcaaaa   71640   ggctgagtgg   aaaagctttg   caagggggaa   aaaaaggcag   ggtctcagct   atgccctgat   71700   tggaggctgc   tggcctaggg   aagttgacct   gaagcacagc   atcttatcgg   attgcttaag   71760   ggtgcatatt   tacctttctc   ctattggtcc   taagttagaa   gcaagggaaa   caattaaagg   71820   agctatcagt   tatgaaccaa   gttctgggtt   tgttgggcta   attgctatgg   tggtattgtt   71880   tggcttcctg   agctggttgc   tgcaggttgt   ggataagagt   tctattttat   gtatatggtc   71940   tggccattgt   ctatatgttc   agtctctcaa   tgcattattt   gtacttattc   ttttgaattt   72000   gcctttacta   attatggtct   ctatattgtc   tctttttata   tacatatttt   ttgcctgttt   72060   aacgtatgat   agatgactaa   aagtgctatt   aaaaactttc   acacttttgg   ttggacgtgg   72120   tggctcatgc   ttgtaatccc   agcactttgg   gaggctgagg   cgggcagatc   actggaggtc   72180   aggagttcaa   gatcagcctg   gtcaatatgg   tgaaaccccg   tctctattaa   aaatataaaa   72240   atcagttggg   catgatggtg   cacgcccgta   ataccagcta   ctcaggaggc   tgaggtggga   72300   ggatcgcttg   aatccaggag   gtggaggttg   cagtgagcag   ggatcacgcc   actgcactcc   72360   tgcactccag   cctgggcagc   agaatgagac   tccatatcaa   aaaaaaaaaa   aaaaaagaga   72420   gaaaagaaaa   gcctttcaca   ctttctctca   tcttttagta   tttttgatca   atgtttgttt   72480   tatacatttc   aatggtttat   tatttggttc   ctaaaggctc   ctgattagca   tatctttaat   72540   gtatgtttgt   gtatgtatgt   tgatctctct   ctctctctct   ctctctctca   tacacacaca   72600   taagtctttt   catttattag   ctttgcttta   aactttgcta   atgtgatatt   aatattatgt   72660   ttatctatct   tctttttatt   gcacttgttt   gccaagcacc   gaactaagca   ttttaaataa   72720   tatcacattt   ttgtaactgt   tttggcattt   atcttcttca   ttgtgacttt   acttgaaata   72780   attatcaatg   tgtatttcta   tatttgagtg   tttcaatact   tatcaagagt   tttcttcccc   72840   attcttggtt   cctgtgtttc   gacacagctc   tcaactttct   gtagtacttt   gggtagtatt   72900   ttttttttca   agaagtcatt   ggataagatt   ataaacacct   ggcatttctt   tatgagcctt   72960   tttcttcact   ccacaacaat   aatgatttag   atgggtatag   aaatcttgaa   ttcataactc   73020   aaatctccat   agatgttgct   ccatttaatt   ctggtatttt   gtgttaccaa   aaaaaaatct   73080   aaagtcagtc   caagatttct   ttcttgatag   gttatccaat   tttgtctttt   gtttctgatt   73140   tttttttttt   tagatttcat   gctttcacca   ggttatgtct   attttttggt   cgtctgttat   73200   gtgagttctc   tgagctgcca   tatgctcttt   tgatctgctg   attaatgtga   attcgtaggt   73260   tattttcttt   atgtctttga   atacagatta   cttaccattt   attttaatct   tatcttcagg   73320   aactctaact   gtctgtctat   atgttagtaa   ttttggcctt   tctgtcatca   tttaaatatc   73380   ttggtcctat   tcttctacgt   gggaagaata   atttatactt   tattaatcaa   taatttgatt   73440   ttgattcaat   gattagcagt   gctaattcta   ctttttattg   tttctgaagt   atcttaaagg   73500   ttgctttgtc   tatgaaaagt   tttaaaacat   tcttcctttg   tattatctgg   tgccttttta   73560   tttctgtcag   cttcttggtg   tttcccttct   cacaacctat   ctaatgtctt   attatctcgt   73620   ctttcacctt   taattccata   ctgtcaaagt   gtccccaaaa   tgtaaaaaga   atgcaaaata   73680   tgcgctaaaa   tttcagggtt   tcttgagcta   aagttttttt   gaatatatat   tctttttctg   73740   atatttttgt   gctcttttta   actcaatgct   aggaaatctc   tctctgtctc   atatgtttca   73800   cccatctact   gaaagttttt   taattggcta   acatctgaat   aaatgcacat   atcttttatt   73860   tggattattc   ctacactagc   tgcacaatgc   catcattgtt   tcctccacca   agaagagctg   73920   ggtgggccat   gttcattgag   aaaatcagtt   tctctcaaac   aggagaaaaa   atggcattgt   73980   cctatcgcta   tgtgaataat   tcaactaaaa   agaataactg   ttctcccagt   aaagcttttt   74040   ctttcttaga   aaaattgggt   ttctttatat   tccatgcctt   tgcttcagag   cacaatatgt   74100   gacctgggaa   aggcatttac   ccttgttaaa   ccttcatttt   ctcatttcta   aaattacaag   74160   ggttatctcc   atgatcttta   ggaagccaga   actgatctgg   ttctaatggt   cctttgattc   74220   taattagaaa   tttccaactg   caaaagttga   gggggattct   tacatcttca   gaagccaact   74280   aattaaacaa   cttttgctgt   aatgagtgtc   agtcaccttt   atgaatcagc   ttgtcactta   74340   ataatagaaa   tagctggctt   ctggcccatg   cagaaggaat   gtagaaaata   gactatagta   74400   acaagctgac   tcccattctc   ctgttgttcc   ctgcctcact   cagtttccac   agtcaactta   74460   ggacaattcc   atactgtgat   tatttactcc   tgctttgctt   tcgtaattca   acaaacagta   74520   ttaaccactt   actatggcga   ggcatggttc   caggtgctaa   ggacccacag   tggatttgat   74580   tacaaattac   aggtaaaata   ataaagcaat   aaaaaggata   attttgcttt   catcatcgaa   74640   agctcagtga   tagagtaagc   tttagacttt   atttgcctca   gcaatgcagc   agtggtagag   74700   ggcagaggag   aagtctcatc   ctaagccttg   ctcccaaggc   tggttatagg   atgctgtatt   74760   agtttgctgg   gactgccgta   acaaatacca   aagattgggc   agcttaaaca   acagaaattt   74820   actttctcac   agttctagaa   gctagaagtc   ctctatcaaa   gtgttggcta   attttatttt   74880   gaggcctttc   tcattggatt   gtagatggct   gtcttctccg   tttttttcac   atggcttttt   74940   ttctgtccat   gtctgtatcc   taatcttctc   ttcttatgag   gacaccacca   gttatactgg   75000   attagggccc   acccagaatt   acctcatctt   tttttcttca   ttatctcttt   aaagtcccta   75060   tctctaagca   tgtcacattg   tgaaggactg   gaggttagga   ctttaacata   tgaatttagg   75120   ggagatgcaa   ttcagcccat   aacaaatgcc   tgctagtaac   tcatgggaac   tctcttccac   75180   atgcaaagag   ataaaatgtg   atttctgcaa   gttttattag   aaatatagga   aggaaccctt   75240   cccagcaggg   cttgacaagc   ctgtgcatgt   gcctcactga   cataactaag   ttatagggtc   75300   atatttgaac   taatccatgg   aacaagacat   gcaattacac   tgatgagctt   aggctgaagt   75360   caaatgtctc   actcctgaaa   attcagactt   ccccaaaaca   catgggctga   acaggtaaga   75420   tggatacttc   aactgaaagc   cagggtactt   aacaaaagca   ggagaaatag   atgctgggga   75480   ggtaatccca   acgtccagtg   caaatgctat   gagggaaaac   gacagagtcc   caaacatgct   75540   ggagcttata   gttagaggaa   ggaaaaatta   gatttttctc   agggctttgg   gattggggca   75600   cctggcagtc   agcgcctaga   ttccaaagca   ggtcagaaga   tgcctctgcc   caagtattgg   75660   aaaccttttg   aatatctttg   agtattaaaa   acattttaac   actgcctgag   aaaagtagta   75720   gttatgaagt   tatacattta   gcctatccca   ttcaactcag   gggattgtac   acagatcaaa   75780   taaattgcaa   ttgtgtttcc   ctccctgata   ttcacatagg   aaggtgagtg   gtgagcagta   75840   aacttcagga   cctttaaagt   tatcattgct   ttcacccttg   caccagcttt   ttgaggtaaa   75900   tattcccatt   ttctaggaga   ggataatgag   gctcagagag   ggtaagtggc   ctccccaggg   75960   tcttctagct   gaattgtaac   cagcttgatc   ttccttttga   attccaccat   cttgcctgtg   76020   agggaaaaag   gaataaggag   ccaggagttg   cagggtcatt   cttctcctca   agaggcttga   76080   gagtaactaa   gtatctgttc   aaaacacttt   tgagaatttg   gcggcctcat   tacgtttctc   76140   aggagtccct   gggggaggtt   acacttttag   gatccccagg   aatagagtga   caaaatacag   76200   ctaccacaac   ttggtcctgt   gcaactgtgc   ccttgctcag   caattactga   ctccattctg   76260   actgaactct   gccaaaaact   cttaacatta   ataattatta   cctgttctca   attttaataa   76320   ttaagagttt   tacaacttcc   agtgactctt   aattcaggtt   tctttggctt   actgttcttg   76380   gacaataaca   gtatggtcac   tgttgcacag   acagaagcta   gacatagtct   gtatagctaa   76440   ctccctcatg   tccttcaaat   ctttcttcaa   atgtcagttt   cttttggggc   cttctgtgac   76500   caacctgatg   aatagagccc   accagcccca   gaccacagcg   ttcctgactt   cctttactct   76560   gcttcacttt   tccctttttc   acagcaattg   tcacattcta   ttctactata   acttacttat   76620   ttattgaatt   taccactgat   tgtttgccta   acctcactat   aagacaggga   tctttgtttt   76680   attctccaag   aacttagaac   agtagttatt   caattaatat   ttgttgaagc   aatgaatgaa   76740   ccaaatgaat   gaatgaaaga   tgaatataaa   aatatttata   tgtcctaaca   ctctactaag   76800   cattatgaca   gaatatccac   caaaaaaaaa   agatgagaag   gagaaaccat   tattcttctg   76860   tagcagggct   aacagaacat   gagcagatgg   caaacgtgag   attatcagca   ccttctgggc   76920   tagttcctgg   aaatcagctg   atttcctgct   gcttccaaaa   attcttaaag   ctttcagact   76980   cctgagcaag   ctcctgcaaa   acacctattt   tggtgctgca   agcacctgca   gtagctctct   77040   agtcattcag   gtacttgttg   accttaggaa   ctcaagccat   agcttccacg   attttcaatc   77100   cctgatccct   cttcagtctc   aaattgctgt   atccaaaacc   cttcctactt   gcaatatatt   77160   agaaaggcca   ccttctgacc   aaatctgaaa   agttgggtag   gtttgtagag   gatgagtttt   77220   tggattattc   ccaattcttt   tgatcctcac   cccaacctcc   taagactaaa   gctacagtgg   77280   cctcccttta   gggaagaaag   cagaggaaag   gagcagagga   attattttct   gtgaattaac   77340   tgtttctaga   ttctaatatt   agtgactaaa   ctgccatttg   tacttcctat   attcctgttt   77400   tagcaagagc   taaaagagtt   tcaaagaagg   taaattgtat   tctaagaaga   gaattcagaa   77460   agtcctttcc   agaatagatg   aatcttaaat   gatgggcagg   tttcccacag   agataggtgg   77520   ggaagccatt   ctaacctaga   aaacaggatg   cctagaagta   tgcagtgttt   tcagaaaatg   77580   gcagacacag   gcttccttgg   ctagagaaca   aagacttcat   acaggagtag   gttcataaat   77640   acaagcacaa   gagaccttta   tagcattggg   gtctagtgat   tctcaattat   tctgcctata   77700   agcatcaact   gcaagaatag   ttaaaattct   attgaagaga   agaaatgttt   cctgctgtgc   77760   cagaaaccct   taagaaaaag   caaaggaatt   tcacagagct   gaagatcaag   cacctgagaa   77820   acaagtttgc   ctcaagagat   gcttccaaag   gcaagttgga   agcttattga   tgaaaaagtg   77880   aaacactatc   acaaggagta   caggcagatg   ggacagaact   gaaattcaaa   tggctaggat   77940   ggcaagaaaa   gctgaccact   tctatgtacc   tgcagaacgc   aaattggcat   ttgtcatcag   78000   gatcagagat   atcaatgatg   tgagcccaaa   ggtccaaaag   gtgttgtagc   ttcttcgcct   78060   ttgtcagatc   ttcaatggaa   gctttgttaa   gctcaacaag   cttagcatgc   tgaggattgt   78120   agaaccatat   attgcacgag   ggtacccaaa   cctgaagtca   ttaaatgaac   taatctacaa   78180   gcattgttat   ggcaagatca   ataagaagag   aactgccatg   atagatacca   atttgattgc   78240   ttgatctggc   ttcatctgcg   tggaggatct   gattcatgaa   atttgtactg   ctggaaacac   78300   ttcaaagaag   caaataactt   cctgtacccc   ttcaaattat   cctctctatg   agatggaatg   78360   aagagaaaaa   ccactcattt   tatagaggtg   gagatgttgg   caacagggaa   gaccagatta   78420   atgggctcat   tagaaggata   aaataaggtg   tctgccatga   ttatttttgt   aatcttgtca   78480   gttaataaac   agtgactgct   ttcgatttga   aaaaaaaata   catagttaaa   atacacctac   78540   acattcacac   caaagattct   gatttaattg   gtatgaagcc   aaaccaagac   attagtactt   78600   ttccagagct   tcccaggtga   ttttaatttg   cagccagtgc   tgagaaacac   tgagacgatg   78660   cacataaaat   gcctagccca   gagcctgctc   aagaaatgtt   agctgttgct   ctctactgtc   78720   actgtttggg   tggttgagac   ttgtgcccag   gcttcctagg   ggagtgttcc   tcccattgcc   78780   ccacagttcc   ctgtaagtgg   tggaaaagga   gaccctatag   cagcacttct   caacaccggc   78840   tacacatttg   aatcaataga   ggagccttgc   caaggtttgc   aaaagatggc   tgggtctgtc   78900   tctaaccaat   tagaatctct   gaaggtggga   cccaggtatc   cccattattt   actttcccca   78960   ttaactattc   cacaaatatc   ctcatatgat   ctgatctcta   tttccccata   atgattgact   79020   ggtatttctc   tttgccattt   cagtattcac   aatctgcttt   gtattaaagc   tgcttattta   79080   cgttatctct   cccactgtga   accactgcag   atcagggact   ggatttgttt   tgttttgttt   79140   tagtttttat   acaattgctt   tccccttgca   ctcagcatac   agcaaagctc   taacacatga   79200   tatatactta   cataatatta   atttaattaa   ttgtcagaaa   ccatagatac   tcaactataa   79260   gactttcctg   ccttctccac   tctacttttc   catttttcat   gttttctttg   ttgttgtgct   79320   tttctatgcc   ttcttcacac   tgtagccaaa   gttattttat   tcaaaattca   aattaattta   79380   attatggaag   ctgggtaatg   agcctaataa   tataaacaca   gtggtgcagt   gagcatgcag   79440   ttagttacaa   tgttccatct   atttttctgt   tgtgtgaagg   catccacaat   ttataaagta   79500   cctggtgatt   caaaaattaa   gaaccactat   tctaagagta   ggtggaggcc   agactatgga   79560   atcttccaac   aaccaggagc   aggagttgga   atggaataaa   ttagagcact   ggatcatccc   79620   tgaaatccct   gagcagagga   acaaccgaaa   cagagatgta   tttaagctta   taagtctaaa   79680   taaaagttgg   gggtttaaaa   aggctactta   attacttctg   agaaaaaata   ttttattcat   79740   ttctcactgg   gcttctataa   tcacctcagc   tctgtctgca   gtttcctttc   ttctgtaatt   79800   atttttctct   ccattttctt   gccctgatag   ccacgtgctc   atttgggtat   ccatcaaaag   79860   actacatttc   agtatattaa   ttctctcact   ggatggtaca   gcttgaattt   ccttctgcta   79920   gtttcattaa   aatctaaatg   gggggaaaag   tcctctacat   tatctaggag   atccatgtga   79980   gctgcccagc   tccattagct   gttgtctgtt   agaggtggat   tacaagcagg   tgtaatctta   80040   aggacaccgc   ccttctgtga   ttgttaagaa   ggaacacaga   ctgagcagag   cggctactgt   80100   tctgatcagg   agccaccacc   actggttcct   acagtataca   gaatttcaat   tgtcagatct   80160   gtcctggagg   aacaatagaa   tacaactgtg   aattaaataa   tttagaaacc   aaggacataa   80220   tctcactttt   ctaaatgagt   ttgcctcttt   taggagtcag   tatattcgta   gttttgctag   80280   cacagtcctg   tgatgtagga   ggaaaatgcc   caaaaatatt   ctagaaaaaa   cattgagaat   80340   tcaagttaaa   atgtcagatg   gagcagcctc   tagaattcct   cctttgtagg   ccatcagtaa   80400   tctcactgaa   aattagacat   gtattattta   taaataataa   tataatatta   tgcatatttt   80460   aaatattaat   gatttaggtc   aaacgcagaa   ggtgaaattt   gtaggtgtca   gaaatgaagg   80520   ccaaatcttt   agtgggaagg   ctgtgggagc   tgaagctctg   cagagcacag   tggggatc   80578     <210> 16 <211> 112 190 <212> DNA <213> Homo sapiens <220> <221> exon <222> (31560) .. (31663) <220> <221> exon <222> (64868) .. (64961) <220> <221> exon <222> (80499) .. (80700) <220> <221> exon <222> (87482) .. (87558) <220> <221> exon <222> (97994) .. (98080) <220> <221> exon <222> (101581) .. (101761) <220> <221> exon <222> (102663) .. (103161) <400>   16   gatcacgagg   tcaagagatc   gagaccattc   tggccaacgt   ggtgaaaccc   cgtgtctatt   60   aaaaatacaa   aaattagctg   tgcatggtgg   aacgcacctg   cagtcctagc   tactcaggag   120   actgaggcag   gggagtcact   tgaacctggg   aggtggaggt   tgcagtgagc   cgagactgcg   180   ccactgcact   ccagcctggc   gacagagcaa   gactccacca   aaaaaacaaa   aaaaaaaaaa   240   aaaaagaggc   atttctaaga   gagagacctt   aaccctaagg   cagacactga   agaggatttg   300   gggcatgaaa   taggagctgc   aggttggaga   acatcacaag   caagggcatc   tgtaacccac   360   cctacccact   ttctcaagag   acagtccaga   gagcgagtaa   ggcacagggg   ccctcttctg   420   catattgaaa   gtgagaagaa   atgcctatgc   gagtgtctag   gcagagagct   tgaaacagcc   480   acttgtgggt   ctctgcacct   gcagcctggt   gcagggatca   gaagaatgta   cttattctgt   540   gcaacaagag   ccatgccttt   gtgacaaagc   ccgtggccca   agaaggccct   gcatttggaa   600   caaggaggca   ccactttgat   ctgctgacag   ctaagtgcaa   ggtggaatca   gagatatctt   660   aatagatgtc   aatgataaca   gatgatacca   agaaccaggc   atcttagaca   cacacacaca   720   tacacacaca   cacacacgtg   cacgagcacc   cacgcacgca   tgtgactgga   taccgcatga   780   gttttctaaa   gcttaaagat   gactacaagg   acaaagagta   aacacttaat   tgactgcaat   840   taagtttttg   atatccagca   gagtaggagt   ttttactagc   aattaacttc   agttttagaa   900   cacgacaaat   cttattttta   ttatacaact   acaaacaaat   atataatgaa   tgctcagatt   960   ccaggaccct   atacctgggg   tgatgggtgg   gacgagttaa   agaagggatc   tggacactgc   1020   tgcctctctg   ccttctttat   atggggaccg   tatcacaaat   gaggctcact   gtgcctccag   1080   tcatgtttac   tgtgcagcaa   ggcttttggc   cagagaacat   aattgacatg   cacagagctg   1140   gagctcttgc   cagctcacag   caaaagggcc   tctaattcag   agaaaaagga   gttggacctc   1200   tgccagtcat   gtacctgtta   tgctttcata   taggcatcag   ttagaccctg   atctcagtct   1260   gacaaaccag   aaaatataaa   accatcagga   tcaaggcata   ctttttactg   ggagtgttag   1320   cctggtaatg   gagaggaatg   ttattgctct   aagcattcgc   ataatatcta   ttgatcaaac   1380   ctctgtgaag   ctgctattct   tcatgccagc   gtctggcggc   aagggaggac   tatttcaggt   1440   ctcaccatca   cagccacttc   acactcaggc   catgccacag   tcaccagcac   agcttgggat   1500   tggggcagga   agtccccagg   gtgaccagac   agaaagcagc   ctcaggacgg   ggccagagca   1560   tgagactagg   ggtggtggca   gctgggtttt   atttactctg   ggctgtgtga   ccttgggcaa   1620   gctgccagcc   cattctgttc   ctctctgcat   gccatacaac   cgggaattcc   tacattactg   1680   agtcactcct   gggccaaggt   ttgatgtccc   tattaaaatg   ctgttgagag   agtaagtggc   1740   ttcagtcact   agccctggag   aatgagttca   ccagtttgat   ttgtttacaa   ctgagaactc   1800   tttcagtttg   tgtgcgtgag   gtgctggggg   agcaggtgga   agaggctttg   ctggagtggg   1860   ggaaatatcc   tgcctgggag   tgggattggc   ggaagggggc   atatttgtca   atcacaatag   1920   aatcctagac   acttatgtct   gggaggcata   gcataggcca   ttgcattcat   cttcccccca   1980   gtacctgaat   ctgtgaccaa   tggaactgag   gatctttact   taacaacctc   tagtgatggg   2040   ataatcacct   ccttctaagg   caagctactt   ggaaactcta   agatattact   gagaaaagac   2100   cctgagccca   gccttgaatt   agtccaaagt   ctaatgcttc   tcccaggtga   cagcccttga   2160   aagagtgtgt   gacaggcgcc   atgccccctg   aatggtctct   tctcccaaat   aaataatacc   2220   tgattctccc   tctgcaatgt   tcatgaccca   aacaaagctg   agcctctgtg   tggccactct   2280   cagtttgatg   ttgtacccca   aaccttcaac   ctcagtctta   atgcctggga   atgggggaat   2340   gtgtggaaaa   aggcatgaaa   cacagtacac   agcaaaatgt   cttaacattt   ctcttgattc   2400   tctagctcct   tatccctttt   cctaaaaatc   tctttctaaa   tctttcaagg   ataaagggaa   2460   gggggtagaa   aggggaagtg   aggagaggta   aaggaagatg   ttcattattg   ggagcctata   2520   atgttccaaa   cacgtgggac   atttaatctt   cacaaagtaa   gtatacccac   cctaatttta   2580   taaggtagaa   actaagaccc   aagatatcta   agtaagttgc   ccaaagtcac   aaagcaatta   2640   cctggcaaac   aatggattca   gataatgact   atatcatagg   ccaagcccaa   ttacactcaa   2700   aataattcac   atcttcctaa   gaggcaagcc   ctgggggccc   aagagggggt   tgttgagtaa   2760   gggcagaaag   tacaagggag   acaagttgcc   aatcacagct   tggccctgat   tattgtaata   2820   actgacacaa   ttaaagttca   atttcgtatc   cttagaaggt   ttccgtcata   ttccaaggct   2880   gatgagaaat   ttccattcta   agaatgggga   tctgttgaat   gttttaaatt   caggcactta   2940   attaaatctt   ttaagatgac   tctttctgtg   tttgccctta   gtcttttatt   tctatccaat   3000   attgttcctg   gaccaaatag   ggtcgggctg   ctgtttcttg   tagcccaata   atgagatgca   3060   gatgaactgg   ggaggaagag   agtttttttt   ttaagtatat   aaaaacattt   attcattaga   3120   aaacaaggag   actggcaaac   atatattcca   aagtgaaagc   agctcaatgc   agttcagtta   3180   ggctaattta   agagaaaggc   cttgcatttt   aaagatcgtg   tatgtatttt   tttttttttt   3240   caaaaaagga   gacaggcaaa   tattctacaa   ggggaacaga   attagaattc   taggtcaccc   3300   tacaagttac   cctgcacagg   gaggaaagga   acaggcaaga   tgacttctca   ggatctgtgc   3360   ctgcgagctg   atgctctgag   aatgggggtt   attttcttgg   gtgtcctgtc   ttctgtcatc   3420   taggctaaaa   aatcttcctc   acttgactca   tcacttgaga   agacaacttt   tggtttcttt   3480   tcggaagctc   gctgctgggt   gttcggatgc   cgatgggtag   tacgacgggg   ctctggtgtg   3540   acgaagtcat   tgtctgaggc   tgcagaagcc   agaggctggt   gcctacaaac   agtggatggt   3600   ttcttggctg   ttggggctct   ctggctacct   gcttggttaa   tctcaagctc   ctcaatccca   3660   tccaaacctc   tggcatgaca   ggcctgaagc   ttttgctcaa   attcatctat   tatagccttg   3720   ccctcaggct   tggccccatg   ttgtagcttg   cccataactg   acaaaaaaga   actgaagatg   3780   gactcatctg   acagtttatc   tccaaaacag   tcaaaattgc   caagcatctt   gcaagggcct   3840   tgctctgtga   ggggcagctg   tgacacacag   taggccaggt   cctgctgcac   ctgctcagtt   3900   taggctgtgt   ggaaccgctg   acacagcttt   tccaccaggc   tttgtctgct   tgtctttggt   3960   gatgatgtag   gagaagaact   gcttcataat   ggtgtgaaaa   ggcacttcct   ccacccccag   4020   ctcggggtct   gacaggtggc   tgatgatatc   tggaaggaga   ctatagattg   cattgccctt   4080   gtggaagagc   tcattgaaga   acttcttggc   cagggcagca   atttgagact   cagggttgat   4140   gagcagcatg   gccatctcac   tcacctgccc   ctttaccttc   accatgtcct   tgaggatcag   4200   gtggagtcat   caccagcccc   actgttttcc   acactggctg   agcagggtcc   tgaaggtgag   4260   catatagatg   aggagtccag   gggtccacca   gattgggaaa   gtggatggcc   agatccccag   4320   tggcaacgat   gagattagac   cggacaatgg   gaagtggaga   cttttctagc   atggtgaaca   4380   gaagatgagg   ctgggagttg   cagaaagtgg   tactgatcat   gcagaacttg   ccaagggtag   4440     gtgaagcagc   tgcagagagg   tctgggttgc   tatagaggcc   tgagttgttg   tagactttaa   4500   gcaagagtgg   aacaaaggca   gccagtgtct   gtttgccatc   caacagttcc   atcttgcaga   4560   agccgcggat   tagttctgcc   tccgtgtcat   ctgctgcccc   aaccagcccc   agctcctcct   4620   ccatagtggt   ctcgaaactt   gtattcttct   ccttgggatc   tttggtcttg   tgctcctgtt   4680   cttcccggag   aactcggcgc   tgacagagct   ctccactcac   tgcctgctcc   aagtggacca   4740   gctgctgcag   agccacatcc   ccagccaggg   acaagaggtt   catcaacagg   aaagtgggga   4800   gcattgtggg   agactccttc   gggtccccct   gactggttct   cttctcttct   agcttctcca   4860   gggcctgttt   tgcacagccc   tgccatatct   gggcacagat   cactgtggga   ctctctgcca   4920   gttggtaaat   gagggtcact   gccacctctt   tgaatgggat   ccagagtggg   tctggtggac   4980   aaagcctttc   gggaccgtct   cccgcagtca   ctcaaacaac   ttgtgttcct   gaggcaactg   5040   gaagtggggg   tgacgtttgc   ccagagaagt   ctttctactg   tcagagatgt   tggcgatggc   5100   atggcacacc   tgctgggcca   gccggtagtc   ctgtggaaac   ttcggaagag   agttttattt   5160   tctgcaaccg   gtgacgggga   gaaggcctgg   aaattattgc   cagaccaact   taaaattaca   5220   aagttttcca   gagcttatat   accttccaaa   ctatatgtct   acgtgtaagg   tatgcattca   5280   tctaaagatg   taaatggtta   acttctttta   atctataacc   aaggtctgag   tcctaaatac   5340   cttcctctgg   agcctcagta   aatttactta   atctaaatgg   gtccaggtgc   tggggggatt   5400   acccttatct   tgtctcctgc   taaattacag   aggttttggg   agttccttca   gacctccaat   5460   aaacttgttt   gtggaggcct   aaggagtttc   cttagacccc   cagtgaaact   tgtttaatcc   5520   taaatgggtc   ctgttaagaa   ttcctttgtt   attttgtcat   gccttaagtc   ccaggaaagg   5580   cctaggtaaa   actcttgatg   ggcttttgtt   acattccagc   cttcgtacag   gggcactggc   5640   ttttaatatt   taacttaacc   actcagtcag   tactgaaaga   gttgtcagtg   acacctggcc   5700   tgccacaatt   atgagtgtcc   agaattttac   tagatgtttg   ggggaagggg   aatgagaagc   5760   ctgaattagg   aatattctcc   acgagtctct   cacaatctag   ttggaaagac   tagtaaactc   5820   aatgacatat   taaatgataa   gtaaatgaca   acctgtggtc   cttacttcat   gcctttgcac   5880   acactgctct   ctctacctgg   atgcccttct   ctcacctgat   ctctccagca   aagcactact   5940   cattcctcaa   gacacaaagc   tgaacaggca   ctctgtgaag   cctgcctagt   ccgcttgctt   6000   ccttcccaga   agattttgcc   aatcatccct   cttttggaca   aatatcgtgc   aacactgtat   6060   tttaattatt   tcccaactcc   ctgaaacact   agaatgtgag   caatttgaag   ataggaatgg   6120   agtttccttc   cttgcattct   cagcacatag   cattgtgtct   gaaaccaaat   agtcagcaat   6180   caatagtaat   tgatgagtgt   ttaaagaagc   actacaggaa   gtggcaaaaa   tcagggtcct   6240   ccagggcaac   acgatgggga   cttcacagag   tgggtggagc   ttgtattggg   ccttagagga   6300   tgactaacat   ttggagatgc   tgatgctgcc   aaaagtagaa   agatcaggag   gaataaaagc   6360   ccagaaatac   gagactgtct   tccccagtgt   tttatggtgg   tcctagattc   aaatatccca   6420   tcatgtcaca   actttgaaat   gaggaaaata   ctttcaccaa   gaagttgaat   gctattctga   6480   attacaaacc   caggtgagtg   ctcatcaaga   acagaagacc   agccaggtgt   ggtggtgcac   6540   gcctgtagtc   ccagctattc   aggaggctga   ggtgggagta   tcacttgagc   ccaggagctc   6600   atagtgcgct   atgtggatcg   gatgtttgta   ctaagtttgg   catcagtatg   gttaccccta   6660   tcccaggagc   aggtgaccac   tagattgcct   aaggaagcat   gaaccagccc   aagttggaaa   6720   cagagcaagt   caaaattccc   atgctgatca   gcagtgggat   tatgccagtg   aacagccact   6780   gcactacaat   gtgagcaaca   taaagagacg   cccctgtcca   aaaaaacaaa   gaaggaaacc   6840   tgcaatccga   gcactttggg   aggctgggga   aggaggattg   cttgaagcca   ggagtccaag   6900   accagccagg   gcaacatggc   gagaccctgt   ctctacaaaa   ataaaaataa   gttaaacata   6960   aataaaaaat   tctaaatgaa   ttttaaaaag   aaagaaaaaa   taatagaagg   ctttggggat   7020   cagggcagcc   caaagggatt   cctggagttt   caaacacaaa   atagacatga   gagacttaat   7080   ctggatgtca   gaccagcact   agaagaaaac   ctggacattt   tgaagccctt   aaattcataa   7140   aactttgaca   gtttcatgaa   catttgttga   aaccttgcca   tgtccaagaa   ggtcacctag   7200   gatcagaact   tccacccatc   tcctcttctc   cccctgtcct   tttctgaggt   ctacaccccc   7260   tttgtagtag   gtggtcaact   ggacacagga   ctaggtttat   caaaacttgc   tctcgggaaa   7320   taaaaataag   acaagaaata   caatatacac   aagaacacat   gttcttgtac   ctcagtctct   7380   gaatagcctg   tctctagact   ctgtttgatg   taagagtggt   tgcttctgaa   aaaagaactg   7440   agtgactaac   aatggttgga   gaagggtgcc   tttcttaatt   ttagttttta   ttatttttat   7500   tgaagtatga   agttcaatcc   ggtgcacgca   agtatcagct   gatgaatttt   cacaaacaga   7560   agataccagt   gtaactagca   cccaaatcaa   gaaatgttac   catctgcctc   tatgccccct   7620   cctaccaatt   cccagtacca   cctcttcccc   ccaccaagag   taagccctga   ggcccagaca   7680   ggcttcacct   aattttatat   tttatttttt   ctttttatga   acagctttat   taaaatattc   7740   acataacaat   acagttcacc   cattaaaagt   ataaaactga   gtggctttta   atatattcac   7800   agatatgtat   aaccatcacc   aaagtcaatt   ttaaaatatt   ttcatcacct   caaaaagaag   7860   gcaaatacct   tttagctata   actcccacct   cacatcctcc   gtaaccctag   ccaactgcca   7920   atctacttcc   tatgtctatg   aaattgccag   ttctccacat   ttcatataaa   tggaatcatg   7980   taatatatta   tcttttgtga   ctggtttatt   tcacttaaca   ctttcaaggt   tcatttttgt   8040   tgtagcacgt   atccatattt   cattttttat   ggctaaataa   tattctatca   tatggatata   8100   gcacattttg   tttatccgtt   catcagttga   tgattatttc   agtggttttt   accttttggc   8160   tcttaggaat   aaaatgctgc   tataaacatt   catgtacaac   tttctgtgtg   gacagttttt   8220   tctcttgggt   atgtaactag   aagcagaatt   gctgggtcat   atggtaacac   tgtggttagc   8280   attctaagca   actacagttt   tatattccca   ccagcagtgt   acaaaggttt   caattcctcc   8340   tcatcctcac   caacatttgt   tgtttgggga   ttgttcatta   caaatacatg   aaacagtggt   8400   taccagagtc   agtcagggtg   ggagggagga   atgcagagat   ttaggtcaag   gatacaaagt   8460   agcaaatacg   tagaatgatt   taattcttta   cataaataga   accttacagt   atgtgtgctt   8520   ttgtttctga   cttcttttgc   tcagcattag   attcaggaga   ttcatcaaca   ttgctgcgta   8580   tagttgcaga   cagtttacta   tccttattga   atggtgttct   actgtgtgaa   catactacag   8640   ttgctttttt   cattccacta   gtggtgggca   tttgggtcat   ttccagttta   gggctactat   8700   gaatactgct   gccttgaaca   gtccaggata   catcattttg   tgagctgctt   atttttatta   8760   tttgtgctac   ctgaattttg   tattatgtgc   aggtattact   ttttttaaat   gtcctaaaaa   8820   cacctttaag   ggcttttaaa   ctaggatcag   aaacttagct   ccttccctac   tgacaagcag   8880   gcagaaaaaa   aaaataatag   taatagctag   tatttactga   atgctcattg   agccacctct   8940   ttagggaaaa   aaatcttttc   atatatgttt   gcttttaatc   ctccaaatta   ctcaaagcca   9000   taggtattct   cattatgccc   atgttagacc   tgcagaaact   gagaccgaag   gaggttaaat   9060   aacttgtgca   aaaccataca   actagcaagt   agggaatcac   agtacaaata   caggtctctg   9120   aatataaagc   ctgtgcattc   accctccaat   tacacagcct   ctgattacac   tcagctggga   9180   gttcagggag   cctgctagac   atggggagga   gaagctgaaa   tgagagcacc   tgaaactgcc   9240   agctcagtcc   agctcaacag   ctcggattgt   tctccccacc   gcccccctac   ttcctgtcct   9300   aattaatcaa   atgctcttga   acatgaaaga   gagtgagatt   ggatttttgt   taagatgaaa   9360   gaaaggaatc   tgttcccatc   tggcaccttc   aaaactctcc   tattcaagcc   agcagggagc   9420   atttatgcaa   cctacatggc   agtttaataa   ttccccaaga   atgccaggag   attaagcaat   9480   gtcacaaccg   atctggatcc   accggcactt   gagaataatg   gaaagagcag   gggtctaagg   9540   tctaggacag   gggtcaacca   aatttctctg   taaaggccca   gatattaaat   attttaggct   9600   ttgcaggtcc   tatgatctct   gtgtcaacta   cccaactctg   gcattgtaac   atgaaagcag   9660   ccatgagcaa   acaggcatgg   ctgtgtaact   aaaacttctt   ttatggcaca   gatatttgaa   9720   tttcatataa   ttttcatgtg   cctgggcata   tatttctttt   ttaaaaaata   ttttctaacc   9780   agtaaaatat   gtaaaagtta   ttcttagttt   gaaagctgca   caaaaatgga   ggacaaagtc   9840   tagcgacctg   aattcaaatc   ctctctctgc   catttgttat   ttctgtggct   ttcgacatat   9900   tacttacatt   cactaagctt   catttctctc   ttgagaaaat   gagcagagca   ggtgccatgc   9960   attcagggcc   tcccttgtgg   ctggcactga   gccacatgct   ttatatgctt   tactgaatta   10020   aatcctacaa   taaccacaaa   aggctgaaat   tatatgattg   ccttcttcag   atagttacga   10080   tgattaagaa   attatgtcta   taattgtgcc   tgagctggcc   tccatagttc   attgagtttg   10140   atgcttcctg   ttgtacgcaa   aaattgccat   gttggttcga   tgagagcatg   cctgcttttt   10200   ctactacctg   tggtgttttg   agtacctata   tttgtattgc   acttacagtg   aaaattatct   10260   aattatatta   ttctatttaa   ttacaattgc   atcaagctcc   aataaacaat   ccaaatttca   10320   ctgaactgaa   agttgagtgc   acactttgaa   tagtcactcc   ttctgttagg   ttggtgcaaa   10380   agtaattgcc   attactttta   gtagcaaaaa   cagcaattac   tttcgcacca   acctaatata   10440   tatgtgcaca   gtaggcagtt   tggctcaggt   acacttattc   aaaacattgc   attaagtaag   10500   ataatacatg   ttaaatacct   agcacacagc   ctgggatgca   ggtgtcggcc   agtgtttgtt   10560   ccccacttcc   ttagtgacaa   agattaggct   tagctacttt   ttgggaaagg   tgagtgccac   10620   cagaaagatg   aggccaaggc   cttatcagac   acactggaac   acagccttcc   caaggaataa   10680   atccagaata   ttttgtgtat   gagttctggc   tctgaagccc   agcctgttct   tacactgacc   10740   ccccagtttc   tgccttgatc   agccacccac   taggctgggc   attgcttcct   ctcactctgc   10800   tatctagtga   cccaacctcg   gcccaaccct   ccagctttct   tctggccatg   cgactgttct   10860   gctagtctgc   tccgcatatc   tgctcctacc   ttccgatgcc   tcctctgagc   tttggctcca   10920   ctccccactc   agcacaaaca   acttactttg   ccccagtatg   gggtacaaca   gctcagactc   10980   ttctagaggt   ctaatcctgg   acacctcgtt   tgctttataa   gcacaatctg   gaaagaagag   11040   tttcattgct   cagagcccct   caggaagcag   aagaggagag   tggtgaagag   agcattcatc   11100   ctctggaatg   accataactg   gtgtttcaat   cccagctccc   ctccatacta   atacttgggt   11160   gagttactca   ccaggtctat   gcttcagttt   cttcatctgt   aaaatggagg   ttgtaaaaat   11220   atcatctacc   ccatagggtt   atggtgatga   ttaaattagt   tcttctatag   aaagtgcctg   11280   cagcagtggt   tagcatgtgg   taggcacttt   gtatgtgtta   tctgttatgt   ctgtttgctc   11340   ttactaacat   ctatatttct   gagtcctaga   cctccccaga   gagtgtgaag   ttcatggtgt   11400   ttcatgggaa   cagatgtaag   cccacttgct   gaatctctct   ctgagctaat   aattaacaaa   11460   tacattgcac   agtagcttac   caggaaagcc   tatactggtc   tgaacaagaa   gaaacttgct   11520   gaacaattct   aaagctcact   tctcaatata   acagcagccc   ttgctcaaca   tgttgcccat   11580   atcttcagta   ccgtgcagag   gtcacagcca   tcatattctg   ccagctcttc   ccctatgggt   11640   tgagcaacca   tatgtgtctt   cccaaccagg   acatttttca   tttttagctt   cttgttttga   11700   cataatttca   ggtttacaga   aaatcttcaa   gagtagtaca   aataccccca   tgtaccctgt   11760   acacatattc   ctgaaatgtt   aatatttttc   atatttgctt   tatcatttcc   tctatcagtc   11820   tatctataat   acctacctat   ctacctatcc   atccatctaa   aaatgtccca   ggtaatcagc   11880   aaacctgctt   taaatgtcac   caattatccc   aagaacatcc   tttttagcac   aaaaaacccc   11940   gaatcatgta   ttgtattcag   ttgccatgta   cgtttagtgt   tctttaactt   agaacagttc   12000   cttggtcttt   ctttgtatct   tatgacatca   acatttttaa   atagcatagg   ttagatattt   12060   tgtggagttt   ctctcacttt   gggtttgtgt   gatgtttcct   tctgaataga   ttaaggttat   12120   gtgcttttga   gagttataca   ccaggaatga   tgctatgttc   ctcttagtat   atagtatcag   12180   cagacacagt   atatcaattt   gtcccattac   tagtgatgtt   attgtcagat   ttctccactg   12240   taaagatact   atttttcctt   ttgcaactga   caagtatttt   aagagtacat   agccatcctg   12300   atactcctta   aactttcact   cacagcactt   accatacagt   gtagattcct   gtctgaatca   12360   atgctataaa   ggtttccaaa   tgatgatttt   ctaatttcat   tatttcttct   atacttatta   12420   gcagtctctc   tactttaagg   aagaaatttc   ccttctctcc   catttttaaa   tttacatcat   12480   aggttcttct   tttattctgt   gggttataat   ttgttgccat   agttatttct   ttttatgcct   12540   agattgctga   ctggggtcag   caagagccct   acaatttggt   gcgtgtttct   gttgacatgt   12600   ttctcagcat   ttttgggcat   gtcctcactt   gccagcacaa   catgatgttc   caggcttatc   12660   ttgtcctttt   cccagcccca   ggccaggaat   cagccatctc   caatgagcca   agtaaaacca   12720   tggcatattt   aagagaaagg   gccacaggca   taatcatggc   gggtataaac   tgaaactgac   12780   ctagaaatgc   tgggtcatat   tcccaatcta   ccactagatg   tccttccaca   cctcaatagt   12840   tcttcacagg   aagcagtcct   ggggcatttg   gaaatggtgg   gatatttgga   gttatcacaa   12900   agactggggc   agcaggggtt   acttctggca   tttagtaagc   acagcccagg   catgccaaat   12960   ttcctgcaca   ggaccatcta   tcctaacaca   gaattgcccc   ataagtagca   atcctgttaa   13020   taaatagtgg   cccaatccct   gaactaatct   atcttgtgtt   ctagtctctc   aaggattggc   13080   ttagttcttt   tcaccttatc   cgccatcagt   taatttaatc   accactgttt   ccatagtgag   13140   aatctaatat   attagactga   tcaagtgtgt   taatcaaata   tattggatta   gctggcatgc   13200   attatagcaa   ataaaatagc   cagtcttcat   tttagatcta   gtttaaagaa   agcctctact   13260   tcttatgtgt   caatttttgc   agaaaacatt   attctagcaa   gttggcaagg   agtctaggac   13320   ttatgttaag   acacaccatc   tcatccccag   ttcttgcagc   tctctgtcta   gttattggcc   13380   ctattcattg   catcagcacc   aacaacagct   acccaggttt   tatagggcaa   gcattttact   13440   aaacatttta   cataaactat   ctcatttaac   cttcacagta   gccctataaa   tcaggcattc   13500   ttattacaat   tttacaaaga   gccctggatt   ccgtgtggtc   agtagcaagc   ccaagatagc   13560   acagctgtca   tgtagcagag   ccaacattca   aaatcccaca   gtcgggctcc   agtagccatg   13620   ctgtgtactg   cccatgcatg   taagaacata   cccaacacaa   tgaaaaataa   gaaatgaaat   13680   agaaagggca   tggaaaatat   caaatcataa   ctgcagagga   aaaaagtcaa   atttaagctt   13740   agagttactg   caaagttact   gcaacccatc   atattttatt   ttgtgcttcc   tagttagtgg   13800   ctaaaactaa   gggcaaaact   gactaaatat   aatttgttgt   aatcttgagg   gaaatgcaca   13860   gcaagatgtt   tcttgatcaa   gaaaactgat   actacggagc   tttatattca   gtactttgat   13920   gaaacagcac   taaaatgtcc   tcaataacat   ctctacaaag   aatcagtgat   tcttagacct   13980   ctaagaggag   ccagtgtgat   gcctaaacat   aggcatgtag   gctacttgac   agacagactc   14040   agcatagcag   tcatcatcac   gtagtaattt   ctagcatcct   agtagctttc   tcactggctt   14100   tagttttgaa   taaatgggat   ttttgctaag   atcttacatt   atgcaacaga   ggcaaccaca   14160   aagaccaccc   ctagaaaatg   aaatgatctt   gggccccttc   ctcccttacc   tcctgcttca   14220   cactgcattc   tttgggaagc   ggatgctgaa   acagagtcag   gtatacaaaa   atctggggag   14280   aagtcactcc   tgtgaaagca   aagggaagaa   gtaagattgg   gctggaggac   ccatcagagc   14340   ctgacattaa   cctgacagtc   tctggcagct   caggaatcct   ttaggatttc   catgttgggt   14400   ataaatgatt   agactcttct   actataactt   agtcattggt   agaggttgta   ccaagaatag   14460   catgatccca   gctaaaaagc   tgaggcaaac   cctgaggaat   ctaacagctg   gaggatgtca   14520   gcaaaccata   tacctcacag   ctggacagca   aggtctttct   tgaaggggga   tctgagcagc   14580   atatctctat   gtctgcctca   tttccctatg   tacaagcgtg   tacctgattg   cagcagaatg   14640   agatgagaaa   caaatatttt   tttgttacaa   tttaccactt   ccatctcagc   ctgctacccc   14700   ttagctctcc   ttgggcaaag   ggaaaaagac   atttgaagag   cctgcctctt   ggataagtga   14760   aaatggaact   ctcctctcct   tctcttggat   ttctaaacat   cccagatcat   ttctgtgaat   14820   tctttttatt   aggaaaatag   catcctttaa   aagagaaaac   aagttgtcta   tgtcttatat   14880   tcagagtcag   tttgagcccc   atgttgtgca   atgatcttgt   gcaatgataa   ttacatgcta   14940   ccatgcctca   agaaatttcc   ccttaagtgg   agtgaggagg   agaaatagtg   gaggctcaat   15000   gtcttgtttt   aagaaaacac   ctgttattgc   tgaacctgat   gcttgtgctt   atgagcacag   15060   gagttttttg   tggcccagca   ataaccaaca   actcaggtag   gccagttgcc   tcatttaatg   15120   agcttcttgg   tttctgaata   caacagaggc   aaagagagta   agttttacga   ggacttgggt   15180   ttccttaatt   agacacaagc   tttttatttg   gttgggggaa   ctaatggaga   aagggagaca   15240   ctagcggcat   ttgtaacttg   ttaagactaa   ttgaaaatct   taggaccaaa   gctttctgtg   15300   aagagagtgc   tctaatgaga   ataaatgtga   tgattattta   cctaattgag   cagtaatgac   15360   tgaggtgtga   gccacttccc   caggattaat   aaagagccgg   ggagttgaca   gtttagacca   15420   ttaggcttag   ctggtacttc   atcttggcga   aggacacagt   gccagaatca   tattaatatt   15480   gtaaaatgag   cccaaaacat   agacatatac   ttttggagaa   ataattttaa   ttggctaagt   15540   acctgacatc   aggttgggcc   attcagtttt   atcattaaac   aaagtttcac   acaagttctt   15600   caagacttgt   gccatttaca   gtgttctcag   cattcagtgt   ttaaaaaaga   aaagtagaag   15660   agggaccttt   atcaatatca   aaaatagttt   tttgtgcaac   caaaagcaag   caagctatta   15720   ttatatatac   acactcgctt   taaaaacaac   actaaagaca   ttgagttgag   ctggaagcat   15780   tgcatactct   gagatacaca   gttgatattg   ttacccacag   tgtcccttca   ggaaagccag   15840   aggtcatctg   caacaccaca   attaaggata   aaaggacctt   atattatcag   aaatttcaat   15900   gtcttcataa   aaattgtgtt   tatattcaat   atctccctta   tttagtgtga   caaatgttca   15960   tgtacgagtc   atctcaagaa   aaagagttac   tcagcttgct   acgtattcat   tattctatgg   16020   ctctgctatg   caggggaaag   tgcaaacagc   aactgagaga   aagcaggaat   ctaagacatg   16080   ggactcaact   tgccctttac   acagggtgaa   tcctacacta   gtgacctttg   tgttttaagg   16140   aagtttgcaa   atatgtttaa   ttggacaaca   tttcttcctt   tatctagcaa   gcactgagta   16200   tcttgccagg   cactatgaaa   aaagataaaa   ctataaacat   gagcaaggca   cagccactgc   16260   caatcagaag   cttacaatct   agcagagatc   ttaaaatgca   gggagctctt   ttagaaatct   16320   gcttagggat   caaaatttcc   aatcacactt   ttccttccag   catctttatt   ctgtaatcat   16380   tattttaatc   ttcattgaac   gagatgatta   atgtggttcc   atctgctgta   tgaaatcacc   16440   catgctacag   tcagccaatc   aagcaacagc   agtaataatg   tactacttca   tttaatggaa   16500   atttaccaag   ctgttactat   gtaatatcac   agttttaggg   cctggagagg   tagagctgaa   16560   ctaaaaataa   taatctctgt   tatcccatag   tatagtggaa   taggaaagaa   atacaaaaaa   16620   aaataatgtg   ttcatgtatt   ataggtacta   taatagtagg   gtaggaagat   acacagaagt   16680   gggaataacc   aagccaatct   cagtactcag   aaaaggtgtt   ataaaggagg   taaccttgaa   16740   ttaaaccttg   aggaaatgga   tattcttcag   gtagaggtgt   ataaagagct   tcccacacca   16800   atggagcagc   ataaagaaat   ggcatggcag   agtgaacagt   tgggtatggc   caaggaaact   16860   ctcaactgca   tcttcttcac   tgcctcaact   gatcactcaa   cgctctccat   tctctcagca   16920   aacaccctga   ctaactaaac   tcccatgact   agcaggatat   tattagactt   aacaaacctg   16980   tcaatttata   tgctttccct   ttagaaagac   ttgacttttt   tttagtgaac   tgcaatagaa   17040   ctaagctcag   gagatgcaac   tatgaaagat   gaaggaacat   tataaaaatc   tctgcaatga   17100   gattctgcct   tagattgctt   tgctggtatg   tcaactgatt   actccaattt   aaagaaaccc   17160   aaactggaaa   ttataaaaga   taaattgtga   gtgtatttta   tgcaaaaaag   aatcaaatca   17220   aatcaaagtc   ttggctctac   attaaacaaa   cgacaaaggc   atgtttgttt   ttatcaaaat   17280   gttttagggt   agatttgtat   gtatgcgtta   tttctttaac   caatgtttta   atcagttaaa   17340   cagctgctat   tattaatgtg   ctggttgaaa   gtgtttttct   taaagtacaa   atttaaaata   17400   gaatgagaaa   agatcagact   tgagaagaag   gaaagtcaca   gtaggacatt   tggaatttat   17460   tctctagacc   agaggtctgc   aaactccagc   ctacaggcca   aatccatcct   gcagcttatt   17520   tttataaata   aagttttatt   ggattacagc   caggcccatc   cagttacaga   attgtctatg   17580   gttgcttcca   ctctacaatg   gcagatttga   gtagctgtta   cagagactat   acgcaagtgt   17640   gcaggagagt   ttagaccaca   cctacaggaa   aggtatgaaa   tactaccctc   cttcctagac   17700   acttccccaa   acaaaaaact   taagtcagtg   tgggaacaaa   cactgttgcc   ccacagagca   17760   ctaataaagg   cagataaatg   actgagaaaa   ccctatccct   tggggacgga   ggccagaaat   17820   ggcaacacca   ataccattgg   aagtctccta   cttctaggta   aggacatgaa   attctctcat   17880   atgcaacacc   catcacagat   acaaggcagg   agtttgattt   tcatggggag   gaggaattgg   17940   aacactgaga   ataccccacc   cctaaggccc   aggcacactg   gcctatatgg   gtctgtggct   18000   gaactgggac   aaaagagaag   ccctaaaacc   accagtagca   agcattgagt   aacaagcagt   18060   agcagtatta   cagcaaaaca   ggcacagctg   aaagcagaga   gtggggcatt   aaaacccttc   18120   agcgtctcag   ctactatgct   aatagaaaat   ccaaagaaaa   atctgaagcc   cataggaggc   18180   tgaggatgac   cctactaaca   acaaaactca   agtctagcta   gactcctaag   cagattgatt   18240   taatacccca   ctctaacaac   cttaaagaag   tgtgcccatt   tccatgcata   aatactatgt   18300   atctcaatat   ttgatatact   ctacacaaaa   tgtttgccat   ttaatcaaaa   tgcatgagac   18360   tcaaaaagaa   aatgcaaccc   actgttaaca   gccaaagcaa   tcaacagaac   caaacagaga   18420   catgactcca   atgttggagc   tattagacac   ggactttaaa   taaaataatt   acaatgaata   18480   agaaaaatac   attaaaaata   cttcatgaaa   gaggatatat   ggatggcaaa   gaagcacctt   18540   taaaagtgtt   caacagttta   tacactattg   atggaaatgt   aaattagttc   ttccactgtt   18600   ggaagcagtc   tggagagttc   tcaaaaaact   taactaccat   ttgaattagc   aatcccatta   18660   ctgggcatat   acccaaacag   aagaaaactg   ttctcccaaa   aggacgcatg   cactcatata   18720   ttcatcatag   cactatgtat   attcacaata   ccaaagacat   ggaatcaacc   tgaatgccaa   18780   tcaatggatt   ggattaaaaa   atggaatact   atgcagtcgt   agaaaagaat   gaaatcatgt   18840   ctttacagca   acttggatgc   atctggaggc   cattatccta   agtgaattaa   tgcagaaaca   18900   gaaaaccaaa   tgccacatgt   tctcatttat   aagtgggagt   taaaccttgg   atacacatgg   18960   acataaagat   ggcaacagta   gaggggaaaa   ggaagaacta   gagcaaggga   tgaaaatcca   19020   gctattgggc   attatgctca   gtacctgtgt   aatgggatca   atcgtacccc   aaatctcagc   19080   accacacaat   atacccatgt   aataagcctg   cacatgtacc   ccctgaatct   aaattaaagt   19140   tgaaatcatt   tctaaaaaaa   gaaaagaaaa   atgaacaaat   tgaaaataaa   aagatgttca   19200   acatgattag   tcattagaga   aatgaaaaat   aaagccatga   tgaaatatca   ctacatatct   19260   agtagaacaa   ctaaaagaaa   acacactgac   agtcctgagt   gctagaaagt   gtgtgcaacc   19320   acagtctctc   tcatacattg   ttggtgggag   tacacagtgg   tgcagcaaca   ttggaaaact   19380   gctgggcagt   tttttatgaa   gttaaacata   tatttactta   ctctgtggcc   caacaattcc   19440   acttctgagt   atttgtccta   gagaaatgaa   aatttatatt   cacacaaaaa   cctgtacatg   19500   aatgtttata   tcagccttgt   ttgaaataac   aaaaaactga   aaataaccca   aacatatttc   19560   agtgagtgaa   taaacatatt   gtaatacatt   tatacaattt   aaaactattc   ggcaataaga   19620   acaaacaaat   gatacacaca   ccttagaaga   atttcaaagg   cattatgatg   aattattttt   19680   ttttttttga   gatggagtct   cactctgttt   cccaggctgg   agtgcagtgg   cgcaacctca   19740   gctcactgca   agctccgcct   cctgggttca   cgccattctc   ctgcctcagc   ctcccgagta   19800   gctgggatta   caggcgccag   ccaccacgcc   cggctaattt   ttttgtattt   ttagtagaga   19860   cggggtttca   ctgtgttagc   caggatggtc   tcgatctccc   gacctcgtga   tccacccgcc   19920   tcggcctccc   aaagtgctgg   gattacaggt   gtgagccacc   gtgcccggcc   tatgatgaat   19980   tttttaaagc   taatttttaa   aggttgcata   ttttgtaatt   ccatttacgt   aagattctta   20040   aaatcacaaa   actataatga   taaagaacac   attagtgttt   gctaggaaat   gggtgggtag   20100   ggagtgtggc   tataaaggga   tagcacaagg   gaatttctcg   acagttgtag   aactgttctc   20160   tatcccgact   gtggtgttag   tgacatgacc   ttatatgtat   gtgagattaa   atttcattga   20220   tatatgacat   gcttgtaaaa   attggtgatg   tgcaaataat   tctatagttt   agttaacagt   20280   attataccaa   tgtcaatttc   ctggttttca   tcattatact   gatattgacc   aggtagagcg   20340   tacatgggaa   acttctgtct   tatttttgca   actttaatat   gggtcttaaa   tgatttcaaa   20400   ataaacactt   taaaacattc   tccctactaa   ataaccatga   ttaataggtt   aaagtttcca   20460   ggggaaaaaa   gtggacaaca   tagatgaaca   gttgagtgat   ttcacagaaa   gataaaaact   20520   ataagcaaga   ttcaagtaga   aatgtgtggg   ggaaataaat   cacaatagta   attcacagat   20580   ttaattctag   gaatcgtaga   ctttacacag   ccagaggaag   aattaggtaa   tctgaagata   20640   gatccataaa   aattgcccaa   agtgaaaatc   agtaagaaaa   ggacaaagaa   agtggatgag   20700   gagaacaaag   tatttaaaaa   tgatgggaca   atatcaaata   gtctaaaata   tatgtcattg   20760   aagttacaga   aggagaagag   agaatgggca   aaagaaatct   ctgaatgata   gctggaattc   20820   tccaaaaata   atgaaagcat   caaatcatag   atccaaaatt   cttaaataac   ctgaagcaga   20880   ataaatttta   aaaagctagc   tataacttat   ccaaagtacc   aaataaaggt   aaagaagaaa   20940   ttgaaggaag   acagtggaaa   atatatgaac   atattaaatc   agagaaattc   ttaaagaata   21000   caaaattaca   gctagataga   actagaataa   gttctagtgt   tccatattac   tgtagaatta   21060   ctatagttaa   caataatata   tggtttcaca   taactaaaag   aatattgcat   gatcctcaca   21120   aaaaaatgat   aaatgctaga   gatgatagat   atgccagtta   tactctgata   actatatgta   21180   ctgagacatc   accatgtagc   cccatgaata   tgtacactta   ttttgtcatt   ttaaaaatta   21240   aattaatata   aagaggtaaa   gacaagaatt   atagctgact   tcttatctga   agcaattaaa   21300   gccagatgat   aaaacatatc   ttcaaagcca   gctgggggga   caactatcac   ccagaattct   21360   acatctagca   aaaatacctg   tttaaaatga   agatacaata   aggactattc   aaaatttttg   21420   taatgggaga   attccttacc   agcagacctg   tactttaaga   aacattaaag   aaagttcttc   21480   tgacagagga   atgtgaaacc   agaccggaaa   ttttatctac   agagagatgt   gaagaactct   21540   gaaaatagta   aaaataaaga   ttaatattaa   aagccacatt   tttcttaatg   ttaaatgact   21600   ctaaaatgta   attgactact   tgaagaaaaa   ttgtagcagt   gagatttata   acccatgtaa   21660   aatttaaatg   tctgataaca   atagcacaaa   gtttatgagg   aatttgaatg   tctaagtctt   21720   cacaatatgt   aagaatttgt   ataatattat   ataaaacatc   taatactcag   tctttacttg   21780   ctgcagggaa   gatcaatttg   gcaggtccag   cactggaatt   accatttaga   ttagttagga   21840   aactgcggaa   gttcaagaaa   gaaatcatca   gtacctagag   tatgccaaat   aatagtaaga   21900   atacagagag   gattcaaatt   tgagatctac   ttaataaggt   aaaataagga   ggatgcggtg   21960   actgagtatt   gagatgttct   gaatgtgagg   tagagggaag   agtctactat   gtattatggg   22020   ttcctggctt   gggtgtccct   agcagacagt   ggtgctatca   agtgagtctg   aggagagagg   22080   aggataggat   cattaatatg   aaaagttaat   gagtttggat   tttaactggg   gtttcaggca   22140   atttacatgt   aatgttatta   ttgatgtgtt   tggattgaaa   tctaccactg   gtttcttgct   22200   tttatttgtt   ccatctatat   ttttcttcct   tttttccttc   tatttcagcc   tggattaatt   22260   gagcttttta   tgatttcatt   ttgtctccac   tattggctta   ttaattatgc   ctctttttct   22320   atttctttta   atgttgtcct   aggcttcaca   atgtacatcc   tctctacatt   cctacagtta   22380   tttaacatac   gtttcttcct   tgaacttctg   catagtttcc   taattaaatg   gtaattccag   22440   tattggacct   aacaaattga   tcctcaccac   agcaactaaa   gaaatctatt   tataatacat   22500   gtttggagac   ataaacattc   aacaactctc   acagtataaa   aaaatacatc   ctcttttaat   22560   tggcatgtga   gacccagaag   tctggattct   tcttgtttct   gcagacccat   ttttgccact   22620   tcctagctcc   tatttaggct   tcagcaataa   caaacagatg   cttaatatac   acaagccaag   22680   tagtttctca   tctccataac   tttgttcatg   ctgttatatg   tgcctagaat   caatattctg   22740   cattcctttc   tcacctgacc   aatcctcact   gaaaagctca   gctattgccc   cattcttgac   22800   ttcttccttg   tccacccgct   tttgtgctcc   aactggaacc   ccattcatgt   ttacaagaaa   22860   cactcaaagg   acaataaagc   ctgttgtgac   tctacttcgg   cttttctgag   tattaaaatt   22920   tcctaagatc   tttcacagga   caaattttta   aaagcaagag   taatacatgg   aggtgattaa   22980   gcctttaaag   tgtacccttt   gttcccaagg   tcttgttaat   tatcatttag   gagcagttta   23040   tctgcaatgt   gctcaattac   ttaggtaact   aaatagcttt   ttactgatca   gcattgaaca   23100   tcatttgctt   aatagcaact   aacaaatgcg   gtggcagaat   tccactgata   gtctactttc   23160   agaaataggc   ttatttaaga   ttgagctcca   ggggaaaaat   agattctaac   agaatagatt   23220   attgttattt   acataaccac   ttaggtctag   agaaacaaaa   taatctttcc   gttttttaaa   23280   attaaggaat   cagtaagcca   aaagagaata   tccaagtaac   tcccaagtcc   attccgttca   23340   ttcatccagt   ccatttgtgc   catgaaaatc   ttcatgacat   tcctatttga   tcaatcacaa   23400   ctcacagctc   tcaatgactg   ctcacatctg   accttctcag   taaggacttt   tcttatagct   23460   tagaacagtc   ataggatttt   tcctctccaa   gcttttgaca   ctttggatta   tagatcccat   23520   gtgcttgcca   atgctttttt   tgttaactgt   agaaaaataa   aaagcaaata   cagacaggcg   23580   aaagactcag   agggtcacat   aacattgtat   ggtgaactat   accatgtgaa   tggtcaagga   23640   aagcttctgg   agcatgggac   atattagcta   ggtgttgaaa   gaaaaatagg   aattcaatag   23700   gctggaaaat   ggggggaggg   tatttcaagc   agagggaaca   cctatctaga   tgcacaaatg   23760   tatgatcaat   atggtgttca   acattgaaat   gccacatggg   gacctgacaa   gaggtagttg   23820   tattagtttg   ttttgtgctg   ctgtgatgga   atgccacaga   ctgtataatt   tataaacaga   23880   aatttattag   gacacagttc   tggaggctgg   gaagtctaag   gtccaggtgc   cagaaagttc   23940   agtgtctggt   taggtcattc   tcttccaaga   tggtgccttg   catgctgcct   agatcctcac   24000   gtggcagaag   gccaaaggga   gaaagaggat   gagcccattc   actttttata   atggcattaa   24060   tctattcatg   agggctcaac   ctaaatgctt   cccattaggc   cccactgtca   cattgggaat   24120   taagtttcca   acacaccaat   tcttgggcac   acattcaaac   tgtagcagta   ggcaaagcca   24180   aatagtggaa   ggcttttatt   ctaagctgtg   ggactcagct   ttattctgtt   aacagtgaag   24240   atgcagaaag   ggcgagatgc   agtggctcat   acctgtaatc   ctagcacttt   gagaggttga   24300   agtgagtgga   tcacttgagc   ccgggagttc   aagaccagcc   tgggcaacat   agtgagatcc   24360   cgtctctatg   aaaattttta   aaaagtaaaa   gttagctagg   tgtggtgtca   cgtactgttc   24420   ccaactactc   aggaggctga   ggcaagagga   tcacttgatc   ctgggaagtt   gaggctgcaa   24480   taagccatga   tcacaccact   gcactccagc   ctgagtgaca   gcaataccct   gtaccacttc   24540   cccaccaaaa   aagatgcaca   aaagggagat   ttatctccct   agttttctaa   aggagcaaaa   24600   gcaacttctg   tgggtggaat   agccctataa   gtagaagggg   ccaaaagata   aaaagggcaa   24660   ccaaagaagg   gaatgagggg   atgagaagaa   gacagggact   catatctgca   cattagtcta   24720   gggaataaag   aagaaaagac   acaacatcaa   tctaaaaaga   aataaatgaa   aacaataaaa   24780   ggcagtgttg   aaggcagtcg   agcactgact   tctatagtat   gggttaaagc   ccagcttcac   24840   ctctaacttt   ctctgaaatc   ttaaaatgtt   ctaacctttc   ttattttgtt   catctataaa   24900   atggagattt   tatctcagtt   tgttgtgaga   ttaaacaagt   taatgattct   aactacttag   24960   aaccttgcca   aggatataat   aactgcttaa   taacccctca   caatcatgtt   ctgtgttcta   25020   gagaagtcag   gttaagggta   gtaattctct   tttaaaccat   gaagtctacc   tagagttatg   25080   taggatgtca   ccattggggg   atgctttgcg   gtaggcacat   gaaacctttc   tgtaccattt   25140   attgtaacat   cttgtgagtc   tctaattatt   tcaaaataaa   agttttattt   tgttttgttt   25200   ttcttaaagc   cttttcaaac   cacaatggca   attcattcaa   gcatgttctc   agcacttgct   25260   gcatgccata   ccctgagctc   cttctgttct   aacattagta   aacctcattt   tccaatattc   25320   agaaagcaat   taaccaaatg   acagtgtgac   tttccatttt   tactattgct   ctttagcaga   25380   aacatagcaa   taaaaataaa   ctattggaaa   tgaaattatg   agtatcctag   attaatcccc   25440   aaagttgcag   tttcttttgt   tgttcagctg   tttgattttc   taagagcctg   aacccagcca   25500   agtgcatact   ctatttctga   gcagagttct   gcccagttct   ctgtctgact   agaattgcac   25560   agaaaccaca   aatctctcat   aaagcctaat   ggtgtttctc   aacagaggca   tgattggaat   25620   tctgagctgg   ataatttaat   actagggtac   agcccatggg   tagtacagaa   aagagtcaat   25680   acttgccttg   tctttgtaca   accaggatgt   atgtctgcat   gacaataatg   aagactcaat   25740   ttcagccccc   attcaccctt   ccacaacagc   aggggcaggg   caccctagag   cataaataat   25800   ctgaacaacc   ctgtgttggg   ggcctgctga   agatgcttaa   catccctggc   cccatccact   25860   aattgtctgc   catcaatcac   aacttgacag   ttacaagtaa   attgcctgtg   ttctccccat   25920   atgcctgcct   ttctcattgt   gtaaaaacag   ctctacctcc   atctgctagt   cactaccaat   25980   cactcttgcc   aagatggtga   ctcctcttct   tgcctgctag   tccctagaca   caaagagtcc   26040   aaagtgccct   ggtggcagcc   atagctttta   gttcaatggt   acctttgcta   tgttccctgg   26100   cagaagtaca   cttcctttgg   tgaccaggac   ctccaagttg   aagagcacag   aggtgcagat   26160   gaagaagtgc   aaagcccgtt   agtgggtcaa   tgggagtgat   gttaacttag   gccatttctt   26220   cctgtacccc   ttgattctaa   gacctatgta   tacttcctgt   tagggaccca   acaccatgaa   26280   gtgggctctg   atttaacaca   tatgctgcac   tttaaaggat   gggaccctaa   actttcagag   26340   cattaccttt   aagtggcact   tcatatgttc   cttcagtagt   tcattctagg   gctctaaaag   26400   tacagctact   tctgtctggt   gaagtatgta   atacaaacag   taggttctag   gatcaagagc   26460   tatcctacac   ccccttcaca   attaagtggg   tcccctgatc   acatactatg   ttgtgggatt   26520   tccattcttg   tttggtaatc   taaaagcccc   cagataacag   ttagctgtgt   tgactatgac   26580   tcagagatca   ggaaaggaaa   gcttataccc   agaataagtg   catcattgga   aggatggaca   26640   gctggctgat   gcaggacaga   ggggaaccaa   tgtagtcaac   ttgccacaaa   tcaaccagtc   26700   gaatctcctg   aagacctagt   aacctattga   gggctccttg   ttggtctctc   ttgttagcaa   26760   gttggacatt   caaaatcagc   agtagctaga   tttaccagga   taattggtag   tccattttat   26820   tggctaatga   gtagtcttgc   tctctgccac   tgtggccact   cgattcatgt   gcccatcatg   26880   ccagtactta   atccaaagac   aaagcctgga   taatgtcaac   tggacaagtt   atcttgtcta   26940   tttgtttttt   cagtgcttct   tgtgtacttg   atgctttcca   attagtgtta   aacgcatgac   27000   acaaaaatct   tcacacattg   tgcccactcc   catagtccca   accacatgtc   tctgtctcag   27060   actcttgttt   ccaatctcca   agtccctttc   attcaggtcc   ctgtccacac   agctagacta   27120   ttggctactg   tgcagaaacc   tatacatata   ttcacacctt   aggccaattt   ttcttccaca   27180   cgaatagata   accatattca   ttgcttgcag   cttcattcat   tgggaagacc   ttccttctcc   27240   actgtcttcc   aaggctaccc   cataattcag   ctgtacttca   gtgatcatct   atttttgctt   27300   tacagctact   taaggaacta   acccattgtc   agccaggatt   ggccttttac   ctccctttca   27360   gctagtcata   gagaaccccc   atatagccat   aagtgtgagc   tatgcaaggg   cactggtaca   27420   accctggtgg   gtgacatgga   ggcatctgtg   ctacttgctc   atgcagcctc   ttcatgccag   27480   tctagtccca   tatgggtttg   atcccagatg   tgccatttcc   attttccaat   agaaggctgt   27540   gggacacgtc   tgagtgtatg   acttagtgca   ttggcaataa   tctagctcat   aatggtcagt   27600   tccaaataca   tttggtgccc   cattatcagc   tgttccatct   tgaccaaggc   tcagcaacat   27660   tctaggaagg   gtttctcaaa   aggtgtataa   ctttgtactg   aaaatggatg   gcattgcttc   27720   agagccctag   gggcctgcat   tctgattctt   tcgcttgggc   atccatcaac   tccaaactgt   27780   atctttcccc   actaacacat   tcagcaccat   ggggtccacc   agatcatatg   cctcaagtag   27840   tagggttgct   tatactgcag   cccatacatg   ctgcagagcc   ctttcttgct   ttcttgctcc   27900   aggcccactc   aaagctgtga   gacttctatg   tagttcagta   tatgggacca   aaaatattcc   27960   tagatgtgga   atgtgttgcc   tccaaaacac   aaagaggcct   accaggtatc   ttgctctatt   28020   tattgagatt   gagaatgtaa   gatacagcaa   tttaactttt   acttgaaaat   gcagtggtgg   28080   gagtggtatt   ctagcagact   ttcactactg   aaccactaaa   aactttaccg   atgtggcatg   28140   ttcctgaatt   ttcacagggt   ttgtctctca   ccctccagag   tgtatgtgtc   ctaccaatgc   28200   ctctaatgta   ttagacactt   cttgttgact   cttaatgaca   tctttgatgt   aaatggatca   28260   atgtgatatt   ctacagaatg   ttcagatggt   ccatgtctct   tcagacctta   ttatgacaga   28320   tgtttgaaga   ggtaacatag   ccatgggcaa   agcagaaaac   gaatattatt   gtctgtttca   28380   tgtgaataca   gactctgatt   ttttggatgt   gtctttgtct   ggttttggta   tcaggataat   28440   attggcctcg   tagaatgagt   ttggaagtat   tccctcctcc   tctatttttc   agaatagttt   28500   gagtaagatg   atattagttc   ttcattaaat   gtttggtata   attcagcaga   gaagccatca   28560   gttcctgaac   ttttctttat   tggaagacat   tttattataa   gttcttgtta   tttgttattg   28620   gtcatagttc   aatcttggta   cattgtatgc   atctaggaat   ttgttccttt   cttctgtaat   28680   ttccaattta   tttacaattt   atttatttga   gtcttctctc   ttctttatta   gtctggctaa   28740   aggtttgtca   attttgcatg   acatttcaaa   aatccaactt   tttgtttcat   tgatattttg   28800   tattgttttc   ttcatctcaa   tttcatttat   ttctgctctg   atatttatta   tttattttct   28860   gctactaatt   ttgagtttgt   tatgctcttc   cttttctagt   tctttaagat   gcatcattaa   28920   gttgtttatt   taaagtttat   cttctttttc   aatgaaggta   ctaatagcca   taaacttccc   28980   tcttagtgat   tttgctgtat   cccatagatt   ttggtatgtt   gtgtttctat   tatcatttgt   29040   ttcaagaaac   ttttaaattt   tcttcttaat   ttcttcattg   acccaccagt   cattcaggag   29100   catatggttt   gatttccatg   tatttgtata   gtttccaaaa   ttcctcttgt   tattgatttc   29160   tagttttatt   ccattatggt   cagagatgat   gcttcatatt   atattatttt   attttgtttt   29220   aatgttttca   ggctttttag   tgatttaaca   tatagtctac   tcttgagaat   gacccatgtg   29280   ctgtggaaaa   taatgtgtat   tctacagctg   ttgtttggtc   tatagcgcag   attaactctc   29340   gtgtttcttt   accaattttc   tgtctggaag   atctgtccaa   tggtgaaatt   gtggtgttga   29400   agtctccagc   tattattgta   tggggtctat   atctctcttt   agctctaaca   atatttaatt   29460   tatatgtctg   ggtgttccag   tgttgggtgc   atatatattt   acaattgtta   tattctcttg   29520   ctgaattgac   ccctttttca   ctatatagta   accttcttta   tctcttccta   tagtttttgt   29580   cttgaaatct   attttgcctg   atataagcat   aactagccct   gctcttttgt   ggtttccatt   29640   gacatggaat   atcttttccc   atccctttat   tttccgccta   catatgcctt   atgtctttat   29700   tggtgaagtg   tgtttcttgt   aagcaacaga   tcattgaatc   ccttttgttt   tttcaatccg   29760   ttcagccact   tggtgacttt   tgattggcaa   gtctcagagt   ctcatccaaa   gtcttcaatg   29820   tacctgggta   ttgctgctgg   ttattctgag   cccagggact   ctttagttag   caggtgatga   29880   atgttgccag   gactatgtcc   ttcccttcaa   ggcagtagtt   tcccttctgg   cctagggcat   29940   gtctagaaat   gccatccagg   agctagagct   tggaaaggcg   gcctctcaga   gctgactagt   30000   gccctctcct   gttgtgactg   tgctggtatc   caagatgtaa   gacaaaatcc   ttcctactcc   30060   ttccttctcc   ttcttctcct   ttccttaagt   ggaagaaaga   gacctctttt   ggagctgtga   30120   gaattgcagc   ctggggttag   gggaggggta   gtgccagaac   tcccttagcc   acccaagctg   30180   gtatctcagt   agtccatgtg   cctccccagt   gtactggctc   tgggcccagt   tcagaactag   30240   aacttgcttg   aaagttgcac   tccttgtggc   ctagactgac   tttcaagtgt   atttagagcc   30300   ctagagcact   ttagcttgca   gtggtaaggc   ttgtgggaac   tcaagttctg   accaccagga   30360   ttggtgattt   ccttttggct   agagctaata   taaatgctgt   ctccatgggg   tggggcatca   30420   gctgagtttg   gcccagtttt   cttttatgct   ataataggac   agcactaagt   tcaatgcctc   30480   acaattgctg   tgcttttctc   ccctggcacc   taggaatgct   ctccacaccc   tgctgccatt   30540   ggtgggggat   gggagagggg   tggcgtcaga   gatttaaaac   tgtttttttt   ttttttctac   30600   ttcttcagtg   cctctttcag   caatatgaag   ttaaaaccaa   gcactatgag   agctcacctg   30660   atttttggtg   cttacaaagc   tacttttttt   gtgtgtagat   agttgctaaa   ttagtgtcct   30720   tgctgaggga   ggaagaggga   acgatcagtg   gaaccttcta   ttctgcaacc   ttgcctgcag   30780   cactcttttg   agaaagattc   tggattacgt   gataaattag   tgatgtctgt   catggagggg   30840   tgaggggcag   tttttagaca   tatagcctat   cttgccattt   ctgctataaa   catctgcaca   30900   gcttgtacaa   tctgtaacta   tagaattgag   tacagatgac   ctgcccacaa   gaggaaaaat   30960   gtcctgatct   tggccttagt   gcctctgtaa   ttcaaccaag   tatattaatt   aaatggatag   31020   atcatacact   tgctgtttcc   caagtgtaga   ggttgaaagt   ggaaaggaga   agaaagcaaa   31080   gaaaactggg   aggaggggca   ctaagactgg   gcataaggag   aaacaagagg   tccaatttgc   31140   aaaataatgg   gtgtagtaag   gggaattttg   aagtattctc   caaagataac   gattttgctt   31200   ttctcaattt   tgtcctcttg   attctcacac   agtgctttct   ttagtacaga   cctacatacc   31260   ccgaaaactt   ctgttctgtg   acttaaaaag   tcaactctat   gattagcatg   attcctcttc   31320   agtcattact   gaaattcttc   ctaggtggtt   acatttaagt   ggtaagaagc   ttcagtaata   31380   taagtcaaag   aactcctacc   ttgctttgca   tttcctcagt   ggccatctga   taaagcagga   31440   aggaaaaaaa   ttaaaaatga   acctccccag   gatttcattt   ctattgtggc   ttagctgcaa   31500   atttgaaaaa   taaaaataaa   aataaaccct   gaatgtaagc   acttgtacgt   ttttgttaga   31560   ttctttcaca   ggactggtca   acatcttgac   agatattaat   tggcgattca   ctggagactt   31620   cacggcacct   gacctggttt   gccgagtggt   ccgctatttg   caggtatgtc   acaccttcca   31680   aatgtgatga   gacaaactga   taccagagtt   agcttttaga   tttatctaag   gaaaatcata   31740   taaaaccttg   cattcctatg   catcaaactg   cccgctttct   cctgggtaga   agggcaatga   31800   aaatttgatt   tttaatgtat   ttcctttgct   aaaaatagta   tctatataaa   gaaaacacaa   31860   acgagccaca   aatgagcaat   gctaaggaca   taaagcattc   atatttcaaa   tgttaaataa   31920   aatgtctggg   tcaatattaa   ggattttggc   atattgtgga   taatctccaa   aaagtttgtt   31980   aattatattt   acctttcact   tctcaaatga   actcaacgct   ggctactgct   cttttttgta   32040   aaccaagggg   aatttgttgg   aacataacca   ttaactattg   aattactgcc   taaattatac   32100   tttttaaagt   atctattggc   atttgaattt   cttcctctca   aatttcctat   tttccttttt   32160   gaatgatttt   ataactgaga   gactttttag   tttttctaag   atctattttt   agattaaaga   32220   tattgatgct   tcaaatgtca   atctgcttca   ttgtaagcca   tatttatgta   tcatatgtat   32280   atttgattta   aaatattaca   tatatacata   tttacatata   tacatacttg   ttaatatgca   32340   ttagatgttt   ataatataga   tttagacact   tcaactaagt   ctggaaggaa   tggattattt   32400   cataactggt   attaggtttg   atgtgctgtt   tctgagttct   taagattatt   ttttattaat   32460   atatttttat   tctgatgcta   gtacttcagg   tctaaattat   tatagcttta   tgtatatata   32520   atgcatttct   agaaagcttt   atcctgctag   attccatttg   ttagtagact   cttttaggat   32580   ctttgtatat   atattcataa   gtaaaatttt   aatttttttc   attggtttta   ttattttcag   32640   gtcatggtaa   tgaagttatt   ccatattcag   aaaatgaata   tggaatagtt   acaatcttct   32700   tttgtgctct   gaatataacc   ttgaaagttt   ggtatgtctg   aaagagcctt   ttctattatg   32760   tacggctagt   attttctcag   gggaaatttt   taaaaacata   tttcaatttc   tttctgagga   32820   tatttgcttc   cttgtaaata   aatgctttca   attgcttatt   atattagcaa   tcaattgctg   32880   atacacaaat   tatgacaaaa   catggtggct   taaaacaata   aacatttatt   atctcacagt   32940   ttctattagt   caagaattca   ggaacagctt   ggctaggcag   atctggataa   aggcttatca   33000   aaaggttcgg   tcagatgctt   tgtggtactg   tagtcatcag   aacgcttggc   tgggactgga   33060   caagatggat   cactcacata   ggtggtaaat   tagtactggc   tttagtggaa   tactcagttc   33120   tccacatggg   cctctctata   ggtacatttt   agtgtcctca   caacatagta   gctggctttt   33180   ccaagaataa   cctattcaag   tgactgaggt   ggaagtggca   acatatttat   gaactagtct   33240   caaaggtcat   acaccatcac   tttcatatga   ttattttggt   tgtacaaatc   aacttggatt   33300   cagtatggga   aaagaagaac   cacaaaacta   ggaagtaagg   attattttcg   acatgttgga   33360   ggttggcaac   cacactatct   tcatattttt   ccaaagcatt   tgataaccta   tggtttctcc   33420   caattgttaa   ctttaaaatg   tcttcttgat   ttagaaatat   tcatgtcagt   aaaatttttt   33480   ctaactgtat   ctacaataaa   tgttaagact   atatataaaa   tatatctttt   gacataggga   33540   aaacgataga   gaaagaaatt   ctgataattc   tctcttctat   aaaggaagca   agaaaattgg   33600   caaaaaatat   cagaatcaaa   tttgtaaaac   tctggaaatt   aaccaaacac   ttgcaattac   33660   tcagggagca   tttattcagg   aaaatggctg   aatctcagtg   agaaaaatga   gctttgcggt   33720   gctttggctt   gcattattct   cagcccccag   tatccaactc   tgtaacagcc   ttaaaaagta   33780   atagctctca   ttcatagtga   aaacaaattg   tctggcagcc   actgtagaaa   gcaaaacagg   33840   gacagagccc   tttcaaattc   ccaagaaatt   gccattattt   gacatgtctg   gcagttctct   33900   ggaagactcc   acttgcaagg   ctgtctttat   tttacttaaa   tcagaactca   cccactgtga   33960   aaaacgtttc   tcctctgggc   attcgtcaaa   acagttacag   gcaattgatt   aactttgctt   34020   atgacaaagg   tattggataa   tagtcaggga   aaacaacaga   ctaaccaaag   cttaagaaca   34080   tacgctgagt   aatgagatat   ccatagggga   tttgaaaatc   tctgacatac   tcccaaaaat   34140   ctagaagacc   aggtatatgc   ccaggactat   gctcatccta   aggaaaaaat   agagaaggca   34200   ctgagctgta   acctgtgaat   gatcttgagg   ctttgtgcaa   gcaggaagtg   agggcaaaga   34260   cagaattgta   agttgcctgg   ctaagtgtca   aaatcatgct   ccaacattca   cacagacttc   34320   ctctgcaatg   aatgggggaa   ttaatagttt   ctagaatgta   agaaaatctc   tgtttaggac   34380   ctatcaccaa   gatcatggag   taggagatat   tcatccctcc   aaagcattca   agtatagaca   34440   gctattcaca   aactaaaata   gcccagagag   gactcaaggg   cccattaaag   aatttgcagc   34500   aacatcgtga   aggaaaaaat   ggagaataac   cacatagaaa   gaattgctgg   tgtgattggc   34560   acacctaaga   ggccatgatt   tggctaggaa   caaagaagaa   aggtggaagc   tatcagtatt   34620   agacacagaa   aggaccaaca   ttgtctctag   tggcctgctc   cacaggagac   actagcatct   34680   tttcccactg   agtaaccaac   agtcattcct   gcaagagaac   cccagagggg   gagatgtggc   34740   tataccttcc   tcctatcccc   caagaagcag   ttgctgtcga   gctgctttgg   gaatggggtc   34800   accacctctc   ccaaccatgt   gtaagctcca   gccctggagc   catggccacc   ctgggaatgc   34860   ccacactcca   gactgatagt   ctgactacac   tgggctcacc   catgttttta   acatcacagc   34920   cagcaccata   gtaagctagt   taacactcca   agccccagtg   ttaaactctg   cctgcatgtg   34980   tccacactcc   agacactgac   ttaaccacca   tagagagcta   gccccatcca   ggcctcagag   35040   ccattataat   tctgtgcata   cctgtgctct   cattcttgtc   tccttgacta   atttatacgt   35100   atactattac   caacattaca   gcaggggcac   ctgtgccctg   ggcaacagct   cagccataga   35160   aagctacgcc   ttgccccaac   cctgaagcca   ctgaaagact   gtgttctcag   ctcccacact   35220   actttgcatg   tgcactcatg   cctcacatac   tataccatgc   agcagaaggg   gcatctgcac   35280   cttgagcacc   aatgtcatta   ccattctaat   cccagagcca   taatctctcc   atgtgagccc   35340   atgcatcaga   cctcagtttc   atggctactc   catgagtgtc   acccatcaga   cactggtgat   35400   actgccacta   agagaggccc   tacaagccag   atctagtacc   aagagaaatc   ttctcagctc   35460   cagcttccct   ggtgggggga   aagatcagaa   gaaccttagc   agccatcacc   actgaagacc   35520   ctatcaattc   ttgctgctac   tgccaatatc   cactgctttg   gccactgagg   atccttatga   35580   tctttatcaa   ctctgacctg   agctgaaaaa   gctacacaga   gactacaaat   ctgcaccatc   35640   actgatgcta   ccccacccaa   caaaccctcc   ctcatagggc   atggtctttc   catagtgaac   35700   tagttcataa   aatctggaag   gtgattcttt   ttggttttgt   tgatgtttat   ctttccctta   35760   tattgatgtg   tgtgcatttg   actgctttac   caaacgcaca   cacatcaaca   taaggcaaaa   35820   ataaacataa   aaaaacagac   ataacatcat   caaaagaaca   cagtaatttt   acagtaactg   35880   gccccaatga   aatagagata   tatgcaatgc   ttgaccaaaa   aaatcaaaat   aattttttta   35940   ggaaagctta   gtgaacttaa   agtacagaga   agcaattcaa   tgaaatcagt   aaaacagtac   36000   aaccaaaaca   aagaattaaa   gagagagact   gaaattattt   tttaattaaa   tagaaattct   36060   ggagctaaaa   aacattataa   atgaaataat   aaatgcacta   gacagcatcc   agcagaaatt   36120   atcaagtaga   caaagaattt   gtgaagtgaa   aaacaggtta   tttgaaaata   tacaactaaa   36180   gaaaaagaaa   gaaaaaagaa   tgagaaaaga   tgaagaaagc   ttaagagatt   tatgaggcag   36240   catcaaaaga   gaaaatgtgc   aatgcattag   tccattctca   tactgctata   agaatgtatt   36300   caagactggg   taatttataa   gggaaagagc   tttagttgac   tcacagttct   acagggttgg   36360   ggaggcctca   ggaaacttat   aatcatggtg   gaagggaaag   caaacatgtc   cttcacaagg   36420   tggcaggaga   gagaagaatg   agaactgagc   aaaaggggaa   gccccttata   aaaccatcag   36480   atctcatgag   aacttactat   catgagaata   ccatggggga   aaccacctcc   atgatttaat   36540   tacctcccac   tgaattcctc   ccatgacatg   tagggattat   ggcaactaaa   attcaagatg   36600   agatttgggt   aggaacatag   ccaaactata   ttatacaagt   tataggagtt   atagaaggag   36660   atgagagaga   caaaaaaaat   atagattatt   taaagaaata   atagcagaat   agtttgcaaa   36720   tctggggaaa   gatacaaata   tccaggtgta   caagaaggtc   agaagtttct   aatcagattt   36780   aatgaaaata   agactacacc   aagacatatt   ataatcaaac   tgtcaaaaat   caaaaacaga   36840   gaaaatcctg   ataacagcaa   gagagaagag   gcaaatcaca   tataagagag   ttataacaag   36900   gctagcagaa   aatttctcag   caaaaacctt   gcagaacagg   agagactaga   agaacatatg   36960   taaggaagga   aaaacaaact   gccaaccaaa   aatatttacc   caacaaagct   gtcctttagg   37020   aatcaagtag   agagaaagat   ttccccagac   aaacaaaagc   tgaaagagtt   catcaccacc   37080   aaacctattt   tccaaaaatt   ctacacagaa   tcttcaagct   aaaggaaaaa   aaaaagagcg   37140   agctaattag   tgacatgaaa   atatataaaa   gtgattttgg   gctgagacaa   tggggttttc   37200   tagatataca   atcatgtcat   ctgcaaacag   ggacaatttg   acttcctctt   ttcctaattg   37260   aatacccttc   atttccttct   cctgcctaat   ggccctggcc   agaacttcca   acactctgtt   37320   gaataggagt   ggtgagagag   ggcatccctg   tcttgtgcca   gttttcaaag   ggaatgcttc   37380   cagtttttgc   ccattcagta   tgatattggc   tgtgggtttg   tcatagatag   ctcttattat   37440   tttgagatac   atcccatcaa   taccgaattt   attgagagtt   tttagcatga   agggttgttg   37500   aattttgtca   aaggcctttt   ctgcatctat   tgagataatc   atgtggtttt   tgtctttggt   37560   tctgtttata   tgctggatta   catttattga   tttgcatata   ttgaaccagc   cttgcatgcc   37620   agggatgaag   cccacttgat   catggtggat   aagctttttg   atgtgctgct   ggattcggtt   37680   tgccagtatt   ttattgagga   tttttccatc   aatgttcatc   aaggatattg   gtctaaaatt   37740   ctcttttttt   tgttgtgtct   ctgccaggct   ttggtatcag   gatgatgctg   gcctcataaa   37800   atgagttagg   gaggattccc   tctttttcta   ttgattggaa   tagtttcaga   aggaatggta   37860   ccagttcctc   agaaaaaatg   ctcaccatca   ctggccatca   gagaaatgca   aatcaaaacc   37920   acaatgagat   accatctcac   accagttaga   atggcaatca   ttaaaaagtc   aggaaacaac   37980   aggtgctgga   gaggatgtgg   agaaatagga   acacttttac   actgttggca   gaactgtaaa   38040   ctagatcaac   cattgtggaa   gtcagtgtgg   cgattcctca   gggatctaga   actagaaata   38100   ccatttgacc   cagccatccc   attactgggt   atttacccaa   aggactataa   atcatgctgc   38160   tataaagaca   catgcacacg   tatgtttatt   gtggcactat   tcacaatagc   aaagacttgg   38220   aaccaaccca   aatgtccaac   aatgatagac   gggattaaga   aaatgtggca   catatacacc   38280   acggaatact   atgcagccat   aaaaaaggat   gagttcatgt   cctttgtagg   gacacggatg   38340   aaattggaaa   tcatcattct   cagtaaacta   ttgcaaggac   aaaaagccaa   acatcgcatg   38400   ttatcacact   tagatgggaa   ctgaacaatg   agaacacatg   gacacaggaa   ggggaacatc   38460   acactctggg   cactgttgtg   gggtgggggg   aggggggagg   gatagcatta   ggagatatac   38520   ctaatgctaa   ttgatgagtt   aatgggtgca   gcacaccagc   atggcacatg   tatacatatg   38580   taactaacct   gcacattgtg   cacacgtacc   ctaaaactta   aagtataata   aaaaaagaaa   38640   gaaaatatat   aaaagtatat   atcactcact   gttaaaagta   agtacattta   caatactcta   38700   ataatgtaat   ggtggtgtgt   aaatcactga   tatcattagg   ataaaggcta   aaagacaaaa   38760   ctattaaaaa   taataatagc   tacaatagag   gtatacctca   gagatattgt   gggttcaatt   38820   ccagaatgcc   taaataaagc   aaatttcaca   ataaagctaa   tcacaatatt   tttgtttccc   38880   agtacatata   aaagttatgt   ttacatttta   ctgtagcctt   ttaaatgtgt   gataatatgt   38940   ctaaaaagta   tacatgttaa   tttaaaaata   cttaatggct   aaaataaaaa   gattaacagt   39000   catctgagcc   ttcatcaaat   attggccttt   ttactgttgg   aaggtcttgc   tttgatgttg   39060   atgactgctg   acttatcagg   gtggtggttg   ccaaaggttg   tggtggctgt   ggcaatttct   39120   taaaatcaga   caacaataaa   gtttgttgca   tcaattgact   tttccttaca   tataaaaaat   39180   tatctgtagt   atgcaatact   gtttgatagc   attttaccca   gagtagaact   tctttcaaaa   39240   ttggagtcaa   ttctctcaaa   tcttgccact   gttttatcaa   atatgtttat   gtactagttt   39300   aaattatttg   ttatcatttc   agcaatgttc   acagcattat   caccaagagt   agattccatt   39360   tcaaggaatc   actttctttg   ctcatccata   ataagcaatt   cctcatccat   tctagtttta   39420   ttaagaaagt   gaagcaattc   agtcacatgt   acacactcca   cttctaattc   tatttctctt   39480   gctatttcca   ccactttcgc   agtacctttc   tccagtgaaa   tcttgaaccc   atcaaagtca   39540   ttcataagag   ttggaatcaa   ccttctacac   tcctgtaaat   gttgatacat   tcaccttctc   39600   ccataaatca   tgaatgttct   taatgacatc   taagtagtga   gtccttctgg   aaggttttca   39660   atttcccttg   tcctgatcca   gcagaggaat   tactatctat   ggcagccatg   gccttacaaa   39720   atgtatttct   taaataataa   cactggaaag   tcaaaattac   tccttgatct   atgggatgca   39780   aaatggatgt   tgtgttagca   gaaatgaaga   caagtttaat   ctttttgttc   atcgtcatca   39840   aagctcttgg   gtaacaatgt   gcactgtcaa   tgagcagtga   tatttggaaa   ggaatctttt   39900   tttctgagca   gtagatatca   acagggagct   taaaatattc   agtaaaccat   gctgtgaaca   39960   gatgtgctgc   cctttaggct   ttgttgtttt   atttatagag   cacaggcaca   gtagattagt   40020   ctaattgtta   agggccctag   gatctttaga   atgatcaatg   agcattggct   tcaactcaaa   40080   gtcaccacct   gcattagctg   ttaaaaagaa   tatcatgatg   tcctttgaag   ctttgaagcc   40140   aggcattgac   ttctcctctc   tagctataga   agcgctagat   ggcatcttct   tctagtattt   40200   tactgtttgt   ttccactgaa   aaactatcgt   ttactgtagc   caccttcatc   agttatctaa   40260   cctcaatatt   ctggataact   taatgcaact   tctgtattat   cacttgatgc   tttaccttgc   40320   atgtttatat   tacagagaca   acttccatcc   ttaaacctca   tgaaacaaac   tatgctagct   40380   tcaaactttt   ctgctgcagc   tttcacatct   atctcagcct   tcatagaggt   gaagagagtt   40440   taagaacttg   ttctggatta   ggctttggct   taatgttgca   actagtttga   tcatatatcc   40500   agaccactaa   aactttctcc   atatcagcag   taagcttgtt   tcattttctt   gtaatttgtg   40560   tgttcactga   agtagtcatt   ttacttttct   tcaagaaagt   ttcccttaca   ttcacaactt   40620   ggctaactgg   cccaaaacta   agcttttggt   ctatcttggt   ttctgacatg   ctttttctcc   40680   ctaagctaag   tcatttctag   cttttgattt   aaagtgagac   atgtgtgact   cttcctatca   40740   cttgaacacc   taaagacaat   ttcagggcta   tttattggct   taatttcaat   attactgtgt   40800   ctcagagaat   aggaacagct   gaggagaggg   agacagatgg   aggaacaacc   agttggtgga   40860   gctgtcagta   tgcacacaac   atttatcagt   taagtttgtc   accttatatg   gatgtgattt   40920   acagcaccac   aaaacaatta   caatagtatc   atcaaacatc   acttgatcac   agatagcctt   40980   aatggacgta   ataataacaa   taatgaaaaa   gctcaaaata   accaaaatat   gacacagaca   41040   tgaaataaac   acatagtgct   ggaaaaatga   cagacttgct   caacacaggg   ttgccacaaa   41100   ccttcaattt   gtaaaaaaaa   tgcactatct   gtgaagaaca   ataaagtaaa   tcatgacaaa   41160   atgaggtata   tttgtagtgt   gttaagggat   atactatata   aaaaatttaa   attgtgacat   41220   caaaaactaa   aatgggggtg   gtagagtaaa   agtgtagagt   ttttgtgtgt   aatcagagtt   41280   aagttatcat   tttaaaatcg   ctcgttataa   gacattcctt   tgtaagtctc   aaggtaacca   41340   caaagcaaaa   cctgtagtag   atacatgaaa   gataaaaagt   aaggaatcaa   agcataccat   41400   tagagaaaag   catcaaccac   agaagaagac   aggaaaaata   gaagaaagga   aaaaaaacac   41460   acaaagcaga   aagagtggaa   gacagaaaaa   aaatatatga   tagccagaat   ataattatta   41520   aaatggcaat   ttagggcaca   tataactgaa   agtaaaaatg   gaaaaaagat   attccatgca   41580   agtggaaacc   aaaatagatg   aggggttgct   acatttacat   cacatagaat   agactttaag   41640   tcaataacag   taaaacaggg   caaataaggt   cattgcataa   taataaacgg   attatttcat   41700   caggaagaaa   taacagttat   aaatatatat   ggaccctaca   ctggatcacc   taaatatata   41760   aagcaaatat   taatagacct   gaagagagaa   atagactgca   ataaaatcat   aggtcttcaa   41820   aatcccactt   ttaataaaca   gattatctag   acagataatc   agtaaagaac   gattggattt   41880   aaactacact   ttagaccaaa   tggacctaat   agacacatat   agaacatttc   acctagtggc   41940   agcagaatat   ataatctttt   gaagcaccca   aggaacattc   tccaggataa   tcatagagta   42000   agccacaaaa   caagtcacaa   caaaattaag   attgaaatat   ataaattatt   ttttcaaaaa   42060   caatgacatg   aaactagtaa   tcaataatat   aaggaatttc   aaaaatgtta   caaacgtgaa   42120   aattaaacaa   gagactcctc   aacaatcaat   aggtcaaaga   agaaattaaa   agcaaaaatt   42180   taaaatatct   taagataaat   gaaaatgaaa   acacatcata   acaaagctta   tgggatgcaa   42240   caaaagcagg   tctaagaggg   gagtttatac   caataaatac   ctacatcaaa   caaaagattt   42300   aaaataaata   acctaatatt   acaacttaag   taactagaag   aagaagaaga   gtcagggttt   42360   ccctttcata   aaaagggaga   aataataaaa   attagagaag   aaagaaaaca   aatggagaat   42420   agaaaaacaa   tagaaaagat   caatgaagag   ttagttcttt   gaaaagataa   aattgacaaa   42480   cctttagcta   gaataactag   gaaaaagaag   aaaactcaaa   taaaattaga   aatgaaagct   42540   gagacattac   aactgatacc   atagaaacaa   aaagcatcat   aacaagcgac   tatgaaaaac   42600   taaatgtcag   ccaggcgtag   tggctcatgc   ctgtaatccc   agcacttcgg   gaggctgagg   42660   caggcagatc   acctgaggtc   aggagtttga   caccagcctg   accaacatgc   agaaaccccg   42720   tctctactaa   agatacaaaa   attagccagg   tgtggtggcg   ggcccctgta   atcccaacta   42780   ctcaggaggc   tgaggcaaga   gaattgcttg   aacccaggag   gcagaagttg   cagtgcgcaa   42840   aaaaaaaaaa   aaaaaaaaaa   aatagataac   ctagaggaaa   tggataagtt   cctagataca   42900   caaaaactac   caagactttg   agtattgaag   gaatggaaaa   tctgaacaca   ccaatactaa   42960   ggagatttaa   tcagtaataa   aatgtttcca   ttccaagaaa   agaccaggat   cagatggctt   43020   cactgcaaaa   ttctaccaaa   cacttagaga   agaactaata   gcaatctttc   tcaaagtctt   43080   cccccaaaaa   agtgaaaaaa   gtgtttactt   ccaaactcat   gttacaaagt   cagcattact   43140   ctgataacaa   agtcagacaa   gatactatga   gaaaattaca   gggcaataac   cctgatgaac   43200   ataggtttaa   aaatcctcaa   caaaatacta   gcaaacatat   ttcaatagca   cacttgaaat   43260   attattcacc   atgattaagt   gtaatttatt   cctggaatgc   aacaatagtt   taacatacac   43320   aaattaataa   atgtgacatg   ccacattaac   aaaatgaaag   ataaaaggca   tatgatcatc   43380   ttaatagatg   cagaaaaaca   tcgtgacaaa   attcaatatc   tttttgtagt   aaaaatgctc   43440   aacaaattag   gtacagaaga   gaagtgcttc   aacaaataaa   gaccacatat   gacaagtcca   43500   ctgctaacat   cacagtcaac   agtggtaaac   taaaagcttt   tcttctaaga   ttaggaacaa   43560   gataagaatg   ctcactattg   caacttctat   tcaacatagt   actggaagtt   ctagccaaag   43620   aagtaaggca   agaaaaaaag   aaaagaaaaa   agaaaggcat   tcacatcaga   aaggacaaac   43680   ttaaattttc   tgtttgaaga   tgacctgatc   ctacatatat   ataaaatata   taatttatat   43740   ataatatgta   ttatgttata   aatatatgtt   atatgttcta   tgttatatat   aatatatgtt   43800   ctatgttata   tataatatat   gttatatgtt   atatgttata   tataatatat   gttatatgtt   43860   atatataata   tatgttatat   gttatatata   tgttataggt   tatatgttat   atatgttata   43920   ggttatatgt   tatgttatat   atatgttata   tgttatgtta   tatatgttat   atgttatatt   43980   atatatcata   tatgttatat   gttatattat   atatcatata   tgttatatgt   tatattatat   44040   atcatatatg   ttatatgtta   tattatatat   gttatatgtt   atattatata   tgttatatgt   44100   tatatgttat   attatatatg   ttatatgtta   tatattatat   tatatatgtt   atatgttata   44160   tattatatta   tatatgttat   atgttatata   tattatatat   aatatgttat   ataatataat   44220   atatatgtta   tatattatat   ataatatgtt   atatattata   tatgttatat   attatattat   44280   atatgttata   tgttatatat   tatattatat   atgttatatg   ttatatatta   tattatatat   44340   gttatatgtt   atatattata   tgttataagt   tatatataat   atagataata   tataaaatat   44400   ataatatata   tacacaaaac   acaaaacacc   accaaaaact   gttaaaacta   acaaatgaat   44460   ttactatagt   tgcaggatac   aaaatcaaca   tgcaaagtca   gtaatgtttc   tatacactaa   44520   caatgaacta   tccaaaaaaa   aatagaatct   atttacaata   actccaaaaa   gaataaaata   44580   cgtagaaatg   aagttaatca   agtaggtgat   agacttatac   aaaaaaacta   taatatattg   44640   atgaaagaaa   ttaaactagg   caaaaattgg   aaaggcatcc   tgttttcata   aatatacaga   44700   cttaatactt   taaagtgtct   ataaatcaaa   gctatctaca   gattcaatgc   attccctatc   44760   aaaatcccaa   cagcatgttt   tacagaaatt   aaaaaaaatc   aaaaattcaa   attaaatcac   44820   aaaagtccca   gaattgccag   tgttaacttg   agaaagaaca   aagctcaagg   catcacactt   44880   cctgcttaca   aaaatatatt   atgaagctac   agtaatcgaa   acagtgttgt   actggcatga   44940   aggcagacat   atagacagat   gaaacagaat   aaagaagcca   gaaataaatc   catgtatgta   45000   tggtccactg   atcttcaaca   gtgatgccaa   aaataaacag   tggtgccaaa   aataaacaat   45060   ggggaaagga   cagtctcctc   aacaaggtgt   tggaaaactg   gacatctaca   tgagaagagt   45120   gcaatttaac   ccttaactta   tacaatacag   aaaaaaatca   actcaaaaca   gattaaagac   45180   ttaaatgtaa   gaccttaaac   tataaaattc   ctagataaaa   acaggggaaa   tgttccttga   45240   catttctctt   ggcagtgatt   tttttggata   agacaccaaa   aactcaagaa   acaaaaccac   45300   aaatagacaa   gtgggacaac   attaaactaa   ataactctgg   tgcaataaag   aaaacaatga   45360   atcaaatgaa   aaggtgacct   atggaaaggg   agtaaatatt   tacaagccat   acatatgata   45420   aagggttaat   atccaaaata   tattagaaac   atctacaact   caatttaaaa   aatagtacca   45480   ggcatggtgg   ctcacgcata   tagtcctagt   actttgggag   gttgaggctg   gtgaatggat   45540   tgagcgcagg   tattcaagac   ccgccaggga   aacatggcaa   acccctatct   ctacaaaaac   45600   acacacacac   acacatatat   atatatgtac   atatatacac   acacacatat   atatgtacat   45660   atatatatat   gtgtgtgtgt   atatatatac   gtatatatgt   gtgtgtatat   atgtgtatat   45720   gtgtgtgtgt   gtgtatgtgt   atatatatat   gtatatatat   atttatatat   atgtatatat   45780   atatatgtat   atatatacgt   atatatatat   atttatatat   atatatatat   attagccagg   45840   catggtgggt   gtgtgtctgt   agtcccagct   actcaggagg   ctgaggtagt   aggattgctt   45900   gaacccgggg   tgtcaaggct   gcagtgagcc   atgacagcac   cattgcactt   cagcctgggc   45960   aacagagaga   gaccctgtct   caaaacaaac   aaacaaaaac   ataggctgtg   tgcagtggct   46020   catgcttata   ataccagcac   cttgggaagc   caaagcagga   ggatttgaga   ccaggagttt   46080   gagaccagct   tgggaaacat   agcaagaccc   tgtctctaca   aaaagtaaaa   ataaaaataa   46140   ataaccaggc   atattggcat   gagttagtag   tcctagctac   atgggaggct   gaagtgagag   46200   gattgcttga   gcccaggagt   tcaagaccag   cctgggcaac   atagtgagac   ccccatctag   46260   gaaaacaaaa   agcacaggca   acaaaagcaa   aagtatacaa   atgggattgc   aggaaactaa   46320   aaagcttctg   cacagcaaaa   gaactatcaa   cagagtaaag   atacagttta   cagaatggga   46380   gactattggc   aaactatgca   tctgatgagg   agttaatatc   cagaatatat   gagaaactca   46440   aacaactcaa   tagcaaaaaa   caaaaaataa   acccaatcca   aaaaatgggc   aaatgacctg   46500   aatagacatt   tctcaaaaaa   tatatataca   aaagaagaca   tacaaatggg   caacatgtat   46560   acgaaaatat   gcttaacatc   cctagtcatc   agggaaatac   aaatccaaac   cacagtaaga   46620   taccacctca   gtctagttag   aatggctact   acagaaagac   aaatgataac   aagtgttggc   46680   aaggatgtgg   agaaaggaaa   ctcttacata   ttgttggtgg   gaatgtaaat   tagaacagtc   46740   actataaaaa   cagtatggaa   gttccttgaa   aaattaaaaa   ttgaactgcc   atatgatcca   46800   gcaatcccac   taatgggtat   aaacccaaaa   gtaatgaaat   cagtatgtca   aagagacatc   46860   tgctgtcctg   tgtttattac   agcactattc   ataatagcca   agatatggca   acaatctatg   46920   tccatcaatg   gatgaatgaa   tttaaaaatc   ttggtatata   cataatatgc   aatactattc   46980   attcataaaa   aagaaagaaa   ttgtttcatc   tgtgacaaca   tagatgaacc   tggagaacat   47040   tgtgtcaagt   gaaataagct   acacacagaa   agaaaactac   tgcatgattt   cactgatatg   47100   tggaatctta   aaaagttgat   ctcatagaag   ctgagagtag   aatggtggtt   agcagaagct   47160   ggtgagggta   agtgaaagag   ggtatgggag   aacactggtc   aatgagtaca   atgttaaagt   47220   taaatagagg   aacgggttct   agtgttctat   tgcacatcag   ggtgaatata   aaaaacaata   47280   gtgtattata   tatttcaaaa   taacttgaag   aaaggatttt   aaatgttctc   accataaaga   47340   aatgataaat   gtttaagatg   atggatattc   taattaccct   gatttgagca   ttacacaata   47400   tatacacata   tcaaaacatc   atatcgtacc   acataaatat   gtacgattat   tctggctcaa   47460   ttgaaaataa   aataaaattt   tagaaacgtt   aagaaaggaa   tattatgaac   aatcttatct   47520   cacagatttt   ataacctagt   taagatggac   caattccttg   aaagatacag   tctattataa   47580   ctcatacagg   gagaaatagg   caatctaaat   actgttaatt   gtattaaagg   aattaattat   47640   tgaataaact   tccaaaacag   aaagcatcaa   gcccagatgg   gttcactgga   gtattccacc   47700   aaacatttag   gaaagaaaac   tacaccaagt   ctctacagtt   gcattgaaaa   tatagaagca   47760   gaggcaatat   tttctaactc   attgtataag   gtcagcatta   cactaatacc   aaaagcagac   47820   aaaaatattg   caaggaaaaa   aacacatcaa   tatatctgat   gaatatagat   gcaatatttc   47880   tcaagaaata   ttagcaagtc   aatccaacaa   atttataaaa   ataattctac   acttgagata   47940   agcctgggca   acatagcaag   acactgtacc   tacaataaat   acaaaaatta   gccaagcatg   48000   gtggtgcata   cctgtactcc   tagctacttg   ggaggttgag   gcaggaggat   tatttgagcc   48060   caggggttag   aggctacagt   gagctatgat   tgtgccactg   cactccagcc   tgggtgacag   48120   agtgagaccc   tatctcttaa   aaaaagaaaa   aagaattata   caccacaaac   aagtaaggct   48180   tatcccaaaa   aagtaagact   ggttcaacac   tggatatcag   ttaatgtaac   ccattatatc   48240   aataggctaa   agaagaaaag   tcacaagatt   ataacaacag   atttttaaaa   agtgtttaac   48300   aaattacaac   acctattatg   atttcaaaaa   aaattcagta   aactaggaat   agaaagtaat   48360   cttcctcaac   ttgaaaaata   acatctacaa   aacagccttc   atctgacatc   ttatttaata   48420   gtgagaaact   atgagctttc   ttgctaagat   taggaagaaa   acaaggattt   ccctctcaca   48480   ttccttttca   tcattgtaca   gtaagtccta   gctagtgcaa   tgagacaaaa   aaggaaataa   48540   aagacacact   aattggagga   gagaaataaa   atgttgcagg   tgacatgatt   gtctatgcag   48600   acaatctaaa   ataattaacc   aaaaaaactc   tcccagaact   aataggcaat   taaagcaaga   48660   ttacagaata   tgttaatata   caaaagttca   ttgctttcct   atataaccgc   aataaacaag   48720   tataatttga   tattaaaaaa   caatattatc   tgcattagca   cacctgaaaa   tgaaataatt   48780   aggtataaat   ttaataaaat   atgtgcaaga   tctatacaag   gaaaactacg   aaacttgaat   48840   gaaagtaatc   aaagaaaaac   taaataaaga   gttattccat   gtatacatag   gaagattcaa   48900   tattgtcaag   gtgtcagatc   ttctcaactt   actctatgta   gattcaacac   agtcccagtc   48960   agtaacccag   caagttattt   tgtggatatc   aacaaactga   ttctaaactt   tatgtgaaag   49020   acaaaacatg   cagaatggtc   agcataatat   tgaaggtgaa   gaacaaattt   gaaggactga   49080   tacttcccaa   aatcaatatt   tactataaag   ctatagtaat   taagacagtg   tgctattagt   49140   gaaagaacaa   acaggtcaat   ggaacagaat   agcaagccca   gaaatagtct   ggcaaaaata   49200   tagtcaacca   atatttacaa   aaaagcaaaa   tacaatggag   aaaaggtcat   aattttaaca   49260   aatggtgctg   aaacaagaag   acatccacac   gcctaataat   aaatctaaac   acagatctta   49320   tatccatgac   aaaaacgaac   ataaaagaaa   acctagatga   ccttgagtat   ggtgatgact   49380   ttttagatac   aacaccaaag   gcatgatcca   tgaaaaaaag   aactgataag   ctggcattta   49440   ttaaaattaa   aattaaaatt   tctgctctgc   aaaaggcaaa   tgagaagcaa   accacagact   49500   aaagagaata   tatttgcaaa   aggcatacct   gataaaagat   tgttattcaa   aatatgcaga   49560   agagctctta   aagcacaata   ggtaaatgaa   caatctgatt   ttaaaatggt   caaaaaaaca   49620   gacacctcac   caaaaaatta   cacagaaggc   aaataagtaa   atgaaaagat   gctcatatac   49680   ttatgaatat   ctttgttatt   agggaattgc   aaattaaaac   aatgatatag   cactgcatat   49740   ttattagaat   aatcaaaatc   taaaacatta   acaacacaaa   atgctattga   gaatgtgaag   49800   caacaggaac   gttcactcat   tgctagtggg   aatacaaaat   agcacagcca   cttcagcgac   49860   agtttagtgg   tttattccaa   aattacattc   ttaccatgca   ttccagcaat   tgtgctcctg   49920   ggtatttacc   caaataagtt   gaaaacatat   attcccacaa   acacctgcat   acaaacgttt   49980   atatcatgtt   tattcctatt   ttccaaactt   ggaagcaacc   aagatgtcct   ttggtaggta   50040   aatggagaaa   taagctgtgg   tacatccata   caatgaaata   ttattcagtg   ctaaaaagaa   50100   atgagctgtc   aagcttattt   aaaggttacg   tgtctcatgc   aaagacatgg   aagaaattta   50160   tgcatattag   taagcaaaag   tgacagtctg   agaagctaca   tactgtagat   tccaacttta   50220   tgacattctg   gaaaaggcaa   aactctggag   aaagtgaaaa   cattagtggt   tgccagggct   50280   tggggagagg   gaggcagagc   acagaggggg   aacaaagggc   agtagaacta   ctctgtacta   50340   tactataaca   gtggacacat   gtctatactt   ttgtcaaaat   acaatgtata   acaccaagag   50400   tgaatcctaa   tgtaaactat   gaactttggg   tgataatgat   gtgtcaatgt   aggttcatcc   50460   actgtaacaa   atataccact   cttgcatgga   agcgtgatgg   tgagtgaggc   tctgtgtatg   50520   ggggacaggg   ggtttatggg   aaatctgtaa   cttctgctca   attatgttat   taatctaaaa   50580   ctgctctaat   aataaaatct   acttttaaac   atgtaataga   attcccaaga   actgtgggac   50640   aattacacaa   agtgtagcac   acacatataa   cagaaatacc   aaaaaaagaa   aagagcagaa   50700   gaagtatttg   aagaaataaa   agatgaaaaa   ttttctaaat   taatggcaaa   caccaatcca   50760   gagatccagg   aagctcagag   aacaccaaag   caaaataaat   gtcagaaaaa   atgtacctct   50820   aggcatatca   tattcaaact   gcagaaaacc   aaagacaaag   ataaaatcct   gatagaagtt   50880   gagggggaaa   cttacagata   aataaacaat   aataagaatt   tcctctgact   tatcagaaat   50940   cagccaagta   agaagatagt   ataataaaat   atttaaagtg   ggaaaagaat   acaaatcccc   51000   ctgacctaga   attctctatc   aagtgaaatt   attcttcaaa   agtgaagaag   aaataaagac   51060   tttctcagac   atacaaaaac   aggaaatttc   tcaaaagtaa   acttttctta   caaaaaatgc   51120   taaaagaagt   tcttcagata   aaggggaaaa   tgataaaata   ataagcacaa   atcaatataa   51180   aggaagagca   attgagaagg   aataaatgaa   ggcaaattaa   agctactttt   cttactctta   51240   attgacctat   agatagttgc   tcgacacaaa   aatagcaata   atgcattggt   aatcattatt   51300   accacttatt   aggataagtg   aactgaatga   caatattaaa   agagatgcag   cccaggcacg   51360   ggtggcccat   gcctgtaatc   ccagaacttt   gggaggccga   ggcaggtgga   tcacttgagg   51420   tcaggagctt   gtggccagcc   tgaccaacat   ggtgaaaccc   cgtgtctact   aaaaatacaa   51480   aaataggcca   gatgtggtgg   cacgcacctg   taatcccagc   tactcgggag   gctgaagcag   51540   gagaatctct   tgaacccgga   aggcgaaggt   tgcagtgatc   caagattgta   ccactacact   51600   ccagcctggg   tgacagagtg   agactctgtc   taagaaaaag   gggcgggggg   ttgaatgcaa   51660   agagaaatac   aatttaactg   aacagctatc   agttaaagtg   aactgaacta   ttaattaaat   51720   ggatctaatt   ggcatttgta   gaatacttca   tccagtaata   gcagaatata   tcttcttatc   51780   aaacttatat   gcgatttcca   tcaagagaaa   cttcatatgg   accataaaac   acattttaac   51840   aatgtaaaaa   aatagaaatt   acacaaagta   tactctcaga   acacacagaa   tagaactaaa   51900   aatcagtaac   agaaggatag   ctgaaaattt   tcaaactgtt   tggagattga   acaacacact   51960   tctaagtaat   aagtgaattg   aagaagatat   ctcaagaaat   attttgaaat   aaatgaaaat   52020   gtaacatcat   agtttgtagg   atgcagtaaa   agcagttctt   tgagtgaaat   ttataacact   52080   gaatggatat   attagaaaat   aaaaaaatgt   gatattaata   aactaatctt   aggccttagg   52140   aaactagaga   aagaaaagaa   atgtaggcct   aaagaaagca   gaataaaaga   aatgataaaa   52200   attggagcat   aaattattaa   attgaagtag   aatttaaatt   ttctctgcag   agtaagtatg   52260   tgaggtaatg   aatatgttaa   ttagctttac   ttagccattc   cacatgtata   cataaacacg   52320   ttatacagca   taaacgtata   ttattttgtt   tttcaattaa   aaatttaatt   aaaatcagaa   52380   aaaattaata   attaaattta   aaacaggaat   tcaatactga   aaaatcaatg   aaaccaaaag   52440   ctgtttcttt   aaaaagataa   taaaattgat   aaatttctag   ccaggctaat   caagaaaaga   52500   agaggattca   acattcttaa   agaaaacaat   tttcaaccca   gaatttcata   tccagccaaa   52560   ctaagcatca   taagtgaagg   agaaataaaa   tattttacag   acaagcaaat   gctgagagac   52620   ttttgtcacc   accaggcctg   ccctaaaaga   gctcctgaag   gaagcactaa   acatgcaaag   52680   gaacaaccgg   taccagccac   tgcaaaacca   tgacaaaatg   taaagaccat   cgagagtagg   52740   aagaaactgc   atcaactaac   gagcaaaata   accagctaac   atcataatga   caggatcaaa   52800   tttacacata   acaatattaa   ccttaaatgt   aaatgggcta   aatgctccaa   ttaaaagaca   52860   cacactggca   aattggataa   agagtcagga   cccatcagtg   tgctgtattc   aggagactca   52920   tctcatgtgc   agagatacac   ataggctcaa   aataaaggga   tggaggaaga   tctaccaaga   52980   aaatggaaag   caaaaaaaag   caggggttgc   aatcctagtg   tctgataaaa   tagactttaa   53040   accaacaaag   atcaaaagag   tcaaagaagg   ccactgcata   atggtaaagg   gatcaattca   53100   acaagaagag   ccaactatcc   taaatatata   tgcacccaac   acaggagcac   ccagattcat   53160   aaagcaagtc   cttagagacc   tacagagaga   cttagactcc   aacacaataa   taatgggaga   53220   ctttaacacc   ccactgtcaa   tattagacag   atcaacgaga   cagaaggtta   acaaggatat   53280   tcaggacttg   aactcagctc   agcaccaagc   agacttaata   gacatctaca   gaactcttca   53340   ccccaaaaca   acagaatata   cattcttttc   agcaccgcat   tgcacttatt   ccaaaattga   53400   ctacataatt   ggaagtaaag   cactcctcag   caagtgtaaa   agaacagcaa   tcacaacaaa   53460   ctgtctctca   gaccacagtg   ctatcaaatt   agacctcagg   attaagaaac   tcactcaaaa   53520   ccgcacaact   acatggaaac   tgaataacct   gctcctgaat   gactactggg   tacatagtga   53580   aatgaaggca   gaaataaaga   tgttctttga   aaccaataag   aacaaagaca   ctatgtacca   53640   gattctctgg   gacacattta   aagtagtgtg   tagagggaaa   tttatagcac   taaatgccca   53700   caagagaaag   caggagagat   ctaaaattga   caccctaaca   tcacaattaa   aagaactaga   53760   gaagcaagag   caaacaaatt   caaaagctag   cagaaggcaa   gaaataacta   agatcagagc   53820   agaactgaag   gagatagaga   cacaaaaacc   cttcgaaaag   tcaatgaatc   caggagctgg   53880   ttttctgaaa   agatcaacaa   aattgataga   ctgctagcaa   gactaataaa   gaagaaaaga   53940   gggaagaatc   aaatagatgc   aataaaaaat   aataatgtgg   atatcactac   caatcccgca   54000   gaaatacaaa   ctaccatcag   agaatactat   aaacacctcc   atgcaaataa   actagaaaat   54060   ctagaagaaa   tggataaatt   cctggacaca   tacaccctcc   caagactaaa   ccaggaagaa   54120   gttgaatctc   tgaatagacc   attaacaggc   tccaaaattg   aggcaataat   taataggcta   54180   ccaaccaaaa   aaagtccagg   accagaccaa   ttcacaactg   aattctacca   aaggtacaaa   54240   gaggagttgg   taccattcct   tctgaaacta   ttccaatcaa   tagaaaaaga   gggaatcctc   54300   cctaactcat   tttataaggc   cagcatcatc   ctgataccaa   agcctggcag   agacaaaaca   54360   aaaaaaagag   aattttagac   caatatcctt   gatgaacatc   gatgtgaaaa   tcctcaataa   54420   agtactggca   aactgaatcc   tgcagcacat   caaaaagctt   atccaccatg   atcaagtcgg   54480   cttcatcccc   gggatggaag   gctggttcaa   catatgcaaa   tcaataaatg   taatccagta   54540   tataaacaga   accaaagaca   aaaaccacat   gattatctca   atagatgcag   aaaaggcctt   54600   tgacaaaatt   caacaaccct   tcatgctaaa   aacactcaat   aaattaggta   ttgatgggac   54660   atatctcaaa   ataataagag   ctatctatga   caaacccaca   gccaatatca   tattgaatgg   54720   gcaaaaactg   gaagcattcc   ctttgaaaac   tggcacaaga   cagggatgcc   ctctctcacc   54780   actcctattc   aacatagtgt   tggaagttct   ggccagggca   attaggcagg   agaaggaaat   54840   aaagagtatt   caattaggaa   aagaggaagt   caaattgtcc   ctgtttgcag   atgacatgat   54900   tgtatatcta   gccccatcat   ctcagcccca   aatctcctta   agctgatagg   caacttcagc   54960   aaagtctcag   gatacaaaat   caatgtacaa   aaatcacaag   cattcttata   caccaataac   55020   agacaatcag   agagccaaat   catgagggaa   cacccattca   caattgcttc   aaagagaata   55080   aaatacctag   gaatccaact   tacaagtgac   gtgaaggacc   tcttcaagga   gaactacaaa   55140   ccactgctca   atgaaataaa   agaggataca   aagaaatgga   agaacattcc   atgctcatgg   55200   gtaggaagaa   tcaatatcgt   gaaaatgccc   atactgccca   aggtatttta   tagattcaat   55260   gccatcccca   tcaagctaca   aatgactttc   ttcacagaat   tggaaaaaac   tactttaaac   55320   ttcatatgga   accaaaaaag   agctcacatc   gcgaagtcaa   tcctaagcca   aaagaacaaa   55380   gctggaggca   tcacactacc   tgacttcaaa   ctatactaca   aggctacaat   aaccaaaaca   55440   gcatggtact   ggtaccaaaa   cagagatata   gatcaatgga   acagaacaga   gccctcagaa   55500   ataacgccgc   atatctacaa   ctatctgatc   tttgacaaac   ctgagaaaaa   caagcaatgg   55560   ggaaaggatt   ccctatttaa   taaacggtgc   tgggaaaact   ggatagccat   atgtagaaag   55620   ctgaaactgg   atcccttcct   tacacctcat   acaaaaatca   attcaagatg   gattaaagac   55680   taaattgtta   gacctaaaac   cataaaaacc   ctagaagaaa   acctaagcat   taccattcag   55740   gacataggca   tgggcaagga   cttcatgtct   aaaacaccaa   aagcaatggc   aacaaaagcc   55800   ataattgaca   aatgggatct   aattaaacta   aagagcttct   gcacagcaaa   ggaaactacc   55860   atcagagtga   acaggcaacc   tacaaaatgg   gagacaattt   ttgcaaccta   ctcatctgac   55920   aaagggctaa   tatccagaat   ctacaatgaa   ctcaaacaaa   tttacaagaa   aaaaacaaac   55980   aaccccatca   aaaagtgggc   gaaggacaag   aacagacact   tctcaaaaga   agtcatttat   56040   gcaggcaaaa   aacacatgaa   aaaatgctca   ccaccactgg   ccatcagaga   aatgcaaatc   56100   aaaaccacaa   tgagctacca   tctcacacca   gttagaatgg   caatcattaa   aaagtcagga   56160   aacaacaggt   gctggagagg   atgtggagaa   ataggaacac   ttttacactg   ttggtgggac   56220   tgtaaactag   ttcaaccctt   gtggaagtca   gtgtggcgat   tcctcaggga   tctagaacta   56280   gaaataccat   ttgacccagc   catcccatta   ctgggtattt   acccaaagga   ctataaatca   56340   tgctgctata   aagacacatg   cacacgtatg   tttattgcgg   cactattcac   aatagcaaag   56400   acttggaacc   aacccaaatg   tccaacagtg   atagactgga   ttaagaaaat   gtggcacata   56460   tacaccatgg   aatactatgc   aaccataaaa   aaggatgagt   tcatgtcctt   tgtagggaca   56520   tggatgaaat   tggaaatcat   cattctcagt   aaactatcgc   aaggacaaaa   aatcaaacac   56580   cgcaggttcg   cactcatagg   tgggaattgg   acaatgagaa   cacatggaca   caggaagggg   56640   aacatcacac   tctggggact   gttgtggggt   gggaggaggg   gggagggata   gcattaggag   56700   atatacctaa   tgctaaatga   caagttagtg   ggtgcaccac   accagcatgg   cacacgtata   56760   catatgtaac   taacctgcac   attgtgtaca   tgtaccctaa   aacttaaagt   ataatagtaa   56820   taaaataaaa   tttaaaaaaa   gatagcatga   gtagaaaaaa   aaaaaagaag   agggaagcca   56880   caagttacta   atatcagaaa   tgaaagaggg   gtcatcacta   ctgattctgt   ggacattaaa   56940   agaatgataa   ataaatacca   taaacaactc   tatgcccaca   agtttgatag   cttagatgaa   57000   atgaacctat   tccttgaaaa   gctcaatctg   ccaaaactca   cataaagtga   aatagataat   57060   ttgaataggc   ctatacctac   taaagaaact   gaattaataa   ttaataatct   tccacaaaag   57120   aaaacaccag   gcccagatgg   gttcattagt   gaattctgtc   aatcatttaa   aaaaaaaatg   57180   gtaacaatta   tctgcagtct   tctctagaaa   acagatgaaa   agggaaccat   tcctaattca   57240   tatacttgct   gacttgttcc   atgaggccag   aaatgcccta   attctaaaac   caatgatatt   57300   aaaagaaagg   aaaactacag   aacaatatcc   ctcatgaaca   taaatgcaaa   aattctcaat   57360   aaatattagc   aacttaaatt   tttgtttaaa   ggtataatca   tgtatagaaa   gaattatagt   57420   ccgtggccaa   gtgagattta   ttccaggcat   gaaaatctgg   ttaaatattt   aaaaatcaat   57480   taatataatc   tatcacacaa   acaagataaa   gaaggaaaat   tatgtgattt   ttccatcaat   57540   ggacacagga   aaagcattta   acaaaattca   acatccattt   ataataaaaa   ctcagcaaac   57600   tggaaacaaa   gagaaactgc   ctcaacttca   ttaacagcat   ctaaaaaaac   catacagtta   57660   acctcatatt   taatggcaag   aaattaggtg   cttttgtcct   aagactggga   acaaggtaaa   57720   gatgtcctct   ctcgccactc   ccattcaaca   ctgaactaga   tgtcttagat   aatttaacag   57780   gacaagaaaa   gtaaataaaa   agtttataca   ttggaaaaga   tgaaataaaa   atgtctttgt   57840   taataaattt   gtaattttct   atattaaaat   ctgaaagaat   agacaagaaa   aataaaaaca   57900   aacaaaaaag   aaacttccag   acctagtagg   caaaaaaatg   gcaacactga   agaatagaag   57960   gttaatatac   aaaagtcagt   tgtttttttc   cagcaattac   caagttgaga   ttgaaattaa   58020   aaacacaata   ccatttatgt   gaacaccaaa   aaatgaaact   taggtgtaaa   tctaacaaaa   58080   tatatacaag   atctattgaa   gacgactaca   aaactgatgg   aaaaaaaagc   aaatatctaa   58140   ataaatgaag   agatgttcca   tgttcatgaa   taggaagact   taatattgtt   aagatgtcta   58200   ttctgcccaa   cgtgatctat   aaattcaata   caattccaat   caaagtccca   acgagatatt   58260   ttatcaatat   aaacaaactc   taaaatcata   tggaaaggca   agagacacag   aataaccaat   58320   gcaatattaa   agaaaaagag   caagtttgaa   caaccaacac   tacctgattt   taagacttac   58380   tataaagctt   cataatcaag   acagcatggt   attggtaaaa   gaatcaacaa   gtagattgat   58440   gaaacagaac   agagagccca   gaaatagaca   tacaaaaaga   tagtcaacta   gtctttgaca   58500   aatgagcaaa   ggcattgaga   gatagccttt   tcaagaatgg   ctgtcttagt   tgaaagagtg   58560   gttgaatatc   cacatgcaaa   aaaaaaaaaa   aaaaaaaaac   ttaatgtaga   ccttatgcct   58620   ttcacaaaaa   ccaactcagt   ttagatcata   gatataacat   aatgctcaat   actattatat   58680   aaaacgacta   gaagataaca   taggagaaga   tttaagggac   ctactctttg   tctatgagtt   58740   tttagataca   acaccagaag   cataatccat   gaaaaatgat   tgaaatgttg   gacattatga   58800   aaatttacaa   ctttgcaaag   gacactgttt   aagagaatga   aaagacagcc   ctcagaatgg   58860   gagaaaatat   ctgcagaata   tatatttcat   aaaggatttg   tatccaaata   tgtacaaaga   58920   actcttaaaa   ctcaacaata   agaaaacaac   tcatttcaaa   aacggccaaa   agatctgaat   58980   aaacccttca   ccaagaagat   ggcaagtaaa   cagatagaaa   gaagctcaaa   atcatatgcc   59040   attagggaat   tgcaaattaa   aacaacagtg   agacaacact   acacatctat   tagataatca   59100   ctaaaattca   aaaaaaactg   aaaataccaa   tggctgatga   ggatgaatat   taacaggaac   59160   tttcattcat   tgctgctggg   aattcaaaat   agtacagtca   ctttggaaga   taatttagca   59220   gtttcttaca   aaactaagca   tggtcttacc   atacaaccca   gcaattgtaa   ggctatgtat   59280   ttactcagtt   gagttgacaa   cttatatcca   cagaagaatg   tttattgtag   ctttattcat   59340   aatcaccaaa   aactggaagc   aaacaagagg   ttcctcaaaa   gctgaatgga   taagctgcgg   59400   tacatccatt   aaatgggata   ttattcaatg   atataacaaa   atgggctatc   cagccatgaa   59460   aagacatgga   ggaatcttaa   atgcccattg   ctaagtgaaa   gaagccagtc   tgaaaaagct   59520   acatactaaa   tgattcaact   atattacatt   ctcaaaaagg   caaaactgta   gaagagcaag   59580   aaaccagtgt   tgccagagat   ttggagtagg   aaagagatga   atagataaaa   gcagtggatt   59640   tttaggacag   tgaaactctt   ctgatattat   aatggtgggt   acataacatt   atgcaatggt   59700   caaaacccat   agaactgtcc   aacacagagt   gaaccttaat   ataaacaatg   gactttagtt   59760   aatgataata   tatccatatt   ggttcatcag   ttttaacaaa   tgaactacaa   gatgttaatg   59820   acaaggaaaa   gtatatagat   gggagggagt   catagggaaa   ccctctgtgt   tgtttgcaat   59880   tttttgtaaa   tataaaattg   ttctgaaaaa   taaagtctat   tttcttaaag   tcagtccagg   59940   gatcctcctt   tatatgaata   caatctggca   ttttgctaag   taaaatttct   ctgtttccag   60000   catttaagtc   tcacttctct   gtgcaagatt   ttctttagtt   ttatatttcc   taggtcattt   60060   ctgtcagaat   taaggggtgg   ggtagtgaga   attcagacat   atgtcctcaa   gctgccatct   60120   gttctccaaa   tggtatccca   tatgttgttg   atgtgtagag   ttttcattgt   tgacatttat   60180   gaattgacta   taattgtact   ttgatttcct   tcttgactcg   agttaattat   gagggtgttt   60240   aaatttccaa   gtggtatttt   ttaaaaaaac   atgttactaa   tttctgcttc   attgctcatt   60300   gttttaaata   tgtgatttgc   gtaagtctta   ctttttagag   ctattgagag   ggtttttttt   60360   cttggtagtt   tagtatatcg   ccaatttttg   aaatgtctag   atattttata   gcaatgtgta   60420   ctattcttat   gttatgaagt   ctattttttt   ctatctgtat   atttttgttt   attcttctgt   60480   aaaactttga   acggtttcat   tatatattta   gcattatgtt   acttgttaca   tataactttg   60540   tgaatatcct   attgccatca   tgtgttgtat   cttttatcaa   tataaataat   cctttatcac   60600   atgaaatgct   gtttgctttg   aatttaattt   gtctaatagc   aatgttgtaa   tgaatgcctt   60660   ctttaatttt   tactttcttg   atataccttt   ggccatccct   ctatttccac   acttagatat   60720   ctaagtgatg   tggtttaaaa   tttttctctt   ttaaacagca   tagaattaga   ttttagggaa   60780   gttttaaaat   aagttatttt   cttaatgaaa   gaccttaagt   cattcacatg   tattgcgaca   60840   aatagtgttt   tggtttggtg   taatttcagt   tccttatttt   agtctttcta   tattaagggt   60900   tttttgtttg   tttgttgcta   tttacttgtt   ttttcttttt   atttgattat   caattgtggt   60960   aggcttgaaa   aaagctcccc   gcccctaccc   cccaccacca   ccaccaccac   aaaatccgta   61020   tcctaatccc   tggaacctgt   gatgttacct   tatataagaa   aagggaactt   cacaaatgta   61080   atcaaattaa   agatcttaaa   actgggagat   tattctggtt   tatcccagtg   attcccacat   61140   gtaatcataa   cagggtggta   gaagggagtg   tgacacacag   acagaagagg   aaaaggcaga   61200   gcttgggccg   atatggtcac   acgccaagga   ctgctggcag   tcacccgaag   ccagaagagg   61260   caaggggcag   attctcccct   agagtctctg   gaggactcat   gggcttgcca   aacttttatt   61320   tttgcccagt   agaactaatt   ttggacttct   ggtctccaga   gctgtgcgag   aataaattta   61380   tgttgtttta   agccataaag   tttgtagtag   tttgttacag   ccaccatagg   aaactaatta   61440   aaatttctcc   atttttattt   tactctatca   tagcttatct   ttaactttta   gctttaatat   61500   atctctatca   aggttaaaaa   acaaaagaga   gacccaaaat   acataacaaa   attgaacatg   61560   atgtctttct   ttgtactctg   tcccttcact   caactccctt   aagtttttcc   tgaaatgttc   61620   catagtcata   tttataatta   ttttgtaagc   atttcatgtg   tttctaactt   cattgcttac   61680   catagttgtt   attattctat   gctcttccct   cttcttgaat   tcttaatttt   aattcaattc   61740   tcaattagct   gaagtatgcc   aggaagacta   taaaatatta   tattttcttc   cttcttatat   61800   atctgagaaa   actgttctgt   gccttggcac   agaaatcata   tttacaaagt   tcttgaatca   61860   caattttttt   tcaaaactct   gaacatgttg   ttatagtgtt   ttctggcatt   actttttgtg   61920   ggaaagaggt   atgataattt   aagatgcatt   tcttttaggt   aacttttaaa   tttttttacc   61980   ctgccttaaa   tgcctgaaga   tggttaatct   ctgaaattaa   atactctttt   gattgtatct   62040   caaagtcagc   atctttttat   caacattgac   tgctcttctg   gacactctta   tttatggctc   62100   ttcctcctgc   tccagattct   gaactgtgcc   tgtcacctcc   ctcaagacag   ccacagagcc   62160   ccaagatcct   tataacctca   ccatgatgtt   ccactccaca   gggttctgcc   attgtgtgct   62220   ttctttttca   tccttctctg   atgccacctc   ctcctctgtt   tgttgccagc   cttaataatt   62280   ttaaattcat   ggatcacagg   tagaaacccc   agttttgtct   aagtagacga   tcgatttttt   62340   ttttcttaaa   gctagtcttc   catttttctc   tttctctatt   tgtcaccttt   tcccaagaga   62400   taattttttc   ttttagtttt   cagaggagcc   ttcaatttct   ttttttaaac   cttttttcta   62460   acttttattt   taagttgagg   ggtacatgtg   caggtttctt   atacaggtaa   acttgtgtca   62520   tgggggtttg   ttgtacagat   tatttaatca   cccaggtatt   tttttttttt   tttttttttg   62580   agacggagtc   tcgctctgtc   gcccaggctg   gagtgcagtg   gcgcaatctc   ggctcactgc   62640   aagctctgcc   tcccgggttc   acgccattct   cctgcctcag   cctcccgagt   agctgggact   62700   acaggcgccc   gccactacgc   ccggctaatt   ttttgtattt   ttagtagaga   cggggtttca   62760   ccgttttagc   cgggatggtc   tggatctcct   gacctcgtga   tccacccgcc   tcggcctccc   62820   aaagtgctgg   gattaagctt   catattcatt   agttattttt   cctgatccta   accctcctcc   62880   caccctacac   cctcctatag   gccccagtgt   gttttgttcc   cctctatgta   tctgtgtgtt   62940   ctcatcagtt   agcttccact   tataagtgag   aatatacagt   attttgtttt   ctgttcttgc   63000   attagtttgc   tgaagataat   gccctccatc   catgtccatc   catgtccctg   caaagcacat   63060   tatctcattc   tttcttacga   gccttcaatt   tctaatcatt   cctcagaacc   tcctccctat   63120   ggtggctttc   acaacattag   tgctgagatg   catttgtaga   aataacaaat   cataaagaac   63180   cacaatgtgc   ttgattgtta   tttgtctgag   ccagagaaag   aacaaaaagg   cacgaattca   63240   gaacaaagat   aactgccaag   aagaaagtga   agtaaatgct   ctccattaga   cttcaatcta   63300   caagttactc   ttctactccc   tctggatttg   aacctaatgt   tacaaagtgg   agatacagcc   63360   agactcttga   tagaaatgag   gctccaagtt   acctgaaaaa   tgtccacctt   gatcatccct   63420   tcaaatatat   ttcatttaat   cacattctaa   atatgctctt   cacataagag   aaattaactt   63480   cttaaatccc   aatttccctt   tctgctaatt   atgcaaaaaa   agattgtaca   cagataatta   63540   aaaggtgtgg   gtaaagttaa   taaaagagac   agtgggaagg   aagaggagac   ccagatcctg   63600   agcctaatga   agtggatgaa   aaaggggagc   tctctgatgg   tgtcagtgct   gatgatgata   63660   aatgattcaa   caccgcacat   aggctcccta   aaaccggaac   ccttctatca   tctcttgact   63720   cccactgata   aaaagcaaat   gaacttttgt   ttctaattat   tctggaatgt   caattgtggc   63780   agaaaaaatt   agtttagctt   taaaagtcaa   ttccaaacaa   atgtaccatg   caaatgtcaa   63840   ggactgatcc   tgaaatatgg   agtttccaag   aggccaaggg   aataagtgag   caagaaagaa   63900   ctgaagcatt   gtcactcatc   atggccagca   gtacaaattc   tagcaccagg   aaccaaactc   63960   tgcttcctgg   tgtggagatg   aaaagggaga   gttccctggt   ccatccatca   caggacatgg   64020   gacaggggta   tggctctctg   ttcagcttcc   tcaagctcaa   actccttata   ggagggggag   64080   catgcagaag   ggtaggtgca   ggagccaggg   caagtgcatt   gaactccggc   cccacagtag   64140   tgtctggggt   gggtgcctgt   gactcctgaa   gccccagtgt   tatagtgctg   ttttaactct   64200   gccatctgca   gatggcttaa   gtggtaacca   gctcagtgcc   ctcttggtgg   ccatgtcctc   64260   gtccataggc   caggaagagt   caggtcacac   acagacttga   aggatgaatg   caggagtttt   64320   attgagtggt   ggaggtggtt   ctcagtggga   tggatgggga   actggaaggg   ggatggagtg   64380   ggaagatgat   cttcccctgg   agtttggctg   tccagggcca   ttctcctgtc   tgattgtccc   64440   cagctgaact   cttcttagtg   ttcagaccct   ccttctcttc   tctccttctc   tgccatgctg   64500   ttctgccatt   cgtctgctca   tctcctcctg   gagcctggga   tttggggttt   atatgggtac   64560   aggatagggg   catgtagctg   gccaaaaggc   aactttttgg   gcacaaaaac   aggaatacct   64620   gttctcattt   aagaccactg   gtttccaggc   ttgagggtgg   ggtctttgct   gggaaactgc   64680   cctctttcac   ccagtatttc   cctttctctt   gtccatgtaa   gagagttatt   tcccttccta   64740   atccaacact   caccccttct   ttggttcacc   tctcagattt   ctaaataaca   caaggtgcta   64800   ttctttccat   tatgtgcctt   ccctcacctc   tctgaagtca   taagcttcct   ctcatttctg   64860   tctttaggtt   gtgctgctct   acgcctctac   ctacgtcctg   gtgtccctca   gcatagacag   64920   ataccatgcc   atcgtctacc   ccatgaagtt   ccttcaagga   ggtgagctgg   ctttaccagg   64980   tgctctttca   taaagaactg   tccttgagtg   agggtaggat   cattttcact   aatattttac   65040   tcttaataat   agtgatcttc   ctcctctttc   cttctctttc   ttctctcttc   attctcctct   65100   tccacctctt   tcctcacttc   atgggtctga   ttgtctaaaa   agatgtacaa   aagaaaactt   65160   agatgcctgg   aaaaatgccg   atggttccca   tggacagccc   agagccacta   aggaaggtgg   65220   gtggctgacc   cactttgtga   ccattcagct   tgatgctgat   gcttccatct   ttttcctcta   65280   gagcagtgtt   gacatgcata   cctgtccaaa   gaaatgaact   ctcccacact   aacttctcat   65340   tttccagaaa   caaacacctt   gttctcctta   tccaaaacca   tagaaataat   tggaaagatg   65400   tttttaattg   atcatcgtat   tgggaaaaaa   aaataaggaa   actctctttg   atccagaaat   65460   ggttagggat   ccctaagaaa   aggaagcaga   tgggtcagag   ggaaggctga   gaccacacag   65520   cccacaaaac   ccctaggaga   tgctgaagtg   ggatgggagg   ccttaggagc   acttcctgaa   65580   caggctgcat   caatgtggag   tgtaggagag   gacccgaggc   aactgacaag   gtgatttctg   65640   aagtacaaca   cagaggatta   atcacagcaa   gtttccctgg   atttgaagag   gcagagaggg   65700   gccctgggga   gcagtaatgc   cctagaggaa   gggtgagtac   atattcacaa   gactagctgc   65760   ctactgctct   gcccacaatc   agagaccact   ctcctccata   caccctgaaa   gcaaatacaa   65820   agccattgtt   tagcaacctg   ttctcccttc   ctccagataa   cttcgaaaac   tgataaagaa   65880   cctaacaaaa   aatcttgacc   acaaaaatga   gtgcaatcaa   atattattta   aaaagtgaca   65940   aaatccaaac   tgtagaaaac   agataatacc   tttaaaatct   cattccacac   aatttggatc   66000   tagggaagca   aatttaatag   aagtagcaga   attttaggat   aaatataatt   aacgtcatca   66060   aggatctaca   agaagtggag   tgaaataggg   taatttcatt   tactattgac   aggggtataa   66120   gtaaacacat   cttattgaaa   tataaaatgt   gcagcccttt   gatatgactt   cttggtatct   66180     gctctggaga   aacacccaca   cgtgtgcatg   tatacatatg   tgatgtatat   tgtggcactg   66240   cctacaatag   caggaattca   tcgctaacct   tggtgtgtct   cagtggagga   atggctaact   66300   agtctatgga   agaagtttca   gtaccttctg   tttattgatt   gcatactagg   tgtcattcat   66360   gattctaact   gattcataca   tattaaatat   tttaatattc   ataatatctc   tgtgaggtgg   66420   gctctattat   tatctgcatt   ttacaggtaa   ggcaatcgtt   gttccaagag   tgaaagtaat   66480   ttgcccaagg   tcacagctag   taagtgcagg   aacaggattt   gaacccacac   agtctgggtc   66540   tagctccagc   gccctcaaat   gctatatagc   ctgcctctca   atgtggaagt   ggtagagcca   66600   tgggcagtaa   tatgcatcaa   catgaacata   tccatattgt   atgctgagtg   acaaaaacaa   66660   tatgcacaaa   aatccatgtg   aacgataaca   ttaaagtcaa   aaactcaaaa   caatgctaga   66720   tattgtgtac   agaaagacag   agaggagcag   aaagagagaa   cacacaaaag   aaaactccag   66780   gaaaaataaa   aaactatttg   caaaaacaca   caaaacttga   catttgacct   cacttctagt   66840   tatgggagtt   agaccagtgg   tgtgaccacc   acaggaactt   atatgcaagg   tggtaatttt   66900   tacatgaaga   acatatcaat   gtgttgtgtg   caattaaaat   gaatttattt   tgatcatgct   66960   tgttatgcct   actgtacagc   ataagacata   tgagtaaaaa   tactaaattg   tatacttaag   67020   aatttgctgg   ggatctgttc   caagatggcc   aaataggaac   agctctggtc   tgcagctccc   67080   agcatgatcg   atgcagaaga   caggtgattt   ctgcatttcc   aactgaggtg   cctggttcat   67140   ctcactggga   ctggttggac   agtgggtgca   gcccatggag   ggtgagccaa   agcagggcag   67200   ggcatcgcct   cacctgggga   gcacaaaggg   ttgggggatg   tccctttcct   agccaaggga   67260   agctggggca   cactgtacct   gggacactcc   tgccaaatac   tgcacttttc   ccaaggattt   67320   agaaacgagc   agacaaggag   attctctccc   atgcctggct   caggaggtct   ggagccttgc   67380   tcactgctag   cacagcagtc   tgaaattgaa   ctgtgaggca   gcagcctggc   taggggagga   67440   gcgtccacca   ttgctgaggc   atgagtaggt   aaacaaagtg   gcccggcagc   tcaaactggg   67500   tggagcccac   tgcagctcag   caaggcctac   tgcctctata   ggctccacct   ctgtgggcag   67560   gacatagctg   aacaaaaggc   agcagacagc   ttctgcagac   ttaaacgtcc   ctgttcgaca   67620   gctctgaaga   gcacagtggt   tcttccagca   tggcgtttga   gctctgagaa   tggacagact   67680   gcctcctcaa   gtgggtccct   gacacctgtg   tatgctaact   gggagacatc   tcccagtagg   67740   ggccaacaga   cacctcatat   aggtgggtgc   ccctctggga   caaagcttcc   agaggaagga   67800   tcaggcagca   atgtttgctg   ttctgcaata   tttgctgttc   acagcctctg   ctagtgatac   67860   ccaggcaaac   agggtctgga   gtggatctcc   agtaaactcc   aacagacctg   cagctgaggg   67920   acctgactgt   tacaacggaa   actaacaaac   agaaaggaat   agcaccaaca   tcaacaaaaa   67980   ggacatctac   accaaaaccc   catctgtagg   tcaccaatat   caaagaccaa   aggtagataa   68040   aaccacaaag   atggggagaa   accagagcgg   aaaagctgaa   aattctaaaa   atcagagcgc   68100   ctcttctcct   ccaaaggact   gcaactcccc   gccagcaatg   gaacaaagct   ggatagagaa   68160   tgactttgac   gagttgacag   aagtaggctt   cagaaggtcg   gtaataacaa   actactcctc   68220   cgagctaaag   gagcatgttc   aaacccattg   caaggaagct   aaaaaccttg   aaaaatggtt   68280   agacgaatgg   ctaaatagaa   taaacagtgt   agagaagacc   ttaaatgacc   tgaaggagct   68340   gaaaaccatg   gcacaagaac   ttcgtgaggc   atgcacaagc   ttcaatagcc   aattcaatca   68400   agtggaagaa   agggtgtcag   tgattgaaga   tcaaattaat   gaaataaagc   gagaagacat   68460   ggttagagaa   aaaagattaa   aaagaaacaa   acaaagcctc   caagaaatat   ggtactatgt   68520   gaaaagacca   aatctacatt   tgattagtgt   acctgaaact   gatggggcga   acagaaccaa   68580   gttggaacac   actcttcatg   atattatcca   gaacttgccc   aaaatagcaa   ggcaggccaa   68640   cactcaaatt   caggaaatac   agagaacacc   acaaagatac   tcctcgagaa   tagcaacccc   68700   aagacatgta   gtcagattca   ccaaggttaa   aatgaaggaa   aaaatgttaa   cggcagccag   68760   agagaaaggt   caggttaccc   acaaagggaa   gcccatcaga   ctaacagcag   atctcttggc   68820   agaaacccta   caagccagaa   gacagtgggg   gccaatattc   aacattctta   aagagaagaa   68880   ttttcaatcc   agaattccat   atccagccaa   actaagtttc   ataagtgaag   gagaaataaa   68940   atcctttaca   gaaaaacaaa   tgctgagaga   ttttgtcacc   accaggcctg   ccttacaaga   69000   gctcctgaag   gaagcactaa   atatggaaag   gaacaaccag   taccagccac   tacaaaaaca   69060   tgccaaattg   taaagaccat   tgatgctagg   aagaaactgc   atcaactaac   gggcaaaata   69120   accagctaac   atcataatga   taggatcaaa   ttcacatata   acaatattaa   ccttaaatgt   69180   aaatgggcta   aatgctccaa   ttaaaagaca   cacactggca   aattggataa   agagtcagga   69240   cccatcagtg   tgctgtattc   aggagactca   tctcatgtgc   agagatacac   ataggctcaa   69300   aataaaggga   tggaggaaga   tctaccaaga   aaatggaaag   caaaaaaaag   caggggttgc   69360   aatcctagtg   tctgataaaa   tagactttaa   accaacaaag   atcaaaagag   tcaaagaagg   69420   ccactgcata   atggtaaagg   gatcaattca   acaagaagag   ccaactatcc   taaatatata   69480   tgcacccaac   acaggagcac   ccagattcat   aaagcaagtc   cttagagacc   tacagagaga   69540   cttagactcc   aacacaataa   taatgggaga   ctttaacacc   ccactgtcaa   tattagacag   69600   atcaacgaga   cagaaggtta   acaaggatat   tcaggacttg   aactcagctc   agcaccaagc   69660   agacttaata   gacatctaca   gaactcttca   ccccaaaaca   acagaatata   cattcttttc   69720   agcaccgcat   tgcacttatt   ccaaaattga   ctacataatt   ggaagtaaag   cactcctcag   69780   caagtgtaaa   agaacagcaa   tcacaacaaa   ctgtctctca   gaccacagtg   ctatcaaatt   69840   agacctcagg   attaagaaac   tcactcaaaa   ccgcacaact   acatggaaac   tgaataacct   69900   gctcctgaat   gactactggg   tacatagtga   aatgaaggca   gaaataaaga   tgttctttga   69960   aaccaataag   aacaaagaca   ctatgtacca   gattctctgg   gacacattta   aagtagtgtg   70020   tagagggaaa   tttatagcac   taaatgccca   caagagaaag   caggagagat   ctaaaattga   70080   caccctaaca   tcacaattaa   aagaactaga   gaagcaagag   caaacaaatt   caaaagctag   70140   cagaaggcaa   gaaataacta   agatcagagc   agaactgaag   gagatagaga   cacaaaaacc   70200   cttcgaaaag   tcaatgaatc   caggagctgg   ttttctgaaa   agatcaacaa   aattgataga   70260   ctgctagcaa   gactaataaa   gaagaaaaga   gggaagaatc   aaatagatgc   aataaaaaat   70320   aataatgtgg   atatcactac   caatcccgca   gaaatacaaa   ctaccatcag   agaatactat   70380   aaacacctcc   atgcaaataa   actagaaaat   ctagaagaaa   tggataaatt   cctggacaca   70440   tacaccctcc   caagactaaa   ccaggaagaa   gttgaatctc   tgaatagacc   attaacaggc   70500   tccaaaattg   aggcaataat   taataggcta   ccaaccaaaa   aaagtccagg   accagaccaa   70560   ttcacaactg   aattctacca   aaggtacaaa   gaggagttgg   taccattcct   tctgaaacta   70620   ttccaatcaa   tagaaaaaga   gggaatcctc   cctaactcat   tttataaggc   cagcatcatc   70680   ctgataccaa   agcctggcag   agacaaaaca   aaaaaaagag   aattttagac   caatatcctt   70740   gatgaacatc   gatgtgaaaa   tcctcaataa   agtactggca   aactgaatcc   tgcagcacat   70800   caaaaagctt   atccaccatg   atcaagtcgg   cttcatcccc   gggatggaag   gctggttcaa   70860   catatgcaaa   ttgataaacg   taatccatca   cataaacaga   accaatgaca   aaaatcacat   70920   gattatctca   atagatacaa   aaaaggcctt   caacaaaatc   caacgccctt   catgctaaaa   70980   actgtcaata   aactaggtat   tgattgaaca   tatctcaaaa   taataagagc   tatttatgac   71040   aaactcacat   ccaatatcat   actgaatggg   caaaaactgg   aagcattccc   tttgaaaacc   71100   ggcacaagac   aaggatgccc   tctctcacca   ctcctattca   acatagtgtt   ggaagttctg   71160   gccagggcaa   tcaggcaaaa   ggaagcaata   aagcgtattc   aattaggaaa   agaagaagtc   71220   aaattgtccc   tatttgtaga   ccacatgatt   gtatatttag   aaaaccccat   tgtctcagcc   71280   caaaatctcc   ttaagctgat   aagcaacttc   accaaagtct   caggatacaa   aatcaatgcg   71340   caaaaatcac   aagcattcct   atacatgaat   aagagacaaa   cagagagcca   aatcatgagt   71400   gaattcccat   tcacaattgc   tacaaagaga   ataaaatacc   taagaatcca   acttacaagg   71460   gatgtgaagg   accttttcaa   ggagaactac   aaaccactgc   gcaaggaaat   aaaagaggac   71520   acaaacaaat   ggaagaacat   tccatgctca   tggataggaa   gaatccatat   agtgaaaatg   71580   gccatactgc   ccaaggtatt   ttatagattc   agtgccatcc   ccatcaagct   accaatgact   71640   ttcttcacag   aattggaaaa   aactacttta   aagttcatat   ggagccataa   aagagcctgc   71700   attgccaaga   caatcctaag   caaaaagaac   aaagctggag   acatcaagct   acctgacttc   71760   aaactgtagt   acaaggccac   agtaaccaaa   acagcaaggt   acttgtacca   aaacagagat   71820   atagaccaat   ggaacagaac   agagccctca   gacataacac   cacacttcta   caaccatctg   71880   atctttgaca   aacctgacaa   aaacaagaaa   tggggaaacg   attccctatt   taataaatgg   71940   tgctgggaaa   cctggctagc   catatggaga   aagctgacac   tggatccctt   ccttacacct   72000   tatacgaaaa   ttaactcaag   atggattaaa   gacttaaatg   ttaaacctaa   aaccataaaa   72060   accctagaag   aaaacctagg   caataccatt   caggacatag   gcatgggcaa   agacttcatg   72120   actaaaacac   caaaagcaat   ggcaacaaaa   gccaaaattg   acaaatggga   tctaattaaa   72180   ctaaagagct   tctgcacagc   aaaagaaact   accatcagag   tgaatgggca   gcctacagaa   72240   tgggagaaaa   tttttgcaat   ctacccatct   gaaaagggct   aatatccaga   atctacaaag   72300   aactcaaaca   aatttacaag   agaaaaaaca   accccatcaa   aaagtgggca   aaggatatga   72360   acagacactt   ctgaaaagaa   gacatctatg   cagccaacag   acacatgaaa   aaatgctcat   72420   catcactgat   catcagagaa   atgcaaatca   aaaccacaat   gaaaaaccat   ctcatgccag   72480   ttagaatagt   gaccattaaa   aagtcaggaa   acaacaggtg   ctggagagga   tgtggagaaa   72540   taggaacact   tttacactgt   tggtgggagt   gtacattagt   tcaaccattg   tggaacacag   72600   tgtggcgatt   cctcaaggat   ctagaactag   aaatactatt   tgacccagca   atcttattac   72660   tggtatatac   ccaaaggatt   ataaatcatg   ctactataaa   gacacatgca   cacgtatgtt   72720   tattgctatt   gcagcactat   tcacaatagc   aaagacttgg   aaccaaccca   aatgtccatc   72780   aatgatagac   tggattaaga   aaatatggca   catatacacc   atggaatact   atgcagccat   72840   aaaaaaggat   gagtttgtgt   cctttgcagg   gacatggatg   aagctagaaa   ccatcattct   72900   cagcaaactg   tcacaaagac   agaaaaccaa   acactgcatg   ttctcactca   taggtgggaa   72960   ctgaacaacg   aaaacacttg   gacgcagggt   ggggaacatc   agacaccggg   gcctgtcagg   73020   gagtgagggg   ctgggggagg   gatagcgtta   gcagaaatag   ctaatgtaaa   tgacaagttg   73080   atgggtgcag   caaaccaaca   tggcacatgt   atacctatgt   atccaacctg   cacattgttc   73140   acatgtacaa   tgataattta   aaaaaaagaa   tttgctaaca   agataaacct   tatgttaagt   73200   gctcttgcca   caaaaggaaa   aaactaaaca   aaacaatact   aaagggggac   aggaagaagc   73260   ctttggaggt   gatggataca   tttatggcat   tgcttgtaat   gatggttaca   ggaatgtata   73320   catatctcca   gacacattaa   gtggtatgca   ttggatatgt   atagcttttt   atatgccaat   73380   tatacatcaa   taaagtggtt   tagaagttaa   ttttaaaaat   agaaaatata   cttgtttcct   73440   gggtttcttc   tcggtcattt   ggtgtgtctg   ttttgtgaag   ggttgagacc   tgtaccaagt   73500   tttctgggca   ctgtaaagcc   ctagttatta   tgatagggga   ggcaggaaat   tgatgaggca   73560   ggggagatag   gagagacatt   tcgtgacata   ggtatgcaaa   aatgtgtcaa   tgaagcatca   73620   aattctgtaa   aatatttgta   gagatttatt   ctgagtgaaa   tatcagtgac   aatggacaaa   73680   gccctcagga   ggtcatgaga   atatgtgccc   aaggtggtca   gggtgcagct   tggttttata   73740   cattttaggg   cgacatgaac   cttcaatcaa   acacatttaa   gaaatacata   gggttggtcc   73800   gaaatggcag   ggttgggtag   tgggggatgg   ggtgggcgaa   gcttccagat   tatagttaga   73860   tttaaatttt   ttctggttga   caattggttg   agtttatcta   aagacctggg   atcaacaaaa   73920   aggaatgtct   gagttaagac   aaagaattgt   ggagaccaaa   cttcttattt   gcaggggaaa   73980   cttttaggtt   tcctcttcag   ggagactatg   ttgtaaaatg   tttctcatca   gacttaaagt   74040   ctgtgttgat   gttaacgctg   gagaggtata   atgagacaca   cctgaacccg   cactgcccat   74100   catggcttgg   aacagcctct   caggttaaat   cttaaaagag   ccttggctga   ggaggaaatc   74160   cattcggaag   gttggggggc   cttataactt   tatttttggt   ttacattcat   ggatttgtga   74220   tacacttttt   tttttttttt   tttttttttt   tagacagagt   ctcactctgt   cgcccgggct   74280   agagcgcagt   ggcgcgatct   cagctcactg   caacttctgc   ctcccaggtt   caagtgattc   74340   ttgtgcctca   gcctcccgag   tagctgggat   tacaggtgtg   catcaccacg   cctaattttt   74400   gtattttagt   agagatgggg   tttcaccgtg   ttggccaggc   tggtctcaaa   ctcctgggct   74460   caagcaatcc   accttggcct   tccaaagtgc   tgggattata   ggcatgagac   actgcacctg   74520   gcccgtgata   cacttattta   tccatctatt   tgtttatgcc   tagccgactt   tagaaagggt   74580   tgaatgtggt   ggacaaaagg   acatcgacgc   caaagtattt   taaggtgaag   aaatataagt   74640   gaaggaaaaa   taagtatagg   aaattgcagg   agggaagtcc   caggaaactt   gttagaggta   74700   cagacttgag   acccatagcc   agagggtgac   atgatgaatt   acatgtttca   ccatattcac   74760   taggttaaaa   aattagctgt   ttgctcgtgg   aaaatatgtc   tagttctggg   agacactgtg   74820   taatatcatc   aagagtgtcc   taaattatca   gaaaggcagc   aattaatatt   atagtgcttt   74880   tctaaagatg   taaaacaact   tattccctca   ctacaaaatt   aagagaggct   caacataagg   74940   tgcttggaaa   atcctcatca   tcagtagctt   ctgggaaaga   tgaaagctta   gaaacacatt   75000   agccagctgc   cttcctctgc   ccaggcccaa   cgaaagtcac   tgaagaaatc   taaaaacagg   75060   gggagacctg   catctggaat   gagagccaag   gaaagaagcc   atccaaatac   cagaaacttc   75120   taagaattcc   cagcacacga   gggagcagat   gaaacacgga   agccaagtgt   ataatgtata   75180   attcagcccc   cgggagaact   ccctggcctt   gtagaggatg   aactgatact   cacaagttta   75240   tagggtgagg   gcagccggtg   agaacgcgac   acgagaggtt   ccaggaagcc   caacgcggga   75300   gtcattttag   taccgcagct   tgtcagaatt   taagggcagc   tgagaggcag   ggttcggagt   75360   ttatgcggct   cagctgagag   aaagccgtag   aaagaagaga   ttccccgaga   gtggcagggc   75420   tctctgggag   agtcatggag   cagggagaaa   ccaggaggga   gtcaaatgta   gctgtgcagg   75480   cacaaggcaa   tggccgctta   actgagggga   ggccatattt   tatttatttt   ttactgctat   75540   ttactgaagc   agtatttttc   ttagccagca   actccagaaa   agttgacaat   tttaaaggtc   75600   ttcaccaaaa   atgaatatga   gaataattag   aaaaatatca   aaataataaa   gagaaatatg   75660   cagtgttata   ttaaaatgta   atataaaact   ttgataatta   aaacaatgct   ggaaccagaa   75720   taacacatat   aaatcataca   aaatttcaaa   tcagttggca   aaaaatgaat   tagtatacgg   75780   tgttgaaata   gctgtctggc   cttttttcag   aatattagat   caccacccac   catagctaat   75840   aacaaagtaa   attttagttg   gattgaaaaa   attaacactt   caaacataaa   aggagtaaaa   75900   gaatatttgg   gtaacacctt   cataatttca   gtgaaggaaa   ggcctttcca   ggaaccatat   75960   caaaagcaga   agcaagaagg   aaaatatagt   tttttactac   ataaaaattt   aaattattta   76020   tgtaaataat   aaatatccat   aaacaaactt   aaaaagcaaa   attataaatg   gaaaatattt   76080   ttatgtaaca   ggcaaagaaa   aatatccttt   ttataaagaa   gtctggcaaa   gaaataaaaa   76140   cattctaatt   gagtaaatgg   gcaaagggca   tgaacagact   aacaagaaac   aaagaaacat   76200   aaatgagacc   aggcgctgtg   gcccacgccg   gtaatcccag   cacttcggga   ggctgaggtg   76260   ggtggatcac   ttgaggtcac   aagtttgaga   ccagcctggc   caacatagtg   aaaccctgtc   76320   tctactaaaa   aaatacaaaa   attagctggg   catggtggca   tgcacctgta   atcccgctac   76380   tcaagaggct   gaggcacaag   aatcgcttga   acctgggaag   cggaggttgc   ggtgagtgga   76440   gatggcacca   ctgcactcta   gccctctagc   attggcaaca   gagcaagact   tcaccagaaa   76500   aaaaaaaaaa   aaaacaacac   cataaatgaa   tagaaaatgt   atagaaatat   tcattttgac   76560   tactaattaa   agaattgcaa   aattgaacat   ttggatacaa   aattttacca   accacataac   76620   aagattaaaa   ggaagaatta   tccaactatt   aggagagtat   aaggaaattg   gtacttttgt   76680   tgttacagtg   agagccataa   atagtaggca   cttttgaggg   aaatgtaaca   atatttgtca   76740   aaatttataa   atcttctgtt   tctaagagtt   tatcctaagg   aataacaaga   actgagaaaa   76800   atagatgaat   ataaattatt   tataataaaa   aatggaaaac   aacttagata   ataatcggct   76860   attgataaaa   ttggagtaca   gtataaccac   cggaaaaaat   gttatttatc   agtgcttttc   76920   caattgcact   tcccacagtg   acccccagag   ggctcatttt   gctggcctgc   tccagggcag   76980   atctgggagc   tgctggctgt   ctcagtttca   accaaagata   cctgagcttt   caacactttc   77040   ctatactggg   ttcgagataa   aattatattt   gaagaaagga   tcttactacc   tgaaaaaaaa   77100   tgccacttgt   acaagttcat   atttcttaac   aggaaaagat   ggtcataaca   ggcttttcag   77160   aggaaaaaaa   ttaggttaca   aggtaggatc   tgtatcatga   tctcatttct   aaatgtgtaa   77220   tctatgtatt   atatatgcat   gcgttatcaa   aatttaagct   tgacgtttct   ggttggtgcc   77280   actatggatt   tttttccttt   tcgcttattt   ttatttgcta   gtgtttctat   aatcaatgtg   77340   tgtaacttat   acgattttat   aaaaaggtaa   atgagttaca   tggaaaagag   gaaggctaag   77400   cggcagagaa   gtacagaaat   ggacaagagc   acatggaaga   gggagtctga   ctttccgggg   77460   gtgagaaaca   tgagacagtc   taccttatat   gcaaacctgc   atttcatctg   ttcgatcctg   77520   gatccttcag   ccctgagaag   tcactcttca   tggcaaatgg   gctcagctgc   ctggaaggcc   77580   atcagtctcc   ctgctgtggc   agctgcgtca   cagcctaaga   gagtggggct   ttcacagtgt   77640   agccccagca   ccagtaagag   aaaaacagct   ccaagcctat   accctgctgg   gcctttgggg   77700   aaacatcttc   ggagaccttg   gtagaattga   ttctaaagta   tgattctata   ctctcttccc   77760   tgtccatccc   tctcaaaagt   gaatgcactg   ttcatccatc   aagtggaaac   tttgtggggc   77820   ccaggagtca   cacgttgccc   ttaagagtgc   cctggaacct   atagaagaag   atgacattag   77880   aattctaacc   attgaaagaa   caagagcaca   gggggcagct   actaatgtca   cccaggacca   77940   tagaaaccca   tcaggcttca   cacccccacc   atcaagaaga   accccctcca   gtcctattac   78000   ctgccctctt   catcccaccc   tgcttctttc   ttaatctgaa   ctcccactgc   ttctctccac   78060   gcctggtttt   tctctgttcc   tttagggacc   agctccccta   tgccatcaaa   ctcctaatgg   78120   gaggctccct   ccacctcctg   cctcctgagg   tggctgattc   ccttaagaca   tctcaagagg   78180   atgctcctca   accttccttt   ttaggtacct   catggcctgg   tggtggtgaa   gacattttcc   78240   taccgttcta   gaccacctcc   aggccttttc   tccttcagta   ttgcatacgc   atgccctctc   78300   tgtttctctg   atgctcatgc   tatcagtcta   gactaccttc   tacaacacct   ccccatatgt   78360   ctgtgcccat   atggccatgg   ccctcatcca   gtgaagatgt   tgacactatt   ctcagtctct   78420   cacctttcat   actgtcctaa   tcacaggtga   ctcgatgtct   acataaacta   agtctagtga   78480   gagggcttca   catgatgact   cacagagctt   gacatgttcc   ctgcatttga   aagcattgtg   78540   gaccaatgac   tacatcacac   ctgagaatgt   tgaagagaag   ctggctggcg   ctgtggtcct   78600   agcccagggg   gctgatgtaa   aacctgcatc   atgaagtttg   cctgaggctg   tgttaaaaaa   78660   gatcctgccc   cagacatttc   tgccagagaa   agaggcctcc   ctgaaggtca   ccagggaggg   78720   tggaaggtca   ctgcccaagt   caaggaagtg   gccaccagca   agaccccaca   ggataatttc   78780   caaaggaccc   agaaaagttc   ctcaggggac   caagaatcag   ctttggatgc   agccaggcac   78840   agagagcaca   aagccctctg   atgacagcag   cagtcaagca   agaactctgt   cttcttcctc   78900   ctcctggttc   cctgtacccc   agctccagag   agaccccaat   gtggatagca   acctggggga   78960   ggaggaggaa   tagcagacaa   ggatagaaag   cagaagggct   ggtacctctg   ctggacttcc   79020   tacttgaagc   aagcccacct   agggagaaaa   gacttaactt   gaaatcttct   tgggaatttg   79080   gggttttttt   attgttatct   aggattttaa   ttactaattt   tcatgtaatt   attaaattga   79140   ccaatgttta   ccacttaatg   tgtcagtcac   ctgtgagtca   tcactaacat   caagggactc   79200   gagtttccaa   atgcaggcag   gaaaacctct   ccccctgtcc   ccagtggatt   accggttgca   79260   tgtaaggaac   aggaaagacc   ttaatcagat   tgctattata   attctacccc   aagactcctg   79320   cttgctcagt   gggccaataa   caactctttt   ctacatgaat   ctaatatctc   ttttcagtgc   79380   atacaattca   ggaagaacag   aaccctagat   attattctcc   taaatatttt   gggtttggat   79440   tttaatagag   tccctagttt   cctctagatt   tttctcactt   cttgtagctc   gaccacagtc   79500   aacacactgt   cgaacaacac   cacctgactc   tacattccaa   ccccacccaa   agacacacac   79560   ctgctttcca   agttgaaagt   aggggagcaa   ggtggcctct   tatttctgga   agaaggtccc   79620   agagccaatg   agcacagaaa   gcccagctag   gtgagggatt   aagatgagag   tttcaacaac   79680   agcttgacag   aaagcatcac   atcataacaa   cgggtgtatt   ttaaaggtac   ttgcttatgt   79740   attaattgtc   tctctccagc   cccatcataa   ccagctgagg   tggccagagc   aggtattttt   79800   tataccaatt   taaagaaaag   atgaggctga   gactgcatgg   tttagccaag   gtcagaatat   79860   tgcaactaaa   gtccagtgtg   cagacatagc   taccaggttt   ctgggctggt   tttgaatgca   79920   ccctactctc   cagaacgaag   ttcagtgttc   tttctctgcc   ttgtaagtcc   acccatgttc   79980   ctaccaaaaa   aaaaaaaaaa   gaaagaagtt   tccttcccat   ttggcaaata   actctcaaaa   80040   actgtcccag   tgctttgcag   tgctgggtac   agtgaaaaga   tgccacgtgg   accaaaggct   80100   ctagccctgc   ctggtaacaa   gagactgcat   atgggtagac   acaactcagc   atttgcaccg   80160   caacttacca   aatgtacatg   gtacaaactg   agctccctaa   tactgcagga   atggatgcgt   80220   cagcatctca   agtcagatgg   gtaaatcagg   aaagaaaaag   caccaacctt   aggccaggca   80280   atgtgctggc   ttctgttcaa   gcttcccttt   gtaatcttca   cattgacaat   gtatggagat   80340   actgtatccc   tgtttacaga   gaaggaaact   gaggctctgg   gaggttcttt   gaggtccagg   80400   gtggctgccc   cacagtgatc   cttttctata   gcagactcca   ttgtgctccc   aaaaatggtt   80460   gccacatacc   cagacattcc   tctcttttct   ttgggcagaa   aagcaagcca   gggtcctcat   80520   tgtgatcgcc   tggagcctgt   cttttctgtt   ctccattccc   accctgatca   tatttgggaa   80580   gaggacactg   tccaacggtg   aagtgcagtg   ctgggccctg   tggcctgacg   actcctactg   80640   gaccccatac   atgaccatcg   tggccttcct   ggtgtacttc   atccctctga   caatcatcag   80700   gtaagaagcc   gtcaggacag   gaccacacgg   gggggtgggg   cggggggggc   tttcctgttt   80760   ctccagctgt   tgctctcctc   cccaacagac   ccatttttcc   tagtgccctt   gagggaactg   80820   accaggccac   aagattttat   gtgtagctac   cataggctaa   gcccaatttc   ttcaaagtag   80880   agtaaggaga   agggaattaa   caattactgg   gtatataggg   caggcattgt   gctaaatagc   80940   taacaattat   cacattagcc   acagtataac   cctgtgaaat   aatttccatt   gccccaattt   81000   aatgaatgat   gaaactcaag   ctcagagtga   ttaagtaact   tattcaacat   caaacaggat   81060   tgatatccag   gtttttctga   aatcatacca   tattatttct   tttatatctc   gcagccttcc   81120   aaatataagg   caaattacca   gtttcaggga   ttttgtaaat   gaaaagatag   acatacttga   81180   aacaaactga   acacaaaact   cagtatgcac   taatatgcta   aactgtctat   gcagtggagt   81240   cagataatgg   cagggaatat   attttggaag   ccaccattga   aggaggaaat   gattcaggta   81300   ggataaggga   gaagtgggga   ggaagcagag   tctgtgatga   ccagaaatca   gagtgagtgt   81360   agaggcagag   tggacaggta   cacagtgcca   gtgggacaga   agttggcagt   gagctcctcc   81420   cagtgactgg   gggaggctca   gttgtgaatg   accttggcaa   aaaagtcata   ggaagtcctt   81480   tgtcctgcag   ccaatggtga   gctgtagagg   gtctttcaac   agagattagc   acaacaacct   81540   ggtcaaggta   gggtctcact   tggccatgcc   atgtggcagg   atcaccctct   gaaatgggct   81600   ctgcatgtgt   agatggggtg   ggagactaca   ggaagagacc   tggcacattg   tgtgtatcct   81660   tgagccatga   tgatgctgca   ccttttgggt   aggtctaagc   ctgccatagt   gtgtcatata   81720   acacattgta   agcaaaggaa   gattatcagc   ctttgttttt   gatagtgtaa   agagagctgt   81780   gataaaaatc   cttgtactta   aaactgtaca   tgagtctctg   gttatttcct   cataactcat   81840   tcttgtggaa   ttactagatc   aaaggtatag   atcaaagatg   atgcacattt   taaaactttt   81900   gatgcagagt   cacaagctgc   ccttgagaaa   agctgtattg   cttcttggcc   tgttagctaa   81960   gatcacttgt   agaaaaggta   tattctgggc   aataccatac   aagggctctt   gttttctgct   82020   cccctgaccc   aatggaaggt   agcactatta   ccttactaaa   tctttgccaa   cctaagagtt   82080   taaaaagtat   ttcattgttt   taatatgcat   ttctttaatt   actaagctgg   tggcatgttt   82140   ttcaagtgtt   ttctgataca   gtgggagtca   ccttttcctg   ctcattattc   atctttagtc   82200   taataaaata   ctatagctgg   gagaaacgtt   atccatcata   tggtcaaacc   cctgaatttt   82260   tcataagagt   agaattaaga   ctaaaagaag   ttaactgcct   tgcccccgcc   ccctcatcag   82320   agattgagat   gggcaagcac   atagtgacct   caaaagccaa   cccagcagcc   tttccaaaat   82380   ggggtgttgt   ccagtcatct   ctgctgaatg   gacaacttaa   caccaacact   ttattttgac   82440   ttattgccct   ccactccatc   aagctgagct   tgagagaaag   tttgaaggag   aaaaggttat   82500   ttagacaact   atccaaggta   agtcacagcg   catgggaagg   ggcccatggc   agaggagaca   82560   ggcaagtgag   ccacagcggt   cagagagcat   ttggaagaaa   gtggcttgtg   tcatcctgag   82620   caactgaggc   atcgatttca   gtggcagcac   atgtcctggc   ttcactgctg   attgatttcc   82680   catcaactca   agaaatgaac   tccagtcaaa   gtccccgctc   agcagagtgg   atgctgcagc   82740   acatggacca   cgcagctcag   gttatcctgt   atctatgatt   tccccattaa   tttgggctgc   82800   tttttccacc   aggacttctc   ctcaaacttg   cctgtgggct   agctagtgtg   tctttttcgt   82860   gtccttgaaa   gggcaaaata   caatgtgttt   gctgattgag   ggaaatgatt   tcaaaaactg   82920   gtttccctca   gctctcacca   agctaaagcc   tctggaaggc   cagatcctag   ctccttgctt   82980   ccagcccgtg   acggtcatga   aaactcagcc   atgacctcca   gccatccttg   agcttctctg   83040   aagcatttcc   tacctgacaa   gaggctcctc   ttccatcacc   ttccctcttt   cgtggctccc   83100   tgctgctgaa   gaaacaactt   ctctcttggg   gactaacatc   ctctacctcc   ttcagcaaca   83160   actatcttct   caaacaagac   caggccctgg   gggggcgagt   tttgtgagga   acttgtttcc   83220   aactagtacc   tggcataggt   gctacaattt   catctcagat   ataaggttca   actcttattc   83280   tgtctttatt   acctatattc   tggccccctg   ggctcaattc   tctgagtaag   gttaaattct   83340   tcatttcaag   cttaaataga   caaggaaaac   agctcaagga   aagtgttcac   tcgtgttcac   83400   catgcctcat   tctccttcat   gcatgctata   tatgctacat   ttaccaaatt   gacagaaaaa   83460   aaaacatatc   ttagttgttt   tattttgcat   ttctttcatt   acaaatgaga   ttgaatatat   83520   tttatatgct   ttttgccatt   tttgtttttt   ttaaagtact   tattaggttg   gtgcaaaagt   83580   aattgcggac   tttgccattg   aaagtaatgc   cacttttctt   gtttaaatgt   ttttattttc   83640   tctattctct   atctttagaa   attattcata   caataacctt   tttctttttg   catatgttgc   83700   aaatatttct   cctaatttgt   catttgcctc   tctattgagt   atggttttat   tttgtttttg   83760   attattaaaa   gtagtgattg   acgtaattaa   gaaaaaaaga   aaaagcacaa   ataactaact   83820   cagtggcttt   tctgccacaa   atacctcatt   gttttactta   ttgcccagct   tcgtatgcat   83880   ttaagtatct   gctaggacaa   gatctacttg   ttttttttca   aagattccct   ggctgcttca   83940   tggtgagctg   ctgatgcctc   acctgcactc   ccagctccag   ccatggaaca   aacacccctc   84000   tggcccatgt   gcatcactcc   catgcctgaa   cactctgcca   agctcacctt   gccttgtttc   84060   acccaactag   ttcacatccc   ttgtgtctca   tttccaatac   caactccaca   gtagagcttc   84120   ccttgccaac   cctggaccaa   gaggcacctc   aggtcctcac   tccctaacct   ccaacctcac   84180   tttgtccatg   tcgtcatcta   gacttcatgg   cagtttgact   gtatctcatg   agtgcaggga   84240   ctgtggcctc   ttcatttctg   gatttcacat   ccagcacaca   ctaaacagtt   ggtgaatatt   84300   agattgaatg   aataacgcaa   taaattgcac   cgaacaacag   atgtgtcatg   tcactctctc   84360   tactctttct   ctccctctct   cattctctct   ctctctgtct   ctcacacaca   tcacacacac   84420   acacacacac   acacacacaa   acacacgtgt   acacatactg   ctaatataga   aatttctggc   84480   tatctaatgc   ctagatcagt   ttttggcata   tgatagctcc   tccataaata   tatgtgagag   84540   gaaagaagaa   agggagaaaa   gaaaggaggg   gggaagaaag   ggagggaggg   aaagagagag   84600   agagagaggc   agttgtattg   ttggtggcaa   actctcatga   gagctgcgta   tgaataaaga   84660   tgtagctgtt   ccatcatcca   ctaaatcagg   ctcccaggag   ttgaaaccat   agtgactgac   84720   agcaaaggca   atacattgtt   tttctgggtc   aaaacaacag   gagttgtttt   gaaattcact   84780   tgacataact   gtctaattgt   tgctggggag   gttattagcc   ctggtgtaat   tgctttactt   84840   tagctgtgct   tattttcttg   ctacctaatt   gatttttcat   tctcttatca   ttttcatctc   84900   tgtaagataa   agatagaagc   agaagagatt   cacaaccgac   aattgcaaaa   aaaaaaagtg   84960   gaaaaagcac   attagtcatg   gatcattgat   ttgtcatatg   tgagaaaaat   tcatttattt   85020   tgcttaggaa   atgggatcac   attgtgggaa   agccccatgc   caggttggtg   cagccctgtg   85080   gatagaatac   atggccatgg   tgggtaaaag   cgccccaggg   acctggccag   cactggccat   85140   gggcataagg   tccaaggtca   aagaggatca   cagggcccac   aataaaagta   gtcatcaaaa   85200   tgaataaaga   gagattgtaa   gggtctccac   taatgctgaa   aacctggatg   gagaggaagg   85260   tacagattat   ggagtcattt   tgcagggaca   atccacaaaa   cttatctgca   atctgattga   85320   tcagataaag   gaaaaaggaa   aagagacaaa   gccaagtgtg   gtgactcaca   cctgtaatct   85380   cagcactctg   ggaggcctag   gcaggtggac   ctcttgggcc   cgggagttca   agaccagcct   85440   ggcagcatgg   tgaaaccctg   tctctataaa   atatacaaaa   attagccggg   catagtggcg   85500   cacactggta   gttccagcta   ttcaggtggc   tgaggtggga   ggtcaaggct   gcagtgaact   85560   atgatcctga   cactgtactc   cagcctgggt   gagagagcaa   aaccctgtgt   ccaaaaaaaa   85620   atggggacac   acagcaacgc   agatgtctct   ttgatgcttg   atcactaagc   tccaagaagt   85680   agtctgattt   ggcaataata   ccctagttat   taggtgttcg   ccatgctaac   agaaaagctg   85740   gtactggttc   tcactcatct   gattggccct   aagtaaaaag   aggggtgact   taggcccaaa   85800   tccctaaagc   ataagacatt   tcttaacttt   tcccctgtgt   acatggttga   taaccataaa   85860   tatttgaatt   attggtcaat   taaatactgg   ggttgcaatt   gacataacca   ttaactttcc   85920   atggtgagtg   taaccttacc   ttgcacagta   tttcactttc   tggggaagaa   gtttccctgt   85980   tctagcaaat   agtcaactta   gggttgaact   cactaataac   atttttctca   ttttttcttt   86040   actgcaggca   tacaaattgc   aaaaatagtc   cctgggcaat   tatataagaa   agaatggaga   86100   tgctccaaaa   aagtccctga   gctttcctgg   agcacttgga   ggggttcact   cagcaagaat   86160   gggaaaaggc   agtaagaagc   agagatttta   acattgccat   taattaagtt   atattctaaa   86220   gatcattaaa   agcaattagt   gctttgtcgg   ctactgcaaa   ctgaagctct   gattttttaa   86280   aataattctc   aagagggaag   atacacagtt   ttaaaataaa   gcattgcttt   gtcccaaagt   86340   ttttagtcaa   gatgcttaat   gtagtcaaaa   tatcaagtaa   tattaaagaa   gagtagtttt   86400   tatatttatc   aaagagtacc   taggatagaa   aacttgctga   cacttttgaa   tccttaacac   86460   tttgttttcc   tgttaggcac   agattaaatc   atgtaagaat   tggtgtgata   tagtataaaa   86520   aataggcaaa   aaagaggaga   aaggaaatta   gtctctacta   attttccagc   accgtacttg   86580   gcactttaaa   tattgtcaca   tctgatactc   aaaaagttcg   agaggtctgt   attgttaaca   86640   ccattttgta   gaacctcaac   tgaggctcag   tgtagtagtt   atctattact   gcattacaaa   86700   tcaccacaaa   cctagtgact   taaatcaaga   tacatttgtt   atctcacagt   ttctgtgggt   86760   caagaatttg   gccgtggctt   aaccgggtcc   tctgctcatg   caaggcagcc   attcaagttg   86820   tttatccagg   gcaggggtca   tcagaggctg   gactgggaga   ggctcttttc   caagctccct   86880   caggtcctta   gcagaattca   ttttcttgca   gctgtgggat   tcatggcacc   ttgcttcttc   86940   aaagccagca   atgcatagag   acaccaacaa   gacaggcact   acattctatg   taatcagtcc   87000   catccccttt   gccacattgc   attgattaga   ggcaagtcac   ctgctcttcc   cacactaaag   87060   gggtgaggat   caacaaagcc   atgaatgcca   gaatcaggga   tcatagatgt   catttaaaag   87120   actgtctaaa   gtcaggactg   tacaacagac   aagtagagga   gacccaagtt   tttctgactc   87180   taaggcctgt   ttttggattc   taatcacagc   tctccctcaa   accatttggg   ctaccttgag   87240   gaaactgcat   aactttacca   ggtcttagcc   tcctcactta   agaaagaagt   aactccatcc   87300   agtggatcct   catggtcact   ttcagttgca   attctatgaa   cagcaagtat   aagggggcag   87360   gcatggttaa   aagatctgtc   ccttcacacc   ttgcactcgg   ggaggtggga   gtggtcttct   87420   ctgtgtcctc   acagtaggga   tccaccagtc   actttaatac   tacctttggt   tctttctgca   87480   gcatcatgta   tggcattgtg   atccgaacta   tttggattaa   aagcaaaacc   tacgaaacag   87540   tgatttccaa   ctgctcaggt   aagtctctac   tctgcatggc   ccaattcttg   gctaatttgc   87600   ttctccagag   gttttcttct   agcaaatgtg   agctagggac   tccttgatac   acaagcatac   87660   aggatcatat   caggacccag   ccggcagacc   agacccacca   aaggctgtca   caggcgggct   87720   ctgtcttccc   ccatgtgacc   tccctgaaat   ctaggagaag   gaagggggca   ctttggacct   87780   ggtcaagcta   tgccagcaga   gagttagcct   ttgacctagg   atccaagggt   taccccagcc   87840   tcagatctag   caaagaagca   ccttggagga   cacagataag   aaaagacaga   ctgtccaagg   87900   aggacatcac   accttcagca   aagatgagca   taggcttttc   tgattggctt   gaattttcag   87960   atacataaag   ctgccatctg   tccatagaac   aatcttggac   ccttagccag   taaaggcacg   88020   gtagtccatg   ctttcccaac   tgacacacca   attcaaacca   gatgaagcct   aagaccacct   88080   actacaactc   ttccccacat   tccttctcag   taaacctgaa   tgcagagcct   ctcgtatgca   88140   atagactctg   agccctggaa   gcctaccctg   cttcccagga   aaacattact   ggacctgacc   88200   aaggtcaacc   tggagaaaac   acagcaaacc   aaggggccaa   aagaagaaag   ggcttctagt   88260   tttctgttgc   tataatgtga   taggtctgca   gaagaagact   ggctccgctc   agcttccaca   88320   cccctcccat   tccttgtgca   cacacataca   cacaatgtag   gctcatatac   acaaatacac   88380   acaccttcac   acactcagtc   tcacacacac   taacaatgca   tactcacact   tatatgcact   88440   cacacaagga   caacaagcta   taccacaaat   atctggaata   agaagagcta   ctttctgttc   88500   cagatgtacc   actaagagct   gtgccacctg   gcaaagttat   ctaatccctg   ggctccaaga   88560   ggttaaaata   aggggataga   ctggattctc   cctaagccct   cctcttttca   gactgtgagc   88620   taaacttttg   ttggaccaag   acagagcttg   ttttctccct   cttaaagcta   ctttttccaa   88680   taagactgtg   aggccaccaa   gaggatgcac   aagtattatc   gggcactgat   aacttgtgtc   88740   tctttcccta   gaagtatgag   tggagataca   taagtctggg   ttcttcacct   gaattcagtt   88800   caattcttct   gtactcttct   gcatgtcagg   agtggtacag   ggactggaaa   tgacaaagaa   88860   tgaatagcat   ggttcctgac   tataggcagc   ttacagtcta   gggccttggg   tcttggccct   88920   ggctacacag   tagaatcgcc   tggggaggtt   ttacaatgca   ccgaaaccca   gacaccattc   88980   cctgagagat   tctgattcac   tccatctgga   gtggggcatg   gattttatta   tttcggaaaa   89040   gctcttcagg   catttttaat   gttcaaccag   gtttgacaat   ctggtccagt   gagacacaga   89100   gtcataggtg   gctgaccttc   acactgactg   accagtgtaa   ccaagagatg   tgcttaggct   89160   gcttggaaat   tttcaaccca   cgaagaccca   gtcattcagc   ttggggctaa   ttccagcccc   89220   catgcagatc   ctagtgcttc   ccttccactg   gtgcagtctc   cacaccctgg   tttttctgga   89280   agagcaaaag   caatcagaga   gaataaacac   aagtgaattc   attctattca   gtgcagttgg   89340   ctttggttaa   aagttagaat   atttgtccaa   aaaaaaaatt   ggatgaagac   atccttcctc   89400   tatcagatat   caaataaatg   cagtaagtac   tataattata   atactaaaat   cattgaaaga   89460   gaattatatg   gacagggaaa   tccaacagaa   cagaaatgcc   aaaggaaacc   tcaggaattt   89520   agaatgtgat   gaagtatttt   aaactgatag   agaaatagaa   actgtttaat   tactaatgtt   89580   gagacaactg   cccatatgga   aacaatcata   tattagattg   ttgtatcata   tgctacaagg   89640   aaatacactc   cttatgaaac   gaagaacaaa   agatagaatg   agggagaaag   aaagagtgag   89700   agagaaagag   tgaggaaaga   aaagaagaaa   gaaagaaaga   aagagaaaga   aagacagaaa   89760   gaagaaagaa   agagagagag   ggagggaggg   agaaaggaag   gaaggaagga   aaagaaagaa   89820   aggaaaggaa   agatgggtag   aagaagagag   gcggggggga   agaaagagaa   agaaaaaaat   89880   ataggaaata   ttgtttatat   tcatgaaaaa   ctgaaaagat   ctgaatatcc   tacaatgagg   89940   gaactgagca   taataattat   atagccatac   aattaaatgc   tacagctcta   aaaattatta   90000   tagagagtgt   acttattgcc   atagaaatat   ttctccaata   tagtacatta   agaaaaagaa   90060   agcagtatgg   tatccctttt   atagatatgt   gcacatattt   ttacttagcg   gaatatatat   90120   gtgcctgtat   agacagatga   tagctagaga   gagagggaga   tggaaaatat   caagaagcta   90180   aatgttatca   atgttattgc   agttatctct   gagtggtcga   attttaggtg   atttttgttt   90240   tcttctgttt   tccacttagc   catgtgttaa   aattctctac   agttacctaa   atcatacaat   90300   tgaaacaatt   gccctgtggc   cctttagcct   gagtaaacag   ccaatattat   cctagatgta   90360   tatatgtata   ttcatttttt   aaaaccatca   agtcaggccc   cacaaaacat   atatttgcca   90420   tgcacttctc   tgtataaatg   tcatgcttag   accctctaag   gcccactcaa   agccatcgtc   90480   cactcatctc   aacacagaca   gacaagtcag   agggacagat   ggagcagctc   ctgtctgcca   90540   ctgctaggtc   ctttccttgt   gttcttccct   ttaactcatt   tcctgtttgc   cccatttcac   90600   ccatggccca   ttgcccattt   cccatttaac   ccatttcccg   tttgagaata   ctgcacaggc   90660   agtgagttgc   actttttttt   ctaaacagta   aatgggttaa   tccacaactc   tttgaggtgg   90720   tgttattaac   tgtctctcac   agacagaaga   caggctctaa   ggttaagcag   tttgcttgtt   90780   ttcacaggta   gcatgtgaca   caactggtat   tcaaatgcac   acatatcgaa   tgcacacata   90840   aagacaagac   tttttaaaaa   ttaaaaacac   aaactcaggg   tcttgggtgt   ggcttccttt   90900   cctggacact   ccctccagag   ctctgccttc   agagagctat   tcctacctca   tctcttgaag   90960   gttttgtttt   gatatattta   aaatctaaag   gctgagtcat   gactggcagt   gtggagacca   91020   attttgtgga   ggtagccaag   gtcttcctgt   actttcagac   gaatcagatt   ggtgtaaacc   91080   cctgtcttgt   ggtttctgcc   aattccatgg   gtctgacatg   ctcctccctg   cttgtccctc   91140   tgagcactca   ctctcactgt   ctcacctcct   acagctctcc   tgttccctcc   acagcagcac   91200   cgcccttgct   caccaccttc   ctcagcatgc   ctccccccag   ggaggctgac   ttcctggctc   91260   tctccaaagt   tcccatgtct   cctgctacta   catatgccag   tccttgcact   cttgggctcc   91320   cctgacttcc   agtccttgaa   actgtacaaa   gccacttgga   ggtctgtttc   ccactgatat   91380   agagtatgcc   attactccct   cccaaaggtg   gaaatggcta   ccgccacttt   ttattcttta   91440     atatagtttt   atatctccat   tatatctatt   tttcagttac   cattagtgac   ctcatcccga   91500   aatgagggca   gctggtattc   tcagaagctt   atatgctctc   tatatcttat   tcgtaaaagg   91560   aacacagttg   atttcccatc   tgtcattaca   gccaagcagc   tctggagttg   gcaggtaaat   91620   cggatctcca   gctgggtccg   catgccagga   ttattgctca   tcaacaaggg   gacattagcc   91680   atgattagtg   tgaacagaag   ctcactttgt   aaacactggc   taaaaagctc   aagccatcct   91740   tagtgcgaga   accactcttc   ttactggcaa   atatgtctat   tttctcccag   tcaaaggctg   91800   acagaaagga   aaagaaaaag   aactgtatca   taaattatct   aaagactttc   ccataaacaa   91860   gcatggaggg   caagacgtgg   aggagatggg   gtttccaatc   ctgtatgtta   tctttcataa   91920   atgagcatat   tattgatgtc   caggaggatt   tacttgtaca   aactagaaca   tttactgcat   91980   gggcaagtgt   acccatggaa   cagacaaatt   tctcgttata   actatgtcta   gctatcatgg   92040   cttggtggat   atgatccatg   ctatatccaa   attccatgtt   tttaattaaa   tgaggcatac   92100   tagagtgaag   aaaaaaagct   atggattagg   aattgggtta   gagccctatt   cactttctag   92160   cttaaaaaat   ttaagagaag   cccattaact   gctgtccata   tcagcttctt   catcagtaac   92220   tgtgaataaa   atggcttctc   tacaattaac   tacatttaat   aaaagatgtg   acagaaaaac   92280   ataaagatac   aaataaatga   agattatgac   atgttcgtgg   atagtaaggt   tcaatatcat   92340   aaagacgttg   aatctcccca   gctgatctac   aagttaataa   atttccaatc   aaaattgaat   92400   tgaaacattt   tattgagctt   cacaagccta   ttttaaaata   gacaaggaaa   attcaagaga   92460   catgagtagc   ctagacattc   ctagagatgc   agaacaaggt   agagaaagtt   cggtttacca   92520   aatattgaaa   tgcattgtaa   agtcagagtg   cggtattggc   acaatagaca   aatggaaacc   92580   tcagtttcct   tgctacctgc   tgcctgccca   caaatccaat   gctacatgtt   tcaggttttc   92640   attaagacag   cactgcatat   gaggtaccaa   tctctacttt   agaagggaaa   cactagcagc   92700   tgtgtaaacc   caaaatatca   gtagccttaa   ccacatagaa   gtttatgtct   taattaaaat   92760   ccagttggca   ataaagacca   gagggaaagg   cattgttcca   cacagtcatt   cagaaattca   92820   agcaggtgga   gattctgcca   cgcttaacag   agggctgcta   aggtactcct   cagcacctcc   92880   ctccagcaag   caggggcagg   ggaagagaga   gagtggagga   ctgctcagga   gaggtatctg   92940   agcaaggttt   ggggtggtag   acattgactc   ttcccagcgt   cccttaggca   gatcttagtc   93000   acatgactac   acactgctgg   aaggcaggct   gggaattgta   gtctagttgt   gtgccctggg   93060   gaaacaggaa   tgggttttgt   gaacagctcg   tctgtctttg   cataattatt   ttaggttgtc   93120   agggctcctg   cttctgcttg   agctgaggtg   ctgaggtata   gaaagggcta   tctggtgggt   93180   taggtgaaat   cttctcatat   ctagctacaa   tttacagttg   gctctgggct   aaaaacctcc   93240   tgaaagggaa   tgcaaaatac   tttcctactt   gaaggttcag   agatatgcag   ccatgcccag   93300   ctaatgggga   gagagggagc   acctcagatg   acaagaaagg   gacaataaga   tccctgaatg   93360   tgaaaacatt   ctggagaata   cagagtcagg   cacaacgagt   gccttaaggt   caaagggaaa   93420   tgtaagttct   gggtcacaga   agaacattcc   cacctaacca   gtctagccac   ttgtcagcat   93480   ctagggagga   aaacgctgtg   tggtgggcta   aagaagctga   aactcttggg   agttcaagag   93540   agtctgctgc   aagctcctgg   aatagcacag   cacagaggag   agaggccatt   cccttgcttc   93600   ctggcagaag   aacttggaga   gcaggtatcc   ccgggaagag   cagcagtgct   gaataaccca   93660   cagcccaggg   acagaactga   ggccctgaga   ggtggagcag   aagagcaaag   caactgctca   93720   gggtcaagag   caataatgaa   gaggactcac   aatgtggctt   tgaagtgact   gtggctgtgg   93780   agcatctgct   gcccagccgg   agcccctcaa   gaggcagtca   ttggcaaaga   agaggtctgg   93840   gtgaagtgac   ctggggcagc   ccagggcagc   tgcagcgagc   ctagaaagca   ggatccagag   93900   agcgtcacca   tagaggcaca   ggcggtggcc   tgggtggatc   ctggaacact   gagacacctc   93960   cttgggtctc   tcagaaaaac   tagagggaca   aagacaccct   gggaagaaac   tgactgcagt   94020   cctgcagcta   aacagatgag   acctaatgtg   cttaaaatac   tgaactgatg   agttcagggg   94080   gtattcaagc   tatattagta   aagagaatgt   tactgttcaa   aatgaatttc   aagtttcaag   94140   aattttagaa   atcaggaaaa   actgctaaaa   ataatgttga   agtttttaaa   aatcaaccta   94200   acatacacaa   agtttctaat   ataggaagca   tatttatttt   aaaatattgc   cagttagact   94260   tccatgtttc   taaacaaatc   acctgcactc   tttataggtc   cacagatgaa   ttccccccac   94320   tacctcccag   gaaacaattc   tgcttccatt   tgaaatccca   aggtaccaat   tgctcccact   94380   tcaggtgctc   agcctgcatg   aagaatgggt   tcccagtggc   tctgtaagag   gcctgtggtc   94440   ttgggaaggt   gaagagccag   aaatggccat   ttggagacta   aattttcagc   atgtgtcccc   94500   attagggata   tactgtggaa   ctaaaacagt   ccacatcagt   gtgcttctcc   cagtcccaaa   94560   tcgaatggac   gtgccacttt   caggctaccc   aagcccgatt   aatgcacagt   gacagctgaa   94620   tttaactgct   tttcactgca   gctcagcacc   aaggcctgag   tagaaataat   ccttttatct   94680   cccaataaag   gagaattacc   agcatttcgt   agctcagctc   tctctgaagg   gccatcttta   94740   gaagctagag   gctttgcatt   tcctggaatg   gtcagattaa   agatgctttt   cactgatctc   94800   tgggcaggta   gatgattata   ctgtcaccca   gttgccctgt   gggaagcact   gcgggtttcc   94860   attttaaagg   gatttcagag   cacaaatgtc   attctcgaaa   acttttctga   aagaccaact   94920   tagtcttcag   agccatactg   tttaggggcc   agctgttgcc   taggaacagc   cagtggggac   94980   actttgaaag   aatataattt   ggtctgctgt   cttctttact   tgcagcccat   gggtataatc   95040   tcacccagat   cattttctgc   tgtacataat   tcttacacag   ataacaacct   tgtataatag   95100   acaatcaaac   gtaataatac   cttctgttca   ctttcataaa   gatggctcac   cttccacctg   95160   aaatactgtt   aaaactgcat   tcatctcaac   tgataagata   tccaacatcc   aagcttgggg   95220   cttggttcct   tgaatatatt   attttatgtt   aatcttcaaa   gtaactctgt   gagttaggtg   95280   ttctacacct   caatttttaa   atgggagaac   tgaagccggg   aactttagac   gatttgccta   95340   agtttacaaa   gttttagtaa   ttggtagaga   cggaggatgc   attccagtct   gatcttaaag   95400   cctgtgtcct   tcccaagatc   ccatcctaga   aatcaaggtt   tctcaccttt   gacacctttg   95460   gcattttgga   ctggataatt   ctttgttgtg   gaggctgccc   tgtgcattga   aggatgttta   95520   gcaacattcc   tggtctctac   ccactagacg   ccagtagcac   acatgcacac   acaattgtga   95580   caattaaaaa   tgtgtccagg   ctttgccaaa   agttccctgt   aaagcaagat   cacccccaat   95640   ggagaatcac   tgctctaaat   gttctgaagc   atttgctttc   ttacaaggcg   gaggttcgtg   95700   gtcccatgaa   acgcacagaa   taatttgaag   accctaatag   tagttaggag   tggccttaca   95760   gggtatgctt   taagaactat   gaattaatta   taattagcag   ttttcctttg   tacttaaaga   95820   gcccctattc   ctgttactct   tgaaaatcat   ttgtcctctg   caatgagtaa   atgttcacac   95880   tacaaaaaca   ctcagaaact   tcaatggcct   gcggccgcct   caggagttca   gcccaagctc   95940   ctcagaaaga   cattcaaggc   ctccccagtt   tgccttcagt   tctctcctcc   actctccacg   96000   catttgtagg   ctacagccaa   gcaatatgac   tcactgtggt   taaaacgcat   cctacatttt   96060   cttgttccat   gtcttggctg   aggatgttca   ctctctgcct   cccacctctc   ctattcccac   96120   gctgcacccc   gcctttgtct   tggaatcctt   cttatgtcat   ttgagttcac   ttgatgctct   96180   ctgcatctcc   cacacagcct   ccccagcaca   gtggtggaag   gatgggtctc   tcctctgtcc   96240   acagttggtt   tctatctgtc   ttagtccatt   ttctgctgct   ataacagaat   accacagact   96300   gggaaattta   tttttaaaaa   aagagattta   tttggcttat   ggttctagag   gctgggaaat   96360   ccaagagcag   ggcacttgca   actggcaaag   gtcatcccat   ggcagaaaac   agagggcgaa   96420   agcgagcaca   caagacagag   agaacgtcaa   gccaaactcg   tccattccat   cagaagccca   96480   cttccacaat   aacacaataa   tagcaataat   ccattcataa   gggccaagcc   ctggtggcct   96540   aatcacctct   caaaggctcc   acctcccaat   actgttacaa   tgacaattaa   atttacacat   96600   gagttttcgc   agggacattc   aaaccatagc   agcatctcgc   tcatctcaac   cagttttatg   96660   gagagtgtgt   gatctttcat   gctggattga   aaattcttag   agaacaagta   ctatatttca   96720   gcccttctag   tcagccctta   tctcttagca   gatgtgctat   acttgtgggg   ttggactaga   96780   tcagattgga   ggtgggattg   aatgaatatt   gcaaaggtaa   acatctgact   ggtttccatg   96840   ggcattaaaa   taaatttcaa   acgtgagagc   tgggcagtgc   agtgaagaca   atggagcaca   96900   gcagtttgcc   tttctagtgc   acagctgccc   tactccatgt   gacttcgacg   cactgagtgg   96960   gcctctgacc   agggccacca   ctagggttgt   gcccattgag   ccccactggg   tggctcctgg   97020   ctgagagttg   tgtggggctg   gcattctctt   catatttccc   tcactgaatg   tgtgctctag   97080   gcaggcctgt   gcttatcaga   gcaagggccc   cttttccaaa   acttcacaaa   ggtggcatat   97140   gggctggtgg   aagccctgac   agtagaaact   cttccatgag   gaaagggaca   ccacatctga   97200   catttttatc   cccaaattta   taggtaatca   ctactttaac   taagacctat   tgcactggat   97260   cctgcctagc   ctaatcttat   ctctttgttt   taccactgct   cgactaatga   gacgtggtta   97320   ttgattcatt   caacatattt   gtgttagtca   ttgaatatgt   gtttaggcta   caggagtgcg   97380   aaaaacaaag   cccctgtcct   catcaatcta   gtagattctt   atatttaccc   ctcagtgttc   97440   accatcagac   aacagtctac   atgaaaaaaa   tgtatatcca   agctcctatt   ttggagagtt   97500   ctgatacccc   tggcttgagc   ttttcttgag   agcgtttttt   tgtttttgtt   tttttgtttt   97560   ttttttgatg   aatgtatttg   tctggggcaa   ccattttaga   aagtaacttt   gcctaggttg   97620   ctaaagtggc   cacatgggtg   tctcagcata   gccagggctt   agagagtagc   actcctcagg   97680   gaagccctca   gctcctgcat   ggtcacatag   tctaatttca   gctgcagctg   ttcaggccat   97740   ctgaccaagt   ctcttgagag   agtctgagca   ttccatacca   ttttcatata   atattaaggt   97800   gtaaactgag   cattctccag   aaaccagccc   agaaccaact   ttaatctaaa   aatttccaag   97860   gaaggccatc   tgggcataaa   tgcacaggat   ataagtgaaa   actggagtct   aacctttaat   97920   gttcctattg   gctgaaacag   aaactgtcac   atcttgaaca   cttcatctta   ctattcccct   97980   cttgtatcta   cagatgggaa   actgtgcagc   agctataacc   gaggactcat   ctcaaaggca   98040   aaaatcaagg   ctatcaagta   tagcatcatc   atcattcttg   gtaagcaatg   tccctccttg   98100   agctgtaaag   tggtatgaac   gtcccaaaag   caagtttgct   cctgggacct   ggtgacagaa   98160   aggaatttgc   cagcaggcac   ttctacatcg   gtctgcttag   aaaggaatgt   gtgaaactca   98220   tcagcccatt   catggttcct   tcaagggctg   acatttggtt   cctaatccct   ctccttatga   98280   atacagctgc   ctgccaaggc   tatgggttga   ataatttgga   gaaaatctga   ttaggaccta   98340   actagctgaa   gtggatgctc   acatcaccag   ctaactcagt   cacttccttt   aagacacact   98400   gttccccagg   gctggggttc   acaaggaaaa   agaagataga   agtgactttc   caccttatct   98460   gaatatttga   tgttaggttc   actacaaatt   tcccccttat   ggcaccttac   ctgtccttcc   98520   accctcctgt   cttctgccct   ttccttttcc   tttctctcta   ctcttaagac   tcataaagtc   98580   accttgtact   atttcctgtc   tgaaatgctc   tgctctccgg   tggtgccccg   gcttacttcc   98640   taatctcatt   tgttccccaa   caacgccacc   tccctgaggg   ctccttgata   agtcttttta   98700   aatatccttt   ttctagtctt   tactctgcct   tatttttctt   cttggttcta   cctcacacta   98760   caggatatag   attgattttc   ttatagtctg   tctcccctgt   tggagtaata   aatctatgag   98820   ggcaatgact   ttggcttttg   tgttacttat   attatcacca   agacctaaaa   tagggtctcc   98880   tacacactgc   tcaattaaca   tttgtcgaat   gaatgaatga   atgaatgaat   gaaactatgc   98940   aagaaggaaa   gggatttgtg   ctattttctt   ttttatttag   cttaaatacc   ccctccttgc   99000   ccctctggca   gctcttgaag   ctccaaggaa   gtatagatct   cccattagct   gcatgtcaaa   99060   gcaccagaac   agtcttctca   cctttctgaa   acgcaactgc   aaaatgccaa   acagaccaca   99120   tgcagcatga   ctcagccagc   aggacgtagg   cacaaaacaa   cactgcaggc   ggaactggga   99180   taaaacatat   acaacccacc   agggctcaaa   aactttctga   gaggcttctg   aagtggatca   99240   aagactcctc   tgtcttctca   gatagtcaga   agcacagatg   caccttccct   ggatgttgag   99300   ttggaaaatg   tatgtttaca   agtgtgtccc   agagcctgca   tagcccttta   tcatcagcaa   99360   ccatactcca   aggccctgtg   ggatcccttc   agtcagcaag   caggaacacc   ctctagaggg   99420   tcagcaaata   ttgacagccc   taccagacca   aaagggccct   ttctatagca   aacactttgt   99480   aatgtccctt   tcccaccttg   aaataaaatg   cctacttata   ctacctacca   tgatcattta   99540   aacattaata   tattgcccaa   aaaataatat   aaaagaggaa   taagaaaata   atctaaaatg   99600   aaacaatact   atttttatta   attaaaatgt   ataccaattt   taatgtgtaa   aaacataagc   99660   acagctacac   tagaagaata   atgaagtagt   cacacctaca   ccgtgacata   ataacagcca   99720   caatgaaaac   caatacaggt   gtattttcat   ggagacataa   aaaaatgtaa   gcggtattac   99780   tgtcagtgac   ttgattttct   cagccggcaa   aaagaaaaaa   tccttgtaaa   attttgaatt   99840   aaacaaagta   atctttcctc   catttacaac   agaattgcat   ttctacaaat   ttcaatattt   99900   atttaaaata   cctgtgttta   tgtgtaaaac   aaagtcaggg   ataactatat   agagggttct   99960   cctctacatg   aatggacatt   caaagtagat   acaaatcttt   gctggctggg   ctgcgtgtac   100020   catgccagag   gactggtatc   cctgtgaaaa   cttgaatcct   ccaacccaac   ttacacacac   100080   atggaaacac   ccaagcacac   aaggccattt   gcccccttag   tgccccaaat   actgttcagc   100140   gccagtgttt   gggttaatta   atcaggatcc   agaacactta   gtgggttttg   tgtttgaatt   100200   gctcctcttt   aatacattgc   caaggtctgg   ggtcgagagg   ggaaacatga   gtcagtttat   100260   cattccaaaa   ttcccaacaa   gttacagaat   ccacattaca   cagttatgac   ggaaagcttg   100320   tccctgcatg   cagacagcca   aggaagccaa   acttctactc   cgagaaaaaa   ttctatggat   100380   gagaaagtgg   gagaaggaga   gtggagaatt   tatggtgaga   aaatgagaga   tgcgtgtatt   100440   tccagtcaac   ttaggagtgt   ttagctctgt   gtcgtagtct   gtgctttcca   gaggaaaatg   100500   agagagagag   agagagaggt   ttattagaag   gaattggctc   aggtgattac   tgaggctgag   100560   aattcccaag   ctgcagttgg   aagctggaga   cccaggaaag   ccaatggtgt   aggttctagt   100620   ccaagtctga   gtccaaaggc   ctgaaaacca   ggagaatgga   tggggtacat   ttcagtttga   100680   ttccaagtgt   gaaggcagga   gaagaccaac   gtcctagctc   gaagacaggc   aagcagagtg   100740   aatcttctct   tacttaacct   ttttctatga   ctcaggcctt   caacagattg   gatgcggccc   100800   acaaacaggg   gagggcaatc   tgcttttctt   actctaccag   atcaaatgtt   actctcctcc   100860   agaaacgtct   tcatacacag   ccagaatagc   gtttaaccga   atatctgagc   actccgcagc   100920   ccagtcaagt   agtctcaaaa   ttaaccatca   cacctgaatc   cccacctttc   atccatttct   100980   tttgagaatt   tggggccaaa   taaactggtg   tctcacacat   actaatggat   ttagagaagg   101040   gcatagtgtg   ggtgttttct   ggagaggcca   acttaggggt   gaaagagaac   actgtcatta   101100   aactggggtg   tgcttagaaa   atattgtgat   caccccttct   ccctgacata   acctaaaacc   101160   agtagtcact   acgtggtcaa   aatgcttcct   gttcttctgt   cttttcctag   agagtcttca   101220   aagtattttg   ctgttttttc   ctgatccctg   tgtttgatga   gaggctatgg   ccatagtcac   101280   ttgttcctgg   ggtttctata   aagaatccac   agcagtagaa   atttgaccct   tctttacaga   101340   tagagaagct   ctacaacctt   tgctcagttt   taggaacatg   tctgtaagat   gtgtccatat   101400   atatgtgtgt   atgtgtttaa   gtgaatatga   catcaatgct   ccaaacaaca   tcaaactcca   101460   atatttctga   gaaattccag   gtcccaaaga   acctctcagg   taaaagtcca   gtcaggacca   101520   accaaaagag   acccctcaac   tgctacctgc   tgtgatgcta   atggctctct   tctcccccag   101580   ccttcatctg   ctgttggagt   ccatacttcc   tgtttgacat   tttggacaat   ttcaacctcc   101640   ttccagacac   ccaggagcgt   ttctatgcct   ctgtgatcat   tcagaacctg   ccagcattga   101700   atagtgccat   caaccccctc   atctactgtg   tcttcagcag   ctccatctct   ttcccctgca   101760   ggtaagggga   gctcttgcat   gggtcagaca   cactgatggc   cattgcactg   ggattctgcc   101820   aacatgtggg   tttctcatgt   ccaaactctc   cagcaggtta   caggatcccc   cttttataga   101880   tgagaggctt   gagagtcaga   aagattccac   taacatggct   gattcaaacc   ctggcccagg   101940   ccacagctga   tgtgcttttt   cctataccac   aacacctcct   atagaaaccg   ccttaatcat   102000   gacacatcta   gaagaccttg   cttttattca   aagcatcttc   acatgaattg   cctcaataca   102060   attctatgac   aataatagga   catggagcct   tattaccatt   ttacagataa   acaaagtaaa   102120   tcttacatgg   ttaatagcat   cattgagttt   atatgaataa   ttagcagagc   tgaaattaaa   102180   tccctcccac   agttattgtc   attcttctgt   ttttctgctg   tttttcagct   gttttcctgc   102240   ataagccaca   gagggtaggt   ggagaattcc   ataggagata   caagaggaga   atgatttaag   102300   tcttgggctt   aatgccagtg   gttgaagttt   atatcttgcc   attgtttatt   cgtagcgatg   102360   tgtgaccttg   ggcaagttat   ttaacatctg   taaacctcag   cttcttcatc   tgtaaaacag   102420   ggctaatagc   agtgtctacc   tgttggcctg   tgatgtctct   gtcctctggg   ctctggtgct   102480   gaccagtgag   aaggagagct   gtgagtcatg   gagaaggaag   gtcagggtat   tacatgtaaa   102540   gtgcctggca   cagttgtctg   acatttaata   catttttcat   aactattgtc   tcttaacatg   102600   tctacttgcc   ttttcattag   gtcaaattat   ttccctccct   gctttatttt   gactttctcc   102660   agggagcaaa   gatcacagga   ttccagaatg   acgttccggg   agagaactga   gaggcatgag   102720   atgcagattc   tgtccaagcc   agaattcatc   tagaccctag   ggcagtgcca   gtgctaggct   102780   gagcaccatc   agctctccca   ggtccttgtc   acctgcttgg   gcacgtgcat   ggaacccgag   102840   ccaacttcac   cccaccctcg   tcattacctg   ggagatgcac   aagacaaatg   ttctaatgac   102900   tgcatgcact   gcttaagtat   tggccaacac   gaactcccca   gttattcatg   ccagccagga   102960   aggaaacgcc   ttccttcccc   accattccca   gccctccttc   ccactggcca   gcacctgaac   103020   ccagtgaaca   caggcattag   tggtccaggg   tcctggcttg   gagccagtga   gtagacaggc   103080   aagcagaggg   gacaaaggta   gctgggttat   acatgaatat   tctcattaca   atagaagaaa   103140   ataaaagact   taattaagcc   catatttttc   ccccagtttt   ggattggagg   ttcagtgctt   103200   gtagccaaga   aagtgtttgt   ccttgaaatg   ccaacaaatt   catttccagg   cattggtgct   103260   ttgcacctct   tctgcagata   gcctcggttt   gcagatgggt   gtgctggcag   cagccaagtg   103320   catctcctat   tccatcaaca   aatgatttgt   ggaaagggca   tgctgttggc   cctccaaaac   103380   aaggtgaaat   gaagggatac   tggttctgat   agaaaacaag   ttgtgttaaa   aagccatggg   103440   cccctgactt   ccagcctctg   agagctggtc   cctagaacac   acatgtcctt   gaggcctttt   103500   tttttttttt   ttttttttga   cagagtctcg   ctctgtcgcc   caggctggag   tgcagtggca   103560   cgatctcggc   tcactgcgag   ctccacctcc   cgggttcatg   ccattctcct   gcctcagcct   103620   cccgagtagc   tgcgaccaca   ggtgcccgcc   accacgcctg   gctaattttt   tgtattttta   103680   gtagagacgg   agtttcaccg   tgttagccag   gatggtctca   atctcctgac   ctcgtgatct   103740   gcctgccttg   gcctcccaaa   gtgctgggat   tacaggcatg   agccaccgcg   cccggcctgt   103800   ccttgaggct   ttctgagacc   tctcactctg   tatttaacac   ttgggccact   ctgggctcca   103860   ttcctcacac   ctcaaggaca   agtcttcccg   tttttaaatc   cctgaacact   ctctttagtt   103920   aggagctagg   ttcctgcggc   ttttccaggc   ttaatgtttc   tctaagatct   ttatcgtttt   103980   tgcctgtcac   agcctttgtc   tagcacctcg   gccaggaagt   gctggtgggg   cttattttga   104040   cagatctgga   gtttggtact   tatgagcata   atccaagtga   aatatgacta   aatgttttct   104100   agtcaatttc   tcctatcaga   tttttccaca   agtggaagtg   aaataatttc   atttgatcat   104160   atatacatcc   cacccacggc   tccggtatgc   acgtgaacac   cttattttag   cttcacgtat   104220   attcttgaca   ttaaaatctc   cttggcaaaa   tcctccttcc   cagaattatt   cagagcttaa   104280   atcttttttt   ttttttttgg   catactgcta   gtggagtcat   taaattgttc   agtatatttt   104340   aatagcactc   cattcctcat   tatggtgtca   actcaaagtc   tgcgtgttta   aggtcatgtg   104400   ggcttttttt   ttttttttca   taaactcttt   tcaattcgat   ctgaccttag   ggaaagtatc   104460   tctttaggag   gaaatcagac   tcctcactca   ccatctctcg   gagaattatt   ctctgaattt   104520   gaggagcctg   aataatctag   ttttatttct   acacaacagt   tttgaaggct   gcctccctct   104580   ctgtccaatc   agctaaactt   gagcaaccct   ggaagctttg   agcatttgag   aaatgctttt   104640   caacatttca   ctttaaatca   agatgtgcca   cagaagaccc   acggaactgt   tgctaaaata   104700   gctggctgca   taggagccaa   ttaacagagc   aaaatctggc   agatgtccta   catggcaaaa   104760   caggaggacc   aggttctctt   gacagatgct   ttaattcttt   gtcaagtctg   gtctgcaaaa   104820   tacattacat   gcctacctca   tccatcagta   gagtctcaaa   gtgaaggaga   gagtggagtg   104880   aggggcgttg   tgatctgtag   aaactgtgaa   catgattttt   tagttgtgat   gctgtctctc   104940   tcatttcccg   agaagactgg   agatcaataa   gataatggta   caggttgctg   caggataatc   105000   taccatttaa   atgaaacttg   ctttatattg   gtaaggattc   tggaattgtc   atttccatgt   105060   atttgctgag   cttggagaaa   catgatttaa   gcattatgaa   aatactggta   attggccggg   105120   cacagtggct   cacacctgta   atcccagcac   tttgggaggc   tgaggtgggc   ggatcgtgag   105180   gccaggagat   cgagaccatt   ctggctaaca   cacggtgaaa   ccctgtctct   actaaaaata   105240   caaaaagatt   agccaggcat   ggtggctggc   gcctgtagtc   ccagctactc   aggaggctga   105300   ggcaagagaa   tggcatgaac   ctgggaggcg   gagcttgcag   tgagccgaga   tcacgtcact   105360   gcactgcggc   ctgggcgaga   gagcgagaat   ccgtctcaag   gaaagagaga   aagagagaaa   105420   gagagagaga   gaggaaggga   ggaagggagg   aaggaaggaa   agaagaactg   gtaattaata   105480   gtcttgtgac   cttgaacaaa   tcatttcaca   cctctgggca   ttgatggtct   catctgcaag   105540   atggggacaa   aatcaggctg   gagcagagga   tcttggtggt   tcctcccaga   cacagggagg   105600   aatctctgag   attgtgtcca   gagactgaga   gcttctggaa   ttctttcccc   aggggaccaa   105660   tgactcagtg   acttaatgct   gagtcatttg   ccctttagag   gtgtgacttc   cctatctatc   105720   actatgaatg   cagagaaata   tcatttccca   actgagaaat   gcagtttgag   ctcttggaga   105780   aaattagaag   cttagtgctg   tgacgttact   gtttgtagcc   tttgatgccc   atgtggaatc   105840   agagaagcca   gcaggacgat   acctaaaata   aaggtgctta   cttcctgcac   caagatgtca   105900   aaaagtctac   ggttacatta   ttgcatggca   tcagggacag   gagaaaattt   aggagagcca   105960   gagtaggtgt   gcttactata   ttcagccata   ctatagaaaa   taagtgactc   tactttactg   106020   aactaaatta   aatatgaatg   gtaaaaactt   ctctacttgc   agggtgcatg   tggaacataa   106080   tttagttgaa   gtttactttc   actaagcaac   atctcccacc   ttttccccca   gaaaatgata   106140   acaccttttt   aatgactttc   atttgtggat   aatcatgaga   aaaaagtggc   ccttttaaat   106200   cagcccaaca   cgcagcttaa   acttcctagc   ttcatttgga   aaattctgta   tactcatata   106260   cagaaatgac   cttgcctctt   ccaactgcat   tcacagtgtc   aatcagaaaa   ggccacaagc   106320   ctgagtctga   gagaaaaaag   gaaaacaaaa   atatctttag   caggaaataa   agtgattctg   106380   ctcttctaga   attatcttag   gtgagctaca   gacctgctgg   caactccaat   tttctcgtca   106440   tcatttgaat   gtacctcatc   ttatatgcag   atcgactagt   gtcccctgaa   gctttccaat   106500   tgcatttaat   tgaagggaga   aggccgtaac   tgtttaggtg   ccaatctctg   tgtttttgga   106560   aacatgaaaa   acccatgcct   catttctctg   caattgcatt   ttaagaatac   attatccgca   106620   acaggctggt   tcagttaaca   ttttaaaagt   gcaattccag   gcttacagaa   tgttaaagtg   106680   gatggagtct   tatatcagct   ggtctcatgg   ttttcagatt   gctcggagac   tccaaagctg   106740   ttccaaggag   gtgactcaat   attaatattt   ttacacggcc   gggcatggtg   gctcacgcct   106800   gtaatcccag   cactttggga   ggccgaggag   ggcggatcac   ctgagttcga   acaggagttc   106860   gagaccatcc   cagtcaatat   ggtgaaaacc   cgtctgtact   aaaaatacaa   aaattagccg   106920   ggtatggtgg   cgggccgcct   gtaatctcag   ctacttggga   ggctgaggca   ggagaatcac   106980   ttaaacctgg   gaggaggagg   ttgcagtgag   ccgagatcgc   accattgcac   tctggcctgg   107040   gcaacaagaa   ggaaactctg   cctggaaaaa   aaaaaaacaa   aaaacaaaaa   aaacattatt   107100   acgcacaaca   atagcataga   aatcgagagg   caattaaatg   tagttagttt   caagttcctt   107160   gaattatctt   gcatgcaaaa   tgtggattat   tagtcattga   tacaccaaga   atgcatgtta   107220   caatttcagg   gtcatcacta   aaatagtaga   aatgcaaaaa   catacatctt   tcagagtaaa   107280   agagaaggaa   agctgagtga   taagtcactc   aaacaatttg   aaagaaagca   agaaaagaga   107340   gaaacaggga   cataaaacag   ctgggataaa   cagaaaacac   ataggaagat   ggtggttttt   107400   ttatccaaac   ttactagtaa   ttataagtgc   aaagagatta   aatgccccag   ttaataatca   107460   aagactatca   gactgatttt   ttaaaaaata   aataactgtc   tgctgtttac   aaaagacatg   107520   cctaaaacac   aaggatacag   aaagtttgaa   aatcaaagaa   tggatcaagg   tagtccatgg   107580   aaactgtagc   taaaagaaag   ctggtaaggc   tatattaata   aagactaaat   accttttaag   107640   gcaaaaagta   ttagtagagt   taaacaggga   cacttcgtaa   tgataaagat   tttagtttaa   107700   taagatgaag   taataattct   aaatttccag   gcatttaaaa   acatggcctc   aaaatatata   107760   cttttaaaag   ttgtcagaac   tatgaaacaa   atagaaaaat   tcacaaccct   aatggggaat   107820   tttatcacac   ttaatgccag   tcattaaagc   agacaaaaat   caggaatgat   ctggataatt   107880   tgagcaacat   gaataacaca   ggaagagcac   tttaccttac   cacccctgaa   cagacatttt   107940   tttcaagcac   acatgaagca   gttaccaaag   ttgaccacat   gctaggccat   aaaacaagct   108000   tcaacacttc   cacgagattg   aattcatata   gtctagtttt   tctgatcatg   ttataattat   108060   gctggaactc   aagagtaaga   ggataactag   aaaattcaaa   taggtttgga   agttaggaaa   108120   tacacttccg   aacaacttat   tgccaaagaa   gaaagcatat   gaaaattaga   aaacaattgt   108180   actgaatgtt   aatgaaagta   ttatgtatta   aatgtatagg   atacaaataa   aacaatactt   108240   aaagaaaaaa   gagaaggatt   aaaaatcaat   aaacaaatta   tctatcttat   gaagttagaa   108300   acatttacca   aattaaactc   aagggaagta   gaaagaataa   aataataaga   ataggagtaa   108360   aattaatgaa   ataagaaaca   aacataataa   aaagcctcaa   caaagatcaa   attctgttct   108420   tgaaaaaaat   taaattaata   aacctctggt   gagactgatc   aaaggaagaa   aaagagagaa   108480   ggcacaaata   aacagtattc   tttaaatgat   atcactatag   atcctagagt   caatcaaaag   108540   ataataaaag   gatattatga   tcaagcttat   gtcaaaacaa   attgaaaata   tagatgaatg   108600   ggaaaacata   taattgaccc   aagaagaaac   aaaatcagaa   tagtagcata   actgctaaag   108660   aaagagaatg   tcttcattgg   tgagctgtac   taaaatttaa   ggacaaagca   acaccaattt   108720   tatagtcttc   agtaaaatag   aaaaagtgtg   acttttcaga   ttatttatga   ggctaaaaaa   108780   acctcaatat   caaatgctac   aaaggcatta   caagaaaaaa   attgcaagtc   aattttgccg   108840   cagaaataca   tgcaaaaatt   ccaaaaaaaa   attaccaatc   aagtccagca   aaatataaca   108900   aggctaacat   atgtgacaaa   actggtttca   ttccaggaat   gcaatgttgg   cttactatta   108960   gaaaatcact   caactcgtta   cattaataga   ttacaggata   aaacaatata   taaactcaac   109020   agatgcagaa   aaaaatgttt   gataaaaatt   tttgagctcc   aattataaga   tattcacaaa   109080   cttgggatag   aagacaattt   cttaaacctt   aaagtttatc   tgcaagaaag   ctacaggaaa   109140   cctatactta   atggtgaagc   tttgaagatg   ttccttttga   aataaagaat   gagtgaagaa   109200   tgccagcatc   aacacttctg   ttcaacattg   taagaaatcc   tagtcaatgt   agtaaaacaa   109260   gaaaaaaaaa   taagataaaa   ggtatatgga   ttcataataa   ataataaaaa   ctgacaatat   109320   agatataaat   atatagctga   atatccaaaa   taatatacag   ataaattatt   agaattaatc   109380   acaaggttta   ataaggttgc   aggatacaaa   aatcaatatt   atttccacat   agcagcaata   109440   aataaaaaat   ttaattttta   aagttttgta   gcaatgaagg   gaaagcttcc   tttttgccct   109500   tggaggcttc   actgaaaaat   caagtcataa   aaaggcagaa   aaataagaga   aaaagcataa   109560   aatttattac   tcctatgcac   acggggaaaa   tcggagtgat   tggccaatac   ttcaatgggt   109620   acagatgctt   gtagagcctt   cttcttaggg   gaaagagaga   tggggaagcg   tggatgattt   109680   taggggaata   ataaatgatt   tttaggaatt   tggtgtgctt   gaagaaaaca   atggtccagg   109740   acagagtctg   ttggactcac   agagcagact   tttgtttgtg   acaaaagtct   gtccaggttt   109800   gttgatagac   tttagtcttc   cttcttgtga   tatgggttca   gttaatgaaa   actcggggaa   109860   gggaccgcaa   gtcattgttt   tcttctttgg   taagtctaga   attaggcaga   taaggaaact   109920   tcatgattga   gaggcaggag   gagggagagg   agaaacaatt   gttctccctg   gtaagagaac   109980   tgaaaactgg   tccagtcttt   atgtagatgg   ggggggaaat   ctcttctagc   atctgctgat   110040   ctctaagggc   cttttaattc   aaaatatatt   ataatgccat   gaagccatat   tttagggtaa   110100   agttccatga   gctcctgcat   tcgttattta   caatagcctt   aaaaaatgaa   attcgtagga   110160   gtaaatttat   aaaaaggtat   gtaaatctct   atggagaaat   tataatacta   ttgagatata   110220   ttctaaaaac   taatgaaatg   cagaaatata   tcatgactat   tgttcagaaa   ctgaatatcc   110280   taagataatc   aattttcaaa   ttgatctctg   aagattcagt   agaatgccaa   gaggaatttg   110340   gtagaacttg   gtaagttgtc   tctaaaattt   atatggaaat   gcaaagggcc   aaaaatagcc   110400   acgacactgt   tgactaaaaa   ggaaaaaaat   gggagagctt   aactaaccac   atatcaaact   110460   tatcatgaaa   gaataataat   gaaagctgtg   gtatgggcgc   taaagtagac   aacttagcaa   110520   atggaataga   aaaggagtcc   agggaaagat   gcccacatat   aggtaaacga   tttgtttcag   110580   tggtaacatt   gcaaagcagt   aaggaaaggg   tgttcttttc   aattatataa   aaaattggac   110640   atttatgtgg   gagaaaattt   actgaaaacc   tactggatta   tatgtcaaat   tggttccagg   110700   tggattatag   acctaaattc   acaagacaat   aggacgctga   aaaaataatg   tgacaaaaca   110760   caaaacactg   tactcatgaa   ggaaaatatt   gatacattta   attatttaaa   agtagaaatt   110820   tctgttcatc   aaaatatatt   aaaaatgtga   aaagacaagc   tatagaatgg   aaaatgacat   110880   ttgcaaaata   tataactgac   caagatctag   tatttgtaat   atataaaaac   tactgcaaat   110940   cagtaagaaa   aagattttaa   aagacagtag   aagacataga   aaaatgggca   agaattaact   111000   tcagaaaaga   agttatccaa   atggacaaaa   aactatgaaa   aggtgtttaa   tctcattaaa   111060   aagcaaggga   aagtaaatta   aatcacaatg   atatatgagt   atgcatattg   gcctaactta   111120   aaacctttga   caatatcaaa   tgttgctgag   gatgtacagc   atcagaaatg   cttatcttgt   111180   gtgaactgga   gaataattag   tacaactatt   ctggaaaatt   atgtgcctta   aactagtaaa   111240   actaaggatt   tgtgtttccc   gtgaccttga   aattccacac   tgagtacaca   ccctacagaa   111300   gtgtgaatta   tgtgcaccat   aatagatatg   aaaaatattt   gtaatagcac   taattataag   111360   agcctcaaat   tggaggcaaa   acaaatgctt   attagcagta   gaatagataa   ataaattatg   111420   gtgtatttca   tacaatggaa   tactttacag   caacaaaaaa   atgaagaaac   tgcatatgct   111480   tgcagcaaca   taaaaaaact   ttaaaaacat   aatataaagg   tcaaagacag   agacattaaa   111540   gataacatat   gatctcattt   atatgaaatt   caaaactaac   caaaattaaa   ttatcatatt   111600   tagcaatgca   cacataggta   attgtattag   tccgtttttc   acactgctga   taaagacata   111660   cctgagactg   gacaatttac   aaaagaaaga   ggtttattgg   acttacagtt   ccacattgct   111720   ggggaggttt   cacaatcatg   gcagaaggca   aggaggagca   agtcacatct   tacatggatg   111780   gcagcagtca   aagagcaagc   ttatgcaaag   aaactcccat   ttttaaaacc   atcagatctc   111840   gtaagaccca   ttcactatca   caagaacagc   acaggaaaga   cctgtcccca   tgattcagtc   111900   atctcccact   gggtcccttc   cacaacatgt   gggaattatg   ggagctacag   gatgagatct   111960   ggggggggga   cacagatcca   aaccatatca   gtgacaaaac   tctaaagcaa   agcaggaaat   112020   cactttttat   aagagtccag   attgaaatat   ctttgtgggg   agagggagga   gatgtacaga   112080   gagaggctgg   cagagtctct   tttttgctct   aggtggcagg   ttcaagggtg   ttcagtttat   112140   tttggaagca   gtgcagagaa   gggagccaga   ctagaaacag   ggaggtgatc   112190     <210> 17 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 17 cgtgaagtct ccagtgaatc gcc 23 <210> 18 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic DNA <400> 18 gatggtgctc agcctagcac tg 22 <210> 19 <211> 880 <212> DNA <213> Homo sapiens <400> 19 agcacgtaga tcctccctgt catcaggcag agctcttcag tgaggtgggc tcagggaggg 60 ctctgtgcct ccgttcagca gagctgcagc tgctgcccag ctctcaggag gcaagctgga 120 ctccctcact cggctgcagg agcaaggaca gtgaggctca accccgcctg agcc atg 177 Met 1 cca gcc aac ttc aca gag ggc agc ttc gat tcc agt ggg acc ggg cag 225 Pro Ala Asn Phe Thr Glu Gly Ser Phe Asp Ser Ser Gly Thr Gly Gln 5 10 15 acg ctg gat tct tcc cca gtg gct tgc act gaa aca gtg act ttt act 273 Thr Leu Asp Ser Ser Pro Val Ala Cys Thr Glu Thr Val Thr Phe Thr 20 25 30 gaa gtg gtg gaa gga aag gaa tgg ggt tcc ttc tac tac tcc ttt aag 321 Glu Val Val Glu Gly Lys Glu Trp Gly Ser Phe Tyr Tyr Ser Phe Lys 35 40 45 act gag caa ttg ata act ctg tgg gtc ttc ttt gtt att gtt 369 Thr Glu Gln Leu Ile Thr Leu Trp Val Leu Phe Val Phe Thr Ile Val 50 55 60 65 gga aac tcc gtt gtg ctt ttt tcc aca tgg agg aga aag aag aag tca 417 Gly Asn Ser Val Val Leu Phe Ser Thr Trp Arg Arg Lys Lys Lys Ser 70 75 80 aga atg acc ttc ttt gtg act cag ctg gcc atc aca gat tct ttc aca 465 Arg Met Thr Phe Phe Val Thr Gln Leu Ala Ile Thr Asp Ser Phe Thr 85 90 95 gga ctg gtc aac atc ttg aca gat att att tgg cga ttc act gga gac 513 Gly Leu Val Asn Ile Leu Thr Asp Ile Ile Trp Arg Phe Thr Gly Asp 100 105 110 ttc acg gca cct gac ctg gtt tgc cga gtg gtc cgc tat ttg cag gtc 561 Phe Thr Ala Pro Asp Leu Val Cys Arg Val Val Arg Tyr Leu Gln Val 115 120 125 atg gta atg aag tta ttc cat att cag aaa atg aat atg gaa tagttacaat 613 Met Val Met Lys Leu Phe His Ile Gln Lys Met Asn Met Glu 130 135 140 cttcttttgt gctctgaata taaccttgaa agtttggtat gtctgaaaga gccttttcta 673 ttatgtacgg ctagtatttt ctcaggggaa atttttaaaa acatatttca atttctttct 733 gaggatattt gcttccttgt aaataaatgc tttcaattgc ttattatatt agcaatcaat 793 tgctgataca caaattatga caaaacatgg tggcttaaaa caataaacat ttattatctc 853 acaaaaaaaa aaaaaaaaaa aaaaaaa 880

[Brief description of the drawings]

FIG. 1 shows a hydropathy plot of KAT0673L polypeptide. This plot was created using analysis software (MacMolly). The predicted seven transmembrane regions (TM1 to TM7) are shown in the figure.

FIG. 2. KAT0673L polypeptide and known GP
It is the figure which aligned the amino acid sequence of CR. Known GPCRs include human vasopressin 1A receptor (V1aR) and mouse vasopressin 1A receptor (V1aR-MOU).
SE), human vasopressin 1B receptor (V1bR), human vasopressin 2 receptor (V2R), human oxytocin receptor (OXTR), human luteinizing hormone releasing hormone receptor (GnRHR), white soccer vasotocin receptor (vas)
otosin). Alignment is CLUSTAL X Mult
iple Sequence Alignment Program ( ftp: //ftp-igbmc.u
-strasbg.fr/pub/ClustalX/ ). KA
The seven predicted transmembrane regions (TM1-TM7) in the T0673L polypeptide are underlined. All GPs
Amino acids conserved in CR are indicated by *, and amino acids well conserved are indicated by: or.

FIG. 3 is a view similar to FIG. 2 and is a continuation of FIG. 2;

FIG. 4 is a view similar to FIG. 2 and is a continuation of FIG. 3;

FIG. 5 is a view similar to FIG. 2, and is a continuation of FIG. 4;

FIG. 6 is a view similar to FIG. 2 and is a continuation of FIG. 5;

FIG. 7 shows the nucleotide sequence of KAT0673L cDNA and the amino acid sequence of the polypeptide encoded by the cDNA. The positions of the primers and the positions of the introns used in the examples are shown together.

8 is a view similar to FIG. 7 and is a continuation of FIG. 7;

9 is a view similar to FIG. 7 and is a continuation of FIG. 8;

FIG. 10 is a view similar to FIG. 7, and is a continuation of FIG. 9;

FIG. 11 shows the nucleotide sequence of KAT0673-3 ′ and the amino acid sequence deduced from the nucleotide sequence. The position of the intron 3 shown in FIG. 8 is shown in the figure.

FIG. 12 is a view similar to FIG. 11, and is a continuation of FIG. 11;

FIG. 13 is a view similar to FIG. 11, which is a continuation of FIG.

FIG. 14. KAT0673L polypeptide and known G
FIG. 3 is a diagram showing a dendrogram created using the amino acid sequence of PCR. Known GPCRs include human vasopressin 1A receptor, mouse vasopressin 1A receptor, human vasopressin 1B receptor, human vasopressin 2 receptor, human oxytocin receptor, human luteinizing hormone-releasing hormone receptor, and white soccer vasotocin receptor. Using. The dendrogram is CLUSTAL X Mu
ltiple Sequence Alignment Program ( ftp: // ftp-igbm
cu-strasbg.fr/pub/ClustalX/ ).

FIG. 15 is an electrophoresis diagram showing the results of examining the expression level of KAT0673L transcript in 35 kinds of human organs by using the PCR method. PCR cycle number is 3
5 The arrow indicates the position of the target amplified fragment (318 bp). The samples in each lane of the electrophoretogram are as follows. nc: Negative control, 1: Adrenal Grand
(Adrenal gland), 2: Brain, 3: Brain, caudate nucle
us (brain, caudate nucleus), 4: Brain, hippocampus (brain, hippocampus), 5: Brain, substantia nigra (brain, substantia nigra), 6:
Brain, thalamus (brain, thalamus), 7: Kidney (kidney),
8: Pancreas (pancreas), 9: Pituitary gland (pituitary gland), 10: Small intestine (small intestine), 11: Bone ma
rrow (bone marrow), 12: Brain, amygdala (brain, amygdala),
13: Brain, cerebellum (brain, cerebellum), 14: Brain, co
rpus callosum (brain, corpus callosum), 15: Fetal brain (fetal brain), 16: Fetal kidney (fetal kidney), 17: Fetal
liver 18: Fetal lung, 1
9: Heart, 20: Liver, 21: Lung
(Lung), 22: Lymph node (lymph node), 23: Mammar
y gland (breast gland), 24: Placenta (placenta), 25: Pro
state (prostate), 26: Salivary gland (salivary gland),
27: Skeletal muscle (skeletal muscle), 28: Spinal cord
(Spinal cord), 29: Spleen (spleen), 30: Stomach
(Stomach), 31: Testis (testis), 32: Thymus (thymus), 33: Thyroid (thyroid), 34: Trachea (trachea), 35: Uterus (uterus), 36: Standard 0.01 fg
(Standard sample 0.01 fg), 37: Standard 0.1 fg, 3
8: Standard 1 fg, M .: Size marker

FIG. 16: KAT067 in various human cell lines, peripheral T cells, and activated peripheral T cells using PCR
FIG. 4 is an electrophoresis diagram showing the result of examining the expression level of a 34L transcript. The number of PCR cycles is 25. The arrow indicates the position of the target amplified fragment (318 bp). The samples in each lane of the electrophoretogram are as follows. nc: negative control, 1: Jurkat, 2: Molt
-3, 3: Molt-4, 4: Hut78, 5: Namalwa KJM-1,
6: Daudi, 7: Raji, 8: HL-60, 9: U937, 10: T
HP-1, 11: IVEC, 12: HUVEC, 13: WM266-4, 1
4: WM115, 15: SK-N-MC, 16: PC-9, 17: HLC
-1, 18: QG-90, 19: PC-3, 20: KATO-III,
21: Capan-1, 22: Capan-2, 23: Colo205, 2
4: SW1116, 25: LS180, 26: T cell (unstimulated),
27: T cell IL-2 (interleukin-2) + PHA (Phyt
ohemagglutinin) + TGF-β (transforming growth factor-β) 2 days (cultured for 2 days), 28:
T cell IL-2 + PHA + TGF-β 4 days, 29: T cell IL-2 +
PHA + TGF-β 6 days, 30: T cell IL-2 + PHA + TGF-β 8
Days, 31: T cell IL-2 + PHA 4 days, 32: T cell
IL-2 + PHA 6 days, 33: T cell IL-2 + PHA 8 days, 3
4: T cell IL-2 + anti-CD3 antibody 2 days, 35: T cell IL
-2+ anti-CD3 antibody for 4 days, 36: T cell IL-2 + anti-CD3 antibody 6
Days, 37: T cell IL-2 + anti-CD3 antibody 8 days, 38: St
andard 1 fg (standard sample 1 fg), 39: Standard 10 f
g, 40: Standard 50 fg, 41: Standard 125 fg,
M .: Size marker

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 45/00 101 A61P 25/00 4B065 A61P 25/00 35/00 4C084 35/00 C07K 14/705 4C085 C07K 14/705 16/28 4H045 16/28 C12N 1/15 C12N 1/15 1/19 1/19 1/21 1/21 C12P 21/02 C 5/10 C12Q 1/68 A C12P 21/02 Z C12Q 1 / 68 G01N 33/15 Z 33/50 Z G01N 33/15 33/53 D 33/50 M 33/53 33/566 C12P 21/08 33/566 (C12P 21/02 C // C12P 21/08 C12R 1 : 91) (C12P 21/02 C12N 15/00 ZNAA C12R 1:91) 5/00 A (72) Inventor Hiroki Kawai 3-6-6 Asahicho, Machida-shi, Tokyo Kyowa Hakko Hakko Kogyo Co., Ltd. (72) Inventor Nishi Tatsuya 3-6-6 Asahicho, Machida-shi, Tokyo Kyowa Hakko Kogyo Co., Ltd., Tokyo Research Laboratory (72) Inventor Yusuke Nakamura 1-17-33 Azamino Aoba-ku, Yokohama-shi, Kanagawa Prefecture (72) Inventor Sumio Kanno Tokyo 4B-13 Minamiogikubo, Suginami-ku F-term (reference) HA12 HA15 4B063 QA01 QA05 QA12 QA19 QQ08 QQ21 QQ33 QQ61 QQ62 QQ73 QQ79 QQ89 QQ91 QQ94 QR32 QR42 QR50 QR62 QR75 QR76 QR77 QR78 QR80 QS03 QS05 QS24 QS25 QS34 QS34 QX07 4B064 CA01 DA14 CA14 DA14 AA24X AA26X AA32X AA41X AA48X AA72X AA79X AA87X AA88X AA90X AA92X AA93X AA93Y AB01 AB05 AC14 BA02 BA03 CA24 CA25 CA44 CA46 4C084 AA17 ZA022 ZB262 4C085 AA13 AA10A50 AA30A30 AA30A30 AA14A50

Claims (35)

[Claims] 1. A G protein-coupled receptor polypeptide having the amino acid sequence of SEQ ID NO: 1. 2. The polypeptide according to claim 1, which has an amino acid sequence in which one or more amino acids have been deleted, substituted or added in the amino acid sequence of the polypeptide set forth in SEQ ID NO: 1. A polypeptide having substantially the same activity. 3. A partial peptide which is a partial peptide of the G protein-coupled receptor polypeptide according to claim 1 or 2, and has a binding ability to a ligand, agonist, antagonist or functional modifier of the polypeptide. 4. A DNA encoding the G protein-coupled receptor polypeptide according to claim 1 or 2. 5. In the DNA of SEQ ID NO: 2,
DNA having a base sequence represented by base numbers 175 to 1287. 6. A DNA which hybridizes with a DNA selected from the DNA according to claim 4 or 5 under a stringent condition, and substantially the same as the G protein-coupled receptor polypeptide according to claim 1. DNA encoding a polypeptide having the same activity as the above. 7. A DNA encoding a partial peptide of the G protein-coupled receptor polypeptide according to claim 3. 8. A DNA encoding a partial peptide of the polypeptide encoded by the DNA according to claim 6, and a ligand, agonist, antagonist or function modifying substance of the polypeptide according to claim 1 or 2. DNA encoding a partial peptide having the binding ability of 9. D according to any one of claims 4 to 8,
A recombinant DNA obtained by incorporating NA into a vector. 10. A transformed cell, transformed plant or transformed non-human animal having the recombinant DNA according to claim 9. 11. Use of the transformed cell, transformed plant or transformed non-human animal according to claim 10, wherein (i) culturing the transformed cell in a medium, and (ii) culturing the transformed cell. Cultivating the transformed plant in the plant, or (iii) breeding the transformed non-human animal to produce the polypeptide of claim 1 or 2 or the partial peptide of claim 3 in the animal. The method for producing the polypeptide according to claim 1 or 2, wherein the polypeptide or the partial peptide is collected from the culture, the plant, or the animal. . 12. An antibody that recognizes the polypeptide according to claim 1 or 2, or the partial peptide according to claim 3. 13. A method for immunologically quantifying the polypeptide according to claim 1 or 2, or the partial peptide according to claim 3, wherein the antibody according to claim 12 is used. 14. A method for determining cancer or dysfunction of the thalamus or cerebellum using the quantification method according to claim 13. 15. A method for determining cancer or dysfunction of the thalamus or cerebellum by measuring the amount of mRNA encoding the polypeptide according to claim 1 or 2. 16. A method for determining cancer, or dysfunction of the thalamus or cerebellum by detecting deletion, substitution or addition of the gene encoding the polypeptide according to claim 1 or 2. 17. The polypeptide according to claim 1 or 2, wherein the polypeptide according to claim 1 or 2 or the partial peptide according to claim 3 is contacted with a test sample, The method for screening a ligand, agonist, antagonist or function modifying substance for a polypeptide according to claim 1 or 2, wherein an agonist, antagonist or function modifying substance is selected. 18. The test sample is contacted with a cell expressing the polypeptide according to claim 1 or 2 or the partial peptide according to claim 3, and the test sample is contacted with the polypeptide according to claim 1 or 2. Peptide ligands, agonists,
The method for screening a ligand, an agonist, an antagonist or a function modifying substance of a polypeptide according to claim 1 or 2, wherein an antagonist or a function modifying substance is selected. (19) The polypeptide according to (1) or (2) or the partial peptide according to (3) and a ligand, and (ii) the polypeptide according to (1) or (2). A peptide or a partial peptide according to claim 3 is contacted with a ligand and a test sample, and the agonist, antagonist or function modifying substance of the polypeptide according to claim 1 or 2 is compared with the test sample. 3. A method for screening for an agonist, antagonist or function-modifying substance of the polypeptide according to claim 1, wherein the method is selected. 20. The method according to claim 1, wherein (i) a cell that expresses the polypeptide according to claim 1 or 2 or the partial peptide according to claim 3 is brought into contact with a ligand; Or a cell expressing the partial peptide according to claim 3 and a ligand and a test sample are compared with each other, and the polypeptide of claim 1 or 2 is compared with the test sample. Agonist,
3. The method for screening an agonist, antagonist or function modifying substance of a polypeptide according to claim 1 or 2, wherein an antagonist or a function modifying substance is selected. 21. A ligand, agonist or antagonist of the polypeptide according to claim 1 or 2, which comprises the polypeptide according to claim 1 or 2 or the partial peptide according to claim 3. Or a kit for screening for a functional modifier. 22. A ligand, agonist, antagonist or function of the polypeptide according to claim 1 or 2, obtained by using the screening method according to claim 17 or 20 or the screening kit according to claim 21. Modifiers or pharmacologically acceptable salts thereof. 23. The antibody according to claim 12, wherein the antibody is an antibody according to claim 12.
A therapeutic agent for cancer or dysfunction of the thalamus or cerebellum, comprising a substance selected from the ligands, agonists, antagonists and function-modifying substances described in 1, or a pharmacologically acceptable salt thereof. 24. When a test sample is brought into contact with (i) a cell expressing the polypeptide according to claim 1 or 2 and (ii) a cell expressing the polypeptide according to claim 1 or 2 with a test sample. And claim 1 or 2 from the test sample.
3. A method for screening a compound that changes the expression level of a DNA encoding the polypeptide according to claim 1 or 2, wherein the method selects a compound that changes the expression of the gene encoding the polypeptide according to 1. 25. The method according to claim 13, wherein the variation in the expression level is measured by the method according to claim 13 or the method according to claim 1 or 2 for quantifying the amount of mRNA encoding the polypeptide. 25. The screening method according to 24. A test sample is contacted with a transformant containing a reporter gene-linked DNA downstream of a region controlling transcription of the gene encoding the polypeptide according to claim 1 or 2, and 3. The method according to claim 1, wherein a compound that changes the expression level of the DNA encoding the polypeptide according to claim 1 or 2 is selected from the sample. A method for screening a compound to be varied. 27. The region that controls transcription is SEQ ID NO: 15.
27. The screening method according to claim 26, which is a region defined by a DNA having a continuous 50 to 5000 bp nucleotide sequence in the DNA represented by the nucleotide sequence at positions 20202 to 25202. 28. A compound or a pharmacologically acceptable salt thereof obtained by a method selected from the screening method according to any one of claims 24 to 27. 29. An agent for treating cancer or dysfunction of the thalamus or cerebellum, comprising the compound according to claim 28 or a pharmacologically acceptable salt thereof. 30. An oligonucleotide having the same sequence as 5 to 60 consecutive bases in the base sequence of a DNA selected from the DNA of claim 4 or 6 and the DNA of SEQ ID NO: 2, complementary to the oligonucleotide. Selected from oligonucleotides having specific sequences and oligonucleotide derivatives of these oligonucleotides. 31. The DN according to claims 4 to 6, and 30.
A transcription or mRN of a DNA encoding the polypeptide according to claim 1 or 2, using a DNA selected from A.
A method for suppressing the translation of A. 32. A gene in which the expression level of the polypeptide according to claim 1 or 2 is altered by deleting or substituting all or a part of the gene containing the DNA encoding the polypeptide according to claim 1 or 2. Deleted or substituted non-human animals. 33. An animal according to claim 32, which is inoculated with a test sample, or a test sample is brought into contact with an organ, tissue or cell of the animal, and the test sample is used to produce cancer or dysfunction of the thalamus or cerebellum. A method of screening for a therapeutic agent for cancer or dysfunction of the thalamus or cerebellum, which comprises selecting a therapeutic agent for: 34. A compound obtained by the screening method according to claim 33 or a pharmacologically acceptable salt thereof. 35. A therapeutic agent for cancer or dysfunction of the thalamus or cerebellum, comprising the compound according to claim 34 or a pharmacologically acceptable salt thereof.