质粒名称: | pJK-caCas9-NatMX-Neut5L-Ade2 drive |
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出品公司: | 爱迪基因 |
目录编号: | 89577 |
存储实验室: | George Church |
载体骨架基本信息 |
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载体名称: | pJK1027 |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 7000 bp |
完整质粒大小: | -- |
载体修饰: | 插入caCas9和Neut5L同源臂 |
载体类型: | 酵母表达,合成生物学 |
筛选标记: | Nourseothricin |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 30 ℃ |
插入基因信息 |
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插入基因名称: | Ade2 gene drive |
物种来源: | C. albicans |
插入大小: | 1000 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | -- |
克隆方法: | Gibson克隆 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | TTGCAATTATTAGAGATCCAGAAAA |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pJK-caCas9-NatMX-Neut5L-Ade2 drive was a gift from George Church (Addgene plasmid # 89577 ; http://n2t.net/addgene:89577 ; RRID:Addgene_89577) |
参考文献部分: | A CRISPR-Cas9-based gene drive platform for genetic interaction analysis in Candida albicans. Shapiro RS, Chavez A, Porter CBM, Hamblin M, Kaas CS, DiCarlo JE, Zeng G, Xu X, Revtovich AV, Kirienko NV, Wang Y, Church GM, Collins JJ. Nat Microbiol. 2018 Jan;3(1):73-82. doi: 10.1038/s41564-017-0043-0. Epub 2017 Oct 23. 10.1038/s41564-017-0043-0 PubMed 29062088 |
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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 pJK-caCas9-NatMX-Neut5L-Ade2 drive 12969 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION caCas9 integrating vector into the Neut5L locus with gene drive for targeting Ade2 locus. ACCESSION . VERSION . KEYWORDS pJK-caCas9-NatMX-Neut5L-Ade2 drive SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12969) AUTHORS Shapiro RS, Chavez A, Porter CBM, Hamblin M, Kaas CS, DiCarlo JE, Zeng G, Xu X, Revtovich AV, Kirienko NV, Wang Y, Church GM, Collins JJ TITLE A CRISPR-Cas9-based gene drive platform for genetic interaction analysis in Candida albicans. JOURNAL Nat Microbiol. 2018 Jan;3(1):73-82. doi: 10.1038/s41564-017-0043-0. Epub 2017 Oct 23. PUBMED 29062088 REFERENCE 2 (bases 1 to 12969) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12969 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1..4104 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 4117..4137 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind 4142..4158 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 4143..4159 /label=KS primer /note="common sequencing primer, one of multiple similar variants" terminator 5095..5292 /label=TEF terminator /note="Ashbya gossypii TEF terminator" promoter complement(5893..6236) /label=TEF promoter /note="Ashbya gossypii TEF promoter" primer_bind complement(7199..7218) /label=T7 /note="T7 promoter, forward primer" promoter complement(7200..7218) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7225..7242) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7225..7241) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7234..7256) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(7470..7489) /label=F1ori-R /note="F1 origin, reverse primer" terminator 7742..7761 /label=SUP4 terminator /note="transcription terminator for the S. cerevisiae SUP4 tRNA gene" misc_RNA complement(7766..7840) /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_RNA complement(7866..7940) /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" primer_bind 9587..9608 /label=F1ori-F /note="F1 origin, forward primer" promoter 9771..9875 /gene="bla" /label=AmpR promoter CDS 9876..10736 /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 complement(10094..10113) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 10907..11495 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 11396..11415 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 11649..11666 /label=L4440 /note="L4440 vector, forward primer" protein_bind 11783..11804 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 11819..11849 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11857..11873 /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)." primer_bind 11862..11884 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 11881..11897 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 11881..11897 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 11918..11936 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 atggataaaa aatactccat aggtttggac attggaacca attcagttgg atgggctgta 61 atcactgatg aatataaggt tccttctaaa aagttcaaag ttttaggaaa tactgataga 121 cactccatta aaaagaattt aattggtgcc ttgttgtttg acagtggtga aacagctgaa 181 gcaactcgtt tgaaaagaac agctagaaga agatatacca gaagaaagaa cagaatttgt 241 tacttgcaag aaatcttttc taatgaaatg gcaaaggttg atgattcttt ttttcataga 301 ttggaagaaa gtttcttagt tgaagaagat aagaaacatg aaagacatcc aatctttggt 361 aatattgtag atgaagtggc ttaccacgaa aaatatccta caatttatca tttgagaaaa 421 aaattagttg actccactga taaagctgac ttaagactta tatatttggc attggctcac 481 atgatcaagt tcagaggtca cttcttaatt gaaggtgatt tgaatccaga taattccgat 541 gtcgacaagt tgtttattca attggttcaa acttacaacc aacttttcga ggaaaatcca 601 atcaatgcct caggtgtgga tgctaaagct attttatctg ctagattgtc aaaaagtaga 661 cgtttggaaa atcttattgc ccaattgcca ggtgaaaaaa aaaatggatt gtttggaaac 721 ttaattgcat tatctttggg tttgacccca aatttcaaat caaactttga tttggccgag 781 gatgctaaac ttcaattgtc taaggataca tacgatgacg atttagataa tcttcttgcc 841 caaattggtg atcaatatgc agatttattc ttggcagcta agaacttgag tgatgcaatt 901 ttgttaagtg acattttgag agtcaacaca gaaattacaa aagctccttt atccgcctct 961 atgattaaaa gatatgatga acaccatcaa gatttaacct tgttgaaagc tcttgtacgt 1021 caacaattac cagaaaagta taaagaaata ttctttgatc aatctaaaaa tggttacgct 1081 ggttatattg atggtggtgc tagtcaagaa gaattttata agtttatcaa gccaattctt 1141 gaaaaaatgg atggtactga ggaattgttg gttaagttga atcgagagga tttacttcgt 1201 aagcaaagaa catttgataa cggttctatc cctcatcaaa tccacttagg tgagttacac 1261 gctatcttga gacgtcagga agatttttac ccttttttga aagacaatcg tgaaaaaatt 1321 gaaaaaattt tgactttcag aattccttat tacgtcgggc cattggctag aggtaattct 1381 agatttgcat ggatgactcg taaatctgaa gagactatca ccccatggaa ttttgaggaa 1441 gttgttgata agggagcctc cgctcaatca ttcatcgaga gaatgaccaa tttcgacaaa 1501 aacttgccaa atgagaaggt gttacctaaa cattctttgt tatatgaata tttcactgtt 1561 tataatgaat taacaaaagt caaatacgtc actgaaggta tgagaaaacc agcattctta 1621 agtggtgaac aaaagaaagc tatcgttgac ttgttattca aaactaatcg taaagtcact 1681 gtcaaacaac ttaaagagga ctattttaag aaaatagagt gtttcgattc cgttgaaatc 1741 agtggggttg aggatagatt taatgctagt ttaggaactt atcacgattt gttgaaaatt 1801 atcaaagaca aagatttttt ggataatgaa gaaaacgaag atatcttgga ggacattgta 1861 ttgactttga ctttgtttga ggatagagaa atgatcgaag aacgtttaaa aacttacgct 1921 cacttattcg acgataaagt catgaaacaa ttaaaaagaa gaagatatac cggttgggga 1981 agattatcta gaaagttgat taatggtatt agagataagc aaagtggtaa aaccatctta 2041 gatttcctta aaagtgatgg ttttgcaaac agaaacttta tgcagttgat tcatgatgat 2101 agtttgacct ttaaggaaga tattcaaaaa gctcaggttt ctggacaggg tgattcattg 2161 catgaacata ttgccaactt agcagggtct ccagctatca aaaagggaat tttacaaact 2221 gttaaagttg ttgatgagtt ggtgaaagtt atgggacgtc ataagcctga aaatatagtt 2281 atagaaatgg ctagagaaaa tcaaacaact cagaaaggac aaaagaattc tcgtgaaaga 2341 atgaaaagaa tagaagaagg aattaaagag ttgggttctc aaatcttaaa agaacaccca 2401 gtcgaaaaca cccagttaca gaacgagaag ctttacttgt actatttgca aaacgggcgt 2461 gacatgtatg tagatcaaga gttggatatc aatagacttt cagattacga tgtcgatcac 2521 atagttcctc agtcattttt aaaagatgat tccattgaca acaaagtttt aacaagatct 2581 gataagaata gaggtaaatc agacaatgtg ccatcagaag aagtggttaa gaaaatgaag 2641 aactattgga gacaattgtt aaatgcaaag ttgattactc agagaaaatt tgacaactta 2701 actaaagctg aaagaggagg attatccgaa ttagataaag ctggattcat aaaacgtcag 2761 ttggttgaaa caagacaaat tactaaacat gtagcccaaa ttttagattc ccgaatgaat 2821 actaaatatg atgaaaatga caaattaatc agagaggtca aagtgatcac attaaagtca 2881 aagttggtgt cagatttccg aaaggatttt caattctaca aggtgagaga gatcaacaat 2941 tatcatcatg ctcacgacgc atatttgaat gcagtggtag gaactgcctt aataaagaag 3001 tatcctaagt tagaatccga attcgtgtat ggtgattata aagtctacga tgtcagaaag 3061 atgatagcaa aatcagaaca agaaattggg aaagccaccg caaagtattt cttttactcc 3121 aacatcatga atttctttaa gaccgaaatc accttggcta acggggaaat cagaaaacgt 3181 cctttgattg aaacaaatgg agaaaccggt gaaattgtgt gggataaggg gagagatttt 3241 gctactgtta gaaaagtttt atcaatgcct caagttaata ttgtaaagaa gactgaagtt 3301 caaactggtg gtttctctaa ggaatccatt cttccaaaac gaaactcaga caagttgatc 3361 gcccgaaaaa aagattggga tcctaagaaa tacggaggtt ttgattctcc tactgtcgca 3421 tattctgtct tggtcgtggc taaagttgaa aagggtaaat ccaaaaaatt gaaatctgtg 3481 aaagaattgt taggtattac catcatggag cgttcatcct ttgagaaaaa cccaattgac 3541 tttctcgaag cgaagggcta taaagaagtg aagaaagatt tgattattaa gttgccaaag 3601 tattctctct ttgaacttga aaatgggaga aagagaatgt tggcttcagc tggagaattg 3661 caaaaaggaa acgagttagc tttgccttca aagtacgtta atttcttata cttagcatct 3721 cattatgaaa agttgaaggg gtcccctgaa gacaatgagc aaaagcaatt attcgtagaa 3781 caacataaac actatttgga tgagataatt gaacaaatct ccgagttttc taaaagagtt 3841 attttggctg atgcaaattt ggataaagtt ttatctgctt ataataagca tcgtgacaaa 3901 ccaattcgag aacaagctga aaatattatc cacttgttca cattgactaa cttgggagct 3961 ccagccgcgt ttaagtactt tgacaccacc attgaccgta agagatatac ttcaacaaaa 4021 gaagtgttag acgccacatt gattcatcag tccataactg gtttatatga aactagaatc 4081 gatttgtcac aattgggggg tgatagccga gcggatccta agaaaaaacg aaaagtctga 4141 ctcgaggtcg acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccgag 4201 tgaaattctg gaaatctgga aatctggttt tgtattcttg ttattcttct ttttgttatt 4261 acatatataa cttgttactt ttttaaaaaa atctttgttt attttataaa tatataaaac 4321 taaatttaag aaaaagagaa aaatgtttta tttgagagat tgaaatttta cttgaattta 4381 gcttagcttt tataaagtat tattatgtaa aaaaacaaaa caaatataca ttaaaaagtt 4441 aagactataa aatagccacc caaggcattt ctatatcttg ttgttgttgt tttcatcttc 4501 tgtatcagag gaacttattt tattattttc gtcacgggta ttttctcttg tttgatgatt 4561 catcccattc attccatcat aaaattctag aaggaccacc tttgattgta aatagtaata 4621 attaccaccc ttatttaatt atttatttaa cttatttatt tatttattat acatatatac 4681 aaatctaata aagtgaaaat ctcccccttc acatttataa ttggccagtt tttttcaaat 4741 aagcattcca accagcatct ctataccttt taccttcaat atctggatct cttcctttac 4801 caaacaatcc tctaccaaca atgataatat cagttccagt gctaacaact tcatcaacag 4861 ttctatattg ttgtcctaat ccatcacctt tatcatctaa tccaactcca ggtgtcataa 4921 taagccaatc aaatccttct tcttgtccac ccatatcacg ttgggcaata aatccaataa 4981 caaattcctt atcggattta gcaatttcaa cagttttttg agaatattct ccatacgaat 5041 tcgagctcgt ttaaacgtcg acactggatg gcggcgttag tatcgaatcg acagcagtat 5101 agcgaccagc attcacatac gattgacgca tgatattact ttctgcgcac ttaacttcgc 5161 atctgggcag atgatgtcga ggcgaaaaaa aatataaatc acgctaacat ttgattaaaa 5221 tagaacaact acaatataaa aaaactatac aaatgacaag ttcttgaaaa caagaatctt 5281 tttattgtca gtactgactc gagttattat ggacatggca tagacatata caaagcttgt 5341 tcaccatcgg aagcagtacc atcgtataaa gcagtatcca aaccacacaa agtgaaaccc 5401 attcttctat aagcatgaat agctggagca ttaacattgg taacttccaa ccacaaatga 5461 ccagcacctc tttctctggc gaattcagta gccaaaccca tcaaagctct accaacacca 5521 tgacctctat gttctggagc aacttcaata tcttcaacag tcaatcttct gttccaacca 5581 gaataagaaa caacaacgaa accagccaaa tcaccatcat caccataagc aacgaaagtt 5641 ctagaatctg gatcaccatc ttcaccagca tcggattcat catcggattc atcatctggg 5701 aaaaccttag tcaatggtgg atcaactgga acttctctca aagtgaaacc atcaccagta 5761 gcagtaactc taaaaacagt atcggtagtg aaagaaccat ccaaagcttc aatagcttca 5821 gcatcacctg gaacagaagt tctgtatcta taagcagtat catccaaagt agtagacata 5881 attgtaggat ccggttgttt atgttcggat gtgatgtgag aactgtatcc tagcaagatt 5941 ttaaaaggaa gtatatgaaa gaagaacctc agtggcaaat cctaaccttt tatatttctc 6001 tacaggggcg cggcgtgggg acaattcaac gcgtctgtga ggggagcgtt tccctgctcg 6061 caggtctgca gcgaggagcc gtaatttttg cttcgcgccg tgcggccatc aaaatgtatg 6121 gatgcaaatg attatacatg gggatgtatg ggctaaatgt acgggcgaca gtcacatcat 6181 gcccctgagc tgcgcacgtc aagactgtca aggagggtat tctgggcctc catgtcgctg 6241 gccgggtgac ccggcgggga cgaggcaagc ttgatatgct aatgatccca ctgatgataa 6301 ttcagctaac atcaataacc ctcttggctc ttggttggtg gtggtttctt tagctccctg 6361 ttttaatcct tcaactactc cattcccagt gacaccatga gcattagtaa tatctgccca 6421 actactaatt ttataaactc caccaatata ttgtttcttc acggtattac caatatcagc 6481 aaattttcta tcttcaaaaa tcataaattg atgtttacgt gaaagttcta ataatggttc 6541 aatagtggat tcataggaaa aatcattgat tatatcaata tgagtcttga ttaagcatac 6601 ataaggaccc aatttatcaa ttaattcaag gaattcctta gtggtatcaa catcaattga 6661 tgcacataaa ttggttttct tcagttccat taatcgaaat aatcgttgtg ctactggtga 6721 ggcatgagtt tctgctctct cactataggt cttagtgttg actgtcatat cttttgtttg 6781 ctatttctat ttctatttct ttcttcaact gtaaaagggg cgatatataa accagtgtgt 6841 atggggttgt tgctcaggaa ttgatcaatg tttaaaagaa tggttgaaat catcattgaa 6901 gaacaaaaaa aaagattttt tctttcatta tgttatctgt ataatatgca ggaaataaga 6961 ttgcaaaaaa atagccgttt ttttggatac tatcaaacaa gaggaatgac taatacagaa 7021 ttagtgttac gaatcaatgg cactacagca actttcaaca aaataatcta cgttcaaaat 7081 ataatatata ctattaaata aaatcaaata ataaaataat tttaaagaaa gtagtaccag 7141 ctcttttttt tgtttccgtt tataccatcc aaatcaattc ctattagtct ccaattcgcc 7201 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa 7261 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa 7321 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 7381 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 7441 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 7501 cacgttcgcc tttaaatgat aattatagta ttcacggata gatctgttag agatgtggtg 7561 gattggtatt tctttctgtg attcaccgaa agtattatca actaagaagg gaaaggcacc 7621 acattttcta atgtctcata tattagttca gttcgatgta tttcttttta tatattaatt 7681 ataattttat tgcaccataa cgtttacttg tttaatatgc tattgatatc tatatgcggc 7741 cagacataaa aaacaaaaaa agcaccaccg actcggtgcc actttttcaa gttgataacg 7801 gactagcctt attttaactt gctatttcta gctctaaaac attaggtcgt atgattgttg 7861 cgcaccaccg actcggtgcc actttttcaa gttgataacg gactagcctt attttaactt 7921 gctatttcta gctctaaaac atttcttcta aagtttctgc caaattaaaa atagtttacg 7981 caagtctcga acttccacca caatttaatt gtaagaacgc tgcgaaaaca tattctccaa 8041 tcctattggg ccgaagtttc agctatgcca aaggcatcaa attttttttt ataaaatttt 8101 ccacttttcc ataccagtga aattatcacg tgatggagtg agtatctata atatttgatt 8161 gtttctttga tgaggcaata ctcgagtata gttattgacc aatgactttt agctatgtgt 8221 agtgcttgta tatgcgtgtg tgatgagaat aagatgaata gattagtaat gtataagttg 8281 attctttttg gtggggggtg ttacttttat tttttctttc catctcattg acaagattgt 8341 accgagtcaa aatgtttggg gtctttttat tgctgcgatg gattgatttt tcactctaag 8401 cctaaaatta aatggctaaa caagtggtat tcaagcacaa ttcagttttc tggatctcta 8461 ataattgcaa ttgcaattgc ttcacatact ctgtttataa ttcagtttct ttctcttgat 8521 gttactgata tattaagtta ataaatacca ataacttttt ttcaaaaatc ttctattttc 8581 ctttcaattg attatttaaa cacgaatcgt taataagctg tgattgcttt ttctctttct 8641 ttttcttttc tttgttcaag tttgaaaaaa aaaaaaaaag gtttctcttg ttcgatgtag 8701 aattcagtgg gttatagttg taatgatata tggaataaaa agaaagaaag agaagagaaa 8761 gagaagaaga gaagaattaa gtaaacgtgt attagaattt aaaaattaaa attcatgata 8821 atctttaata ataattaact aagaatcaca aaattgaaaa tttgaaagaa aaaagatcac 8881 tccaaggtcg tgtcaagtga ctagaaattc ttaattaaaa tagttataac aaaaaaagaa 8941 atttgtatat aacaattaag taaaagaaaa aaaagcaatt cgctttgtat aataataccc 9001 agaaaaatat attcatatat tcataaatca aacatgaaat gaagaatgct gaatcacttg 9061 ataggattta gttccattat ggtactaaga agtaatatta cttctatgaa aaggttgaat 9121 ccgagatcaa aattcatctt ttcatatact ctggagaaaa aaaaaaaagt aattgtagta 9181 agaatgacaa gtatcagttg agaaaaaaag taacaaagag aaagctcgga ggaggctccc 9241 caaaggtttt atcaccaatg gtggtaccac taacccaaga acagaaaaag cattgatatt 9301 ttgaaaaata caatactcat cttttaaccc aaaccttttc ttgtccttta aaacttttat 9361 ctcatgtctg tccaaagtca tattgtctta ctactttctc tccttcaacg tccttccggc 9421 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 9481 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga 9541 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 9601 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg 9661 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt 9721 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc 9781 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 9841 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg 9901 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg 9961 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc 10021 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca 10081 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac 10141 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa 10201 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg 10261 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt 10321 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg 10381 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc 10441 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga 10501 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta 10561 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc 10621 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg 10681 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt 10741 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa 10801 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 10861 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 10921 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 10981 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 11041 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 11101 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 11161 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 11221 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 11281 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 11341 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 11401 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 11461 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 11521 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 11581 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 11641 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc 11701 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 11761 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 11821 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 11881 caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac 11941 aaaagctggt accagagcta ttaagatcac cagcctcgtt tataataaac ttagtctaat 12001 atttcttttg gaccaaaata gaaactgttc tcaggtggca aaaaacagct taaagtctgt 12061 gttaaaaaaa gatctcggct atgccaatca aaaaggagtg acattattat gtaaaagaag 12121 aaatgttgta attccgcgaa acaacaattt taatgcttac tgtgccgcca ccaatatcct 12181 atggccaagg ggcctttaaa catgccacgg aaatatacag tcaaagtaaa cctactatgt 12241 ctttagagcc ttcaggatca attgttttta aatgaatgac tgcaaaacta gaaaagtagc 12301 ctcttacata agcaagtcat ttgtacaccg agagtttagt aaccagtgaa gcaagtatat 12361 ctggcaaacc tgaaacttat ctgtgaatta taaagctcat aagtaaaaag tgtagaaaag 12421 aagataaggc agaccaaata caaatttgca acagtacaac gaatatttca gcgtcaaaac 12481 tagagaataa taaagaaaac gatcttttca aaaagaaaaa accttttagt tttcctttgt 12541 tgttgttgtg ggtgtgtgct atttatatta tatagtttac tcataatacc ataaaatatt 12601 cggtttgatt aggttatttt aataagctaa tttgtttcta atcgtgtaat ttatgctgtg 12661 tatattaagt agtgtgtgca ctgcccaaaa atgtttgttg tttatagtcg gttaaagaga 12721 aaaaagaaaa aaagatccat acacacacgt taattagttg ttcaacgtaa tacactcata 12781 ttttgttctt atttgctttc ggtcgctgtt ctcaccaaga tttattgcca acgaaacaat 12841 ttttttttat atattttcag atttttcttt ttttcctttc ctttcctttt ctaattttca 12901 ctcctggttt tctttctttc ttagaaacat tatctcgata ttaatattaa aaaaatataa 12961 tcattcaaa //
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