CN109295090A - A kind of application of cell cycle regulating protein CREPT in the maintenance of animal embryonic stem cell versatility - Google Patents

A kind of application of cell cycle regulating protein CREPT in the maintenance of animal embryonic stem cell versatility Download PDF

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CN109295090A
CN109295090A CN201811195565.4A CN201811195565A CN109295090A CN 109295090 A CN109295090 A CN 109295090A CN 201811195565 A CN201811195565 A CN 201811195565A CN 109295090 A CN109295090 A CN 109295090A
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crept
animal
stem cell
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embryonic stem
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CN109295090B (en
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常智杰
朱丙陶
丁莉丹
王银银
任芳丽
杨柳
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Tsinghua University
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Abstract

The invention discloses a kind of application of cell cycle regulating protein CREPT in the maintenance of animal embryonic stem cell versatility.Present invention discover that reducing in embryonic stem cell, animal embryonic stem cell proliferation and versatility maintenance are affected after the content of cell cycle regulating protein CREPT, proliferation, self-renewal capacity and the chimeric ability of animal embryonic stem cell is caused to be suppressed (embryonic stem cell that CREPT is knocked out cannot be fitted in offspring embryo tissue), so that embryo cannot be divided into.Show, cell cycle regulating protein CREPT can maintain the versatility of animal embryonic stem cell, the non-human animal embryo's stem cell model that can have proliferative capacity decline and/or self-renewal capacity decline and/or chimeric ability to decline feature by reducing its expression preparation.

Description

A kind of cell cycle regulating protein CREPT is maintained in animal embryonic stem cell versatility In application
Technical field
The present invention relates in technical field of cell biology, a kind of cell cycle regulating protein CREPT is dry in animal embryo Application in cells pluripotency maintenance.
Background technique
Stem cell (Stem cell) i.e. undifferentiated cell, it be one kind can self-renewing (self-renewal) and It is differentiated to form the multipotential cell of functional cell.According to different classification standards, stem cell can be divided into different types, such as root According to stage of development difference locating for stem cell can be divided into embryonic stem cell (Embryonic Stem Cell, ESC) and at soma it is thin Born of the same parents (Somatic stem cell)；The differentiation potential according to possessed by stem cell can be divided into myeloid-lymphoid stem cell (Totipotent Stem Cell, TSC), multipotential stem cell (Pluripotent stem cell), specially can stem cell (Multipotent stem ) and unipotent stem cell (Unipotent stem cell) cell.Embryonic stem cell is a kind of height neoblast, Ke Yifen Change forms the various types of cells of human body, has the totipotency of development, treats disease for medical graft and brings convenience.Embryo is dry The totipotency of cell imparts it in huge plasticity developmentally, and exactly this plasticity to people in the control broken up to it It brings challenge in system, how according to the intention effectively control stem cell of people to be directed differentiation to current urgently to be solved ask Topic, the solution of this problem will further promote the application of embryonic stem cell.
The maintenance of embryonic stem cell versatility be a polyprotein participate in comprehensive regulation process, including transcription because Son, adaptin, chromatin remodeling enzyme and chromatin Structure regulatory protein etc..Many tumor-related genes, as c-Myc, Klf4, STAT3 etc. plays critical function in the maintenance of embryonic stem cell versatility.CREPT(Cell-cycle Regulated and Expression-elevated Protein in Tumor) it is the new gene (Gene that applicant clones and names in recent years ID:58490), it can promote the proliferation and transfer of tumour, be a new tumor-related gene.
Summary of the invention
The technical problem to be solved by the present invention is to how prepare to have proliferative capacity decline, self-renewal capacity decline And/or the animal embryonic stem cell model of chimeric ability decline.
1) and/or 2) and/or 3) in order to solve the above technical problems, present invention firstly provides with following featured animal embryo The preparation method of tire stem cell model:
1) proliferative capacity declines；
2) self-renewal capacity declines；
3) it is fitted into ability decline；
The described method includes: reduce or inhibit in the in vitro animal embryonic stem cell that sets out the content of CREPT protein and/ Or activity, obtain proliferative capacity and/or self-renewal capacity and/or chimeric ability compared with the animal embryonic stem cell that sets out The animal embryonic stem cell model of decline；
The animal is non-human animal or people.
The animal embryonic stem cell that sets out expresses the CREPT protein.
In the above method, the CREPT protein can be following A1), A2) or A3):
A1) amino acid sequence is the protein of sequence 3；
A2) by amino acid sequence shown in sequence 3 in sequence table by one or several amino acid residues substitution and/or Deletion and/or addition and protein with the same function；
A3) in A1) or the obtained fused protein of N-terminal A2) or/and C-terminal connection label.
In order to make A1) in protein convenient for purifying, amino acid sequence shown in sequence 1 can be formed in by sequence table The upper label as shown in the table of amino terminal or carboxyl terminal connection of protein.
Table: the sequence of label
Label Residue Sequence
Poly-Arg 5-6 (usually 5) RRRRR
Poly-His 2-10 (usually 6) HHHHHH
FLAG 8 DYKDDDDK
Strep-tag II 8 WSHPQFEK
c-myc 10 EQKLISEEDL
HA 9 YPYDVPDYA
Above-mentioned A2) in CREPT protein, to have 75% or 75% with the amino acid sequence of protein shown in sequence 3 The above identity and protein with the same function.It is described that there is 75% or 75% or more identity to be with 75%, have 80%, with 85%, with 90%, with 95%, with 96%, with 97%, with 98% or with 99% identity.
Above-mentioned A2) in CREPT protein encoding gene can by will in DNA sequence dna shown in sequence 2 lack one Or the codon of several amino acid residues, and/or carry out the missense mutation of one or several base-pairs, and/or its 5 ' end and/ Or 3 ' end connect the coded sequence of label shown in table and obtain.Wherein, shown in DNA molecular coded sequence 3 shown in sequence 2 CREPT protein.
In the above method, the animal can be non-human mammal or people.The non-human mammal further can be small Mouse.
In the above method, the content and/or activity for reducing CREPT protein in the animal embryonic stem cell that sets out can lead to The expression for crossing the encoding gene for CREPT protein described in animal embryonic stem cell that sets out described in reduction or inhibition is realized.
In the above method, the volume for CREPT protein described in animal embryonic stem cell that sets out described in the reduction or inhibition Code gene expression can by knock out described in set out CREPT protein described in animal embryonic stem cell encoding gene or its One segment imports the substance reality for inhibiting the encoding gene of the CREPT protein into the animal embryonic stem cell that sets out It is existing.
It is described reduction or inhibition described in set out CREPT protein described in animal embryonic stem cell encoding gene expression The exon of the encoding gene of the CREPT protein can specifically be knocked out.The exon of the encoding gene of the CREPT protein It can be the third exon of the encoding gene of the CREPT protein.
The encoding gene of the CREPT protein can be DNA molecular shown in sequence 2 in sequence table, the CREPT albumen The genome sequence of the encoding gene of matter is classified as DNA molecular shown in sequence 1 in sequence table.1-459 of sequence 1 are first The sequence of exon, the 7500-7692 sequences for second exon, 14910-15043 are third exon Sequence, 19423-19532 be the 4th exon sequence, the 21591-21717 sequences for the 5th exon Column, the 30156-30331 sequences for the 6th exon, the 46434-49711 sequences for the 7th exon.
In the above method, the encoding gene for knocking out the CREPT protein or its any segment can utilize Cre- Loxp method is realized.
The Cre-loxp method is that the side of destination gene expression is reduced or inhibited using Cre recombinase and the site LoxP Method.
The substance of the encoding gene for inhibiting the CREPT protein can be the reduction CREPT protein expression quantity Nucleic acid molecules.
The animal embryo prepared the present invention also provides the preparation method using the animal embryonic stem cell model is dry thin Born of the same parents' model.
The present invention also provides the CREPT protein or biomaterial relevant to the CREPT protein to regulate and control Application in animal embryonic stem cell proliferative capacity and/or self more sexuality and/or chimeric ability；The animal is inhuman dynamic Object or people；The biomaterial is any one of following B1) to B9):
B1 the nucleic acid molecules of the CREPT protein) are encoded；
B2) contain B1) expression cassettes of the nucleic acid molecules；
B3) contain B1) recombinant vectors of the nucleic acid molecules or contain B2) recombinant vector of the expression cassette；
B4) contain B1) recombinant microorganisms of the nucleic acid molecules or contain B2) recombinant microorganism of the expression cassette or Contain B3) recombinant microorganism of the recombinant vector；
B5) contain B1) the transgenetic animal cell systems of the nucleic acid molecules or contain B2) transgenosis of the expression cassette Animal cell line；
B6) contain B1) the transgenic animals tissues of the nucleic acid molecules or contain B2) transgenosis of the expression cassette is dynamic Object tissue；
B7) contain B1) transgenic animal organs of the nucleic acid molecules or contain B2) transgenosis of the expression cassette is dynamic Sundries official；
B8 the nucleic acid molecules of the CREPT protein expression quantity) are reduced；
B9) contain B8) expression cassettes of the nucleic acid molecules, recombinant vector, recombinant microorganism, transgenetic animal cell system, Transgenic animals tissue or transgenic animal organ.
In above-mentioned application, B1) nucleic acid molecules can be following b11) b12) or b13) or b14):
B11) coded sequence is the cDNA molecule or DNA molecular of sequence 2 in sequence table；
B12) DNA molecular shown in sequence 1 in sequence table；
B13) and b11) or b12) nucleotide sequence that limits has 75% or 75% or more identity, and described in encoding The cDNA molecule or DNA molecular of CREPT protein；
B14) under strict conditions with b11) or b12) or b13) nucleotide sequence hybridization that limits, and described in encoding The cDNA molecule or DNA molecular of CREPT protein.
Wherein, the nucleic acid molecules can be DNA, such as cDNA, genomic DNA or recombinant DNA；The nucleic acid molecules can also To be RNA, such as mRNA or hnRNA.
Those of ordinary skill in the art can easily adopt by known method, such as the side of directed evolution and point mutation Method is mutated the nucleotide sequence of coding CREPT protein of the invention.Those by manually modified, have and this The nucleotide sequence 75% of isolated CREPT protein or the nucleotide of higher identity are invented, as long as coding CREPT Protein and there is CREPT protein function, is derived from nucleotide sequence of the invention and being equal to sequence of the invention Column.
Term " identity " used herein refers to the sequence similarity with native sequence nucleic acid." identity " includes and this hair Amino acid sequence shown in bright coded sequence 3 composition protein nucleotide sequence have 75% or higher or 85% or Higher or 90% or higher or 95% or higher identity nucleotide sequence.Identity can with the naked eye or computer software It is evaluated.Using computer software, identity between two or more sequences can be indicated with percentage (%), can be with For evaluating the identity between correlated series.
In above-mentioned application, the stringent condition can be as follows: 50 DEG C, in 7% lauryl sodium sulfate (SDS), 0.5M NaPO4Hybridize in the mixed solution of 1mM EDTA, is rinsed in 50 DEG C, 2 × SSC, 0.1%SDS；May be used also are as follows: 50 DEG C, 7%SDS, 0.5M NaPO4Hybridize in the mixed solution of 1mM EDTA, is rinsed in 50 DEG C, 1 × SSC, 0.1%SDS；May be used also Are as follows: 50 DEG C, in 7%SDS, 0.5M NaPO4Hybridize in the mixed solution of 1mM EDTA, at 50 DEG C, 0.5 × SSC, 0.1% It is rinsed in SDS；May be used also are as follows: 50 DEG C, in 7%SDS, 0.5M NaPO4Hybridize in the mixed solution of 1mM EDTA, at 50 DEG C, It is rinsed in 0.1 × SSC, 0.1%SDS；May be used also are as follows: 50 DEG C, in 7%SDS, 0.5M NaPO4In the mixed solution of 1mM EDTA Hybridization, rinses in 65 DEG C, 0.1 × SSC, 0.1%SDS；It can also are as follows: in 6 × SSC, the solution of 0.5%SDS, at 65 DEG C Hybridization, then with 2 × SSC, 0.1%SDS and 1 × SSC, it is primary that 0.1%SDS respectively washes film；It can also are as follows: 2 × SSC, 0.1%SDS Solution in, hybridize at 68 DEG C and wash film 2 times, each 5min, and in 0.5 × SSC, the solution of 0.1%SDS, at 68 DEG C Lower hybridization simultaneously washes film 2 times, each 15min；Can also are as follows: 0.1 × SSPE (or 0.1 × SSC), 0.1%SDS solution in, 65 DEG C Under the conditions of hybridize and wash film.
Above-mentioned 75% or 75% or more identity can be 80%, 85%, 90% or 95% or more identity.
In above-mentioned application, B2) described in the nucleic acid molecules containing coding CREPT protein expression cassette (CREPT gene table Up to box), it is the DNA for referring to express CREPT protein in host cell, which not only may include that starting CREPT gene turns The promoter of record may also include the terminator for terminating CREPT genetic transcription.Further, the expression cassette may also include enhancer Sequence.
The recombinant vector of the CREPT expression casette can be contained with existing expression vector establishment.
In above-mentioned application, the carrier can be plasmid, sticking grain, bacteriophage or viral vectors.
In above-mentioned application, the microorganism can be yeast, bacterium, algae or fungi.
In above-mentioned application, the transgenetic animal cell system, transgenic animals tissue and transgenic animal organ are not wrapped Include propagation material.
In above-mentioned application, the animal can be non-human mammal or people.The non-human mammal further can be small Mouse.
In the present invention, when the animal embryonic stem cell that sets out is human embryo stem cell, the animal embryo that sets out is dry Cell is commercialized embryonic stem cell.
The present invention by gene editing scientific discovery reduce CREPT gene expression after animal embryonic stem cell proliferation and it is more Energy property maintenance is affected, and proliferation, self-renewal capacity and the chimeric ability of animal embryonic stem cell is caused to be suppressed (embryonic stem cell that CREPT is knocked out cannot be fitted in offspring embryo tissue), so that embryo, therefore, CREPT cannot be divided into It is a key protein in embryonic stem cell versatility maintenance process, animal embryo can be prepared by reducing the expression of CREPT Tire stem cell model provides new thinking for the research and application of embryonic stem cell.
Detailed description of the invention
The recombination schematic diagram of recombinant DNA molecules shown in Fig. 1 sequence 4.
Fig. 2 is the influence that the knockout of CREPT gene maintains embryonic stem cell proliferation and versatility.Utilize gene editing skill Art grows counting experiments (A) and Clone formation (B) experiment after mouse embryo stem cell knocks out CREPT respectively, as a result such as Fig. 1 institute Show, knocks out proliferation and self-renewing that CREPT gene inhibits embryonic stem cell.
Fig. 3 is that flow cytometry detection knocks out chimeric ability of the embryonic stem cell of CREPT gene in mice embryonic.
Specific embodiment
The present invention is further described in detail With reference to embodiment, and the embodiment provided is only for explaining The bright present invention, the range being not intended to be limiting of the invention.Experimental method in following embodiments is unless otherwise specified Conventional method.Material as used in the following examples, reagent, instrument etc., are commercially available unless otherwise specified. Quantitative test in following embodiment, is respectively provided with three repeated experiments, and results are averaged.In following embodiments, such as without special Illustrate, the 1st of each nucleotide sequence is the 5 ' terminal nucleotides of corresponding DNA in sequence table, and last bit is the 3 ' of corresponding DNA Terminal nucleotide.
Embodiment 1, the knockout of CREPT gene cause embryonic stem cell versatility to be lost
The present embodiment discovery CREPT gene (Gene ID:70470, US National Biotechnology Information center (NCBI)) exists It maintains to play an important role in versatility of mouse embryo stem cell and cell Proliferation.The sequence of CREPT gene is sequence table Middle sequence 1,1-459 of sequence 1 are the sequence of first exon, the 550-7499 sequences for First Intron Column, the 7500-7692 sequences for second exon, the 7693-14909 sequences for second introne, the The 14910-15043 sequences for third exon, the 15044-19422 sequences for third introne, the The 19423-19532 sequences for the 4th exon, the 19533-21590 sequences for the 4th introne, the The 21591-21717 sequences for the 5th exon, the 21718-30155 sequences for the 5th introne, the The 30156-30331 sequences for the 6th exon, the 30332-46433 sequences for the 6th introne, the The 46434-49711 sequences for the 7th exon, the CDS sequence of CREPT gene are sequence 2 in sequence table, coded sequence 3 Shown in CREPT protein.
One, CREPTFlox/FloxThe preparation of mouse
CREPT usedFlox/FloxMouse derives from Britain MRC Harwell Institute, which passes through gene editing Technology obtains the two sides of the 3rd exon of CREPT gene of LoxP integrations to C57BL/6N mouse.
Recombinant DNA molecules electricity shown in sequence 4 is transferred to the ES cell of C57BL6/N mouse, is weighed by G418 screening The correct ES cell clone of group (i.e. in two articles of chromosomes of the C57BL6/N mouse 8948- of CREPT gene shown in sequence 1 The DNA fragmentation of 19283 interdigits replaces with recombinant DNA molecules (its sequence is sequence 4 in sequence table)), recombination form such as Fig. 1, Fig. 1 In, 1-6 respectively represents first to the 6th exon of CREPT gene.Correct ES cell clone injection C57BL6/ will be recombinated N Mouse Blastocysts obtain Chi-meric mice, and then, Chi-meric mice is mated with C57BL6/N mouse, choose in filial generation containing the small of recombinant fragment Mouse can as stablize the mouse of Genetic Recombination segment, the mouse of homozygous recombinant fragment be obtained after selfing, finally, should Mouse and FRT tool mouse (i.e. B6/JNju-H11em1Cin(CAG-FLPo)/ Nju, Tsinghua University's animal house save, big from Nanjing Model animal research institute is learned, details are shown in http://www.nrcmm.cn/strain/2063) mating obtains CREPTFlox /FloxMouse.
Wherein, 1-5472 of sequence 4 and 13323-17292 position homology arm sequences, 5537-5584 and the 12441-12488 are the site FRT, and 10584-10617,12495-12528 and 13266-13299 are LoxP Site, 12945-13078 are the 3rd exon of CREPT gene.
Finally, CREPTFlox/FloxMouse is by CREPT gene shown in sequence 1 in two chromosomes of C57BL6/N mouse The DNA fragmentation of 8948-19283 interdigit replaces with the mouse that recombinant DNA molecules shown in sequence 5 obtain in sequence table.Sequence 5 1-5472 and 6419-10388 be homology arm sequence, 5537-5584 be the site FRT, 5591-5624 It is the site LoxP with 6362-6395,6041-6174 are the 3rd exon of CREPT gene.
Two, the preparation of CREPT gene knockout embryonic stem cell
1, the CREPT for obtaining step 1Flox/FloxMouse and EIIa-Cre tool mouse (i.e. B6.FVB-TgN (EIIa- Cre) C5379Lmgd, E2a-Cre, Tsinghua University's animal house save, and derive from The Jackson Laboratory, believe in detail Breath is shown in https: //www.jax.org/strain/003724) hybridizing, obtains first-filial generation, i.e. CREPT hybrid mice.
2, by CREPT hybrid mice and wild type C57BL6/N mouse, (it is limited that tonneau China experimental animal technology is tieed up in Beijing Company) hybridization, then, then the CREPT hybrid mice selfing in offspring is chosen, takes 3.5 days embryos, separate blastaea inner cell mass Carry out Colony Culture；Cultural method is as follows: a. puts to death 3.5 days pregnant mouse, takes out uterus, there is the injection of insulin of PBS using suction Device goes out blastaea from uterus；B. the blastaea gone out is transferred to containing M2 culture medium (SIGMA, M7167) using pipettor Trophocyte on.The cell monoclonal grown is embryonic stem cell.
3, the monoclonal that picking blastaea inner cell mass is grown is identified in genomic level and protein level respectively:
Identification in genomic level: extracting the genomic DNA of monoclonal, utilizes primer pair (CKO-F:5 '- GCGCATAACGATACCACGATATC-3'；CKO-R:5'-GTCAGCCACAATCCTAAAGAAC-3'；CFlox-F 5'- TCTTGGTCTTTGCTCCTCCCAG-3 ') it is identified, the results show that CKO-F and CKO-R can be amplified and be carried out band and be The cell that CREPT gene third exon is knocked, clip size 207bp；CFlox-F and CKO-R can amplify band Cell be wild-type cell (i.e. the third exon of CREPT gene is not knocked), clip size 278bp.Selection CKO-F and CKO-R can amplify band and cell that CFlox-F and CKO-R cannot amplify band is denoted as CREPT gene knockout Cell.
Identification on protein level: the egg that genomic level is accredited as the monoclonal of CREPT Knockout cells is extracted White matter detects CREPT protein level using western blot, utilizes the CREPT mouse monoclonal antibody of anti-CREPT protein (preparation method is shown in Characterization of a monoclonal antibody against CREPT, a novel Protein highly expressed in tumors, Monoclonal Antibodies Immunodiagnosis Immunotherapy.2014Dec；33 (6): 401-8.doi:10.1089/mab.2014.0043.) it is used as primary antibody, horseradish mistake Oxide enzyme marks goat anti-mouse IgG (Beijing Bioisystech Co., Ltd of Zhong Shan Golden Bridge, article No. is ZB-2305) to be used as two It is anti-, detect CREPT protein expression situation in monoclonal cell, the ES not being knocked using the third exon of CREPT gene Cell is as control.Choosing western blot can't detect the cell of CREPT protein signal, which is CREPT gene Knock out embryonic stem cell.
Three, the phenotypic evaluation of CREPT gene knockout embryonic stem cell (ES cell)
Research object is the CREPT gene knockout ES cell that step 2 obtains, and is denoted as KO group ES cell.It is obtained using step 2 To genomic level identification in the third exon of CREPT gene be not knocked and western blot can be detected The cell of CREPT protein signal is denoted as WT group ES cell as control.
1, influence of the CREPT gene knockout to embryonic stem cell proliferation ability
1) by equivalent WT group ES cell and KO group ES cell count passage into the 6 orifice plates of the pretreated bottom of gelatin, often Hole is added about 5 × 104A ES cell, ES culture medium is added, and (ES culture medium is by Knock out DMEM (Gibco, 10829- 018) following substance is added in obtain: 20% (percent by volume) serum (Gibco, 10091-148), 2mM GlutMAX (L- third Aminoacyl-L-Glutamine dipeptides, Gibco, 35050-061), 0.1mM NEAA (seven kinds of amino acid, Gibco, 11140-050, often The concentration of kind of amino acid in the medium is 0.1mM), 100 μM of mercaptoethanols (SIGMA, M7522), 10ng/mL LIF (Millipore, ESG1107), the concentration of each substance are the concentration in ES culture medium) in the cell training of 37 DEG C, 5%CO2 It supports and is cultivated in case (SANYO), the volume that culture medium is added in every hole is 2mL.Every group of 6 parallel holes.
2) (0.25g pancreatin is molten using 0.25% trypsin solution (Gibco, 25200-056) after cultivating 24 hours The solution obtained in 100ml PBS) digestion ES cell, it is resuspended using ES culture medium after gently blowing and beating and counts cell number, it is continuous to count Number 3 days.
3) the ES cell count data of WT and KO group is counted, growth curve is drawn in analysis.
The results show that the knockout of CREPT gene obviously inhibits the proliferation (A in Fig. 2) of ES cell.
2, influence of the CREPT gene knockout to embryonic stem cell self-renewal capacity
1) logarithmic growth phase WT group and KO group ES cell, with 0.25% trypsin solution (Gibco, 25200-056) It digests and gently blows and beats, make single cell suspension, make viable count.
2) WT group and KO group ES cell are passaged to 6 orifice plates, every group of 6 parallel holes according to 500, every hole cell.
3) fresh culture is replaced daily, is cultivated one week.
4) cell clone grown is dyed using alkaline phosphatase (AP) staining kit after a week.
5) six orifice plates dyed are placed under inverted microscope, observe number of cell clones, takes pictures.
Above-mentioned steps used medium is ES culture medium.
The results show that the knockout of CREPT gene inhibits the self-renewal capacity (B in Fig. 2) of ES cell.
3, CREPT gene knockout is fitted into the influence of ability to embryonic stem cell in mice embryonic
(1) then the slow virus for packing mCherry fluorescence infects WT group and KO group using the slow virus of mCherry fluorescence ES cell carries out fluorescent marker.Method is as follows: by pLVX-IRES-mCherry, (Clontech Products, article No. are 631237) with packaging plasmid (http://www.addgene.org/11795/)；Packaging plasmid be psPAX2 (http: // Www.addgene.org/12260/) and pMD2.G (http://www.addgene.org/12259/)) cotransfection and 293T it is thin In born of the same parents (ATCC, CRL-3216), culture solution, as viral supernatants are collected after 48 hours.It is thin that ES is infected with the viral supernatants of collection Then born of the same parents go out the ES cell of the mCherry positive by selected by flow cytometry apoptosis to get the ES cell marked to mCherry.
(2) ICR Mouse Blastocysts obtain: mating, examine with ICR hero mouse using after the super ovulation of Estrogenization stimulation ICR female mice Bolt is the acquisition that positive female mice can be used to blastaea.Wherein, ICR female mice and ICR hero mouse are that the experiment of Beijing dimension tonneau China is dynamic Object Technology Co., Ltd. product.
(3) after examining bolt 3 days, the ICR female mice for seeing bolt is put to death, blastaea is isolated；Meanwhile preparing mCherry fluorescent marker WT group and KO group ES cell suspension.The WT group of mCherry fluorescent marker and KO group ES cell suspension the preparation method is as follows: The ES cell that 0.25% trypsin solution digestion step (1) obtains is resuspended cell with M2 culture medium (SIGMA, M7167), obtains To cell suspension.
(4) the ES cell suspension of mCherry fluorescent marker is injected into the blastaea that ICR female mice is isolated by micromanipulation In chamber, then, there is the blastaea of the ES cell of mCherry fluorescent marker to be transplanted to replace-conceive female mice injection.
(5) it after transplanting 10 days, puts to death mouse and separates embryo, digested after grinding with 0.25% trypsin solution, with containing tire Embryonic cell is resuspended in the culture medium of cow's serum.
(6) flow cytometry contains the cell positive rate of mCherry, and for statistical analysis.
As a result as shown in figure 3, the results show that transplanted injection mCherry fluorescent marker WT group ES cell blastaea after, embryo The ratio of cell in fetus cells containing mCherry is (8.68 ± 3.3) %, and has transplanted injection mCherry fluorescent marker After KO group ES cell blastaea, the ratio of the cell in embryonic cell containing mCherry is (1.28 ± 0.3) %.What CREPT was knocked out Embryonic stem cell cannot be fitted in embryonic tissue, show that the knockout of CREPT gene can influence embryonic stem cell in Development of Mouse Embryos Ability is fitted into tire.
<110>Tsinghua University
<120>application of a kind of cell cycle regulating protein CREPT in the maintenance of animal embryonic stem cell versatility
<160> 5
<170> PatentIn version 3.5
<210> 1
<211> 49711
<212> DNA
<213>mouse (mus musculus)
<400> 1
ctcactgcgc cggcgcgcgg gcgcgagcgg ttaaagcgca ttttcggtgt tcgaggacct 60
ggatttgttc cctcggcctg gccggctaga agtcagggac caaggatact gccgcctcct 120
aaggatcgtt ttcctccggc tccgaagcct ctcggcgccc ggctgtggcc cttgcccttt 180
gacccccgtg gcggcggggt gagaggtcgg gtggccatct tgtggcggcg gcgcgggcgg 240
ctgttactgc ggagacctgt cccctccccc cacctggtac cccgagctgt ctgtcagtcg 300
gcggagagcc ctgcagcctg tcagcggagg cctgggccga gtgcggtcgc tttccccgtc 360
tcatcgggcg gcccgggctg ctccccacca ccgccaccat gtcctccttc tcagaatcgg 420
cgctggaaaa gaagctctca gagctgagca actctcagca gagcgtgcag accctctccc 480
tgtggctcat ccaccaccgc aagcacgcag gacccatcgt ctccgtgtgg caccgcgagc 540
tccgcaaagg tgatcacccg caccccggct ctcccatggc tgtgttccct catccactct 600
cccgcagccc tgtggcttct taagccttgt tttgcaggcc tggggctctg atggagggtc 660
tgaggttgga aacggtgtta ttttatgact tggagaggga aactgaggcc cggtaaggta 720
tgggagattc gtctgaagtt ccaagggcgc aaatgcccag acttgaagtt actcagctcc 780
ttccctcatc aggcttcccc ctcacacctt acgacagttg tttttgtttt tgttttaaat 840
cgtggctgtg tttggagctg ggagccccgt gttgggcaat tggaatacga tccagccgtc 900
tctggtcaga ctgcccacat taagacctgg tgcaaacgtg ttctagctaa gtgatcctgg 960
gaacgtttct tcatatcctt gtgccggtgt attctaatct gtaaaacgga tacaaactta 1020
cctgtacttc caaagccttg actccgtgtt agttactatc catagacttg agaacaggat 1080
ggtaaattgg gtagccttct gcccttgcca ctaaattcag actctcacga tattccccga 1140
gcgctacctg tttcttgcca ggcccggtgt tggatgcaag ttgtttctgg ctactgtctg 1200
gaagttaagt ctatctgtca ctgaacccaa cagtttaacc tctatgtcct ttattcctta 1260
tatagaaaat ggagatgatg gtagtttatg ttattgtggc tttattatgt gctgctatct 1320
ttatgcgggt tgcttttgtt tttgctttgg gacagaacct ttagccttta caggtctgga 1380
tctccctatg tagaccaggc tggcctcgaa ctcagacccg cctgtttctg cctcccaaat 1440
gctgggatta gtgacttgtg ccaccatact ggatggttct ttgttttttt tttgtttgct 1500
tgttttttgt tttgttttgt ttttttaaat tcttgggttt tttttttttg aagtaggaat 1560
tttacagatg gggaaaacac agggctcaac aaagccgatt ctggtaagaa gccaggctaa 1620
tccaggaagc attttagtgt gctgtgggca cagtgcacgc tttcatagaa gtggtgagtt 1680
aggactggtg aacgccctgt actgcttggt ctttgccttc aggttgctac atccaggtgt 1740
ggagcagcgg gcaggtacag tgctaactca cactgtgctg gcttcctaag cagaagactg 1800
aagagtcgaa gcttggctgt tgtagacgag gaaactgaaa tggagaaatg gagagtgcga 1860
tagtcaaagt ctttgaactg atgaggctcc atggcaggca ccaaactcca gggtttttca 1920
ggagactgga ctctagactt cttgaagagc ctcccagtta tgatttgcta ttctgacata 1980
ggatttcctt cagccctctg gggcaattag gtagagccta tctacagcat ctgtggagac 2040
aggaaaaaac tgtttgtgac ctttgctgaa ttccctttgg cagagtctct tgatcatctt 2100
gaaatcatca gcccatctca gtctggtgaa gccttccttc ccaatttctc acatttattt 2160
atttatttag agtcctagaa aaatagccat ttagagaagt cttttgttgc tgaagctcct 2220
ttattgataa tcttatctgg tgggccttac atagcaagta ggctggcatt cttatcccca 2280
tttcatagaa gatacagcaa gaccagtggc ttgttcagca gtctaaactg ggacatgggc 2340
ccagggagga ctgctaccaa gacttaatgc tttttcaggt acctagtgcc tcagtgacct 2400
taagtccatt ctatcctctg ctatcctatc ccaccacttg cctttacttc attttacttc 2460
tatctctttc tctaggtttg ttttgtcagg gccctgtaat gcacttagaa ttgtgcctat 2520
cacacagtta acaattagtt tagtagttta gttgtacttc tgataatgtt ttcatgtttg 2580
agtaaagaga ggaccctaaa tgggattggt ttaactatct gtggcaattt cagagctgtg 2640
tctctgctgt ccacaaggca gcacataggt tatttctcca ctggtgaact gagccaattc 2700
aagccagatt agattatatt aaaggctggt ttattgggaa gctgctctag ggcagtttgg 2760
ttttggacct gagacaggaa ctgaggtgca tattttacat atcaaaacag acgtggcctt 2820
caggttctcc cagcatccct cagtccctac ctggcattcc ctgtcccaac cctgaacttt 2880
ccagaccagc ctctgtgctg cccttctcct aaaggctcgt gcctattaat ccagacgttt 2940
tagtctctct ctcctctttc tctgcttgag cacactttct ccacccaccc ccaaccccat 3000
agtgacttcc ctggccttcc ttggggccag tgaatttttc ctgtggggaa cagtttccca 3060
ataaaccagc ctttaatata ttctaatctg gcttaaattg gctcatttca caagcagaga 3120
aataacctaa caacagtcac tatctacatg tggctattgg acatttaaaa tggtgacaaa 3180
ggaataaata tatctaatcg acgattcagt tcctcggttt tattagctct tcattaatta 3240
agtgttgact ggccaccgtg gttgatggct cccaggttga atctagcaga actagagttc 3300
ttccccaagc agtaagatat gtattcatct cctacccagc ttcccttgtg aactgataac 3360
acccctacat taaaatagac aagggttttg tactttgaca ctgaaactga caagcccctt 3420
gcaattgaaa taagccatct gtgcctatga agaagttttc cttgtcttag acattatccc 3480
agtctttgag ttatcactaa agtacttaca aaaattattt cctctcattt aggaatcact 3540
tctgttttta atgtgttaat tagttttagt attaattagt acttttagag tgtaaataga 3600
acttctttat ggaatagttg agtattaaca gtgctattag ctgcttgctt gttatatctg 3660
tctatctgtt ttgaggcagg ctcttagtat gtaaccctgg ctgttctggt acttgctttg 3720
ttgaccatac tggcttcaaa ctctcagagg cctacctgtc tttgcctccc aaatgctagg 3780
attaaaggtg tgtgtggcta agatgaattt ttttaacgtt tgttactgtg gccacaatgt 3840
aggtaaatag tttctttttc cagaatactt tgtagtcttt ctctacagat caacatattt 3900
gactagccac tcttttaaac tttactcagc aaggaggggg gaagataggt tttgtcatat 3960
tgtcacccag agttggagta gtgggaagtg tgtactgtgt gtcggctgtg tcacagtgct 4020
tctacacaca gggtacttgt atttctcttt aaggtgccca atgtgattta ggccagaaga 4080
aggcattgga tcccttggaa ctgtagttaa taggtaattg tgagctgcca catgggttct 4140
gggaactaaa ttttggtcct ctggaaaagc agccagttgc tctttttctg agatagggtc 4200
tcactgtgta gccctggctg gctgggaact gacagagaga gagatctgcc ttcctctgcc 4260
tcccaagtgc tgtgatcaaa ggcatatgtc aagcccagct gtaggagtga gttattgttg 4320
ttgttgttgt tttaatcaaa ataattgttg ttatttctta aaaaaaattc tggcggggca 4380
atggtggtgc acgcctttaa tcccagcact tgggaggcag aggcaggcag atttctgagt 4440
tcgaggccag cctggtctac agagtgagtt ccaggacagc caggactaca cagagaaacc 4500
ctgtcttgaa aagccaaaca acaacaacaa caacaaaaaa ctgaaaggaa agctgtagta 4560
accctgatct gagtgaccac agcatacatg ctgagattag gtgcctatga gaactaagac 4620
ctttaatatg tttaagtaga gtgctatatt gctcttttct attttcactc ttcattttat 4680
tttgtttaat tcttatgaca ttttaattgc atcattttta aaattagttc ctttattttt 4740
ttgagattat aatataatca tggtttctcc cttcaaacct tcccatatac cccttcttgc 4800
cttctttcaa attcattccc cccctttttt gtttttgttt tgttttgctt taatttgttt 4860
ttttgagaca gggtttgtct gtatagccct ggctgtcctg gaactcactc tgtagacgag 4920
gctggcctcg aactcagaaa tcaacttgcc tctgcctccc aagtgctggg attaaaggcg 4980
tgtgccacca cacccagtcc cccctccttt ttaaattaat tgttattata cacacacaca 5040
cacacacaca cacatagtct ctataatgtt actggtatgt gtgttttcag ggctgaccag 5100
gctgaccatt tggtatttga taaacaattg gtgtgttcct ccctggggaa gactgtttct 5160
ccctctctca acatttcttg gttgcctgta gttctttgtg tagggtggag gactttcttc 5220
tgtccatgtt agcttatcta ttttgttctt aatttagatt gtcatgttga tgagactatg 5280
agtgtagctt ccaactctaa aaatctttct gctctttctc atgctccctg agccttgggt 5340
gaaagggttg ttttgtagat gtattcattg agactggatt ccacaactac attttgattg 5400
tgtaatggtc tctatctatt gcaaagaaaa ggtcttttga caagtatgag gactacactt 5460
ttcaggccct ggagaaaaga tacacaccta taagagtgtt cttaactact gaatccccag 5520
tcttttgctg tagtgctgta tttgtaacag taagacactg tatcaactat tgatactttt 5580
ttttgagggg ggcgttaaga cagggtttct ctgtatagct ctggctgtct tggaactcac 5640
tttgtagacc aggctggcct ctcagaaatc tgtctgcctc tgcctcccaa gtgctgggat 5700
taaagagtgc tgggattaaa ggcatgcacc accatgccgg gctaactact gatactttta 5760
tcttgacttt ttttttttct ttttttgaga cagtttcaat atgttgctct gactgccttg 5820
gaacttgctt gcactatgta gaccaggctg gcctcaaact catagagatc tatctattgc 5880
taccttccca gttctgggat caaagacatg aaccactatg cctggctttg ttttgctttt 5940
gaataatcat taagataagt ataggctttt cacaatcttc tcagttattt atctaaatta 6000
ggttattact agatggctaa tgaatgtagt tcaatatctc cctgtctaaa tgctcagcca 6060
gtcatattga gaccctgtct caaaacaaaa aaacagaaca ctataatgtg cttttaaaac 6120
cctatttcta attattttgt agctttaaaa tatcaagtct tctgcctgct gtgcttttta 6180
aaagtgtttt tttttctttc tttttttgag agcagtggag cgaagactag gtacacagtt 6240
gtgtataaat tggcataaat actgctagag ttttcaatat agctctgaag aaaacttagt 6300
gttttgaaac attttattat attcatattt atttctttta aaagaaaact gagaccaaaa 6360
atggtagtga tataacttta gggttgagaa cttatgtgca tagaagccag aaaattcagt 6420
aatgttcccc caagcaactg taattcaggt gggttacgtg acgcttttat ggtttaatat 6480
atgataccat atagagtaat agaaaacagt acattttgta tgtgtaacaa agaagaattg 6540
tgggttttgt aggagctata attttgcata ttctgatttg agatgtgtat aaattttcaa 6600
cagtggtaca gcattttttc acgtttgtgt ttgagataaa cgtttattgg tatatgtgga 6660
gaatcttctg gactgtaaag tcagtgtcac tggctggcta tggctattgg catctcaaaa 6720
aaagattctt cgttgagagc atcagttgca agattgtcct tcaaagttga atcctaactc 6780
ccgtgtttac tctagaatgt ttggtaatga aaactgtatt tgaagtagac ttcctcatag 6840
tacagtaaat gaacactttg gtttagcatt aagattgaca gcttatcttt aagacttttt 6900
ttgaactatc tttatttttg atgccaatta attaccctgg ttaaagtaac acagaaatga 6960
agctaatgtt ttcttggttt ttttttcttc caaaattata ttcccagatg tgcaagttag 7020
acttcaaaga attcctttat tttattccat tgttttattc agataactac ctaactctct 7080
gaggtacact cacattagta ttctttttgt gtgcagggag aggggtaggg gagacagaca 7140
gacagacaga cagacagcat ttgcttgctg tgtgcaggaa catatgtgtg tggagcccag 7200
aagttgatgt ttagtcacta tccaccttat ataccagggt aggacctctc atgtaaaccc 7260
agagctcacc aatttaggtg agctagttgc cagcttgctt cagagaaccc acctccacct 7320
tatattttac ttgggtacta gggaccaaaa tttcagtctt catgtttgta tgacaagtga 7380
caagtaccta cctgctaagc tgtctcctca tatctaatgt tctttaataa ttgcctaaca 7440
atcactaatt agagtttttc gctataagtt aaattgtcct ttcttctttt ttttcctagc 7500
caaatcaaat agaaagctta cttttctata cttagcaaat gatgtcatcc aaaacagtaa 7560
aaggaaagga cctgagttca caagagaatt tgagtctgtg cttgtggatg ctttttctca 7620
tgttgccagg tatgttttct atcgggggct tttatatttt ccaccttttt gttctgttgt 7680
ttttgtatat cctttgcaaa tggacaatgt tgtactttac agagtgctgt gaagaccaaa 7740
ttatgctttt atgtttttct gggcccttaa ctatcatctc agacatgtaa acactttgaa 7800
acactaaaga gattcattct ggtaccagaa acacgttttt ctcctctgaa ttcactcact 7860
ggatttcttt ttcaagtaca ttttccatgg ctgagttgtc tcattagttt atgatcaaaa 7920
accagagcta gtactgcaaa ggagcagttc actctccttc gaagtgtctc ttcaatagcg 7980
gagactggtt tgagacttta atttccccct ctttctagtc tgctaaaaat agaagtcatt 8040
gctgtcatac aaaagtagtc atataatgtg tggggaagca gatgcttgag ccgcatggtt 8100
gggaatctgc aacttggact gcagagtgct gcaccatgga gcacacaggc agctggatag 8160
aaagctcaca cctgctgagc ttccatagca ctcgggcttg tttgtaggtg tgagttagat 8220
tcccaggagg aaaatttagt tttttgttcg tttttgtttt ttgttttgac agactattat 8280
gttcgtaagt atgcacaaaa atacatagat ctttgcaaaa gaaaccatga agagtggaaa 8340
tgagactagt gtattaaaca gaaatgtttg gagtctaaga aggtgttgtg tgggaattga 8400
agtgagtatt ctttggagga tttgaaatgt ttaaaggaat aaaattagta gtccaaaatg 8460
ttaaaaacca cattgagctt tctgaagctg gaggacttgc ttgaacccgc cgtgctggtt 8520
ctgatgacca cgggcactga agcggcagag tgcattaggg acgtgaagaa aacgactgac 8580
tcttcaaaca gctgctaagc tgagagaagc tatctgaaat tggagtctgg ggttggctgt 8640
tttgataagg cttagctggg tgccttagtg tgacactgtt ctgtctcacg tgccatgtga 8700
gtagccttgg gtgtataagt taagtttctg agcccaactt tcctatcttg ttaaaaggca 8760
gggagctttt agagtccagc tcactgtttt actcatccat tttcatgtcc tgcactccag 8820
gcaggaaagg gctgcctttt cctagctctt gggccaggca ccttgtagtg cttacagaca 8880
aggagtgcac tcacatagtc tctcacaggg tgtcccaaga cgcggcagca ctgcatctgt 8940
cctggtgcta ataggggtga gtaaatattc tttgagtcag ttaggataat cgtgactaac 9000
tggggaaaag atgtggttta agggtactat ctctgcccta tagggataat aaactaacac 9060
ttttataact cttcgctgtt ctgaattaga aaagggtaag tagaaagggc tacaaggtgg 9120
ataccaagat tccgagggct tacctagcat gtgggcacat cttaaaacca tacagaagcc 9180
aggcatagta acacacttct agaattccat cactcagagg ccagcccagg ctatgtagtg 9240
aaaccttgtc acaaagaaaa caaagagatg taaacaggag acattgggaa accatctggg 9300
cacattctac ctgatgtttc ccagctcctg tccatatgcc aggctgttta aggtccagtt 9360
tggaccatgt ttcaaggctt ctttccttca tcctatcatt agaacagaaa gtacaatagc 9420
tactttaata ctgttgtaac tcatcaacca agagtaactt agtaagtgtt cttggtgtgc 9480
ttttgctgtg accagatggt gaaccaaggt agctgaccca ttaattcccc aaataattag 9540
caagattttt atgatgcttc tctactttca tccccaagcc aaaaaaaaag acaaaaaaaa 9600
aaaaaccctt tgaattcaga aagttcatag ggtagatttg atttgagtag tggtttaaat 9660
aactagacta tgaacatatt atgcttatgc ataccttgtt ttctttaaag tttggaggta 9720
gttacacatt cattattcaa cttgtgtttg ttctatgcca cacactaatc cagagcttgg 9780
ggaacagcaa ggaggaaggc agataagaac agtgacgtga acaagacact ttgaaatgat 9840
atggacaatt aattagaata gtgttctagg gtagactctt ctccatacat cttgggtaag 9900
aaggtgacat ttgagaacct aaggtggaaa taatcgtgac atcttaagag gagcaggatg 9960
agatggatat ggtaaacaaa gtaagatgtg gccccagatg agatcagaga gaggaaagga 10020
gtggttccat agtccctggg agtctattct gatagggtgt cagtcaagaa ctgtaagcca 10080
gcgaatgaaa acatttctgt agacattata aaaagcccag gtaaaattag agaaagaaaa 10140
caaaacataa aacaaaacct aaccatataa ctaggatctt tccccctcat aatctatttg 10200
ttaattgtgc tgccagggac agttcatgaa aagtttaacc atttgtaaaa tcatgagtaa 10260
gcacttttgg taaaattgta cctgagccag gtggtggtgg tggcgcacac ctttaatccc 10320
agcacttggg aggcagaggc aggcagattt ctgaattcaa ggccagcctg gactatagag 10380
tgagttccag gacagccagg gctacacaga gaaaccctgt ctcaaaaaac aaaaaaaaaa 10440
aaaaaattgt acctgaatga aaagttcact ccattagcag atatccctct ttttagtcaa 10500
ctattctaat atatgataag gcaataagtg tatatctttt tattatattg tagagaaagt 10560
ttttattttt gttaagtttg tttgtttgtg ggttttggag atagggtttc tctgtgtagc 10620
cctggctgtt ctggaactta ctctgtagac caggctggcc tcctaaatac tgggactaaa 10680
ggtgtgcacc accacctcct ggcaaattaa tactttttaa aggtttctat tagatgcctt 10740
taaaaagaag agatgacaga gtgggtaaag gccagactag ctgaatcagc aatctctggg 10800
tacaattgag agaccctgtc tcagtttgta aggtagagaa caacacacac acacattcac 10860
atacttgcaa accatatata tgaaaggccc gtgtagagaa tgggttgtga agggcaggga 10920
aacctgagct cagaaggagt gttgctgtgg tgttggctaa ggtgagattg gttaagacat 10980
ttgcagtgag aaggctctct ccatttattg aatgcttatt ttgtagttgg agtcttcacc 11040
cttgtaaaat gtacatgatg tcttgaaaag ggagctggta ttgaggcaag aaagagattt 11100
ttagtgtgag tcgatggctg gtctgaaccg aggactggga agctttggtg gaggaagtaa 11160
tttttgtttt gttttggcca tgttaagttt gagacggctg gttaggtgtt tcaagtggca 11220
attgtgggtg gacacatggt taaactagta tcaagttcac agaaacttag tctttgttat 11280
tttatgtata tgagcattat gtctacatgt gtgcctgctt gatcacatgt gtgcctagtg 11340
gccacagagg ccacacaggg catcagattt cctggaactg gagttgcagg cagttgtgag 11400
attccatgtg gattccagga atcaaatctg ggtcctctga aatagcagca agtgctgtgg 11460
ttgctgagcc atttctccag gacccctctc cccctaattt ctattttttt aaaaaataaa 11520
ccctaaaatt tcatttgtca tttgtgtgta tacacactca tgtgtatgta catgtgctat 11580
agcaaacaca cagagttcca gaagtcaggt cactccttcc accatgtgga atcccaggaa 11640
tttaactcag gttgtcaggc ttggtggcct taattcctga ttctagactt aatttcttga 11700
gaggtaaatc aggagttatt gccaaataca tgataattcc ttattgtttg tgggtcttta 11760
tacttctccc aggtgggtaa tttcctggga ggtatcatgg ttgttaggtt ttattgcgct 11820
cagggataca gaacagtcag caaaggggag aggtgcatgg gtaattgtga caggaggtgg 11880
gaagggactc agcacccaga gcctcctcta ggatttgtgg agggtgcttt tgtcactgtt 11940
cctgcagagt aagtaggcat tcacttagag ccatgttgtt cacacaagtg aggcacccag 12000
atcatttggg agtggagagt gtcctgatcc ccacttccac cctcacccca atctccaggt 12060
ccgctcctgt cttagctttc tgtcacggtg gaagaatgcc agagagaaac aagggagtaa 12120
ggatgtagct gggctcatgg ttgcaaaggc ttcagttcat gctgccagct cctcctcttc 12180
tgggctgcag ggaagcagag catcacagca gaaagtgtgt gattgagcct cttgatggct 12240
tagtggcaaa tattttaaaa acacttttca cttagatagc tctcatatac acattattat 12300
ttatttgttt gttttgtttt ggagacaagt tttcatgctg cccaaactag ccttcaaaga 12360
atttgtgtgg tcaggaatgg ctttgaatgg attctccagc cttccccctt gagtcctgag 12420
attgcaggca tggaccacca tgcccagttt atgcactgct ggggatcaaa ctcagggctt 12480
tgtacattcg aggcaagcac attgccaaat gctatacatc cccagcctca agaatctcag 12540
tgaacctggg aaagtagtgc cccatctttt gtcaactgct ttcctgctct agccacatct 12600
gattttaggt accacaatga tgtctacttc accgcagtgc cttacttgaa aaccagtttg 12660
tttaatttat gacttgaatt actttgatgc aaaagagttg gaagtcaagt atatttatat 12720
aaaaatttta aaaattcatt taaaaaaaga ttgattcagc caatatggtg tctgtgaaaa 12780
caagcccctt ctcggaggct tcaagttacc ttcagatggg aagaataaac aggtctcctg 12840
tgcagcaggt gagagggatg ggtcgtgtga aacccatgaa ggagcaagaa gtgccagtct 12900
ttgggactgg ggagctgctg ctggagagac aggaaggaca gacctgaggc cacctgaggc 12960
atagctgact taggctctaa gcctgatgac cagagtggat aggtgaggtc agaaagcctc 13020
aagggctttg tatggacttt gttctgagaa gaatggaaag aaacaagaga tgccttccca 13080
tggcagctag aattacttta ttcaaactgg agagtaggct gtaggagtgc aagactggaa 13140
gccgggagag cagtaatgag gctatcagag aaaccagcgg agggctgccc cttgggccgg 13200
gttgttggca gaggacaggg tgagagtaac taatttctgc atacactgtg aagaaagaga 13260
gtgtcttagc ataaaccata gagtgcttgc ttatagctga gctctttaag atgtagcaga 13320
agggtttccc tgtttcccta ggacttacct ctgagttaga aaaaaaatta acttatacat 13380
ttcctgactt agtactttga tctcatttca gtggacttgc tactagtaat ttgtaagtag 13440
gacacttggc cagaccaggc tgagtgttag cataagcttc acagaagtgc tgtcagtgca 13500
gaaggtgggt atagttcctg tctgttgaat ttggtattct tgatgttgtt agtaaagata 13560
cttgacttca accctaagat gtggaggttg tttgccctcc cctttttggc atagttttat 13620
atatctgagc ctggcctcaa actcactgaa gctgaggata cgtgaccttg aatttctgat 13680
gccgttcact tccccagtgc taggttacag tgttgaggat tgaacccaga gctaggcaag 13740
cattttgcca actgagctat agttcaggtc tctctctttt ttgaaagtta tgttttattt 13800
gggaagaagc atggcctact tgtaaggggt cattgaaaaa cttgcaggaa ttggttttct 13860
ttttttactg tgtggtccag gaatggaaat caggttgtca ggcttggagg caagcttatt 13920
tatttgctga gccgtcttgt gagtctcagg atgtcccttc ttaaggtgct cagtgttaga 13980
tgttggcttg tatgcagtct tactaaacag agagcgcctg ggagtatata tgtggcaggg 14040
gcctctcttc agaaagtctc atcctgcttg tctgctgccc tctcaggtat tatggaccag 14100
aaaaagattg cctgattttt tctttcagat aaatattaca aaggactgca ttatttccag 14160
agctttccaa gattatccat tagatacatg agtggtgaca cagaactcag actaaatcca 14220
gtgttaaagt tgttataaag ttgttatggt tgagacactt ggcacatatt gtacacacat 14280
atttatgcat gtagaggtgt ggaatgttaa tatagtgaat ttaaacacat gtgctttaaa 14340
aatcttaaag gtgtggctaa ttttaatgtt cagtatatgt ataggattgt atttaaattg 14400
atgccccccc cccacacaca cacacacaca cacagggttt ctctgtgtag ccctggctgt 14460
cccggaactc actttgtaga ccaggctggc ctcaaactca gaaatccgcc tgcctctgcc 14520
tcccaagtgc tagtattaaa ggcatgcgcc atcaccacca cccagcaaat tgatgtcttc 14580
taaaaagcta aattgtcaaa atcaatatgg tcagttttgt tttgttttct gtctgttaaa 14640
accagtcata caggctttcc ccctatttct tccaccttgc aaaggggaaa aagattccct 14700
gggtcgttgg atgcctcttc ctggttcaac atgggaggaa tgtagtttct caagactgag 14760
catccaagta atacagagtg taccacagtt ttgttttaag tgtgtaaatc attttggcag 14820
ttgttgttgt tgttttaaag taaatctgtc ctaattaaaa acaaaaaatg taagcttata 14880
tatatatctt ttcatttttt tctggacaga gaggcagatg aaggctgtaa aaaaccttta 14940
gaaagattgc tgaacatctg gcaagaacga agtgtgtacg gcggcgagtt catacagcag 15000
ctgaagctgt ctatggagga ctccaagagc cctcccccca aaggtagaac atcaccacat 15060
gtttacagct cttggtcttt gctcctccca gccccacgtg aaaagctgca gtgggggcta 15120
cagtggctgc tgtgatccac cagccgcttt atgactgcag agggatgctc ctgagtgatg 15180
gtgagaactt acacagtggt gcaagcggcc aggcctggtt ctttgctcac gatgccatgc 15240
tcgcacttcc tcactttctg tgtgttcttc atggcccttg ttgctctttc agggactgtc 15300
cttgtgcagg gaagggaaca accaagttct ttaggattgt ggctgactaa cgagtagctc 15360
atcctggccc tgtcgtggac ctcagcgggg cctgggatag gtctcttgcc ctttcagaac 15420
tcagctttat tagtaataag tctgggttta tatagtttaa tgactatcag attactgtct 15480
atattaatta ttgattttcg tctttactga gcccagggta tctttgtgaa gtatatatgt 15540
gaattctaaa attcttaaga gtccgtgtta tttatatctt taaagagaaa gttcagacat 15600
ttggtttttc ggttgctggc tagccatgga tcactagtgg catgacatgc ttcaccctgc 15660
tctgcctttg ttcatatgta ctgctgagga gtaagtctga gtacccagga ctaaccatct 15720
tgtttaaatg agcccgcttt gagattcttc ttccaaccca tctcctagtg atgcttctgg 15780
ctctctggcc aggctgagtt tctgagtgtc ccttgtagat gctttgccag tcctttttct 15840
ctgcacatgt gcattcctca ccttttcact tgtggcaata cattcttaaa gaatgccact 15900
ctggctccca agccttcctt gctgtgggcc atgtagcctc ctgttgtctg gtactgggta 15960
cacactcctg ctgtccgacc tcccaggact agcacaggtg gcagtcctat gcctcaaccc 16020
tttatttgta ctcttcacag tctttcccag gtctaaaatt gcatatcctt cacagtaaac 16080
tggaaggctt tctaagaaag tgcttttgag gcttctggcc actgttgttt tacctaaagc 16140
tgctaaaaca tgctcctgaa aagcaagctt tttcctgctt taggttctac tagtgacagc 16200
attgtggcct ttgggaattt cactctacag ttttgaaaaa atatctagca tttttgccta 16260
taccctatac tgcattcagt agtgcttggg gaaagcaagg aatctcttgc cttaaaggaa 16320
gcttgaaact tatgttggaa tgtccgatca ggcattgcta catatgtggt gctgtgctag 16380
ttgctttgca aagcgcagag gagtaaaaaa cacctgtgct ccccagggag ccatcacaga 16440
tatggcagtg ttggaagtgg ctcctttggg gtttggatct tgtttgcagg ctagtggtga 16500
catgtgctgg tgtcatttgt ctaaagagtg tttttctttc ccttcatctt ttaaagtcag 16560
atactcaaca gactccttgg gtattctggt cttggcagct cattcttgtc ttctcaggac 16620
ctgagtgatc aagatcgttt atacgattct ttcaaatacc catgactttc caaaaggaag 16680
tcattttcaa atctgaatcg ttcttttatg ttgataaatg atatgtatac atgaactaga 16740
gttacagtca gttgtgagct gcagttcagt tgataggaac agagtctggg tcctctgcaa 16800
gggcagccag tgctttttcc acttgacatc tcactcactg ccctcccggt ctcagtggga 16860
cagccagttc tcttaaccac taagctccct cttcagttcc acaacactga aaattttatt 16920
ttgctcttat gttcatactc tggtgaactc tttcacatgc actgctctgc tatcgtggct 16980
aagtgtgtgc aggttgtacc acatgccaca ttaagtagct actgcgcaca cagttctgtg 17040
aggttcatat gtaggccata tgctagtggg cttttcactt aactatatac cagtctttgc 17100
ttttcaaaac tataaaaatt tattgtgtga atggaaacta tgcaaaaaga acttaatcca 17160
agtagaaaat ttcatttctg tgcgtttttg ttttgtcagc taagatgctg cctacaaatg 17220
tatagtgaac agaaaaagag taaaactaat agttgctttg ctatgcacag tggtacattc 17280
ctgaagttct aggactcagg agacagaggc aggaggattg ctagtttgag accagcctga 17340
gccagcttga gctgcagagc aaaggctctc tctcagtaaa ccagtatcta ggatgctgag 17400
taatagggtc agcctggaag ggcaaggccc taaattccat tgtcagcact agagaaaaac 17460
cgaaaataaa cattgaaata aagtaacttt cactgtaaca aaatatatgg agagtagttt 17520
gaactgtctt gccttcggtg ttttaaaata ctaagagtta atttttattt ttaatattta 17580
ataatttgaa ggctggggct ttcacaaagg gctccacatg tgtcagccaa gggttctgcc 17640
agtgagctaa cacagtgagc taacatcccc ttttgactat ttacttttta cttaattgtc 17700
tcgaacttct gtccagctaa gatttcaagt ttttgtcact agatatactc tgaaagttgc 17760
tgtggtgtca agtgtttttg tggggtcact tcttccacca tcatcctctc ttcccttttg 17820
ctgctgagtc ggcatcacca ggcccctctt ccttctgcct ctcaagcact tgctccccag 17880
ctcctctcag tggtgacctt cagattcagt tcctccatgg cttccatcgc agcttctttg 17940
ttgtgctttt atcgttatgg ccagttctct acttattatt gttatgaaat gtcagatggt 18000
ttctgtcact gagaaatggg atggcacagc tgaggaactg atgtgcctgt cactgacaga 18060
catggcagac aagtagggat tggttcttga tttgtgtttt gggaactgaa atattagaaa 18120
actcacgtaa tcccaggact cagagttata gcctaggcgg tgagagctgc agtgtgctgc 18180
tgaggactgt ggcttcactg acctgagaga gtcctctgca ctagcattgt gtgaaacaag 18240
gatagcctcc attgtaaatg tggctgccta ctttctcctt ctctccaaca agcaaaacca 18300
taaggaaaaa aaaagcaatt catgctattg cctggccttc ctgtcacaac ttagctaatt 18360
aatgtgtaag atggaaagtg ggagctgcat ccttagttcc agaagagact taagtgcaca 18420
tgtgcacacc ctccctgggg agttactctg tacgtgtagc aagttccaag gcaggtgcct 18480
ggtatcccct ttttgacaga taactgtact tgcatacagt ggaagaagta ctggtgaggc 18540
tgtcatgcca gagagttgtt taacctccct gggtcttttt cctcttgtaa tgtaatgctc 18600
tgtcaaaatt gttggaaaga tgaaggtaaa gaaaccattg caatgccctg cacatacttg 18660
gtgtccaata tttctgttaa atgcagattt atcatttaga tttatttccc tcaattgttt 18720
ttcttgtttt gtgttgccca atgtgatgca atagaaatga ttttagaatt aacacatcta 18780
tcataatcat ataaaagcac aggggacaga taataaagaa tgcaacttgc caggctcaaa 18840
agtagttaat tctagaatgg aattgtaaat aatttcttcc agtggtacaa tcacatttct 18900
cagtaatagt tttaaatttt atagtaaaat ttagtttatt ctggttttaa agaaagagtt 18960
taaaggttac ttgtacttga agtctttgac agtatagttg atgaatctct cccttctatt 19020
tattttctat aatgatcctt ggaaaccata aaactgtttc ctgtgcagtg tattctctgt 19080
cgcctcgtac atgttgcttt acttcattgt attgcctcat gcgtcttacc atcctaatgt 19140
ttacttcttt tgaagggaag aaggttctaa ttatgataat tggggcatac cacttccagt 19200
agcatttcct tggactcaaa gtttgggaac tttttgtctc ttcagtaaca taaggagcta 19260
taccaattta aaataactgg caagacagtt agcatgaagc catttctgaa aatgaacact 19320
ccctttcccc tcggtgtttg cctgtggtca ttgtgcagag cactggatgc ttatgtaagt 19380
aggacatgca gctcacactt gtctctttat tttccctagc agcagaagag aagaagtctc 19440
taaaacgaac ttttcagcag atacaagagg aggaagatga tgactaccct ggaagctact 19500
ctccccaaga cccttctgca ggccctctct tggtatgtga tgccaggcac aaagtgactc 19560
caggtggcct ttagctgaag atcatgacca caaggtggtg ctgttaccag tggtgtggct 19620
tatcaaaggg cagcagatgc agtagcgcac tcagagattc ccagacacct gacttcaccc 19680
tgccttttac acctccactt tacaatgggt taatgtttat aaaatgttcg gacttgactt 19740
tgtcttttaa aagttaaaaa tactctaaaa gtagatttaa ggatataata aagatgttct 19800
gaggcatttt gctatgcatg ttgctaagaa cagtcttttt tttaaagatt tatttactta 19860
ttttatgtat atgagtatac cattgctttc tttagacaca ccagaagagg gcattatttc 19920
ccactacaga tggccatgag ccaccatgtg gttgctggga attgaactca ggacctctgg 19980
aagagcagtc tatgcagtta accgctgagc catctctcca gcccactaat agcattcttg 20040
tttataagca gataacttac tattatctga ggacagtctt gtagccactg atagtaaatt 20100
tactgcactc tctttattat tactttattg caatagggag tttgccttta aggatacagt 20160
tgcctcatat agtcccttat gagacaagct tgatttggtt catctgttta ttgtaaaaaa 20220
aaaaaaaaaa agagccacaa aacaaacaaa cagattaagt agcatttatg taacttgcca 20280
agtatcttaa tttgaattaa gatctgtcat aattcagagt cattgtgctt ttttttattt 20340
gttttatttg taatgttttg agatagagtc tcactgtgta atgccagctg actttaaact 20400
tgagattttc ctgctgtggc cagattacag ctgtacactt gtactgggga gtctgaggct 20460
ggaggattat gagtttaaga tcagcctggc tatacagtga gacttcctgc ctcagcttcc 20520
catagctggg atgacaggtg tatgccgtca cacttgtctg tctgagactg ccttgtgtta 20580
atcttaactg tattctacat ggtcatttgt ggtcaggaca taggataggt aaataggtca 20640
tatgagtggt catatgtggt catatgagtg gtcatttgtg gtcaggacat aggataggta 20700
aataggtcat atgagtatat gcgtgtgcac tgtacactca acgatggatg agacaggatt 20760
aagtaataga aatgttaggt cagatctttg catctaaaac ctttaaagag gaagaccgtg 20820
gccgatcaac tttcagggca atctgactat tgccgacaga gccagctgtc cttaggtcat 20880
tgtgctctgc cacctgtcag caccctatgc tttctcgtgg ttttttttgt ttgtttgttt 20940
tggttttttg tttttttttt tttttgagac agggtttctc tttgtagccc tggctgtcct 21000
ggaactcact ctgtagaaca ggctggcctc aaactcagaa atccgcctgc ctctgcctcc 21060
caagtgctgg tttgtaaaac ccttttgata gacacaggat atgcttttct tgtctttaac 21120
tcttaatgag ttaatttatt tgacatttta atattggcag ggatttttat tttgtaaaac 21180
aaactctaga aagaatttaa tcagccagca cagctgttca gcatctggca tttgtgcctt 21240
tttttctata ttatagactt ttatactgta aatagtgttt catttgaaat taagcagtat 21300
ggactagaat ctgtattctt aagttgccaa aaatgtgctc tttaattttt ttttttaaac 21360
agaattctgt agaggtttat ttatgtgttt gtagacttaa aataatttta tgtcatttag 21420
aatccatatc atttccattc acaaagttct ggagctaaca cagttgctta tgtagctttt 21480
ggcttgtttc taatgctcag aatggtgatt tttgtcattt gcctctagct gtctttcttt 21540
aaccccttta tgtcttagtg gataatagta tttgcctttg ccttttgtag actgaggagt 21600
taatcaaagc tttgcaggat ctggaaaatg ctgcgtcagg ggatgctact gtccgacaga 21660
agatcgcttc cctgcctcag gaagtgcaag acgtgtcgct gctagagaaa attacaggtg 21720
agaaggtagc ttagctgaaa gataagaaag aaggcgctat ctagagtcat gcttatgtct 21780
gtaggtagaa aatttctgtg tgatgttggg tactctgctg ccttggtgtg tgaagccatt 21840
tataggctaa acaagaaagc tctcagaggg gctgggcagg tagctccaca ttgagtagtt 21900
cctgctccat ttccacacct gcaccgggca gctaacacag ggcatctgat gtattctttt 21960
ggtctccatg gacactcact cactcacaca tacggcatat ttatatacat actcatacat 22020
tatatatata tactcacaca tatacggcat atttatatac atactcatac attatatata 22080
cactcacaca tatacggcat atttatatac atactcatac attatacata tactcacaca 22140
tatacggcat atttatatac atactcgtac attatatata cactcacaca tatatggcat 22200
atttatatac atactcatac attatatata cactcacaca tatacggcat atttatatgc 22260
atactcatac attatatata tactcacaca tatacggcat atttatatac atactcatac 22320
attatacata tactcacaca tatacggcat atttatatac atactcatac attatatata 22380
tactcacaca tatacggcat atttatatac atactcatac attatatata tactcacaca 22440
tatacggcat atttatatac atactcatac attatatata tactcacaca tatacggcat 22500
atttatatgc atactcatac attatatata tactcacaca tatacggcat atttatatac 22560
atactcatac attatacata tatactcaca catatacggc atatttatat acatactcat 22620
acattatata tatactcaca catatacggc atatttatat acatactcat acattatata 22680
tatactcaca catatacggc atatttatat acatactcat acattatata tacactcaca 22740
catatacggc atatttatat acatactcat acatgtaaat aaaataatac aaataaatct 22800
ggatcttgag gccagagaga cggctaagtg gttaagaacc cttgctgccc tttcagagga 22860
cctggttccc agcaccagac acagttgatg tacttacata tatccagaca gacactcata 22920
caaataacaa acaaaaattt taactaaata gataaaaata aaccacaaca aaaagaaact 22980
tttgaaacac actgagtagt gatttgacta gttagggcag aaatgctggg ctgccatcac 23040
tctctgaata agatcatttg gcactcagtg caaattgtgt atatgtggag cagtggaggc 23100
caggaagggc tagactgtga gctgctaaca gaaatatttg gtacacggct gtgttgttta 23160
atgtattgca cactgacaag gtgattcggt gtcgagagtt gttattgagt tcatcacagt 23220
cactcccagt taaacaggag ttggaggaga gagtgaagta tgaactaatc ttttagagag 23280
cgtgctctga actctacata cgcctcgtga gtctctgaga attgtgttct catgtgtagg 23340
aagggattga gactctgtaa agttagctat cttggccaag atcacacggt cttctcactg 23400
caaaagccca atcctttcat cccattctct ccaaaggtca tttccccctc catctctctt 23460
ctttttaaat tggatatctt atttatttac atttcaaata ttatcccctt tctcagtttc 23520
tcctctgtaa cttccccacc cccaccccat cccattccct cttactctgc ttctatgagg 23580
gtgctctccc acttacccac tcctgcctca ccaccctagc attcctctat actggggcat 23640
caagccttca caggaccaag ggcctctcct cccattgatg ccatataagg cccctttagc 23700
tccttcagtc cttcccttaa ctcctccatt ggggtccatg tgctcagtcc gatggttgtc 23760
tgcgagcatc cgcatctgta ttggtcaggc tctggcagag cctctcagga gacagcttta 23820
tcagactcct gtcagcaagc acttcttagc atcagcaata gtgtgtggaa ttggtgtctg 23880
catgtgggat ggattcccag ctggggcagt ctttggatgg cctttctttc agtctctgct 23940
ccactcattg tccctgtatt tcctttagac aggagcaatt ctgggttaaa attttggaga 24000
tgggtgggtg accctatccc tcaaccaggg catggatatg gtctcaacag gttctccctc 24060
ccctttgtaa ggtattttag ctaatgtcat ccccgtggga tcctgggatg ctcttgcttt 24120
cctggcatct gggactttct ggttgctacc cccagttccc cattccccat tgctacacac 24180
ctctgttcaa tttcctgacc ctctatatat ctcctccatc ttctccctta cctgattctg 24240
cccctctttt cccctccccc tttcatcttc ctcccaagtt cctcccaccc tctactccca 24300
ttgattattt tgtttcccct tctaagtagg actgaagcat ccactctttg gtctttcttt 24360
cttttgagct ttatgtggtc cgtgagttgt atcatgtgta ttccacactc tgcctaatac 24420
ccacttgtca gtgagtacag accatatgtg ttcttttgtg actgaatggc tactccatct 24480
tctttcttgt gaccatttgt ttggaaattg ttttctagcc ttttactctg aggtagtgtc 24540
tgtctttgtc actgaggtgc atttcctgta tacagcaaaa tgctgggtcc tgtttacgtt 24600
tccagtctgt tagtctatgt ctttttattg gggaattgag tccattgata ttaagagata 24660
ttaaggaata gtgatcgctg cttcctgtta tttttgttgt tagaggtgga attattttgt 24720
gagcttatct tcttttgggt tttttttgaa agattacttt tttgcttttt ctagggtgta 24780
gtttccctcc ttgtgttgga gttttccatc tattatcctt tgtagggctg gatttgtgga 24840
aagatattgt ataaatttgg ttttgtcatg gaatatcttg gtttccccat ctatgttaat 24900
taagagtttt gctgggtata gtagcctggg ctggcatttg tgttctctta gggtctgtat 24960
gacatctgac caggatcttc tagcttttat agtctctggt gagaagtctg gtgtaactct 25020
gataggcctg cctttatatg ttacttgacc tttttctctt actgctttta ctattctttt 25080
tttgttttgt gcatttgttg ttttgactat tatgtgacag gaggactttc ttttctagtc 25140
cagtctattt gtaggcttct tgtatttcat gggcatctct ttctttatgt tcagtacgtt 25200
ttcttctata attttgttga agatatttcc tggccctttg agttggggat cttctctctc 25260
ttctatactt attatcctta agtttggtct tctctttgtg tcctcgattt cctggatgct 25320
ttgagttagg atctttttgc attttgcata ttctttggct gttgtatcag tgttttctat 25380
ggtatcttct tctcctgaga ttctctcttt taatatcttt tgtattctgt tggtgatgct 25440
cacatttatg attcctgatt tctttcctca gtgttctaac tccagagttg tcttcctttg 25500
tgatttcttt attgtttcta tttccatttt tagatcctgg atggttttgt tcaattcctt 25560
cacctgtttg gttgtgtttt cttgtaattc tctaagggaa ttttgtgttt cctcttttag 25620
ggtttctacc tgtttacctg tattcttctg tattccttta agggagttat ttatgtcctt 25680
cttaaagtcc tctatcatca ccatgaaatg tgattttaaa tcagagtctt gcttttctgg 25740
tgtgtggtgg gagaactggg ttcttttttt tttttttttt tttttttaaa gatttattta 25800
tttattatat gtaagtacac tgtagctgtc ttcagacact ccagaagagg gcgtcagatc 25860
ttgttacaga tggttgtgag ccaccatgtg gttgctggga tttgaactct ggaccttcgg 25920
aagagcagtc gggtgctctt acccactgag ccatctcacc agccccgaga actgggttct 25980
tatgatgcca agtagccttg gtttctgttg cttatgttct tgcacttgtc tctcgccatc 26040
tgcttatctg tggtattagc tggtctgtct ctgactgtgg cttgtccctc ctgtaagctt 26100
gtgtgtcagc actcctggga gaccagctct ctctgggaga aatttgggta tggagagcag 26160
tgacacaggg tcagctccat ggtacagaca gaaactggaa ggatcctgtc cctggctgtt 26220
ccttggttcc tgtgtcctga tggctttgtg cgagtcactc ttgggccagg aatttgagca 26280
gaagtggtgg tctcacctgg ctctcaggtg tgtcagcact cttgggagac caactctctc 26340
ctggtggtat ttgggtatgg cacagggtca gctccgggtg cagaaggaaa ccggaagctc 26400
ccctccatct ctttaagctg gctgcttctt tgtcttcttc catgttcaca tagaggaagt 26460
caaagttact aaagcaatag cttccatcat ttggcatata tatctttgag gctcactgta 26520
gagtaggcgt tcctggtgtc atgcccatag cctttctggt attaaatatt aaggaggagg 26580
cagagttggt gggcagtgaa ttcctgaaga tcagagaaag ctttgtgctg tgggagtctt 26640
cgagcttccc ttgcttgggg tttgtattct actacaagcc tgtcccaatc cttcactgaa 26700
aataccgatc tcactcccag agccccacct tttcttctac tggtcttcag ctgatgaata 26760
atccagcact atgtaagaca ccttcctgtt tttttaataa agatttattt attttatgta 26820
tatgagtata ctgtagctgt acagatggtt atgagccttc atgtggttgt tgggaattga 26880
attttaggac ctctgcttgc tctggtcaat ctctcactcg ctctggtcgg cccagctcgc 26940
tccggtcaac tctgcttgct cagtccctgc tcgctcccca caaggattta ttattatacc 27000
taagtacact gtagctgtct tcagacacac cagaaaaggg cattagatct cattacaggt 27060
ggttgtgagc caccttgtgg ttgctgggat ttgtactcag gaccttcgga agagcagtca 27120
gtgctcttac ccactgagcc atctcgccag ctcatataac accttcctac taagaataca 27180
gacctggaaa cgaggctgat tcttgtcttg tctcctgctg tctgagaacc agtgtaggtg 27240
ggaagagctc aaggattttc taggtgattg gaaagccatg cactgagaat ttggaaggct 27300
cattggggtg aacttgttca cataggagag aggggctggt tacagggact gtaagtgctt 27360
cttgagattg ctgttaaagg gagcagagag gtgaattagc agttggggag tgctagctgg 27420
taggaagaca gggaaagctt gggggactct cagaaactgg gtgtttgagt gagtcagcta 27480
ggaatgcagg gttctccagc ccaagtggag gactggtctc aggaagctca ccaaggcaca 27540
gggaaggcag accaaagcac agccattgtt ggccccaggc aagatctgct tctacatttg 27600
gaatgcatct ttttgtagtg ccaacccacc tttatagttg aaatgatttt tttttcctac 27660
aatatattct gatcacagtt tcccctcatc tcctctgagg gcctccctaa ccctccacca 27720
atcaaatcca ctctttctct attttgaaag caaacaggta aataaaagaa caaataaacc 27780
agaatttaaa caaaaacaga cacacaagcc ctataaaaaa cagaaaatca gaaactataa 27840
tatataagca aagaccagta aaatagataa aagttaccca aacaaagcat ttcaagacag 27900
aacatctcct aaagtaccat taaatatgtt ttgtgttagc catgtactgc tgtccttaaa 27960
tgtgatttgg tttgcatgtc cagtgagatt ccattggaga aactaatatt ttttctttgt 28020
aaacatttga cacttggagg tttttaggcg tcagggctct tgtctacttc ctctctcagc 28080
actgggaccc catctggctt ggacctatgt gggttcaatg catgtctctc tgagttcatc 28140
catgtatggg tcctgttacg tttggaagac actatttcat tggtgttctc catccccaca 28200
gctctcagag cccttagggc agggaaatga agatatccca ttcagacctg ggtgttccaa 28260
atctctgact ctcagcacac ggcccagttg tgggtctaag ttctcttgca ggaggaagtg 28320
tctgtgatga tggctgagca agactctgat ctcgattcca gcccagtgtg gcttggtgaa 28380
ttcttgagtg ataaacaagc tatgtttctg ggttgcaatt acttgtataa atgatctggt 28440
ttccttatac aaattcttga agttgagaat tgacttttta aaaaaatctt atctcttaaa 28500
aattggtgtt tcttacctcc ccccagccct ctaggtactc tagcatctta attacttact 28560
gttataaaga gatcaatagt caaggactga ccaagctgaa agcagccaca ctcaagctta 28620
ttcgcaggca gcaccgccag tctggagcac tgcactggtc ccctgccctc tggcacattc 28680
ccttgtccca ccttgtcttg tttaccccct cccaagtgtg tgttgttctc tcaccctgat 28740
gccagagcgt gatctttggc ctaggtccca gttgggaagc cctcagcagt tcatttccta 28800
gtgttcatag agtccagaaa ggaccagatg gctgttctat gctggaaatt cctaaattcc 28860
agtgccttga aaaatactga gggctctcag atttgtgtca ctgaattgtc tttgaaactg 28920
ctggcagctc tgatgtcctt tgtttcaaaa cataaacctg ctattttcac ccaaaaggcc 28980
ttattttatt ttatttattt atttttaata attatgtgta tgtgtgtact tgcatgaatt 29040
tatatgtacc acacacatgt atggaggtgt cgattgcatt ggactctctg gattgtgagc 29100
tacctgatgt aggtgttggg aaccgaaccc atgttctcta caacagcaat acgtgctctt 29160
aaccactaag ccatctcttc atccctccaa aatacctttt taaagcaaga tgcatgtatt 29220
tacacttctg ccaccaccac acagttgcag cattctttat ggatctaagt tcttccttgt 29280
ctttgcgaat ttgctacacc agtgttgtta ccacttcttt cccaggatac ccgtccttac 29340
acttcagggt ttgttctttt tctgttccta acgtgtaggt gactgccatt attcagacat 29400
gaaatcagtc tggtccatga tgaccagcac tttgaaaata caacacatcg taagaaaagg 29460
ctaatttttt ctcctttcct tctctgtttg gatcttgcta tagaacccag gctgactgga 29520
attggtggca gccctcctat gtgtgtgtgc taccatacat gctgctgctt gtttgtgttt 29580
ttacaaactg tttccaccgt gggtcatgtc tatgcacact tgaaaggccc tgctttttca 29640
agtgcaggag acaagcttgt tcccaggtac taggtactag gtattaggct ctctttctct 29700
ggtggacaga gtcctgcaga tcagcttcta agatcacatt ctgaagctct cacaaaactt 29760
ctgtcttctc tcagtctctg tacatctaag acgtgagtgc ttctttagaa cagtagggaa 29820
agtattctta atgtagtaat ataagcaaac atacttcatc tcagaggcaa gagaaatcat 29880
ttcacagatg actctgagta cttttctgct ggttttgttt ggagagtttt tgctggaagc 29940
tcttcagcct gccagctttc ccgccctgtc cagttgtctc tcacttcagg tagttcagaa 30000
atgcatcttc ttgacagagt tggtgcctgt ggattcttgg ctaaccaaca ctgttggttt 30060
ttttcttgga ctctgaacct tctctcagga atgtttgcaa tcatacctat gtgatttgta 30120
tgttgtctct gttttctgct cctatttgta ctcagacaaa gaggcagctg aacgtctttc 30180
aaaaacagta gatgaagcat gtctgttact agcggaatat aacgggcgcc tggcagcgga 30240
actggaagac cggcgccagc tggctcggat gctggtggaa tacacccaga accagaaaga 30300
ggttttgtca gaaaaagaga aaaaactaga agtgagtaca ttgcagggca gatccgactg 30360
cctgaactgc ccccccccct cccaatctca tgttattcct ggatcaacct ttttatcctc 30420
tgaaaacaaa cgagacagga gcattcatcc agtagccata attaccagag cctcgataga 30480
agtcaaagtc aacttttttt atgttcagga aagaaaaatt catattgtga aaaccggggc 30540
ctcttagtca ttgcatttag gatcgtaaaa ttgcttcagt gtagtgttac agttaaggaa 30600
ttataaattg aggtagttgg ggtagcaaaa ccaaaggata cttaattggt tatatttggg 30660
aatctggatt gttttgacgt ttacttctgg catatgcacc tgcctgcaag ccctacctgt 30720
acatatttta aaagtttaat cttgctatgt agcctgactc aaatgttggg caggaactca 30780
tttttcctga gttgaggcca tgttcctcag tgtctccagc aaggaggggt ttggagtcgt 30840
cccctcactg tctggcttct taattcagct taccttcttt tccctgggga gcagagccag 30900
cagtgtggct gtctgcggtt tcctgtccct cttgttagtc tgtatttctt agctgctgct 30960
ttgtttcttc tctctgtgta aaagatactt agaaaagact aggaagggag cagtggggct 31020
gagctggaga agccataatg accacaggaa acgcctcaga gagtctcctc agtagacaag 31080
ggagtagaag accacccttc ttccagtggg gaaaagttct gtggcacaga gggagggaat 31140
actgagctag ctccagtgat aggcattcac cattctacag tcagctgact ggagaggcag 31200
taccatatcc ttccctacaa cgattatggg agttgtaatg gctagaggtg cgggggactt 31260
caaacgcctg gccagcttga gtgtggcaga taggagacag ctgaacagta gtgtggcttt 31320
catgtagatt cagcactgac cacaagtcct aaagggaact tcatcagcag gagcagacgg 31380
ccatttatct gcagcgctca tctttcactg aggaagcatt ttcactgtac cctgggacta 31440
ataagagatt gtaattaatg attctggttg taagtcatcg tgccatctgt tgaactgccc 31500
cttggcgaca cattaagggc tttctctgat ctgtcagaca ttgattatca tgagcagctg 31560
ttgttgggta gaacgtggcc agccaccata gaggacagtc tttgagaagg agactgttgt 31620
agagccagtt aacccttcgt tctgtttctt ccccatttat aattaggtat ctgtcggatg 31680
ggggataggg ctgggctttt gcagaatagg tagcttactg gataccccaa gaagttatga 31740
ggtacagaag atcagaggtc agtcattttg tagctttggc agctttattg ttgctagttc 31800
accgaatagc atgtcattgt tgaggatgga ggaggaagat cctggcatcg caaggcagtt 31860
agagctcagg gttcctgtcc caaaccatat caaccagctg gctgggctgg accattcatt 31920
gtgaaaataa aatgaagttt gctgcaattt aaaaagcaaa tgtctagata aaatataaaa 31980
ggctgaggta tggcattagc tctttatccg tgcagacatg aacagtctgc tcttctgaat 32040
ttaagatttc ctctcaccca ttatagtagg ctgatcttct tccattctgt cctgatggtg 32100
atgataatgc ccagagtgca gcctgaggaa aggcttggtg ctgcatattc ctccaccttc 32160
cttccatagc acaaacagtt cttgacagtg ccaaattgag aggccggtta aatctgtttg 32220
gaaacctttc attgtcttcg gagtataaat gctacagttg atggactcct ggacatagag 32280
ttgttttctt gggagtctct ctccacaatt ctaattagct aacagtcaag tttttctgta 32340
ttttattctt ttagctttgt ttatccttca gatagcaagc atctctctca ttgagaactt 32400
catgttgggt ttctagtgac ttctgaacct tgaaggggtg gggcactaga tcagaatttg 32460
gagggaaggg ggacccattt tggctcacag aaggaaattg catagtctgt gtcaacatca 32520
tagatgaaca tgaaaccaga cagctaatgg agttaagaca atcaacaatt ttagttgtgg 32580
tttaagatac tgagagtaaa gcctttttgt aaaaggaact ggcgattgga gaagagagat 32640
gtgtgtataa gaggggagca gccagaggcc tggacagttt gtgaagagac atagcatgga 32700
tggacctagg cagtagagac cctgtgggga actggggtct cgtgtgcagg tggggtctcg 32760
tgtgcaggga gaaaccagct cacctgctca catggacaca ttgcaaagat gggtccgatg 32820
cacgcccggc tggaggtgct gctctgagca aggctttccc tgaggacagg agcctgtgtc 32880
ctttttctcc ttaacttccc tccagagaga gaactgcagc aagcaaagac ttaatggaga 32940
gtgtgggaag gcttagagga aggagcatct tagagctcac catgtttcta ttgaaacaga 33000
ccaagagaaa tgtatgaata acattaattt gactatatgt ttattaatga tgtatgcaca 33060
gtgtcaaatg cacaagtact ttctaaacac cagccacctt agccaacctc cttaggtctg 33120
gtgggcattg tgctctttga ctgtggtcat agcactagga gcatacttgt tctcagaaaa 33180
tgcttgctct tgactccgat tccttagggg aagtcatctt ttcctataga gagagctgtt 33240
tctaggtcca ttaagtgatg tgagagattt gatttcatat ctggttgcag agaaggtatt 33300
agtaaaggtg actgtgggag aaaatcatac ggtttcccct ttgtgcagct gcccttcctt 33360
tttcctttaa gtggaattag gattcattta gaaatgatgg cttttttaag ccattttatg 33420
cttctgagtg tcatggttgg cactattgta ccactcttct cctaaactac aagcaaggtc 33480
aaagggacag ctaatgcaaa tctgcagtac tttgtgccga aaacctagaa atagaaaaga 33540
agactggaag ggactggtgt tactcacagc tacagtgtgg ggtctaaagc aaggtggact 33600
gaaaagggta aaagccaaag gtaccagaac agcttccaga catgactgtc agaacaaggg 33660
aacttttgtt tcttgctgac aaaggtgact cttgggccaa gcaggtgctt aaagctcaca 33720
agtcttagtc tgagcaggtt gatcagagtt cctgtcctgt cattttggat gaaatatgca 33780
tatgttcatc agagctatca ggatgggatg ggggaaaggt ggaagagaga agttgcttgt 33840
acaagcttta aagtaaaaca aaacaagaag tcaggtgcct gcagtctcac ctcccagcac 33900
agcagcttca tacattaaac catggtgtgt gtccatctag accagaaggg gctggctatg 33960
gtatacagac tgccataagg acagagctct aaatgaattc ctggctagag atttgtttgt 34020
aggttgaggt gaaattgcaa aacataaagt gaaccatttg caagagcgta catttcagtg 34080
acatttagta cttcctagta cagagcagct atcctctgta aagactcaaa cattttcatc 34140
actgcagagg ggaaccccat ccctgttagg cagtcactcc ctgtgccctc accctctctg 34200
ctcctgggaa ccactaatat gctttctgtc tggacagtgc ttctcctggg gattttacat 34260
actcgtgtct cttctttcag ttaggtaata atttgaggtt tatccacaaa gttccatgta 34320
ctagtactgt gttcctactg aggataaata ttagtgcaaa tattgataat tattgcaata 34380
gtagtagcta gtacctgtta tatggatctg ccacaatgtg ttcattatta ttgaatggca 34440
tttgagtcac atctactttt tggttacagt gaacaatgaa taatgcttct gtagatagtc 34500
acataacaag tttttgtaga catgttttca tttctcttgg gtatgtgtta tactagggtt 34560
gggattgcca gatcaaattg caatagtgat tctgtattta accttttgac actcacatgc 34620
caaacggttg cccatgatgg ctgcttagtt tctgcttcag gtcttcacat tcctgccaat 34680
acttaccact tttttcccct aaccagcctg ggactgtgaa cttgtacctg tggtgattga 34740
ttggtgtttc tcagaatagc agtagtgatg agcatcattg cctaatccag gaccttaggt 34800
gtccttgtgg caagcatgtc tattcaagtc gctttgtacg ctttcaagtt gggttatttg 34860
cagtttgagt ctgtgtgttc tttatgtatt cagatagttg tgttcttacc agacaacatt 34920
tgcataaatt ttttcctact gtgaaggtta tttttattta tttatttttt aaattatttt 34980
tattttaatg agtttcaatt ttactctaat ttttcctgtt ggtcttgttt tggatgttgc 35040
atctaagaaa ccattggcca gttgtaagac atgaagattc ctcattgtgc tttttaggag 35100
ctttagtgtc tgacatgagt tttgtatatg gtgtgaggaa gagagtcagc tcctgataag 35160
agaagctgtt catttgccca gcaggatttg ttgaaatgac tgctttttcc tccaactgaa 35220
tgtgtgacac cctgacaccc ttgtcaaaaa atgattggcc atagatgtat gcattctccg 35280
gtgtgtacca cggatgtctg catgccttcc gatggcacct cactttagtg tccagctctg 35340
taggaagttt gaaagtgaga agcacatgtc ttcagtctgt ttccccctgt gttgttttgt 35400
gtaagagctc ttaaaattct gtatcaattt aggacttgcc atcacctttt taatttctgc 35460
aaaagaaaag aaatccattt gagttttacg agtagttcca tttaatccct atggtggtgg 35520
ggtcttgtct cttaggccgt gaataaaagc tagatggcct tacactaagc ccactttaat 35580
ctctgcagca gaattttaat aagcatagca gagtgtcact tactggtgtg tttggcactg 35640
tgtaggcacc gtaagtctcc ctggtcaagc cagtggtact cctcatctct agaaggcatg 35700
catatgtacc ctctgcaaag ttaccagggt acccacatta caaagcagag tgacacaggt 35760
gcctggatat ccatgtaacg gaattattga gctaaaggga ttttaaaggt cttgtggttt 35820
gatttatgaa ttggtgaagg aactcggacc cagaggagga agctagctac agggcaggaa 35880
tagaatacag gacacatatc tggtaacttt tgtaagtttt tgttcttgaa acttcattcc 35940
atgagacaaa cagcaggaag acatcttatt cttctacgag agtagccaga gttgagtcat 36000
ttagacggtt ccaaagcatc ctgttgtgga acaatgataa ccttatgcag ggaggtagca 36060
gtgactgcag gcaagcatcc cagaatgact tgtgagtgtt gaatctaccc tgccccttgc 36120
tgccctctat ctattgccta tgcgcatgct ctggaggcaa gggagaaatg acctgatgaa 36180
ctcgggctgt ggctgtttct ctgcacttgt cacctcttcc tggcctgtaa tgaatcctta 36240
agaatttgac ttcagacaac acagctggga gggcagaatg atgtagaatc cagcattgct 36300
gtgtgactct ttgtatttct gttcactgca ggttttaaca actcgggaaa ttctttggct 36360
ccagtcaagg aatttctaaa ccacactgat ggcctgttgg ttttggcagc atctcttcct 36420
cagggtgcag atgcttagaa accattgttt caacagtggg acaaacagta ggtggagata 36480
gagagttctt ttgcccagct ttggaatata ttcttcctct ttcacacatg atcaaaacca 36540
gtatcttatc tcactgtccc ttatagtaaa tgcaagcaat cataaaagaa ttagtcttgt 36600
cttttataaa ggaattaatc tctaggttta tataaggagc tcaaggtcat agtgtcagtg 36660
agagatctcc tgaccttagt gaatgtgact agacctgcct cacacggagc ttgtcaggac 36720
agtcaagtgg ccctgaatac ctgcacttgc agggcacagc tctgacctca ctagacgagt 36780
gttgtcagct ctaagggcag ttagaacgga tactgaagca ggcgattttg ctcgctcctg 36840
ccaggtgatc acagggtctg gtacttggat cctttccata gcaggcttgt tccctgcctt 36900
gtcacctgct ttgttaggga agtctcctcc atccccactc cttcatgttt tcctccaatc 36960
tgtgaggaca ccttgttgag ttggtatttg aagttccatg agtggcatgg ggctggacat 37020
tggggtattg gactcctctt ggtcctttct cttcgccaac cctaatagtt ctggccagat 37080
ctacatctag caagatatga gtggtatgtg tgagtgcaag cgcatatagg gctcagggtt 37140
gaaacccagg acctcactca tgcttggcaa gtgctgtgtc attgagctac acccaaccca 37200
gtactggcag ccttgaaccc ccctctgagc agccctgcag agatggagta ggtggcacat 37260
acagtgtgca ggtttcaagc cggctctacc tctcccccaa ttcttgctat ttagccattc 37320
ttactccgaa tattctgttc tttctttttt ttttttctcc tctttataaa aataataaac 37380
cagagaaaac ctggatgata tatataaaga aaaaaatcat catctcatta ctttataact 37440
acaaagaatt agttttaatt taatttcttt ctgttaaatt taaaatggag agccaagcat 37500
ggtggaacag acctgtaatt ccaacagtga gaagttgagg caaaaggatc ctaagttgca 37560
ggccattgca aacctgggct atatgtggtg aggcacattt ctccccacaa agagaaattt 37620
gtaaaataag atgtgatgtg tagagtttct aagtcttggg tttgtttgtt tgtttgttta 37680
ctttgcataa ctactttcct tgctaaaaga agctgtgagc agttcggatt cctccgctcc 37740
cttgtgagcg tttctctctt cactcttcct ctccatttta ttctgtacct cttttggtaa 37800
ctggtgtttg gatgattttc ctgtagattt ctagagctgt ttctgtgtgt ccttctgcat 37860
actggagggt gtctgtttca gggtcttccc cctactccac ctctgtctta ggagctgtca 37920
ttctcttctg aaacttacag atgcagctcc ccacccccaa gttaaggaga gtgtttgttt 37980
tgttttgttt gtaaggagtg aatagtgaag agttaattag gcagagtggg ctggcttctg 38040
gcttttaagt gatcttgttt gcattagtga cagctgtggg gaatggcaga tacttagaac 38100
actgacttga gtgagcataa ttttaagagt agggttcaga tgacagacat ttgagctgtg 38160
gtagctaact gtttgcccca gtgtgtgcta ttttagtctg tgtgaatgcc ctgctgttaa 38220
gctgggcctc tactctcact tcagagctgg tgctttggga aaggggaagc tgcaggatgg 38280
ttgaatgctt tgcagcctag aagttacatc tctgtttctt tttgtaatgc aggtatacac 38340
agctgtaatc tctgaattca gaaggctgag gaaggaggat tgtgatatga gctcaagggt 38400
agctttaagt cttaagatga gactacctta aaaaaaaaaa aagtaaggag tggggcagat 38460
ggctcagtgg tcccaagaag tgtgcactgc tctcgcagag cacccatttc atgacctagt 38520
acctacgtgg ggttcacaac cacctcctgt tctttgagag gacccgaatt cagttcccag 38580
catccattgt ctggcagctc ataacagcct gtgatgtgag ctccagcaga ctcccagtct 38640
ggctttgact tcccaggaat acctgcacgc atgtgcactc ctccctgtcc caccatatat 38700
ataatttaaa agtaataatt aaaaatttaa attaagtaaa acctgcctaa aactcctgct 38760
ctagaggatc tgaccccctc ttctgccttc cttgggcaac tacactcata tacccacacc 38820
acccagacat ataattttaa aactaaatga aatagttcaa cagaaggaag agcttaaaac 38880
aaggccagag ctagtctttt tgtgtaggac aacctcctgg ttactacagg gagtaaacat 38940
ggaccttaag atggctccgt ataccaccag gccaccgtgg ctccgcctac ctccttgttc 39000
attcctttca tggtttgggt cctttatctt gaagtattat ttggaaaaag gtgtgcttaa 39060
gtttcttttt taaatttagc atacatagta aatttcaaca cggtaaaatt acccagcaag 39120
gaaggcctct gagtaccctt gagactagca tagtggtcat gagtgggtgg gtccagttct 39180
ccatgctgtc cctggtggtt gctttgttca tagtgtttac ttctaatggg catagcctgg 39240
ctgggagttg tttccatagc attagacttg ccccactgtg ataccttggt gggagctgca 39300
tggcctcttc cagaccgtca gggtcttacc attgtggcct tttagctctg atgtggtttt 39360
tgttgttgtt aaaatttatt tattttttat ttatgtctat gcgtgtgcat gtgtgtgtgt 39420
gtgtgtgtgt gtgagtttat gtgcattatg tgtatgcagt cgtccttgga agccaaaaag 39480
aaagcatcag atcttaagag ctgaagttct aggtggttgt gaggcacctg atgtgggtgc 39540
tggggactaa acccctgtcc tggaagagca gcagttcctc tcaactgctg ggccacttct 39600
ccacttctta cttgattttt gtcatttgta aagtaacagt tttagttctg gggtagttct 39660
ctgtcttatg actccacctt ctgaccctct ctgtagcatt taatactctg tcttcacaga 39720
acctgttgtt gccacaacct cctagttttc tctgttgcac ctcctggttt ctcagattga 39780
cctgtgatgg tttgtatatg cttgacccag ggagtggcac tattaggagc tgtggccttt 39840
tgggaatagg tgtgtcactg tgggcatggg ctttgagacc ctcatactag ctgcctaaaa 39900
gccagtcttc tagctacctt cagaacaaga tgtagaactc tcagcttctt ctgctctgta 39960
caggcctgga tgctgccatg cttcctgcca tgatgataat ggactaaaac tctgaaaagc 40020
cagccccatc taaatgttgt cctttataag aattaccttg gtcatggtgt ctctacacag 40080
taatggaaac cctgactaag acaccctctt tagctcagca atcagggctg atcataacct 40140
gatggagttc tgagcatggt taagagtctt atctttggca gggaacttcc tctagtctga 40200
ggctgttccc atatggtctg ggctcacttt gtagaccagt tcttcctttc tccaggccta 40260
taggtagatg atttggaatg tggtaccact tagttgaaaa attctgcttt tgtttttggg 40320
tttttttgtt ttttttgttt tttttttttt gttttttgtt tttttttttt tttttttttt 40380
ttggtttcta tgtgtcatcc tggctgacct ggaactcact ttgtagacca ggctggcctc 40440
aaactcagaa attcacctgc ctctgcctcc tgagtgctgg gactaaaggt gtgcgccgcc 40500
acgcccggct ctgtttttgt ttttgttttc tttgttttgt tttgtttttg ctttccctcc 40560
tagatttgga ggttggcttt tgagaagaaa gggtgcctct agtcaaaggg ccttgagtgt 40620
atactcaagg cagtgtctga taagtttgct tgactttaag gacagtcaaa tagttcatta 40680
gaagaggatt ttgtatttca ttaataattc tttttcttca aatctacatg tctctttaac 40740
atataaatta gaaagaataa atagtgccta taatgtaata gtaactagga gggctcagtg 40800
ttctttttgc ctcggatctg ttcattcata ccagttctgt ccaagccatt tccatttagt 40860
catccgcagc acagccgtgg tttctattcc tgttacaggc ctgtaatttc aggggaagat 40920
ggagactagc ctgaactaca taaaaccttg tcttaaaaaa ataatttagt attttgattt 40980
tcctggtatt ctatattgtg aagagtcccc ttgcacaagt atttgcactt tgaaggacag 41040
gagtcttctc actgtatttt atagatttct attaaattgt tgatgtgggg cttgtttgtt 41100
tgcttgcttt tgtttttgtt tttacctctg aggaactgga ggttaaatat gggaccttga 41160
atatgacact accaccgagc tgtgtctcct gctctctttt tcttttcttt tttattttga 41220
agcaggattt tactaggtta cccaggctgg ccttggaatt ccttcttcaa ttgaggctag 41280
ccttaaactc ctttgtagtt caacctgacc ttgaacttag gatcctgtta tttcagactc 41340
ccaaatagct gggattatag gcctaccact tcaaattgta aatgtctagg gtttttttgt 41400
tttgttttgt tttttcacca cattaaaatt atcaggaatt gcctagtctt aaaagtattt 41460
tgtttttatc cgttacaatg ttctcagtgt ctgtatagac atgtaacttt gtatgcttgc 41520
cctgtttatg aaaatcccta tcagatatgt gtgttggatt ttttgtttgt ttgtttgttg 41580
taaatatttt ttatgggtat gcatgttttg tctccatata catctgtgca tcatgttcac 41640
gcctggaacc tacagtggcc agaggagtgt gtcagattcc ctggagttag agttacagat 41700
ggttatgagc cactatgttg gtgctaggaa ttgaacccag atcctcagaa agagcagcag 41760
gtgctcttaa cctccaagcc atccctcgag ccccaagact ggggtttttg tcgaaacact 41820
tgagggtccc cagaggaaaa agaagggccc agaaaaagtt tgtgtatgtt ccagtgttta 41880
agaagcccac tgtcatgtga cagatggatg ggtgtccaga ggacattcct ctgtactcat 41940
ttgccatttg tccttttaga aggttctctg tgcatgcttg tcaatggctt tagtgattaa 42000
gctgaccagt ccgcagacgt ggggatgcag ccacagtgtc ttcttctcct tgtatcttcc 42060
ccagcgtctt actcagcagc tccagccctg ctctgatccc cagaatgtat ccttctgaaa 42120
tgtggtagct gccctggagc tggcagggag agagggacag gttagtgacg atttttgttc 42180
tacatggatc tgtgacagaa aatttgaagc agcattctta tttccagtat gaagatatag 42240
gtgctgttct ttcagccaga tgatggctga acagatgtgc ttgctgaggt ttctgattgt 42300
aagagcaggt cctgtgagca ttgaaggatg agccttctga ggcctgagag ggcctcggct 42360
aggttagacc acatccaggt aaaagtttac ttcctttttc ttttcccact tcctggtcct 42420
ggcattggag ctgtgtcctt gtgcacgcta ggtaagtgct gcactccaga ggctcaaaag 42480
ccagcttctc tatgttacaa agtagctctt ggagttacat tcagtgtggt accaccatta 42540
tttaaagcaa ggcccttcat gaactgcatt gcacatgatt ttcacctagg aggagctgtg 42600
ccctatgtcc catatttatc acagcaaaat atgataaagg agacagccat ggtgccccat 42660
agctgctttt cagcattcag gccaccccag tgttagtact tatataaaca ctctagccag 42720
gggaagagtg ggtttgaaaa agcttatatt cgggtcagca ttttttaggt gagttgctag 42780
aggaaacaca attttgtttg tttcaaaaat gaaattgctg tagaaaggtc actgtcattt 42840
taacctctta ccccagagct catctgtagc atgtcaggtc tggctcactg ccctggccat 42900
aaaaggcaaa gaatgtggaa ggccctgctc cctcaacgct ttgttcactg tgccatgatc 42960
cctgagttca cagtgttagc cagtgctgtt agtgcacttt ctcctgtact ttgtggctgt 43020
gaggcaagac tccaaggctt tgccaagagc ataaatccta gacattatct cagaacacac 43080
cttaggctat ctgtatttga aacattgctt ggcagccaca gagactctag aagctcatca 43140
cagtcccctg gatcttgact ccggtggtgg cagaaggtca gatgtgatgt gacccgtttc 43200
agggttctgc actgatgttc agtgggcagg ctggcggctg cagctaggga ctgaagtatg 43260
cccaagtagg aagcatgtgg gtgggactgt tgggtttgcc taggcttctt agggaaggca 43320
ccagcgaaga atgagagcaa caaagacctt gtattgtagg tggagggtgg ggggcagctg 43380
ccagagagac cacggctagt aaactcactg cagcaacagc atcctggggt gttgatggtg 43440
tgactcctaa tctcttaaat gcatggacca aacagggagg catgagtagt agactcccat 43500
gcctgggtca ggatcacaca caagcatagg agactctgaa taaatagcct gtggattctg 43560
tccttgccct gtttaagtgg cttttggagt gctaaagagc caggaaaact tcatcctttg 43620
ggaatcagag tgctcctagt gggcctactc acaggaaact gctgggtgat agtcactgtt 43680
ctatcactgt gaagagatgt catgaccaag gcaccgctta aaagagaaag catttaactg 43740
ggggtttgtc tacagtttca gaggcttagt tcattatggc agcgaacatg gtggcacaca 43800
tgacactaga gcaatagcta agagatatat cctaatccac aggcagagag aactgggcct 43860
agcatgggcg tttgaaacct caaagtccac ccgcagtgag acacttcccc tgcaaaggct 43920
acaccgacaa taaagccaca cctcctaacc ctcctctact agtgccactc tctagtgacc 43980
agggattcaa atatatgagc ctgagagggc catttttatt cacatcgcca caggtctcat 44040
tgtggaggag gcaggagttg tgacgcacac tgttctccaa gggagccttt cagcaacagc 44100
tcgaggacca cacactgcta agaggaaagg catgagagtc tttgattttc ctgagctctt 44160
attgccctgc ttaaaaccag gacactggga agttccctcg gggctggagt ggggagtaca 44220
ctcccctacc ttggggagtg tcaaaacagg aaagaataga aatgtgaact gaactgaagt 44280
tacacatggg gtagttttgc ttgggcttgt tcttgaatgc tttgatagac cgtgacagca 44340
tttatagttg agttcatttg gaaagcctga gtcttcaaca agggaggaga tggaatctgc 44400
tcagtgacca taagagggct agctacagcg aggcctgtcc ctgtctgtca ctaagctggg 44460
ctgttgagct gccccgtgag gcccttgttc taaggcagtc ttgactgcag taggaacttg 44520
gaggccaggc ctgcttgccc tgttgattat ttaaacctct tggcccacaa ggctttcatt 44580
tgttctgctt ttgaacagga tctctctgtg tagcccaggt tggccttgaa ttcgtgatct 44640
tcctgcttca gccctgcttc caagtgctat gattagtcat gtgttacatc tagaaagaga 44700
ccattggcct gatgtctaag acaagtgcca cgtaaatatt ggtgcaggca gaggcagggg 44760
gccttggcct tgtagagccc aggcttaaat gtggcttccg ttgcaggaat gaaagtgttg 44820
cctgaagtcc aagtttgagg aagattccta tacgtatgtt tgttctctgt gttgctaatt 44880
tatctcgcta ctgtgctggg cactgtgggg tgtggcggat atggagtgag gcgcgtgtgt 44940
ccagagcttc ctggtttgat ggaagagtcc tctttaatag ttttccatac atcctcaaga 45000
ttgtgtcctg tgtgcaaggc atttggcatt gaagaactga gaaattactg ctgggtgtag 45060
tggtgcacac ctttaattcc agtacttggg aggcagaggt gtgtgggtct cttgtgaatt 45120
cagggacagc ctggtttccg taggaaatgt gtcttaacta ggatggaaga gccagaacca 45180
ttgctaatcc ctggcctctg catcttccca gtgctttggg tgcacacctt taggaaagtc 45240
tacgcagcgt gggttgggcc ttgaaaacct ccttgtcctt acaaaatgtt tggggggggg 45300
tgggggtaag cctctagtgt ttctaagcct cagacctcac atagcccagt gttttctcat 45360
acagatttat tagaagtgtt gtgagctaag gacagaaaac cctgccccat ttctatcatt 45420
ggaagtgtaa actaaaggaa gctagaagtc cgatgggact gaggttaaca cagagcagct 45480
actgggatct ctggtcgacc agcaaggggc agggcctgca gagcaacggt gtggatggac 45540
ccagcaaggg gtgggggctg cagagcaacg gtgtgggtgg acacctgctc tttgggaact 45600
aagtggattt tctcaggcag gtggagctca accagtaggt agaggggctt ttgctagaac 45660
ccagagaagc aagagatcct ccaaggcttt gggctctgac tcgtggtaac agggcacttt 45720
gtaccttctc agtggcgagg gtacagggag agtagattct aggtgaggct ggcagcctcc 45780
tagagaaatt agcacagaga cttcaaggac tacagaccca ggcggtggtg gcaggtgagt 45840
gttttgagca tgtagagaac ttgcttgtga gtgattgatc tggggtggtc agtaagcttt 45900
taacacaaaa actgtgcttt catccctcct aacgcggaac ccttaataca acttctcttg 45960
ttgtggtgac cccaaccata aaattatttt gttgctactt ttataactaa ttttcatact 46020
gttatgaatt ataatgtaaa tatttttaga gatagaggtt tgccaagggg gtcgcaatcc 46080
acaggttgaa aactgttggg ctagatgaag ctctcaaaag tctcagcagc atgtgagcac 46140
ctatatattt attcagctgt tgagggcatt cagcaaatcc gtcagctgct gtgttcttcc 46200
ccactccatg tgctgggctg catggccttt cacaggctct cagcattatg gcaaactcat 46260
tgtttctttg atacagctgg acctttgggt gcttgactga tgcagtgtga tgctggtcct 46320
gaggagccct gccagtctgg ttggggatac aaaacaagtg ggtgggacta gctgattggc 46380
atgaagcagc ccagcagtgt aactcaggct ctcttttccc tttgtctttc taggagtata 46440
aacagaagct tgctcgagta acccaggtcc gcaaggaact caagtcccac attcagagct 46500
tgccagacct ttcgctgttg cctaatgtca cagggggcct ggcacctctg ccctctgctg 46560
gtgacctctt ttcaactgac taggacaggt atcatgcccc aggttcccat gtgtgccaga 46620
gaaaagaagc aagtcactgc tggagacagt cccctgcccc agctccatct cttcagccag 46680
catctgcctc agtaatgaag cggggaactc tgacttctta ctctcctccg gctcccctct 46740
gagcatggtt cagagccatg cagatggact cagttttgat tcatatttgc taagaacaca 46800
gactctccct cccactcatg tctcttgcca ctgctgcttt tccatctttc gctctcttac 46860
acccataagc catgaaaatc atgtgtactt ctggaagctt tcaagagaag cttgcctcag 46920
tgacagcaca tgccatggaa gtctgagctg gccttgacca ggttcagaac aggctcccaa 46980
tgaggactca ggctgcaggg attgggcttt cctggccacc attgctcaca aggtcggcag 47040
catagctttg ccacaaagag tttgccaaat tgccgacatt tcctgatcta aacacttctg 47100
ttagagaagt gttgtcatgg ttaggtgtgt ttatttttcg agcattctag agaatccaac 47160
agtaagattc gaagctgatc tgagacataa aagttatagc gcaggtacag agtggcaggg 47220
ctcacaagtc gtttgccagg cttctcacca cagccttccc tgccacattg ctccagggtc 47280
cccagtttca gagcaccttc ctaaagcagg accctctgtc ccctggggct gggaagggag 47340
ctgttgggat agcatgccca ggctgataac tgctccacag ctggagtctg ggctctcgtg 47400
catgttgctc tcaccctgaa gcttacaata acgccaccat gtggacgcat ccacagcatg 47460
gtgactggac ttgaacatct ctacagatag gacactgagc gtgccaggca gttcctgcca 47520
ctatatgcat ccttgctccc atgcagagag aggagtactc aaacaactca cctccagcag 47580
aggaagggag tggagggaga ctaatttggg gttgaattcc attatcatgc cagctgacat 47640
tatgactgaa tgatgtagtt gcgattttta cgtctagtaa agagtgggcc tggccctgga 47700
aatgactgta atctggcagg cttatatttg acattggaaa gggcagaaac cattttgtcc 47760
tactagtgtg ataggaattg tcgcccagag ctgaagtcca aaagccataa aaaggaattc 47820
agtgaagggg cctgagaagc ttcgttgctt taagttgctg ttttcaggat caccaaaaaa 47880
aaaaaagcac acatcgtttt tacacgttaa agcaaatctg tgtgtcacca aataggcagg 47940
ctttggttga agaagccgcc ccacctgcta tgagatggga ctcagttcca ctctgtccag 48000
cctggtcttc gtggaagctg ttattcacag agtgcaggct gctgtttgct tgttcactct 48060
gccttttctg tcagcatcct tttctccaga ggttacagga cagcactgac tcaggtgaaa 48120
cccctgtctc tgctgcggaa cacctcccta cacaccacgc cactttactg tcacatgcac 48180
acaccactct gaacaccctc cagacactgc tttcccacaa aacagaccaa aagactgtag 48240
cagctctgct ctgatgacaa cttagtgggg tcagctcaat ggtgtttatt ctttggagtc 48300
tctgaccagc agtcagtgcg taactcttac agcaggatca agacgtgacc agtagcgatt 48360
atgtaaatga atgtgttggc ctcattacac cacttccagc agatttcctg tgaggaagtt 48420
agtttcgttt taatgaaatg cttcagtttt taatcttcct tgttgtgtgc ctccaccatg 48480
cccccagtct cccaactgtg tgcgaacttc ccttgtttaa cttaagtgaa acttttgtcc 48540
ctgagagttg atctggcggg tggggtgggt gacttctggt tttgtgtttc tttagggcat 48600
ccgtaggcct caaggatctt tcctttaggg tcatattcct cagaaagtct tcagtctttc 48660
tttgtttttg ttttgttttt cttaaaggat atttttaaag cttaaatttg tatattaatt 48720
taggacttta tttagaagta taggctgtca ttggtggcag cagtatattc tgaaatgtct 48780
catagatata tatttttgaa taaagatggt gttgttgaac aacattctgt gatgtttctt 48840
ctccctgcct cgtgcagtga ttagtgatca tcaatggagg gcgctctgtc tgcacagaat 48900
cttcccctgg gggctggtag tggtacatac gcaagacaac tggttgaact tactctccaa 48960
gtctctcttg gagtttgatt ggccagttag cagtctatac tgtagcagag agcgccaact 49020
gggtttcatc cttgtatagg ctgaagactg gttggggggt ggggaggact tgtgatgtcc 49080
ccccacacac agaggatgtc ttgagaggcc tttggttgca gtcttttcag tcacaccagc 49140
aggccttgtg ggaattgttt gatttgcagt atggtgaagc tgctcttgga aaaacggaaa 49200
ataaccaaac atcatctttt ttaagtgttt tcatggactt ctcttctgct gaggttgaca 49260
tccaatggaa gaccctgatg actgagtact ctggagacag ggtgctctga agcagctgct 49320
gtggtccgga atggttgaga tctcccacta agttatggtg gggttacctg tttctcatga 49380
tgcaaaggac ttccttttta gtaatggtgg tccatcaaga ccacctgcag ctgaagtcca 49440
agttcactat caggaaagtg tgtgagtttt gcctgcttgt atgtatgtgc accatgtaca 49500
tgccaggtgc ctgtagaggc cagaaggtgg cattggattc cttggaactg gaattacaaa 49560
tggttgtgtg ctggggatca aacctgggtc ttctgcaaga gccacaaaca ctctttactg 49620
ttgagccatc tctccagccc ttctcccaac cccccacagt gttctggttt aataaaggtt 49680
gagtgtaaag tacgaatcta agtaaacaag a 49711
<210> 2
<211> 981
<212> DNA
<213>mouse (mus musculus)
<400> 2
atgtcctcct tctcagaatc ggcgctggaa aagaagctct cagagctgag caactctcag 60
cagagcgtgc agaccctctc cctgtggctc atccaccacc gcaagcacgc aggacccatc 120
gtctccgtgt ggcaccgcga gctccgcaaa gccaaatcaa atagaaagct tacttttcta 180
tacttagcaa atgatgtcat ccaaaacagt aaaaggaaag gacctgagtt cacaagagaa 240
tttgagtctg tgcttgtgga tgctttttct catgttgcca gagaggcaga tgaaggctgt 300
aaaaaacctt tagaaagatt gctgaacatc tggcaagaac gaagtgtgta cggcggcgag 360
ttcatacagc agctgaagct gtctatggag gactccaaga gccctccccc caaagcagca 420
gaagagaaga agtctctaaa acgaactttt cagcagatac aagaggagga agatgatgac 480
taccctggaa gctactctcc ccaagaccct tctgcaggcc ctctcttgac tgaggagtta 540
atcaaagctt tgcaggatct ggaaaatgct gcgtcagggg atgctactgt ccgacagaag 600
atcgcttccc tgcctcagga agtgcaagac gtgtcgctgc tagagaaaat tacagacaaa 660
gaggcagctg aacgtctttc aaaaacagta gatgaagcat gtctgttact agcggaatat 720
aacgggcgcc tggcagcgga actggaagac cggcgccagc tggctcggat gctggtggaa 780
tacacccaga accagaaaga ggttttgtca gaaaaagaga aaaaactaga agagtataaa 840
cagaagcttg ctcgagtaac ccaggtccgc aaggaactca agtcccacat tcagagcttg 900
ccagaccttt cgctgttgcc taatgtcaca gggggcctgg cacctctgcc ctctgctggt 960
gacctctttt caactgacta g 981
<210> 3
<211> 192
<212> PRT
<213>mouse (mus musculus)
<400> 3
Met Ser Ser Ser Ser Ala Lys Lys Ser Ser Asn Ser Ser Val Thr Ser
1 5 10 15
Trp His His Arg Lys His Ala Gly Val Ser Val Trp His Arg Arg Lys
20 25 30
Ala Lys Ser Asn Arg Lys Thr Tyr Ala Asn Asp Val Asn Ser Lys Arg
35 40 45
Lys Gly Thr Arg Ser Val Val Asp Ala Ser His Val Ala Arg Ala Asp
50 55 60
Gly Cys Lys Lys Arg Asn Trp Arg Ser Val Tyr Gly Gly Lys Ser Met
65 70 75 80
Asp Ser Lys Ser Lys Ala Ala Lys Lys Ser Lys Arg Thr Asp Asp Asp
85 90 95
Tyr Gly Ser Tyr Ser Asp Ser Ala Gly Thr Lys Ala Asp Asn Ala Ala
100 105 110
Ser Gly Asp Ala Thr Val Arg Lys Ala Ser Val Asp Val Ser Lys Thr
115 120 125
Asp Lys Ala Ala Arg Ser Lys Thr Val Asp Ala Cys Ala Tyr Asn Gly
130 135 140
Arg Ala Ala Asp Arg Arg Ala Arg Met Val Tyr Thr Asn Lys Val Ser
145 150 155 160
Lys Lys Lys Tyr Lys Lys Ala Arg Val Thr Val Arg Lys Lys Ser His
165 170 175
Ser Asp Ser Asn Val Thr Gly Gly Ala Ser Ala Gly Asp Ser Thr Asp
180 185 190
<210> 4
<211> 17292
<212> DNA
<213>artificial sequence
<400> 4
taataggggt gagtaaatat tctttgagtc agttaggata atcgtgacta actggggaaa 60
agatgtggtt taagggtact atctctgccc tatagggata ataaactaac acttttataa 120
ctcttcgctg ttctgaatta gaaaagggta agtagaaagg gctacaaggt ggataccaag 180
attccgaggg cttacctagc atgtgggcac atcttaaaac catacagaag ccaggcatag 240
taacacactt ctagaattcc atcactcaga ggccagccca ggctatgtag tgaaaccttg 300
tcacaaagaa aacaaagaga tgtaaacagg agacattggg aaaccatctg ggcacattct 360
acctgatgtt tcccagctcc tgtccatatg ccaggctgtt taaggtccag tttggaccat 420
gtttcaaggc ttctttcctt catcctatca ttagaacaga aagtacaata gctactttaa 480
tactgttgta actcatcaac caagagtaac ttagtaagtg ttcttggtgt gcttttgctg 540
tgaccagatg gtgaaccaag gtagctgacc cattaattcc ccaaataatt agcaagattt 600
ttatgatgct tctctacttt catccccaag ccaaaaaaaa agacaaaaaa aaaaaaaccc 660
tttgaattca gaaagttcat agggtagatt tgatttgagt agtggtttaa ataactagac 720
tatgaacata ttatgcttat gcataccttg ttttctttaa agtttggagg tagttacaca 780
ttcattattc aacttgtgtt tgttctatgc cacacactaa tccagagctt ggggaacagc 840
aaggaggaag gcagataaga acagtgacgt gaacaagaca ctttgaaatg atatggacaa 900
ttaattagaa tagtgttcta gggtagactc ttctccatac atcttgggta agaaggtgac 960
atttgagaac ctaaggtgga aataatcgtg acatcttaag aggagcagga tgagatggat 1020
atggtaaaca aagtaagatg tggccccaga tgagatcaga gagaggaaag gagtggttcc 1080
atagtccctg ggagtctatt ctgatagggt gtcagtcaag aactgtaagc cagcgaatga 1140
aaacatttct gtagacatta taaaaagccc aggtaaaatt agagaaagaa aacaaaacat 1200
aaaacaaaac ctaaccatat aactaggatc tttccccctc ataatctatt tgttaattgt 1260
gctgccaggg acagttcatg aaaagtttaa ccatttgtaa aatcatgagt aagcactttt 1320
ggtaaaattg tacctgagcc aggtggtggt ggtggcgcac acctttaatc ccagcacttg 1380
ggaggcagag gcaggcagat ttctgaattc aaggccagcc tggactatag agtgagttcc 1440
aggacagcca gggctacaca gagaaaccct gtctcaaaaa acaaaaaaaa aaaaaaaatt 1500
gtacctgaat gaaaagttca ctccattagc agatatccct ctttttagtc aactattcta 1560
atatatgata aggcaataag tgtatatctt tttattatat tgtagagaaa gtttttattt 1620
ttgttaagtt tgtttgtttg tgggttttgg agatagggtt tctctgtgta gccctggctg 1680
ttctggaact tactctgtag accaggctgg cctcctaaat actgggacta aaggtgtgca 1740
ccaccacctc ctggcaaatt aatacttttt aaaggtttct attagatgcc tttaaaaaga 1800
agagatgaca gagtgggtaa aggccagact agctgaatca gcaatctctg ggtacaattg 1860
agagaccctg tctcagtttg taaggtagag aacaacacac acacacattc acatacttgc 1920
aaaccatata tatgaaaggc ccgtgtagag aatgggttgt gaagggcagg gaaacctgag 1980
ctcagaagga gtgttgctgt ggtgttggct aaggtgagat tggttaagac atttgcagtg 2040
agaaggctct ctccatttat tgaatgctta ttttgtagtt ggagtcttca cccttgtaaa 2100
atgtacatga tgtcttgaaa agggagctgg tattgaggca agaaagagat ttttagtgtg 2160
agtcgatggc tggtctgaac cgaggactgg gaagctttgg tggaggaagt aatttttgtt 2220
ttgttttggc catgttaagt ttgagacggc tggttaggtg tttcaagtgg caattgtggg 2280
tggacacatg gttaaactag tatcaagttc acagaaactt agtctttgtt attttatgta 2340
tatgagcatt atgtctacat gtgtgcctgc ttgatcacat gtgtgcctag tggccacaga 2400
ggccacacag ggcatcagat ttcctggaac tggagttgca ggcagttgtg agattccatg 2460
tggattccag gaatcaaatc tgggtcctct gaaatagcag caagtgctgt ggttgctgag 2520
ccatttctcc aggacccctc tccccctaat ttctattttt ttaaaaaata aaccctaaaa 2580
tttcatttgt catttgtgtg tatacacact catgtgtatg tacatgtgct atagcaaaca 2640
cacagagttc cagaagtcag gtcactcctt ccaccatgtg gaatcccagg aatttaactc 2700
aggttgtcag gcttggtggc cttaattcct gattctagac ttaatttctt gagaggtaaa 2760
tcaggagtta ttgccaaata catgataatt ccttattgtt tgtgggtctt tatacttctc 2820
ccaggtgggt aatttcctgg gaggtatcat ggttgttagg ttttattgcg ctcagggata 2880
cagaacagtc agcaaagggg agaggtgcat gggtaattgt gacaggaggt gggaagggac 2940
tcagcaccca gagcctcctc taggatttgt ggagggtgct tttgtcactg ttcctgcaga 3000
gtaagtaggc attcacttag agccatgttg ttcacacaag tgaggcaccc agatcatttg 3060
ggagtggaga gtgtcctgat ccccacttcc accctcaccc caatctccag gtccgctcct 3120
gtcttagctt tctgtcacgg tggaagaatg ccagagagaa acaagggagt aaggatgtag 3180
ctgggctcat ggttgcaaag gcttcagttc atgctgccag ctcctcctct tctgggctgc 3240
agggaagcag agcatcacag cagaaagtgt gtgattgagc ctcttgatgg cttagtggca 3300
aatattttaa aaacactttt cacttagata gctctcatat acacattatt atttatttgt 3360
ttgttttgtt ttggagacaa gttttcatgc tgcccaaact agccttcaaa gaatttgtgt 3420
ggtcaggaat ggctttgaat ggattctcca gccttccccc ttgagtcctg agattgcagg 3480
catggaccac catgcccagt ttatgcactg ctggggatca aactcagggc tttgtacatt 3540
cgaggcaagc acattgccaa atgctataca tccccagcct caagaatctc agtgaacctg 3600
ggaaagtagt gccccatctt ttgtcaactg ctttcctgct ctagccacat ctgattttag 3660
gtaccacaat gatgtctact tcaccgcagt gccttacttg aaaaccagtt tgtttaattt 3720
atgacttgaa ttactttgat gcaaaagagt tggaagtcaa gtatatttat ataaaaattt 3780
taaaaattca tttaaaaaaa gattgattca gccaatatgg tgtctgtgaa aacaagcccc 3840
ttctcggagg cttcaagtta ccttcagatg ggaagaataa acaggtctcc tgtgcagcag 3900
gtgagaggga tgggtcgtgt gaaacccatg aaggagcaag aagtgccagt ctttgggact 3960
ggggagctgc tgctggagag acaggaagga cagacctgag gccacctgag gcatagctga 4020
cttaggctct aagcctgatg accagagtgg ataggtgagg tcagaaagcc tcaagggctt 4080
tgtatggact ttgttctgag aagaatggaa agaaacaaga gatgccttcc catggcagct 4140
agaattactt tattcaaact ggagagtagg ctgtaggagt gcaagactgg aagccgggag 4200
agcagtaatg aggctatcag agaaaccagc ggagggctgc cccttgggcc gggttgttgg 4260
cagaggacag ggtgagagta actaatttct gcatacactg tgaagaaaga gagtgtctta 4320
gcataaacca tagagtgctt gcttatagct gagctcttta agatgtagca gaagggtttc 4380
cctgtttccc taggacttac ctctgagtta gaaaaaaaat taacttatac atttcctgac 4440
ttagtacttt gatctcattt cagtggactt gctactagta atttgtaagt aggacacttg 4500
gccagaccag gctgagtgtt agcataagct tcacagaagt gctgtcagtg cagaaggtgg 4560
gtatagttcc tgtctgttga atttggtatt cttgatgttg ttagtaaaga tacttgactt 4620
caaccctaag atgtggaggt tgtttgccct cccctttttg gcatagtttt atatatctga 4680
gcctggcctc aaactcactg aagctgagga tacgtgacct tgaatttctg atgccgttca 4740
cttccccagt gctaggttac agtgttgagg attgaaccca gagctaggca agcattttgc 4800
caactgagct atagttcagg tctctctctt ttttgaaagt tatgttttat ttgggaagaa 4860
gcatggccta cttgtaaggg gtcattgaaa aacttgcagg aattggtttt ctttttttac 4920
tgtgtggtcc aggaatggaa atcaggttgt caggcttgga ggcaagctta tttatttgct 4980
gagccgtctt gtgagtctca ggatgtccct tcttaaggtg ctcagtgtta gatgttggct 5040
tgtatgcagt cttactaaac agagagcgcc tgggagtata tatgtggcag gggcctctct 5100
tcagaaagtc tcatcctgct tgtctgctgc cctctcaggt attatggacc agaaaaagat 5160
tgcctgattt tttctttcag ataaatatta caaaggactg cattatttcc agagctttcc 5220
aagattatcc attagataca tgagtggtga cacagaactc agactaaatc cagtgttaaa 5280
gttgttataa agttgttatg gttgagacac ttggcacata ttgtacacac atatttatgc 5340
atgtagaggt gtggaatgtt aatatagtga atttaaacac atgtgcttta aaaatcttaa 5400
aggtgtggct aattttaatg ttcagtatat gtataggatt gtatttaaat tgatgccccc 5460
cccccacaca caaaggcgca taacgatacc acgatatcaa caagtttgta caaaaaagca 5520
ggctggcgcc ggaaccgaag ttcctattcc gaagttccta ttctctagaa agtataggaa 5580
cttcgaaccc tttcccacac caccctccac acttgcccca aacactgcca actatgtagg 5640
aggaaggggt tgggactaac agaagaaccc gttgtgggga agctgttggg agggtcactt 5700
tatgttcttg cccaaggtca gttgggtggc ctgcttctga tgaggtggtc ccaaggtctg 5760
gggtagaagg tgagagggac aggccaccaa ggtcagcccc ccccccctat cccataggag 5820
ccaggtccct ctcctggaca ggaagactga aggggagatg ccagagactc agtgaagcct 5880
ggggtaccct attggagtcc ttcaaggaaa caaacttggc ctcaccaggc ctcagccttg 5940
gctcctcctg ggaactctac tgcccttggg atccccttgt agttgtgggt tacataggaa 6000
gggggacggg attccccttg actggctagc ctactctttt cttcagtctt ctccatctcc 6060
tctcacctgt ctctcgaccc tttccctagg atagacttgg aaaaagataa ggggagaaaa 6120
caaatgcaaa cgaggccaga aagattttgg ctgggcattc cttccgctag cttttattgg 6180
gatcccctag tttgtgatag gccttttagc tacatctgcc aatccatctc attttcacac 6240
acacacacac cactttcctt ctggtcagtg ggcacatgtc cagcctcaag tttatatcac 6300
cacccccaat gcccaacact tgtatggcct tgggcgggtc atcccccccc ccacccccag 6360
tatctgcaac ctcaagctag cttgggtgcg ttggttgtgg ataagtagct agactccagc 6420
aaccagtaac ctctgccctt tctcctccat gacaaccagg tcccaggtcc cgaaaaccaa 6480
agaagaagaa ccctaacaaa gaggacaagc ggcctcgcac agccttcact gctgagcagc 6540
tccagaggct caaggctgag tttcagacca acaggtacct gacagagcag cggcgccaga 6600
gtctggcaca ggagctcggt acccggaaga tctggactct agagaattcc gcccctctcc 6660
ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt 6720
ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg 6780
ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg 6840
tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc 6900
tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca 6960
aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag 7020
ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa 7080
ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt 7140
tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt 7200
ttcctttgaa aaacacgatg ataagcttgc cacaaccatg gaagatcccg tcgttttaca 7260
acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc 7320
tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg 7380
cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag 7440
ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat 7500
gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc 7560
gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag 7620
ctggctacag gaaggccaga cgcgaattat ttttgatggc gttaactcgg cgtttcatct 7680
gtggtgcaac gggcgctggg tcggttacgg ccaggacagt cgtttgccgt ctgaatttga 7740
cctgagcgca tttttacgcg ccggagaaaa ccgcctcgcg gtgatggtgc tgcgctggag 7800
tgacggcagt tatctggaag atcaggatat gtggcggatg agcggcattt tccgtgacgt 7860
ctcgttgctg cataaaccga ctacacaaat cagcgatttc catgttgcca ctcgctttaa 7920
tgatgatttc agccgcgctg tactggaggc tgaagttcag atgtgcggcg agttgcgtga 7980
ctacctacgg gtaacagttt ctttatggca gggtgaaacg caggtcgcca gcggcaccgc 8040
gcctttcggc ggtgaaatta tcgatgagcg tggtggttat gccgatcgcg tcacactacg 8100
tctgaacgtc gaaaacccga aactgtggag cgccgaaatc ccgaatctct atcgtgcggt 8160
ggttgaactg cacaccgccg acggcacgct gattgaagca gaagcctgcg atgtcggttt 8220
ccgcgaggtg cggattgaaa atggtctgct gctgctgaac ggcaagccgt tgctgattcg 8280
aggcgttaac cgtcacgagc atcatcctct gcatggtcag gtcatggatg agcagacgat 8340
ggtgcaggat atcctgctga tgaagcagaa caactttaac gccgtgcgct gttcgcatta 8400
tccgaaccat ccgctgtggt acacgctgtg cgaccgctac ggcctgtatg tggtggatga 8460
agccaatatt gaaacccacg gcatggtgcc aatgaatcgt ctgaccgatg atccgcgctg 8520
gctaccggcg atgagcgaac gcgtaacgcg aatggtgcag cgcgatcgta atcacccgag 8580
tgtgatcatc tggtcgctgg ggaatgaatc aggccacggc gctaatcacg acgcgctgta 8640
tcgctggatc aaatctgtcg atccttcccg cccggtgcag tatgaaggcg gcggagccga 8700
caccacggcc accgatatta tttgcccgat gtacgcgcgc gtggatgaag accagccctt 8760
cccggctgtg ccgaaatggt ccatcaaaaa atggctttcg ctacctggag agacgcgccc 8820
gctgatcctt tgcgaatacg cccacgcgat gggtaacagt cttggcggtt tcgctaaata 8880
ctggcaggcg tttcgtcagt atccccgttt acagggcggc ttcgtctggg actgggtgga 8940
tcagtcgctg attaaatatg atgaaaacgg caacccgtgg tcggcttacg gcggtgattt 9000
tggcgatacg ccgaacgatc gccagttctg tatgaacggt ctggtctttg ccgaccgcac 9060
gccgcatcca gcgctgacgg aagcaaaaca ccagcagcag tttttccagt tccgtttatc 9120
cgggcaaacc atcgaagtga ccagcgaata cctgttccgt catagcgata acgagctcct 9180
gcactggatg gtggcgctgg atggtaagcc gctggcaagc ggtgaagtgc ctctggatgt 9240
cgctccacaa ggtaaacagt tgattgaact gcctgaacta ccgcagccgg agagcgccgg 9300
gcaactctgg ctcacagtac gcgtagtgca accgaacgcg accgcatggt cagaagccgg 9360
gcacatcagc gcctggcagc agtggcgtct ggcggaaaac ctcagtgtga cgctccccgc 9420
cgcgtcccac gccatcccgc atctgaccac cagcgaaatg gatttttgca tcgagctggg 9480
taataagcgt tggcaattta accgccagtc aggctttctt tcacagatgt ggattggcga 9540
taaaaaacaa ctgctgacgc cgctgcgcga tcagttcacc cgtgcaccgc tggataacga 9600
cattggcgta agtgaagcga cccgcattga ccctaacgcc tgggtcgaac gctggaaggc 9660
ggcgggccat taccaggccg aagcagcgtt gttgcagtgc acggcagata cacttgctga 9720
tgcggtgctg attacgaccg ctcacgcgtg gcagcatcag gggaaaacct tatttatcag 9780
ccggaaaacc taccggattg atggtagtgg tcaaatggcg attaccgttg atgttgaagt 9840
ggcgagcgat acaccgcatc cggcgcggat tggcctgaac tgccagctgg cgcaggtagc 9900
agagcgggta aactggctcg gattagggcc gcaagaaaac tatcccgacc gccttactgc 9960
cgcctgtttt gaccgctggg atctgccatt gtcagacatg tataccccgt acgtcttccc 10020
gagcgaaaac ggtctgcgct gcgggacgcg cgaattgaat tatggcccac accagtggcg 10080
cggcgacttc cagttcaaca tcagccgcta cagtcaacag caactgatgg aaaccagcca 10140
tcgccatctg ctgcacgcgg aagaaggcac atggctgaat atcgacggtt tccatatggg 10200
gattggtggc gacgactcct ggagcccgtc agtatcggcg gaattccagc tgagcgccgg 10260
tcgctaccat taccagttgg tctggtgtca aaaataataa taaccgggca ggggggatct 10320
aagctctaga taagtaatga tcataatcag ccatatcaca tctgtagagg ttttacttgc 10380
tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt 10440
tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 10500
cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 10560
atcttatcat gtctggatcc ggaataactt cgtatagcat acattatacg aagttatgtt 10620
taaacggcgc gccccggaat tcgccttctg caggagcgta cagaacccag ggccctggca 10680
cccgtgcaga ccctggccca ccccacctgg gcgctcagtg cccaagagat gtccacacct 10740
aggatgtccc gcggtgggtg gggggcccga gagacgggca ggccgggggc aggcctggcc 10800
atgcggggcc gaaccgggca ctgcccagcg tgggcgcggg ggccacggcg cgcgccccca 10860
gcccccgggc ccagcacccc aaggcggcca acgccaaaac tctccctcct cctcttcctc 10920
aatctcgctc tcgctctttt tttttttcgc aaaaggaggg gagagggggt aaaaaaatgc 10980
tgcactgtgc ggcgaagccg gtgagtgagc ggcgcggggc caatcagcgt gcgccgttcc 11040
gaaagttgcc ttttatggct cgagcggccg cggcggcgcc ctataaaacc cagcggcgcg 11100
acgcgccacc accgccgaga ccgcgtccgc cccgcgagca cagagcctcg cctttgccga 11160
tcctctagag tcgagatccg ccgccaccat gattgaacaa gatggattgc acgcaggttc 11220
tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 11280
ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 11340
cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc 11400
cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 11460
gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 11520
gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 11580
cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 11640
tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 11700
cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 11760
ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 11820
gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 11880
gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 11940
gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc 12000
gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 12060
ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 12120
cgcggggatc tcatgctgga gttcttcgcc caccccccgg atctaagctc tagataagta 12180
atgatcataa tcagccatat cacatctgta gaggttttac ttgctttaaa aaacctccca 12240
cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt 12300
gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 12360
ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg 12420
atccgggggt accgcgtcga gaagttccta ttccgaagtt cctattctct agaaagtata 12480
ggaacttcgt cgagataact tcgtatagca tacattatac gaagttatgt cgagatatct 12540
agacccagct ttcttgtaca aagtggttga tatctctata gtcgcagtag gcggacccag 12600
caaattgatg tcttctaaaa agctaaattg tcaaaatcaa tatggtcagt tttgttttgt 12660
tttctgtctg ttaaaaccag tcatacaggc tttcccccta tttcttccac cttgcaaagg 12720
ggaaaaagat tccctgggtc gttggatgcc tcttcctggt tcaacatggg aggaatgtag 12780
tttctcaaga ctgagcatcc aagtaataca gagtgtacca cagttttgtt ttaagtgtgt 12840
aaatcatttt ggcagttgtt gttgttgttt taaagtaaat ctgtcctaat taaaaacaaa 12900
aaatgtaagc ttatatatat atcttttcat ttttttctgg acagagaggc agatgaaggc 12960
tgtaaaaaac ctttagaaag attgctgaac atctggcaag aacgaagtgt gtacggcggc 13020
gagttcatac agcagctgaa gctgtctatg gaggactcca agagccctcc ccccaaaggt 13080
agaacatcac cacatgttta cagctcttgg tctttgctcc tcccagcccc acgtgaaaag 13140
ctgcagtggg ggctacagtg gctgctgtga tccaccagcc gctttatgac tgcagaggga 13200
tgctcctgag tgatggtgag aacttacaca gtggtgcaag cggagatggc gcaacgcaat 13260
taatgataac ttcgtatagc atacattata cgaagttatg gtctgagctc gccatcagtt 13320
caagggaaca accaagttct ttaggattgt ggctgactaa cgagtagctc atcctggccc 13380
tgtcgtggac ctcagcgggg cctgggatag gtctcttgcc ctttcagaac tcagctttat 13440
tagtaataag tctgggttta tatagtttaa tgactatcag attactgtct atattaatta 13500
ttgattttcg tctttactga gcccagggta tctttgtgaa gtatatatgt gaattctaaa 13560
attcttaaga gtccgtgtta tttatatctt taaagagaaa gttcagacat ttggtttttc 13620
ggttgctggc tagccatgga tcactagtgg catgacatgc ttcaccctgc tctgcctttg 13680
ttcatatgta ctgctgagga gtaagtctga gtacccagga ctaaccatct tgtttaaatg 13740
agcccgcttt gagattcttc ttccaaccca tctcctagtg atgcttctgg ctctctggcc 13800
aggctgagtt tctgagtgtc ccttgtagat gctttgccag tcctttttct ctgcacatgt 13860
gcattcctca ccttttcact tgtggcaata cattcttaaa gaatgccact ctggctccca 13920
agccttcctt gctgtgggcc atgtagcctc ctgttgtctg gtactgggta cacactcctg 13980
ctgtccgacc tcccaggact agcacaggtg gcagtcctat gcctcaaccc tttatttgta 14040
ctcttcacag tctttcccag gtctaaaatt gcatatcctt cacagtaaac tggaaggctt 14100
tctaagaaag tgcttttgag gcttctggcc actgttgttt tacctaaagc tgctaaaaca 14160
tgctcctgaa aagcaagctt tttcctgctt taggttctac tagtgacagc attgtggcct 14220
ttgggaattt cactctacag ttttgaaaaa atatctagca tttttgccta taccctatac 14280
tgcattcagt agtgcttggg gaaagcaagg aatctcttgc cttaaaggaa gcttgaaact 14340
tatgttggaa tgtccgatca ggcattgcta catatgtggt gctgtgctag ttgctttgca 14400
aagcgcagag gagtaaaaaa cacctgtgct ccccagggag ccatcacaga tatggcagtg 14460
ttggaagtgg ctcctttggg gtttggatct tgtttgcagg ctagtggtga catgtgctgg 14520
tgtcatttgt ctaaagagtg tttttctttc ccttcatctt ttaaagtcag atactcaaca 14580
gactccttgg gtattctggt cttggcagct cattcttgtc ttctcaggac ctgagtgatc 14640
aagatcgttt atacgattct ttcaaatacc catgactttc caaaaggaag tcattttcaa 14700
atctgaatcg ttcttttatg ttgataaatg atatgtatac atgaactaga gttacagtca 14760
gttgtgagct gcagttcagt tgataggaac agagtctggg tcctctgcaa gggcagccag 14820
tgctttttcc acttgacatc tcactcactg ccctcccggt ctcagtggga cagccagttc 14880
tcttaaccac taagctccct cttcagttcc acaacactga aaattttatt ttgctcttat 14940
gttcatactc tggtgaactc tttcacatgc actgctctgc tatcgtggct aagtgtgtgc 15000
aggttgtacc acatgccaca ttaagtagct actgcgcaca cagttctgtg aggttcatat 15060
gtaggccata tgctagtggg cttttcactt aactatatac cagtctttgc ttttcaaaac 15120
tataaaaatt tattgtgtga atggaaacta tgcaaaaaga acttaatcca agtagaaaat 15180
ttcatttctg tgcgtttttg ttttgtcagc taagatgctg cctacaaatg tatagtgaac 15240
agaaaaagag taaaactaat agttgctttg ctatgcacag tggtacattc ctgaagttct 15300
aggactcagg agacagaggc aggaggattg ctagtttgag accagcctga gccagcttga 15360
gctgcagagc aaaggctctc tctcagtaaa ccagtatcta ggatgctgag taatagggtc 15420
agcctggaag ggcaaggccc taaattccat tgtcagcact agagaaaaac cgaaaataaa 15480
cattgaaata aagtaacttt cactgtaaca aaatatatgg agagtagttt gaactgtctt 15540
gccttcggtg ttttaaaata ctaagagtta atttttattt ttaatattta ataatttgaa 15600
ggctggggct ttcacaaagg gctccacatg tgtcagccaa gggttctgcc agtgagctaa 15660
cacagtgagc taacatcccc ttttgactat ttacttttta cttaattgtc tcgaacttct 15720
gtccagctaa gatttcaagt ttttgtcact agatatactc tgaaagttgc tgtggtgtca 15780
agtgtttttg tggggtcact tcttccacca tcatcctctc ttcccttttg ctgctgagtc 15840
ggcatcacca ggcccctctt ccttctgcct ctcaagcact tgctccccag ctcctctcag 15900
tggtgacctt cagattcagt tcctccatgg cttccatcgc agcttctttg ttgtgctttt 15960
atcgttatgg ccagttctct acttattatt gttatgaaat gtcagatggt ttctgtcact 16020
gagaaatggg atggcacagc tgaggaactg atgtgcctgt cactgacaga catggcagac 16080
aagtagggat tggttcttga tttgtgtttt gggaactgaa atattagaaa actcacgtaa 16140
tcccaggact cagagttata gcctaggcgg tgagagctgc agtgtgctgc tgaggactgt 16200
ggcttcactg acctgagaga gtcctctgca ctagcattgt gtgaaacaag gatagcctcc 16260
attgtaaatg tggctgccta ctttctcctt ctctccaaca agcaaaacca taaggaaaaa 16320
aaaagcaatt catgctattg cctggccttc ctgtcacaac ttagctaatt aatgtgtaag 16380
atggaaagtg ggagctgcat ccttagttcc agaagagact taagtgcaca tgtgcacacc 16440
ctccctgggg agttactctg tacgtgtagc aagttccaag gcaggtgcct ggtatcccct 16500
ttttgacaga taactgtact tgcatacagt ggaagaagta ctggtgaggc tgtcatgcca 16560
gagagttgtt taacctccct gggtcttttt cctcttgtaa tgtaatgctc tgtcaaaatt 16620
gttggaaaga tgaaggtaaa gaaaccattg caatgccctg cacatacttg gtgtccaata 16680
tttctgttaa atgcagattt atcatttaga tttatttccc tcaattgttt ttcttgtttt 16740
gtgttgccca atgtgatgca atagaaatga ttttagaatt aacacatcta tcataatcat 16800
ataaaagcac aggggacaga taataaagaa tgcaacttgc caggctcaaa agtagttaat 16860
tctagaatgg aattgtaaat aatttcttcc agtggtacaa tcacatttct cagtaatagt 16920
tttaaatttt atagtaaaat ttagtttatt ctggttttaa agaaagagtt taaaggttac 16980
ttgtacttga agtctttgac agtatagttg atgaatctct cccttctatt tattttctat 17040
aatgatcctt ggaaaccata aaactgtttc ctgtgcagtg tattctctgt cgcctcgtac 17100
atgttgcttt acttcattgt attgcctcat gcgtcttacc atcctaatgt ttacttcttt 17160
tgaagggaag aaggttctaa ttatgataat tggggcatac cacttccagt agcatttcct 17220
tggactcaaa gtttgggaac tttttgtctc ttcagtaaca taaggagcta taccaattta 17280
aaataactgg ca 17292
<210> 5
<211> 10388
<212> DNA
<213>artificial sequence
<400> 5
taataggggt gagtaaatat tctttgagtc agttaggata atcgtgacta actggggaaa 60
agatgtggtt taagggtact atctctgccc tatagggata ataaactaac acttttataa 120
ctcttcgctg ttctgaatta gaaaagggta agtagaaagg gctacaaggt ggataccaag 180
attccgaggg cttacctagc atgtgggcac atcttaaaac catacagaag ccaggcatag 240
taacacactt ctagaattcc atcactcaga ggccagccca ggctatgtag tgaaaccttg 300
tcacaaagaa aacaaagaga tgtaaacagg agacattggg aaaccatctg ggcacattct 360
acctgatgtt tcccagctcc tgtccatatg ccaggctgtt taaggtccag tttggaccat 420
gtttcaaggc ttctttcctt catcctatca ttagaacaga aagtacaata gctactttaa 480
tactgttgta actcatcaac caagagtaac ttagtaagtg ttcttggtgt gcttttgctg 540
tgaccagatg gtgaaccaag gtagctgacc cattaattcc ccaaataatt agcaagattt 600
ttatgatgct tctctacttt catccccaag ccaaaaaaaa agacaaaaaa aaaaaaaccc 660
tttgaattca gaaagttcat agggtagatt tgatttgagt agtggtttaa ataactagac 720
tatgaacata ttatgcttat gcataccttg ttttctttaa agtttggagg tagttacaca 780
ttcattattc aacttgtgtt tgttctatgc cacacactaa tccagagctt ggggaacagc 840
aaggaggaag gcagataaga acagtgacgt gaacaagaca ctttgaaatg atatggacaa 900
ttaattagaa tagtgttcta gggtagactc ttctccatac atcttgggta agaaggtgac 960
atttgagaac ctaaggtgga aataatcgtg acatcttaag aggagcagga tgagatggat 1020
atggtaaaca aagtaagatg tggccccaga tgagatcaga gagaggaaag gagtggttcc 1080
atagtccctg ggagtctatt ctgatagggt gtcagtcaag aactgtaagc cagcgaatga 1140
aaacatttct gtagacatta taaaaagccc aggtaaaatt agagaaagaa aacaaaacat 1200
aaaacaaaac ctaaccatat aactaggatc tttccccctc ataatctatt tgttaattgt 1260
gctgccaggg acagttcatg aaaagtttaa ccatttgtaa aatcatgagt aagcactttt 1320
ggtaaaattg tacctgagcc aggtggtggt ggtggcgcac acctttaatc ccagcacttg 1380
ggaggcagag gcaggcagat ttctgaattc aaggccagcc tggactatag agtgagttcc 1440
aggacagcca gggctacaca gagaaaccct gtctcaaaaa acaaaaaaaa aaaaaaaatt 1500
gtacctgaat gaaaagttca ctccattagc agatatccct ctttttagtc aactattcta 1560
atatatgata aggcaataag tgtatatctt tttattatat tgtagagaaa gtttttattt 1620
ttgttaagtt tgtttgtttg tgggttttgg agatagggtt tctctgtgta gccctggctg 1680
ttctggaact tactctgtag accaggctgg cctcctaaat actgggacta aaggtgtgca 1740
ccaccacctc ctggcaaatt aatacttttt aaaggtttct attagatgcc tttaaaaaga 1800
agagatgaca gagtgggtaa aggccagact agctgaatca gcaatctctg ggtacaattg 1860
agagaccctg tctcagtttg taaggtagag aacaacacac acacacattc acatacttgc 1920
aaaccatata tatgaaaggc ccgtgtagag aatgggttgt gaagggcagg gaaacctgag 1980
ctcagaagga gtgttgctgt ggtgttggct aaggtgagat tggttaagac atttgcagtg 2040
agaaggctct ctccatttat tgaatgctta ttttgtagtt ggagtcttca cccttgtaaa 2100
atgtacatga tgtcttgaaa agggagctgg tattgaggca agaaagagat ttttagtgtg 2160
agtcgatggc tggtctgaac cgaggactgg gaagctttgg tggaggaagt aatttttgtt 2220
ttgttttggc catgttaagt ttgagacggc tggttaggtg tttcaagtgg caattgtggg 2280
tggacacatg gttaaactag tatcaagttc acagaaactt agtctttgtt attttatgta 2340
tatgagcatt atgtctacat gtgtgcctgc ttgatcacat gtgtgcctag tggccacaga 2400
ggccacacag ggcatcagat ttcctggaac tggagttgca ggcagttgtg agattccatg 2460
tggattccag gaatcaaatc tgggtcctct gaaatagcag caagtgctgt ggttgctgag 2520
ccatttctcc aggacccctc tccccctaat ttctattttt ttaaaaaata aaccctaaaa 2580
tttcatttgt catttgtgtg tatacacact catgtgtatg tacatgtgct atagcaaaca 2640
cacagagttc cagaagtcag gtcactcctt ccaccatgtg gaatcccagg aatttaactc 2700
aggttgtcag gcttggtggc cttaattcct gattctagac ttaatttctt gagaggtaaa 2760
tcaggagtta ttgccaaata catgataatt ccttattgtt tgtgggtctt tatacttctc 2820
ccaggtgggt aatttcctgg gaggtatcat ggttgttagg ttttattgcg ctcagggata 2880
cagaacagtc agcaaagggg agaggtgcat gggtaattgt gacaggaggt gggaagggac 2940
tcagcaccca gagcctcctc taggatttgt ggagggtgct tttgtcactg ttcctgcaga 3000
gtaagtaggc attcacttag agccatgttg ttcacacaag tgaggcaccc agatcatttg 3060
ggagtggaga gtgtcctgat ccccacttcc accctcaccc caatctccag gtccgctcct 3120
gtcttagctt tctgtcacgg tggaagaatg ccagagagaa acaagggagt aaggatgtag 3180
ctgggctcat ggttgcaaag gcttcagttc atgctgccag ctcctcctct tctgggctgc 3240
agggaagcag agcatcacag cagaaagtgt gtgattgagc ctcttgatgg cttagtggca 3300
aatattttaa aaacactttt cacttagata gctctcatat acacattatt atttatttgt 3360
ttgttttgtt ttggagacaa gttttcatgc tgcccaaact agccttcaaa gaatttgtgt 3420
ggtcaggaat ggctttgaat ggattctcca gccttccccc ttgagtcctg agattgcagg 3480
catggaccac catgcccagt ttatgcactg ctggggatca aactcagggc tttgtacatt 3540
cgaggcaagc acattgccaa atgctataca tccccagcct caagaatctc agtgaacctg 3600
ggaaagtagt gccccatctt ttgtcaactg ctttcctgct ctagccacat ctgattttag 3660
gtaccacaat gatgtctact tcaccgcagt gccttacttg aaaaccagtt tgtttaattt 3720
atgacttgaa ttactttgat gcaaaagagt tggaagtcaa gtatatttat ataaaaattt 3780
taaaaattca tttaaaaaaa gattgattca gccaatatgg tgtctgtgaa aacaagcccc 3840
ttctcggagg cttcaagtta ccttcagatg ggaagaataa acaggtctcc tgtgcagcag 3900
gtgagaggga tgggtcgtgt gaaacccatg aaggagcaag aagtgccagt ctttgggact 3960
ggggagctgc tgctggagag acaggaagga cagacctgag gccacctgag gcatagctga 4020
cttaggctct aagcctgatg accagagtgg ataggtgagg tcagaaagcc tcaagggctt 4080
tgtatggact ttgttctgag aagaatggaa agaaacaaga gatgccttcc catggcagct 4140
agaattactt tattcaaact ggagagtagg ctgtaggagt gcaagactgg aagccgggag 4200
agcagtaatg aggctatcag agaaaccagc ggagggctgc cccttgggcc gggttgttgg 4260
cagaggacag ggtgagagta actaatttct gcatacactg tgaagaaaga gagtgtctta 4320
gcataaacca tagagtgctt gcttatagct gagctcttta agatgtagca gaagggtttc 4380
cctgtttccc taggacttac ctctgagtta gaaaaaaaat taacttatac atttcctgac 4440
ttagtacttt gatctcattt cagtggactt gctactagta atttgtaagt aggacacttg 4500
gccagaccag gctgagtgtt agcataagct tcacagaagt gctgtcagtg cagaaggtgg 4560
gtatagttcc tgtctgttga atttggtatt cttgatgttg ttagtaaaga tacttgactt 4620
caaccctaag atgtggaggt tgtttgccct cccctttttg gcatagtttt atatatctga 4680
gcctggcctc aaactcactg aagctgagga tacgtgacct tgaatttctg atgccgttca 4740
cttccccagt gctaggttac agtgttgagg attgaaccca gagctaggca agcattttgc 4800
caactgagct atagttcagg tctctctctt ttttgaaagt tatgttttat ttgggaagaa 4860
gcatggccta cttgtaaggg gtcattgaaa aacttgcagg aattggtttt ctttttttac 4920
tgtgtggtcc aggaatggaa atcaggttgt caggcttgga ggcaagctta tttatttgct 4980
gagccgtctt gtgagtctca ggatgtccct tcttaaggtg ctcagtgtta gatgttggct 5040
tgtatgcagt cttactaaac agagagcgcc tgggagtata tatgtggcag gggcctctct 5100
tcagaaagtc tcatcctgct tgtctgctgc cctctcaggt attatggacc agaaaaagat 5160
tgcctgattt tttctttcag ataaatatta caaaggactg cattatttcc agagctttcc 5220
aagattatcc attagataca tgagtggtga cacagaactc agactaaatc cagtgttaaa 5280
gttgttataa agttgttatg gttgagacac ttggcacata ttgtacacac atatttatgc 5340
atgtagaggt gtggaatgtt aatatagtga atttaaacac atgtgcttta aaaatcttaa 5400
aggtgtggct aattttaatg ttcagtatat gtataggatt gtatttaaat tgatgccccc 5460
cccccacaca caaaggcgca taacgatacc acgatatcaa caagtttgta caaaaaagca 5520
ggctggcgcc ggaaccgaag ttcctattcc gaagttccta ttctctagaa agtataggaa 5580
cttcgtcgag ataacttcgt atagcataca ttatacgaag ttatgtcgag atatctagac 5640
ccagctttct tgtacaaagt ggttgatatc tctatagtcg cagtaggcgg acccagcaaa 5700
ttgatgtctt ctaaaaagct aaattgtcaa aatcaatatg gtcagttttg ttttgttttc 5760
tgtctgttaa aaccagtcat acaggctttc cccctatttc ttccaccttg caaaggggaa 5820
aaagattccc tgggtcgttg gatgcctctt cctggttcaa catgggagga atgtagtttc 5880
tcaagactga gcatccaagt aatacagagt gtaccacagt tttgttttaa gtgtgtaaat 5940
cattttggca gttgttgttg ttgttttaaa gtaaatctgt cctaattaaa aacaaaaaat 6000
gtaagcttat atatatatct tttcattttt ttctggacag agaggcagat gaaggctgta 6060
aaaaaccttt agaaagattg ctgaacatct ggcaagaacg aagtgtgtac ggcggcgagt 6120
tcatacagca gctgaagctg tctatggagg actccaagag ccctcccccc aaaggtagaa 6180
catcaccaca tgtttacagc tcttggtctt tgctcctccc agccccacgt gaaaagctgc 6240
agtgggggct acagtggctg ctgtgatcca ccagccgctt tatgactgca gagggatgct 6300
cctgagtgat ggtgagaact tacacagtgg tgcaagcgga gatggcgcaa cgcaattaat 6360
gataacttcg tatagcatac attatacgaa gttatggtct gagctcgcca tcagttcaag 6420
ggaacaacca agttctttag gattgtggct gactaacgag tagctcatcc tggccctgtc 6480
gtggacctca gcggggcctg ggataggtct cttgcccttt cagaactcag ctttattagt 6540
aataagtctg ggtttatata gtttaatgac tatcagatta ctgtctatat taattattga 6600
ttttcgtctt tactgagccc agggtatctt tgtgaagtat atatgtgaat tctaaaattc 6660
ttaagagtcc gtgttattta tatctttaaa gagaaagttc agacatttgg tttttcggtt 6720
gctggctagc catggatcac tagtggcatg acatgcttca ccctgctctg cctttgttca 6780
tatgtactgc tgaggagtaa gtctgagtac ccaggactaa ccatcttgtt taaatgagcc 6840
cgctttgaga ttcttcttcc aacccatctc ctagtgatgc ttctggctct ctggccaggc 6900
tgagtttctg agtgtccctt gtagatgctt tgccagtcct ttttctctgc acatgtgcat 6960
tcctcacctt ttcacttgtg gcaatacatt cttaaagaat gccactctgg ctcccaagcc 7020
ttccttgctg tgggccatgt agcctcctgt tgtctggtac tgggtacaca ctcctgctgt 7080
ccgacctccc aggactagca caggtggcag tcctatgcct caacccttta tttgtactct 7140
tcacagtctt tcccaggtct aaaattgcat atccttcaca gtaaactgga aggctttcta 7200
agaaagtgct tttgaggctt ctggccactg ttgttttacc taaagctgct aaaacatgct 7260
cctgaaaagc aagctttttc ctgctttagg ttctactagt gacagcattg tggcctttgg 7320
gaatttcact ctacagtttt gaaaaaatat ctagcatttt tgcctatacc ctatactgca 7380
ttcagtagtg cttggggaaa gcaaggaatc tcttgcctta aaggaagctt gaaacttatg 7440
ttggaatgtc cgatcaggca ttgctacata tgtggtgctg tgctagttgc tttgcaaagc 7500
gcagaggagt aaaaaacacc tgtgctcccc agggagccat cacagatatg gcagtgttgg 7560
aagtggctcc tttggggttt ggatcttgtt tgcaggctag tggtgacatg tgctggtgtc 7620
atttgtctaa agagtgtttt tctttccctt catcttttaa agtcagatac tcaacagact 7680
ccttgggtat tctggtcttg gcagctcatt cttgtcttct caggacctga gtgatcaaga 7740
tcgtttatac gattctttca aatacccatg actttccaaa aggaagtcat tttcaaatct 7800
gaatcgttct tttatgttga taaatgatat gtatacatga actagagtta cagtcagttg 7860
tgagctgcag ttcagttgat aggaacagag tctgggtcct ctgcaagggc agccagtgct 7920
ttttccactt gacatctcac tcactgccct cccggtctca gtgggacagc cagttctctt 7980
aaccactaag ctccctcttc agttccacaa cactgaaaat tttattttgc tcttatgttc 8040
atactctggt gaactctttc acatgcactg ctctgctatc gtggctaagt gtgtgcaggt 8100
tgtaccacat gccacattaa gtagctactg cgcacacagt tctgtgaggt tcatatgtag 8160
gccatatgct agtgggcttt tcacttaact atataccagt ctttgctttt caaaactata 8220
aaaatttatt gtgtgaatgg aaactatgca aaaagaactt aatccaagta gaaaatttca 8280
tttctgtgcg tttttgtttt gtcagctaag atgctgccta caaatgtata gtgaacagaa 8340
aaagagtaaa actaatagtt gctttgctat gcacagtggt acattcctga agttctagga 8400
ctcaggagac agaggcagga ggattgctag tttgagacca gcctgagcca gcttgagctg 8460
cagagcaaag gctctctctc agtaaaccag tatctaggat gctgagtaat agggtcagcc 8520
tggaagggca aggccctaaa ttccattgtc agcactagag aaaaaccgaa aataaacatt 8580
gaaataaagt aactttcact gtaacaaaat atatggagag tagtttgaac tgtcttgcct 8640
tcggtgtttt aaaatactaa gagttaattt ttatttttaa tatttaataa tttgaaggct 8700
ggggctttca caaagggctc cacatgtgtc agccaagggt tctgccagtg agctaacaca 8760
gtgagctaac atcccctttt gactatttac tttttactta attgtctcga acttctgtcc 8820
agctaagatt tcaagttttt gtcactagat atactctgaa agttgctgtg gtgtcaagtg 8880
tttttgtggg gtcacttctt ccaccatcat cctctcttcc cttttgctgc tgagtcggca 8940
tcaccaggcc cctcttcctt ctgcctctca agcacttgct ccccagctcc tctcagtggt 9000
gaccttcaga ttcagttcct ccatggcttc catcgcagct tctttgttgt gcttttatcg 9060
ttatggccag ttctctactt attattgtta tgaaatgtca gatggtttct gtcactgaga 9120
aatgggatgg cacagctgag gaactgatgt gcctgtcact gacagacatg gcagacaagt 9180
agggattggt tcttgatttg tgttttggga actgaaatat tagaaaactc acgtaatccc 9240
aggactcaga gttatagcct aggcggtgag agctgcagtg tgctgctgag gactgtggct 9300
tcactgacct gagagagtcc tctgcactag cattgtgtga aacaaggata gcctccattg 9360
taaatgtggc tgcctacttt ctccttctct ccaacaagca aaaccataag gaaaaaaaaa 9420
gcaattcatg ctattgcctg gccttcctgt cacaacttag ctaattaatg tgtaagatgg 9480
aaagtgggag ctgcatcctt agttccagaa gagacttaag tgcacatgtg cacaccctcc 9540
ctggggagtt actctgtacg tgtagcaagt tccaaggcag gtgcctggta tccccttttt 9600
gacagataac tgtacttgca tacagtggaa gaagtactgg tgaggctgtc atgccagaga 9660
gttgtttaac ctccctgggt ctttttcctc ttgtaatgta atgctctgtc aaaattgttg 9720
gaaagatgaa ggtaaagaaa ccattgcaat gccctgcaca tacttggtgt ccaatatttc 9780
tgttaaatgc agatttatca tttagattta tttccctcaa ttgtttttct tgttttgtgt 9840
tgcccaatgt gatgcaatag aaatgatttt agaattaaca catctatcat aatcatataa 9900
aagcacaggg gacagataat aaagaatgca acttgccagg ctcaaaagta gttaattcta 9960
gaatggaatt gtaaataatt tcttccagtg gtacaatcac atttctcagt aatagtttta 10020
aattttatag taaaatttag tttattctgg ttttaaagaa agagtttaaa ggttacttgt 10080
acttgaagtc tttgacagta tagttgatga atctctccct tctatttatt ttctataatg 10140
atccttggaa accataaaac tgtttcctgt gcagtgtatt ctctgtcgcc tcgtacatgt 10200
tgctttactt cattgtattg cctcatgcgt cttaccatcc taatgtttac ttcttttgaa 10260
gggaagaagg ttctaattat gataattggg gcataccact tccagtagca tttccttgga 10320
ctcaaagttt gggaactttt tgtctcttca gtaacataag gagctatacc aatttaaaat 10380
aactggca 10388

Claims (10)

1) and/or 2) and/or 3) 1. having the following preparation method of featured animal embryonic stem cell model:
1) proliferative capacity declines；
2) self-renewal capacity declines；
3) it is fitted into ability decline；
The described method includes: reducing or inhibiting the content and/or work of CREPT protein in the animal embryonic stem cell that sets out in vitro Property, obtain proliferative capacity and/or self-renewal capacity and/or the decline of chimeric ability compared with the animal embryonic stem cell that sets out Animal embryonic stem cell model；The animal is non-human animal or people.
2. according to the method described in claim 1, it is characterized by: the CREPT protein is following A1), A2) or A3):
A1) amino acid sequence is the protein of sequence 3；
A2) amino acid sequence shown in sequence 3 in sequence table is passed through to the substitution and/or missing of one or several amino acid residues And/or addition and protein with the same function；
A3) in A1) or the obtained fused protein of N-terminal A2) or/and C-terminal connection label.
3. method according to claim 1 or 2, it is characterised in that: the animal is non-human mammal or people.
4. method according to claim 1 to 3, it is characterised in that: the reduction is set out in animal embryonic stem cell The content and/or activity of CREPT protein pass through the CREPT albumen described in animal embryonic stem cell that sets out described in reduction or inhibition The expression of the encoding gene of matter is realized.
5. according to the method described in claim 4, it is characterized by: the animal embryonic stem cell that sets out described in the reduction or inhibition Described in CREPT protein encoding gene expression by setting out CREPT egg described in animal embryonic stem cell described in knocking out The encoding gene of white matter or its any segment import the inhibition CREPT protein into the animal embryonic stem cell that sets out Encoding gene substance realize.
6. according to the method described in claim 5, it is characterized by: the encoding gene for knocking out the CREPT protein or Its any segment is realized using Cre-loxp method.
7. utilizing the animal embryonic stem cell model of the method preparation any in claim 1-6.
8. CREPT protein as claimed in claim 1 or 2 or biomaterial relevant to the CREPT protein are in regulation animal Application in embryonic stem cell proliferation ability and/or self more sexuality and/or chimeric ability；The animal be non-human animal or People；The biomaterial is any one of following B1) to B9):
B1 the nucleic acid molecules of the CREPT protein) are encoded；
B2) contain B1) expression cassettes of the nucleic acid molecules；
B3) contain B1) recombinant vectors of the nucleic acid molecules or contain B2) recombinant vector of the expression cassette；
B4) contain B1) recombinant microorganisms of the nucleic acid molecules or contain B2) recombinant microorganism of the expression cassette or contain B3) the recombinant microorganism of the recombinant vector；
B5) contain B1) the transgenetic animal cell systems of the nucleic acid molecules or contain B2) transgenic animals of the expression cassette Cell line；
B6) contain B1) the transgenic animals tissues of the nucleic acid molecules or contain B2) the transgenic animals group of the expression cassette It knits；
B7) contain B1) transgenic animal organs of the nucleic acid molecules or contain B2) the transgenic animals device of the expression cassette Official；
B8 the nucleic acid molecules of the CREPT protein expression quantity) are reduced；
B9) contain B8) expression cassettes of the nucleic acid molecules, recombinant vector, recombinant microorganism, transgenetic animal cell system, turn base Because of animal tissue or transgenic animal organ.
9. application according to claim 8, it is characterised in that: B1) nucleic acid molecules are following b11) or b12) or B13) or b14):
B11) coded sequence is the cDNA molecule or DNA molecular of sequence 2 in sequence table；
B12) DNA molecular shown in sequence 1 in sequence table；
B13) and b11) or b12) nucleotide sequence that limits has 75% or 75% or more identity, and encodes the CREPT The cDNA molecule or DNA molecular of protein；
B14) under strict conditions with b11) or b12) or b13) nucleotide sequence hybridization that limits, and encode the CREPT egg The cDNA molecule or DNA molecular of white matter.
10. application according to claim 8 or claim 9, it is characterised in that: the animal is non-human mammal or people.
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1807623A (en) * 2005-12-28 2006-07-26 清华大学 Positive regulation gene for cell cycle and its coded protein and uses
CN101113170A (en) * 2007-03-05 2008-01-30 北京赛尔泰和生物医药科技有限公司 Preparation of new gene cell cycle related tumour high-expression gene monoclonal antibody and its application
CN102559601A (en) * 2012-01-13 2012-07-11 清华大学 CREPT (Cell-cycle Related and Expression-elevated Protein in Tumor) antibody for identifying tumor cells or tumor tissues

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1807623A (en) * 2005-12-28 2006-07-26 清华大学 Positive regulation gene for cell cycle and its coded protein and uses
CN101113170A (en) * 2007-03-05 2008-01-30 北京赛尔泰和生物医药科技有限公司 Preparation of new gene cell cycle related tumour high-expression gene monoclonal antibody and its application
CN102559601A (en) * 2012-01-13 2012-07-11 清华大学 CREPT (Cell-cycle Related and Expression-elevated Protein in Tumor) antibody for identifying tumor cells or tumor tissues

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