质粒名称: | pHSB04X |
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出品公司: | 爱迪基因 |
目录编号: | 117258 |
存储实验室: | Sheng Yang |
载体骨架基本信息 |
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载体名称: | pCas |
载体出品公司: | Sheng Yang, Addgene Plasmid #62225 |
载体抗性: | 卡那霉素低(25 ug/ml) |
空载体大小: | 4476 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | -- |
筛选标记: | -- |
高拷贝/低拷贝: | 低拷贝 |
生长条件: | 30 ℃ |
插入基因信息 |
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插入基因名称: | Cas9, promotor PslpA-guide-RNA, homologous arms of Lb_1019,Cas9 (4107 bps), promoter PslpA-XbaI-guide-RNA (324 bps), homologous arm (1992 bps) |
物种来源: | Cas9 (Streptococcus pyogenes M1 GAS) |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | Cas9:AE004092 (SPy_1046) |
启动子 | -- |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | XbaI(未破坏) |
3' 克隆位点: | ApaI (未破坏) |
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: | pHSB04X was a gift from Sheng Yang (Addgene plasmid # 117258 ; http://n2t.net/addgene:117258 ; RRID:Addgene_117258) |
参考文献部分: | Development of a RecE/T-Assisted CRISPR-Cas9 Toolbox for Lactobacillus. Huang H, Song X, Yang S. Biotechnol J. 2019 Jul;14(7):e1800690. doi: 10.1002/biot.201800690. Epub 2019 May 20. 10.1002/biot.201800690 PubMed 30927506 |
买家导航
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 pHSB04X 14339 bp ds-DNA circular SYN 06-JUN-2019 DEFINITION Genome editing for Lactobacillus brevis ATCC367. ACCESSION . VERSION . KEYWORDS pHSB04X SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14339) TITLE RecE/T-Assisted Genome Editing in Lactobacillus JOURNAL Unpublished REFERENCE 2 (bases 1 to 14339) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 6, 2019 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14339 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(319..1134) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 1042..1061 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 1126..1145 /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1431..5537 /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" misc_RNA complement(12358..12433) /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" CDS complement(12786..13736) /codon_start=1 /gene="rep101 encoding an A56V mutation" /product="temperature-sensitive version of the RepA protein needed for replication with the pSC101 origin (Armstrong et al., 1984)" /label=Rep101(Ts) /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI" rep_origin 13784..14006 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" ORIGIN 1 ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt 61 ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag 121 taaggcagcg gtatcatcaa cggggtctga cgctcagtgg aacgaaaact cacgttaagg 181 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 241 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt acgtttccaa 301 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt 361 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa 421 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg 481 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa 541 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca 601 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc 661 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca 721 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt 781 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt 841 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat 901 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc 961 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt 1021 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat 1081 gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc tcataacacc 1141 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc 1201 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttcctgca gcgaaaagcc 1261 ctgacaaccc ttgttcctaa aaaggaataa gcgttcggtc agtaaataat agaaataaaa 1321 aatcagacct aagactgatg acaaaaagag aaaattttga taaaatagtc ttagaattaa 1381 attaaaaagg gaggccaaat ataatgaaaa atatgaatga caatgatgtt atggataaga 1441 aatactcaat aggcttagat atcggcacaa atagcgtcgg atgggcggtg atcactgatg 1501 aatataaggt tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc cacagtatca 1561 aaaaaaatct tataggggct cttttatttg acagtggaga gacagcggaa gcgactcgtc 1621 tcaaacggac agctcgtaga aggtatacac gtcggaagaa tcgtatttgt tatctacagg 1681 agattttttc aaatgagatg gcgaaagtag atgatagttt ctttcatcga cttgaagagt 1741 cttttttggt ggaagaagac aagaagcatg aacgtcatcc tatttttgga aatatagtag 1801 atgaagttgc ttatcatgag aaatatccaa ctatctatca tctgcgaaaa aaattggtag 1861 attctactga taaagcggat ttgcgcttaa tctatttggc cttagcgcat atgattaagt 1921 ttcgtggtca ttttttgatt gagggagatt taaatcctga taatagtgat gtggacaaac 1981 tatttatcca gttggtacaa acctacaatc aattatttga agaaaaccct attaacgcaa 2041 gtggagtaga tgctaaagcg attctttctg cacgattgag taaatcaaga cgattagaaa 2101 atctcattgc tcagctcccc ggtgagaaga aaaatggctt atttgggaat ctcattgctt 2161 tgtcattggg tttgacccct aattttaaat caaattttga tttggcagaa gatgctaaat 2221 tacagctttc aaaagatact tacgatgatg atttagataa tttattggcg caaattggag 2281 atcaatatgc tgatttgttt ttggcagcta agaatttatc agatgctatt ttactttcag 2341 atatcctaag agtaaatact gaaataacta aggctcccct atcagcttca atgattaaac 2401 gctacgatga acatcatcaa gacttgactc ttttaaaagc tttagttcga caacaacttc 2461 cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa cggatatgca ggttatattg 2521 atgggggagc tagccaagaa gaattttata aatttatcaa accaatttta gaaaaaatgg 2581 atggtactga ggaattattg gtgaaactaa atcgtgaaga tttgctgcgc aagcaacgga 2641 cctttgacaa cggctctatt ccccatcaaa ttcacttggg tgagctgcat gctattttga 2701 gaagacaaga agacttttat ccatttttaa aagacaatcg tgagaagatt gaaaaaatct 2761 tgacttttcg aattccttat tatgttggtc cattggcgcg tggcaatagt cgttttgcat 2821 ggatgactcg gaagtctgaa gaaacaatta ccccatggaa ttttgaagaa gttgtcgata 2881 aaggtgcttc agctcaatca tttattgaac gcatgacaaa ctttgataaa aatcttccaa 2941 atgaaaaagt actaccaaaa catagtttgc tttatgagta ttttacggtt tataacgaat 3001 tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc agcatttctt tcaggtgaac 3061 agaagaaagc cattgttgat ttactcttca aaacaaatcg aaaagtaacc gttaagcaat 3121 taaaagaaga ttatttcaaa aaaatagaat gttttgatag tgttgaaatt tcaggagttg 3181 aagatagatt taatgcttca ttaggtacct accatgattt gctaaaaatt attaaagata 3241 aagatttttt ggataatgaa gaaaatgaag atatcttaga ggatattgtt ttaacattga 3301 ccttatttga agatagggag atgattgagg aaagacttaa aacatatgct cacctctttg 3361 atgataaggt gatgaaacag cttaaacgtc gccgttatac tggttgggga cgtttgtctc 3421 gaaaattgat taatggtatt agggataagc aatctggcaa aacaatatta gattttttga 3481 aatcagatgg ttttgccaat cgcaatttta tgcagctgat ccatgatgat agtttgacat 3541 ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg cgatagttta catgaacata 3601 ttgcaaattt agctggtagc cctgctatta aaaaaggtat tttacagact gtaaaagttg 3661 ttgatgaatt ggtcaaagta atggggcggc ataagccaga aaatatcgtt attgaaatgg 3721 cacgtgaaaa tcagacaact caaaagggcc agaaaaattc gcgagagcgt atgaaacgaa 3781 tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa agagcatcct gttgaaaata 3841 ctcaattgca aaatgaaaag ctctatctct attatctcca aaatggaaga gacatgtatg 3901 tggaccaaga attagatatt aatcgtttaa gtgattatga tgtcgatcac attgttccac 3961 aaagtttcct taaagacgat tcaatagaca ataaggtctt aacgcgttct gataaaaatc 4021 gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa aaagatgaaa aactattgga 4081 gacaacttct aaacgccaag ttaatcactc aacgtaagtt tgataattta acgaaagctg 4141 aacgtggagg tttgagtgaa cttgataaag ctggttttat caaacgccaa ttggttgaaa 4201 ctcgccaaat cactaagcat gtggcacaaa ttttggatag tcgcatgaat actaaatacg 4261 atgaaaatga taaacttatt cgagaggtta aagtgattac cttaaaatct aaattagttt 4321 ctgacttccg aaaagatttc caattctata aagtacgtga gattaacaat taccatcatg 4381 cccatgatgc gtatctaaat gccgtcgttg gaactgcttt gattaagaaa tatccaaaac 4441 ttgaatcgga gtttgtctat ggtgattata aagtttatga tgttcgtaaa atgattgcta 4501 agtctgagca agaaataggc aaagcaaccg caaaatattt cttttactct aatatcatga 4561 acttcttcaa aacagaaatt acacttgcaa atggagagat tcgcaaacgc cctctaatcg 4621 aaactaatgg ggaaactgga gaaattgtct gggataaagg gcgagatttt gccacagtgc 4681 gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa aacagaagta cagacaggcg 4741 gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga caagcttatt gctcgtaaaa 4801 aagactggga tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct tattcagtcc 4861 tagtggttgc taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt aaagagttac 4921 tagggatcac aattatggaa agaagttcct ttgaaaaaaa tccgattgac tttttagaag 4981 ctaaaggata taaggaagtt aaaaaagact taatcattaa actacctaaa tatagtcttt 5041 ttgagttaga aaacggtcgt aaacggatgc tggctagtgc cggagaatta caaaaaggaa 5101 atgagctggc tctgccaagc aaatatgtga attttttata tttagctagt cattatgaaa 5161 agttgaaggg tagtccagaa gataacgaac aaaaacaatt gtttgtggag cagcataagc 5221 attatttaga tgagattatt gagcaaatca gtgaattttc taagcgtgtt attttagcag 5281 atgccaattt agataaagtt cttagtgcat ataacaaaca tagagacaaa ccaatacgtg 5341 aacaagcaga aaatattatt catttattta cgttgacgaa tcttggagct cccgctgctt 5401 ttaaatattt tgatacaaca attgatcgta aacgatatac gtctacaaaa gaagttttag 5461 atgccactct tatccatcaa tccatcactg gtctttatga aacacgcatt gatttgagtc 5521 agctaggagg tgactgaagt atattttaga tgaagattat ttcttaatct aggctcgaga 5581 tctatcgata agcttaagtg attagtcaaa gaatggtgat gacaattgta aattctattt 5641 aatcactttg actagcaaat actaacaaca agacacacac accaaaaatc aaaaattcac 5701 tacttttagt taaaaaccac gtaaccacaa gaactaatcc aatccatgta atcgggttct 5761 tcaaatattt ctccaagatt ttcctcctct aatatgctca acttaaatga cctattcaat 5821 aaatctatta tgctgctaaa tagtttatag gacaaataag tatactctaa tgacctataa 5881 aagatagaaa attaaaaaat caagtgttcg cttcgctctc actgcccctc gacgttttag 5941 tagcctttcc ctcacttcgt tcagtccaag ccaactaaaa gttttcgggc tactctctcc 6001 ttctccccct aataattaat taaaatctta ctctgtatat ttctgctaat cattcgctaa 6061 acagcaaaga aaaaacaaac acgtatcata gatataaatg taatggcata gtgcgggttt 6121 tattttcagc ctgtatcata gctaaacaaa tcgagttgtg tgtccgtttt agggcgttct 6181 gctagcttgt ttaaagtctc ttgaatgaat gtatgctcta agtcaaaaga atttgtcagc 6241 gcctttatat agctttcttt ttcttctttt tttactttaa tgatcgatag caacaatgat 6301 ttaacactag caagttgaat gccaccattt cttcctggtt taatcttaaa gaaaatttcc 6361 tgattcgcct tcagtacctt cagcaattta tctaatgtcc gttcaggaat gcctagcact 6421 tctctaatct cttttttggt cgtcactaaa taaggcttgt atacatcgct tttttcgcta 6481 atataagcca ttaaatcttc tttccattct gacaaatgaa cacgttgacg ttcgcttctt 6541 tttttcttga atttaaacca cccttgacgg acaaataaat ctttactggt taaatcactt 6601 gatacccaag ctttgcaaag aatggtaatg tattccctat tagccccttg atagttttct 6661 gaataggcac ttctaacaat tttgattact tctttttctt ctaagggttg atctaatcga 6721 ttattaaact caaacatatt atattcgcac gtttcgattg aatagcctga actaaagtag 6781 gctaaagaga gggtaaacat gacgttatta cgccctatta aacccttttc tcctgaaaat 6841 ttcgtttcgt gcaataagag attaaaccag ggttcatcta cttgtttttt gccttctgta 6901 ccgcttaaaa ccgttagact tgaacgagta aagcccttat tatctgtttg tttgaaagac 6961 caatcttgcc attctttgaa agaataacgg taattaggat caaaaaattc tacattgtcc 7021 gttcttggta tgcgagcaat accaaaatga ttacacgtta gatcaactgg caaagacttt 7081 ccaaaatatt ctcggatatt ttgcgaaatt attttggctg ctttgacaga tttaaattct 7141 gattttgaag tcacatagac tggcgtttct aaaacaaaat atgcttgata acctttatca 7201 gatttgataa tcatagtagg cataaaacct aaatcaatag cggttgttaa aatatcgctt 7261 gctgaaatag tttcttttgc cgtgtgaata tcaaaatcaa taaagaaggt attgatttgt 7321 cttaaattgt tttcagaatg tcctttcgtg tatgaacggt tttcgtctgc atacgttcca 7381 taacgataaa cgttgggtgt ccaatgtgta aatgtatctt gattttcttg aatcgcttcc 7441 tcggaagtca gaacaacacc acgaccgcca atcatgcttg attttgagcg atacgcaaaa 7501 atagcccctt tgcttttacc tggcttggta gtgattgagc gaattttact atttttaaat 7561 ttgtacttta acaagccgtc atgaagcaca gtttctacaa caaaagggat attcattcag 7621 ctgttctcct ttcctataaa tcctataaaa taggttgttt aattaacttg gtttgctttt 7681 tcattcaact gtttcaatat tgcatgtttt gaaaaagatt tttttccttt ataagtcaat 7741 ttttttccac taatcgaata aattattttg ttattttcta ttaacttata tatataatct 7801 tccccctccg aagaaaaata cttatctgat tttgtttcta agtagatatt tctcttttct 7861 aactctttct taaacgtttc tagtgtatag atatttgcta attttcttat ctccaataaa 7921 ctatttttta tataagtttt acattcatca tgattcatac aaactccacc ttctataaat 7981 gaatacaaaa aaagcaatca aacgatttcc gattgattgc ttaacaattc ttaaattcag 8041 tagcttagat acttgaaaac tctctgattt ccctatataa tgatagtacg gttatatacc 8101 gtcttcaaac aaagttaatt aaataacttc ttacgaggga agagttcatc tgactaactg 8161 ataagcgttg gtttggcaat cttatcgggc tatgcattta taaaatgtcg tcaaacattt 8221 tataaatgtg tcatggctct tttttcgttt ctattcagtt cgttgtttcg ttatatctag 8281 tataccgctt ttaaaaaaaa taagcaacga tttcgtgcat tattcacacg aagtcattgc 8341 ttttttcttc ttccatttct aaatccaatg ttacttgttc tgattctgtt tctggttctg 8401 gttctgttgg ctcatttggg attaaatcca ctactagcgt tgagttagtt aactttgcaa 8461 tttgttctag tgtttttatg gttggatctg attttcctga ttctattcgt gaataatttg 8521 atctactcat ttctaattct tggggtaccg ccagcatttc ggaaaaaaac cacgctaagg 8581 attttttcta taaaaagagc cgttatatta agaataaaac ggctctttta tacgtaaagg 8641 acgtaaattc atttgcccag tgtcatgtaa tccttcaaat ttgtattctc caagaaaatt 8701 gatatgttcc catcctaacg gccacgcata tggcattaaa tcttctctaa attctcctct 8761 tgcttttaat tcttctacgg ctttttccat atatacagtg ttccacacac ttatagcgtt 8821 aataattatg tttagtgcac tagctctttg taactggtct tggagagcac gttctctaaa 8881 ttctccacgt tgtccaaaaa atataattct agctaatgca ttgattgctt ctcctttatt 8941 taaacctttt tgaacccgtc tccttacggc tttattagat atgtaatcca gcgtaaagag 9001 ggttttctcg attcgtccca tttctccaag tgctgttgcg agtttatttt gtcttgcata 9061 tgatccgagc ttccccatga taagagcgct agggacctct ttagctcctt ggaagctgtc 9121 agtagtatac ctaataattt atctacattc cctttagtaa cgtgtaactt tccaaattta 9181 caaaagcgac tcatagaatt atttcctccc gttaaataat agataactat taaaaataga 9241 caatacttgc tcataagtaa cggtacttaa attgtttact ttggcgtgtt tcattgcttg 9301 atgaaactga tttttagtaa acagttgacg atattctcga ttgacccatt ttgaaacaaa 9361 gtacgtatat agcttccaat atttatctgg aacatctgtg gtatggcggg taagttttat 9421 taagacactg tttacttttg gtttaggatg aaagcattcc gctggcagct taagcaattg 9481 ctgaatcgag acttgagtgt gcaagagcaa ccctagtgtt cggtgaatat ccaaggtacg 9541 cttgtagaat ccttcttcaa caatcagata gatgtcagac gcatggcttt caaaaaccac 9601 ttttttaata atttgtgtgc ttaaatggta aggaatactc ccaacaattt tatacctctg 9661 tttgttaggg aattgaaact gtagaatatc ttggtgaatt aaagtgacac gagtattcag 9721 ttttaatttt tctgacgata agttgaatag atgactgtct aattcaatag acgttacctg 9781 tttacttatt ttagccagtt tcgtcgttaa atgcccttta cctgttccaa tttcgtaaac 9841 ggtatcggtt tcttttaaat tcaattgttt tattatttgg ttgagtactt tttcactcgt 9901 taaaaagttt tgagaatatt ttatattttt gttcatgtaa tcactccttc ttaattacaa 9961 atttttagca tctaatttaa cttcaattcc tattatacaa aattttaaga tactgcacta 10021 tcaacacact cttaagtttg cttctaagtc ttatttccat aacttctttt acgtttccgc 10081 cattctttgc tgtttcgatt tttatgatat ggtgcaagtc agcacgaaca cgaaccgtct 10141 tatctcccat tatatctttt tttgcactga ttggtgtatc atttcgtttt tctttttgtg 10201 cgcttcttga taaaagggat agtaattcat tcctggttgc aaattttgaa aaccgctacg 10261 gatcgcatct ttttctaaac tagggcccaa ccgcatcacg attaaacaga cgttcggact 10321 aacttagtcg cgaccgtttt ttattggcgt ggtaaatcct atcatgacat gactgacagt 10381 tatcgaaagt gaaagaagtc tagcgctgtt tgggcagtca gttattaaaa aaacgtagca 10441 tttcaagaat ttcaggcgtt ttagtggctt ctaatgaatg ggaccagtaa actaacccta 10501 atcattcaaa ataggagcaa aaataatgcg caaatttttg ggttttctgt tcttgctggc 10561 gctaggggcc tgctttttct taccccaagt acaaactgag cttccaaaga ttctaaaaca 10621 atcgcaattg accgttgccc aagcggctaa aaacatcggg gttcagttgc cgctggtcgg 10681 ccgacaagag gcggctggtg ccacgccagt cgagtcgatt gttcgtccgc aaacgttaaa 10741 gaaaacgtac tattatcatt tttcggcgga tgttccagta aaagttcaac aagtatttga 10801 acgggccatc gctgtttata atgagaccgg tattgttgac ttagtggcag gtcaggccac 10861 tgccgaacaa aatgaaatta cgatttttgt ttatcagaaa acgatgacgg ctaaacaagt 10921 taattatttg gaattaggta agggtggtcc ggagattcag caactaacgg gatttcaagg 10981 ttacacggct aatcatgcgc gtgcgggact gaacgtgaag tatggtcaaa gcattaagga 11041 atctgtggcg atccacgaat tgggccatgc gttaggactc gatcacagtt cgagtcggca 11101 atcagtgatg tatccaattg atcaggggct aacccagctg tctgctggtg atttaaaggc 11161 actgaagtat ttgtattagg atgccgcgta aaaaaggcca cgacatgtca ttatcttttg 11221 aggaccaata actaaaaaaa gcaattctga aaatgacttt ttcagaattg cttttttaat 11281 tcaaatatta atttaagtcg aaattacttc gtcatgaaaa aaactccttt tcttttcgtc 11341 tacaagaatc agtatactgc aaaggaagta atattcaagt gaaattttgc acaaaaagta 11401 ttacgttttt cctaagatga acgaaaaaag cttatataac agcagtcatt ggttgatctg 11461 aaaagtatat aaatgtgaag caaatgacta atcacttttt cagaagcaat attatcgtcc 11521 gaatgcaacc ggttgttaat ttttcaaaag acagcgacta acagtgatga ataggtaatg 11581 tttcagttaa atgtgagtgg gtagggagtt ttggcataca attatgagaa aatatgataa 11641 aattaaatgt tgaaatgaga atcggacgtc cattgttagt gtttttgctg acagtgggtg 11701 ccgcgaatta gccagaggag ttaacgtgta atgagccgaa ttttaattat tgaagatgaa 11761 aaaaacctag cccggttcgt tgaactggag ttaaagcatg aagggtacga aaccaacgtg 11821 cagtttaatg gtcgaaccgg cctggaagca gccttgtcac aggattttga tgtgattttg 11881 ctagacctaa tgttgcctga attgaatggg atcgaagttg ctcgccgcgt gcgtgaggtt 11941 aagagtacgc caatcattat gatgacggcg cgtgattctg tgattgatcg ggtttctgga 12001 ttagatcatg gcgcagacga ctatatcgtt aagccatttg caattgaaga actgttagcg 12061 cgtgttcggg cgctattgcg gcgaattcac ctagagggtg agcagaagag taccaaacag 12121 acgacggtta agttcaagga tcttacaatt gaaaaggaaa accgcatcgt acgtcggggc 12181 gacgaagtca ttaacctgac taaacgcgaa tatgaattgt tattagcctt gatggagaat 12241 atcaatgttg ttttggcgcg agatgtcctg ttaaccaaag tgtggggtta tgaatcagaa 12301 gtagaaacca acgtggttga tgtctacgtg cggtatcttc gcaagggccc aaaaaaagca 12361 ccgactcggt gccacttttt caagttgata acggactagc cttattttaa cttgctattt 12421 ctagctctaa aactgaaagt cggcgaaact aagtctagaa tagtataacc gaagttagcg 12481 cttttgttaa gaaatttatt cgataacatc agcggacccg atcagctatc ccctaactac 12541 gaagataagt atagcgattt ccttaaggag atacaaggat tacgccctga ttgtcgttgg 12601 attgaaacaa ttatgggtat cctgtaatgt tattgttaca taactaaaag tctactaacc 12661 tgattaaagc ctttgtaagt cgacgtatgg acagttttcc ctttgatatg taacggtgaa 12721 cagttgttct acttttgttt gttagtcttg atgcttcact gatagataca agagccataa 12781 gaacctcaga tccttccgta tttagccagt atgttctcta gtgtggttcg ttgtttttgc 12841 gtgagccatg agaacgaacc attgagatca tacttacttt gcatgtcact caaaaatttt 12901 gcctcaaaac tggtgagctg aatttttgca gttaaagcat cgtgtagtgt ttttcttagt 12961 ccgttatgta ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc attcattttt 13021 atctggttgt tctcaagttc ggttacgaga tccatttgtc tatctagttc aacttggaaa 13081 atcaacgtat cagtcgggcg gcctcgctta tcaaccacca atttcatatt gctgtaagtg 13141 tttaaatctt tacttattgg tttcaaaacc cattggttaa gccttttaaa ctcatggtag 13201 ttattttcaa gcattaacat gaacttaaat tcatcaaggc taatctctat atttgccttg 13261 tgagttttct tttgtgttag ttcttttaat aaccactcat aaatcctcat agagtatttg 13321 ttttcaaaag acttaacatg ttccagatta tattttatga atttttttaa ctggaaaaga 13381 taaggcaata tctcttcact aaaaactaat tctaattttt cgcttgagaa cttggcatag 13441 tttgtccact ggaaaatctc aaagccttta accaaaggat tcctgatttc cacagttctc 13501 gtcatcagct ctctggttgc tttagctaat acaccataag cattttccct actgatgttc 13561 atcatctgaa cgtattggtt ataagtgaac gataccgtcc gttctttcct tgtagggttt 13621 tcaatcgtgg ggttgagtag tgccacacag cataaaatta gcttggtttc atgctccgtt 13681 aagtcatagc gactaatcgc tagttcattt gctttgaaaa caactaattc agacatacat 13741 ctcaattggt ctaggtgatt ttaatcacta taccaattga gatgggctag tcaatgataa 13801 ttactagtcc ttttcctttg agttgtgggt atctgtaaat tctgctagac ctttgctgga 13861 aaacttgtaa attctgctag accctctgta aattccgcta gacctttgtg tgtttttttt 13921 gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa gataaaaaga 13981 atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta ttacaaaagg 14041 atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa aggcttaagt 14101 agcaccctcg caagctcggt tgcggccgca atcgggcaaa tcgctgaata ttccttttgt 14161 ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca 14221 cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa 14281 ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcg //
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