质粒名称: | CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1 |
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出品公司: | 爱迪基因 |
目录编号: | 138418 |
存储实验室: | Jian Xu |
载体骨架基本信息 |
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载体名称: | pLVX-EF1a-zsGreen1 |
载体出品公司: | Takara Bio |
载体抗性: | 氨苄青霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 慢病毒载体 |
筛选标记: | zsGreen1 |
高拷贝/低拷贝: | -- |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | dCas9 |
物种来源: | Streptococcus pyogenes |
插入大小: | 4401 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | EF1a |
标签/融合蛋白: | BioTAP-tag(插接件上的C端子) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | -- |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1 was a gift from Jian Xu (Addgene plasmid # 138418 ; http://n2t.net/addgene:138418 ; RRID:Addgene_138418) |
参考文献部分: | Multiplexed capture of spatial configuration and temporal dynamics of locus-specific 3D chromatin by biotinylated dCas9. Liu X, Chen Y, Zhang Y, Liu Y, Liu N, Botten GA, Cao H, Orkin SH, Zhang MQ, Xu J. Genome Biol. 2020 Mar 5;21(1):59. doi: 10.1186/s13059-020-01973-w. 10.1186/s13059-020-01973-w PubMed 32138752 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1 13286 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CAPTURE2.0 vector containing dCas9-CBio, IRES and zsGreen1. ACCESSION . VERSION . KEYWORDS CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13286) AUTHORS Liu X, Chen Y, Zhang Y, Liu Y, Liu N, Botten GA, Cao H, Orkin SH, Zhang MQ, Xu J TITLE Multiplexed capture of spatial configuration and temporal dynamics of locus-specific 3D chromatin by biotinylated dCas9. JOURNAL Genome Biol. 2020 Mar 5;21(1):59. doi: 10.1186/s13059-020-01973-w. PUBMED 32138752 REFERENCE 2 (bases 1 to 13286) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13286 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2028..2143 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" promoter 2338..3519 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2568..3510 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3467..3487 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 3558..3567 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3573..3593 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3594..7694 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=dCas9 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 7704..7724 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" misc_feature 7972..8544 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 7995..8546 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(8139..8156) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" CDS 8546..9241 /codon_start=1 /product="Zoanthus green fluorescent protein" /label=ZsGreen1 /note="mammalian codon-optimized" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH AIASGSALP" misc_feature 9255..9843 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(9308..9328) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(9726..9737) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 10050..10683 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(10811..10827) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10811..10827) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(10824..10846) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 10835..10851 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(10859..10889) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10904..10925 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(11042..11059) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(11213..11801) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(11293..11312) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(11972..12832) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 12595..12614 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(12833..12937) /gene="bla" /label=AmpR promoter polyA_signal 12985..13119 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(13022..13041) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 13076..13095 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg 2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc 2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct 2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt 2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt 2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatctatttc cggtgaattc 3541 ctcgagacta gttctaggcc accatggcta gccccaaaaa gaagaggaaa gtggacaaga 3601 agtattctat cggactggcc atcgggacta atagcgtcgg gtgggccgtg atcactgacg 3661 agtacaaggt gccctctaag aagttcaagg tgctcgggaa caccgaccgg cattccatca 3721 agaaaaatct gatcggagct ctcctctttg attcagggga gaccgctgaa gcaacccgcc 3781 tcaagcggac tgctagacgg cggtacacca ggaggaagaa ccggatttgt taccttcaag 3841 agatattctc caacgaaatg gcaaaggtcg acgacagctt cttccatagg ctggaagaat 3901 cattcctcgt ggaagaggat aagaagcatg aacggcatcc catcttcggt aatatcgtcg 3961 acgaggtggc ctatcacgag aaatacccaa ccatctacca tcttcgcaaa aagctggtgg 4021 actcaaccga caaggcagac ctccggctta tctacctggc cctggcccac atgatcaagt 4081 tcagaggcca cttcctgatc gagggcgacc tcaatcctga caatagcgat gtggataaac 4141 tgttcatcca gctggtgcag acttacaacc agctctttga agagaacccc atcaatgcaa 4201 gcggagtcga tgccaaggcc attctgtcag cccggctgtc aaagagccgc agacttgaga 4261 atcttatcgc tcagctgccg ggtgaaaaga aaaatggact gttcgggaac ctgattgctc 4321 tttcacttgg gctgactccc aatttcaagt ctaatttcga cctggcagag gatgccaagc 4381 tgcaactgtc caaggacacc tatgatgacg atctcgacaa cctcctggcc cagatcggtg 4441 accaatacgc cgaccttttc cttgctgcta agaatctttc tgacgccatc ctgctgtctg 4501 acattctccg cgtgaacact gaaatcacca aggcccctct ttcagcttca atgattaagc 4561 ggtatgatga gcaccaccag gacctgaccc tgcttaaggc actcgtccgg cagcagcttc 4621 cggagaagta caaggaaatc ttctttgacc agtcaaagaa tggatacgcc ggctacatcg 4681 acggaggtgc ctcccaagag gaattttata agtttatcaa acctatcctt gagaagatgg 4741 acggcaccga agagctcctc gtgaaactga atcgggagga tctgctgcgg aagcagcgca 4801 ctttcgacaa tgggagcatt ccccaccaga tccatcttgg ggagcttcac gccatccttc 4861 ggcgccaaga ggacttctac ccctttctta aggacaacag ggagaagatt gagaaaattc 4921 tcactttccg catcccctac tacgtgggac ccctcgccag aggaaatagc cggtttgctt 4981 ggatgaccag aaagtcagaa gaaactatca ctccctggaa cttcgaagag gtggtggaca 5041 agggagccag cgctcagtca ttcatcgaac ggatgactaa cttcgataag aacctcccca 5101 atgagaaggt cctgccgaaa cattccctgc tctacgagta ctttaccgtg tacaacgagc 5161 tgaccaaggt gaaatatgtc accgaaggga tgaggaagcc cgcattcctg tcaggcgaac 5221 aaaagaaggc aattgtggac cttctgttca agaccaatag aaaggtgacc gtgaagcagc 5281 tgaaggagga ctatttcaag aaaattgaat gcttcgactc tgtggagatt agcggggtcg 5341 aagatcggtt caacgcaagc ctgggtacct accatgatct gcttaagatc atcaaggaca 5401 aggattttct ggacaatgag gagaacgagg acatccttga ggacattgtc ctgactctca 5461 ctctgttcga ggaccgggaa atgatcgagg agaggcttaa gacctacgcc catctgttcg 5521 acgataaagt gatgaagcaa cttaaacgga gaagatatac cggatgggga cgccttagcc 5581 gcaaactcat caacggaatc cgggacaaac agagcggaaa gaccattctt gatttcctta 5641 agagcgacgg attcgctaat cgcaacttca tgcaacttat ccatgatgat tccctgacct 5701 ttaaggagga catccagaag gcccaagtgt ctggacaagg tgactcactg cacgagcata 5761 tcgcaaatct ggctggttca cccgctatta agaagggtat tctccagacc gtgaaagtcg 5821 tggacgagct ggtcaaggtg atgggtcgcc ataaaccaga gaacattgtc atcgagatgg 5881 ccagggaaaa ccagactacc cagaagggac agaagaacag cagggagcgg atgaaaagaa 5941 ttgaggaagg gattaaggag ctcgggtcac agatccttaa agagcacccg gtggaaaaca 6001 cccagcttca gaatgagaag ctctatctgt actaccttca aaatggacgc gatatgtatg 6061 tggaccaaga gcttgatatc aacaggctct cagactacga cgtggacgcc atcgtccctc 6121 agagcttcct caaagacgac tcaattgaca ataaggtgct gactcgctca gacaagaacc 6181 ggggaaagtc agataacgtg ccctcagagg aagtcgtgaa aaagatgaag aactattggc 6241 gccagcttct gaacgcaaag ctgatcactc agcggaagtt cgacaatctc actaaggctg 6301 agaggggcgg actgagcgaa ctggacaaag caggattcat taaacggcaa cttgtggaga 6361 ctcggcagat tactaaacat gtcgcccaaa tccttgactc acgcatgaat accaagtacg 6421 acgaaaacga caaacttatc cgcgaggtga aggtgattac cctgaagtcc aagctggtca 6481 gcgatttcag aaaggacttt caattctaca aagtgcggga gatcaataac tatcatcatg 6541 ctcatgacgc atatctgaat gccgtggtgg gaaccgccct gatcaagaag tacccaaagc 6601 tggaaagcga gttcgtgtac ggagactaca aggtctacga cgtgcgcaag atgattgcca 6661 aatctgagca ggagatcgga aaggccaccg caaagtactt cttctacagc aacatcatga 6721 atttcttcaa gaccgaaatc acccttgcaa acggtgagat ccggaagagg ccgctcatcg 6781 agactaatgg ggagactggc gaaatcgtgt gggacaaggg cagagatttc gctaccgtgc 6841 gcaaagtgct ttctatgcct caagtgaaca tcgtgaagaa aaccgaggtg caaaccggag 6901 gcttttctaa ggaatcaatc ctccccaagc gcaactccga caagctcatt gcaaggaaga 6961 aggattggga ccctaagaag tacggcggat tcgattcacc aactgtggct tattctgtcc 7021 tggtcgtggc taaggtggaa aaaggaaagt ctaagaagct caagagcgtg aaggaactgc 7081 tgggtatcac cattatggag cgcagctcct tcgagaagaa cccaattgac tttctcgaag 7141 ccaaaggtta caaggaagtc aagaaggacc ttatcatcaa gctcccaaag tatagcctgt 7201 tcgaactgga gaatgggcgg aagcggatgc tcgcctccgc tggcgaactt cagaagggta 7261 atgagctggc tctcccctcc aagtacgtga atttcctcta ccttgcaagc cattacgaga 7321 agctgaaggg gagccccgag gacaacgagc aaaagcaact gtttgtggag cagcataagc 7381 attatctgga cgagatcatt gagcagattt ccgagttttc taaacgcgtc attctcgctg 7441 atgccaacct cgataaagtc cttagcgcat acaataagca cagagacaaa ccaattcggg 7501 agcaggctga gaatatcatc cacctgttca ccctcaccaa tcttggtgcc cctgccgcat 7561 tcaagtactt cgacaccacc atcgaccgga aacgctatac ctccaccaaa gaagtgctgg 7621 acgccaccct catccaccag agcatcaccg gactttacga aactcggatt gacctctcac 7681 agctcggagg ggatgaggga gctcccaaga aaaagcgcaa ggtaggtagt tccgccggaa 7741 aggccggtga aggtgaaatc cctgcccctc ttgctggtac cgtttctaag atactggtaa 7801 aagaaggtga cactgttaaa gctggtcaaa cagttctggt gctggaggct atgaaaatgg 7861 agacagaaat taacgctcct actgacggaa aagttgaaaa ggtgttagtt aaggaaagag 7921 atgctgttca aggtggtcaa ggtctaatca agatcggcgt ttaagatccc gcccctctcc 7981 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt 8041 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg 8101 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg 8161 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc 8221 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca 8281 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag 8341 ttggatagtt gtggaaagag tcaaatggct cacctcaagc gtattcaaca aggggctgaa 8401 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt 8461 tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt 8521 ttcctttgaa aaacacgatg ataatatggc ccagtccaag cacggcctga ccaaggagat 8581 gaccatgaag taccgcatgg agggctgcgt ggacggccac aagttcgtga tcaccggcga 8641 gggcatcggc taccccttca agggcaagca ggccatcaac ctgtgcgtgg tggagggcgg 8701 ccccttgccc ttcgccgagg acatcttgtc cgccgccttc atgtacggca accgcgtgtt 8761 caccgagtac ccccaggaca tcgtcgacta cttcaagaac tcctgccccg ccggctacac 8821 ctgggaccgc tccttcctgt tcgaggacgg cgccgtgtgc atctgcaacg ccgacatcac 8881 cgtgagcgtg gaggagaact gcatgtacca cgagtccaag ttctacggcg tgaacttccc 8941 cgccgacggc cccgtgatga agaagatgac cgacaactgg gagccctcct gcgagaagat 9001 catccccgtg cccaagcagg gcatcttgaa gggcgacgtg agcatgtacc tgctgctgaa 9061 ggacggtggc cgcttgcgct gccagttcga caccgtgtac aaggccaagt ccgtgccccg 9121 caagatgccc gactggcact tcatccagca caagctgacc cgcgaggacc gcagcgacgc 9181 caagaaccag aagtggcacc tgaccgagca cgccatcgcc tccggctccg ccttgccctg 9241 aacgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc 9301 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg 9361 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc 9421 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg 9481 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg 9541 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga 9601 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct 9661 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg 9721 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc 9781 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc 9841 cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa 9901 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg 9961 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct 10021 tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca 10081 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta 10141 cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc 10201 agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc 10261 tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag 10321 ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg 10381 atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg 10441 cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta 10501 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 10561 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 10621 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 10681 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 10741 cagagagtga gaggccttga cattgctagc gtttaccgtc gacctctagc tagagcttgg 10801 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 10861 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 10921 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 10981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 11041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 11101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 11161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 11221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 11281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 11341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 11401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 11461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 11521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 11581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 11641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 11701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 11761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 11821 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 11881 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 11941 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 12001 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 12061 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 12121 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 12181 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 12241 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 12301 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 12361 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 12421 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 12481 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 12541 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 12601 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 12661 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 12721 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 12781 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 12841 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 12901 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 12961 ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa 13021 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 13081 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac 13141 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt 13201 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt 13261 taaatatgta ctacaaactt agtagt //