质粒名称: | pCAS9-TPC |
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出品公司: | 爱迪基因 |
目录编号: | 61478 |
存储实验室: | Holger Puchta |
载体骨架基本信息 |
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载体名称: | pPZP221 |
载体出品公司: | -- |
载体抗性: | 大观霉素 |
空载体大小: | 8400 bp |
完整质粒大小: | 14000 bp |
载体修饰: | -- |
载体类型: | 植物表达,CRISPR |
筛选标记: | Basta |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | Cas9 |
物种来源: | A. thaliana (mustard weed) |
插入大小: | 5600 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | PcUbi4-2 |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | EcoRI(未破坏) |
3' 克隆位点: | SacI (未破坏) |
5' 测序引物 | M13 rev |
3' 测序引物 | M13 fw |
文献引用方法 |
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材料和方法部分: | pCAS9-TPC was a gift from Holger Puchta (Addgene plasmid # 61478 ; http://n2t.net/addgene:61478 ; RRID:Addgene_61478) |
参考文献部分: | Both CRISPR/Cas-based nucleases and nickases can be used efficiently for genome engineering in Arabidopsis thaliana. Fauser F, Schiml S, Puchta H. Plant J. 2014 Jul;79(2):348-59. doi: 10.1111/tpj.12554. Epub 2014 Jun 17. 10.1111/tpj.12554 PubMed 24836556 |
买家导航
pCAS9TPC载体基本信息
pCAS9TPC载体图谱质粒图谱和多克隆位点信息
pCAS9TPC载体简介
pCAS9TPC载体序列
LOCUS pCAS9-TPC 14017 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Binary expression vector with A.th. codon-optimized Cas9 for conventional cloning. ACCESSION . VERSION . KEYWORDS pCAS9-TPC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14017) AUTHORS Fauser F, Schiml S, Puchta H TITLE Both CRISPR/Cas-based nucleases and nickases can be used efficiently for genome engineering in Arabidopsis thaliana. JOURNAL Plant J. 2014 Jul;79(2):348-59. doi: 10.1111/tpj.12554. Epub 2014 Jun 17. PUBMED 24836556 REFERENCE 2 (bases 1 to 14017) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14017 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1187..1816 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 2250..3317 /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV MRYRNLIEGEASAGS" rep_origin 3383..3577 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" primer_bind complement(3813..3835) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 3921..4061 /label=bom /note="basis of mobility region from pBR322" primer_bind 3976..3995 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(4076..4093) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4247..4835) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4327..4346) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5079..5870) /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" misc_feature 6398..6422 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" protein_bind 6599..6620 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6635..6665 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6673..6689 /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 6678..6700 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6697..6713 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6697..6713 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 7714..11817 /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 11830..11850 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 13018..13569 /codon_start=1 /gene="Streptomyces hygroscopicus bar" /product="phosphinothricin acetyltransferase" /label=BlpR /note="confers resistance to bialophos or phosphinothricin" /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF WQLDFSLPVPPRPVLPVTEI" primer_bind complement(13668..13685) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(13668..13684) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(13677..13699) /label=M13/pUC Forward /note="In lacZ gene" misc_feature 13880..13904 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" ORIGIN 1 agtactttga tccaacccct ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag 61 ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg 121 cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac 181 tagaaccgga gacattacgc catgaacaag agcgccgccg ctggcctgct gggctatgcc 241 cgcgtcagca ccgacgacca ggacttgacc aaccaacggg ccgaactgca cgcggccggc 301 tgcaccaagc tgttttccga gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc 361 aggatgcttg accacctacg ccctggcgac gttgtgacag tgaccaggct agaccgcctg 421 gcccgcagca cccgcgacct actggacatt gccgagcgca tccaggaggc cggcgcgggc 481 ctgcgtagcc tggcagagcc gtgggccgac accaccacgc cggccggccg catggtgttg 541 accgtgttcg ccggcattgc cgagttcgag cgttccctaa tcatcgaccg cacccggagc 601 gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac cctcaccccg 661 gcacagatcg cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg 721 gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa 781 gtgacgccca ccgaggccag gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc 841 gacgccctgg cggccgccga gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg 901 acggccagga cgaaccgttt ttcattaccg aagagatcga ggcggagatg atcgcggccg 961 ggtacgtgtt cgagccgccc gcgcacgtct caaccgtgcg gctgcatgaa atcctggccg 1021 gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa gaaaccgagc 1081 gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg 1141 cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga aggttatcgc 1201 tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc 1261 cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg gcagtgcccg 1321 cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg accgcccgac 1381 gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc 1441 ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc tgattccggt 1501 gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg ttaagcagcg 1561 cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg 1621 cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc ccattcttga 1681 gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca caaccgttct 1741 tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg ctgaaattaa 1801 atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta 1861 agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag cctggcagac 1921 acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac caagctgaag 1981 atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata catcgcgcag 2041 ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg ctaaaggagg 2101 cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg 2161 aacgggcggt tggccaggcg taagcggctg ggttgtctgc cggccctgca atggcactgg 2221 aacccccaag cccgaggaat cggcgtgacg gtcgcaaacc atccggcccg gtacaaatcg 2281 gcgcggcgct gggtgatgac ctggtggaga agttgaaggc cgcgcaggcc gcccagcggc 2341 aacgcatcga ggcagaagca cgccccggtg aatcgtggca agcggccgct gatcgaatcc 2401 gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc gattaggaag ccgcccaagg 2461 gcgacgagca accagatttt ttcgttccga tgctctatga cgtgggcacc cgcgatagtc 2521 gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg 2581 tgatccgcta cgagcttcca gacgggcacg tagaggtttc cgcagggccg gccggcatgg 2641 ccagtgtgtg ggattacgac ctggtactga tggcggtttc ccatctaacc gaatccatga 2701 accgataccg ggaagggaag ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg 2761 acgtactcaa gttctgccgg cgagccgatg gcggaaagca gaaagacgac ctggtagaaa 2821 cctgcattcg gttaaacacc acgcacgttg ccatgcagcg tacgaagaag gccaagaacg 2881 gccgcctggt gacggtatcc gagggtgaag ccttgattag ccgctacaag atcgtaaaga 2941 gcgaaaccgg gcggccggag tacatcgaga tcgagctagc tgattggatg taccgcgaga 3001 tcacagaagg caagaacccg gacgtgctga cggttcaccc cgattacttt ttgatcgatc 3061 ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca 3121 gatggttgtt caagacgatc tacgaacgca gtggcagcgc cggagagttc aagaagttct 3181 gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg 3241 aggcggggca ggctggcccg atcctagtca tgcgctaccg caacctgatc gagggcgaag 3301 catccgccgg ttcctaatgt acggagcaga tgctagggca aattgcccta gcaggggaaa 3361 aaggtcgaaa aggtctcttt cctgtggata gcacgtacat tgggaaccca aagccgtaca 3421 ttgggaaccg gaacccgtac attgggaacc caaagccgta cattgggaac cggtcacaca 3481 tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac 3541 ttattaaaac tcttaaaacc cgcctggcct gtgcataact gtctggccag cgcacagccg 3601 aagagctgca aaaagcgcct acccttcggt cgctgcgctc cctacgcccc gccgcttcgc 3661 gtcggcctat cgcggccgct ggccgctcaa aaatggctgg cctacggcca ggcaatctac 3721 cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat caaggcaccc 3781 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg 3841 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg 3901 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat 3961 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg 4021 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc 4081 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 4141 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 4201 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 4261 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 4321 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 4381 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 4441 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 4501 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 4561 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 4621 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 4681 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 4741 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 4801 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 4861 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg catgatatat 4921 ctcccaattt gtgtagggct tattatgcac gcttaaaaat aataaaagca gacttgacct 4981 gatagtttgg ctgtgagcaa ttatgtgctt agtgcatcta atcgcttgag ttaacgccgg 5041 cgaagcggcg tcggcttgaa cgaatttcta gctagacatt atttgccgac taccttggtg 5101 atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga 5161 tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc 5221 ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact 5281 gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg 5341 ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga 5401 accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct 5461 tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga 5521 atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga 5581 atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca 5641 ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc 5701 cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact 5761 gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca 5821 actacctctg atagttgagt cgatacttcg gcgatcaccg cttcccccat gatgtttaac 5881 tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctccataa catcaaacat 5941 cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg taccccaaaa 6001 aaacatgtca taacaagaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc 6061 ggtcaaggtt ctggaccagt tgcgtgacgg cagttacgct acttgcatta cagcttacga 6121 accgaacgag gcttatgtcc actgggttcg tgcccgaatt gatcacaggc agcaacgctc 6181 tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca tccgtgtttc aaacccggca 6241 gcttagttgc cgttcttccg aatagcatcg gtaacatgag caaagtctgc cgccttacaa 6301 cggctctccc gctgacgccg tcccggactg atgggctgcc tgtatcgagt ggtgattttg 6361 tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa 6421 acaaattgac gcttagacaa cttaataaca cattgcggac gtttttaatg tactgaatta 6481 acgccgaatt gctctagcca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 6541 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 6601 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta 6661 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta 6721 cgaattcaaa aattacggat atgaatatag gcatatccgt atccgaatta tccgtttgac 6781 agctagcaac gattgtacaa ttgcttcttt aaaaaaggaa gaaagaaaga aagaaaagaa 6841 tcaacatcag cgttaacaaa cggccccgtt acggcccaaa cggtcatata gagtaacggc 6901 gttaagcgtt gaaagactcc tatcgaaata cgtaaccgca aacgtgtcat agtcagatcc 6961 cctcttcctt caccgcctca aacacaaaaa taatcttcta cagcctatat atacaacccc 7021 cccttctatc tctcctttct cacaattcat catctttctt tctctacccc caattttaag 7081 aaatcctctc ttctcctctt cattttcaag gtaaatctct ctctctctct ctctctctgt 7141 tattccttgt tttaattagg tatgtattat tgctagtttg ttaatctgct tatcttatgt 7201 atgccttatg tgaatatctt tatcttgttc atctcatccg tttagaagct ataaatttgt 7261 tgatttgact gtgtatctac acgtggttat gtttatatct aatcagatat gaatttcttc 7321 atattgttgc gtttgtgtgt accaatccga aatcgttgat ttttttcatt taatcgtgta 7381 gctaattgta cgtatacata tggatctacg tatcaattgt tcatctgttt gtgtttgtat 7441 gtatacagat ctgaaaacat cacttctctc atctgattgt gttgttacat acatagatat 7501 agatctgtta tatcattttt tttattaatt gtgtatatat atatgtgcat agatctggat 7561 tacatgattg tgattattta catgattttg ttatttacgt atgtatatat gtagatctgg 7621 actttttgga gttgttgact tgattgtatt tgtgtgtgta tatgtgtgtt ctgatcttga 7681 tatgttatgt atgtgcagcg aattcggcgc gccatggata agaagtactc tatcggactc 7741 gatatcggaa ctaactctgt gggatgggct gtgatcaccg atgagtacaa ggtgccatct 7801 aagaagttca aggttctcgg aaacaccgat aggcactcta tcaagaaaaa ccttatcggt 7861 gctctcctct tcgattctgg tgaaactgct gaggctacca gactcaagag aaccgctaga 7921 agaaggtaca ccagaagaaa gaacaggatc tgctacctcc aagagatctt ctctaacgag 7981 atggctaaag tggatgattc attcttccac aggctcgaag agtcattcct cgtggaagaa 8041 gataagaagc acgagaggca ccctatcttc ggaaacatcg ttgatgaggt ggcataccac 8101 gagaagtacc ctactatcta ccacctcaga aagaagctcg ttgattctac tgataaggct 8161 gatctcaggc tcatctacct cgctctcgct cacatgatca agttcagagg acacttcctc 8221 atcgagggtg atctcaaccc tgataactct gatgtggata agttgttcat ccagctcgtg 8281 cagacctaca accagctttt cgaagagaac cctatcaacg cttcaggtgt ggatgctaag 8341 gctatcctct ctgctaggct ctctaagtca agaaggcttg agaacctcat tgctcagctc 8401 cctggtgaga agaagaacgg acttttcgga aacttgatcg ctctctctct cggactcacc 8461 cctaacttca agtctaactt cgatctcgct gaggatgcaa agctccagct ctcaaaggat 8521 acctacgatg atgatctcga taacctcctc gctcagatcg gagatcagta cgctgatttg 8581 ttcctcgctg ctaagaacct ctctgatgct atcctcctca gtgatatcct cagagtgaac 8641 accgagatca ccaaggctcc actctcagct tctatgatca agagatacga tgagcaccac 8701 caggatctca cacttctcaa ggctcttgtt agacagcagc tcccagagaa gtacaaagag 8761 attttcttcg atcagtctaa gaacggatac gctggttaca tcgatggtgg tgcatctcaa 8821 gaagagttct acaagttcat caagcctatc ctcgagaaga tggatggaac cgaggaactc 8881 ctcgtgaagc tcaatagaga ggatcttctc agaaagcaga ggaccttcga taacggatct 8941 atccctcatc agatccacct cggagagttg cacgctatcc ttagaaggca agaggatttc 9001 tacccattcc tcaaggataa cagggaaaag attgagaaga ttctcacctt cagaatccct 9061 tactacgtgg gacctctcgc tagaggaaac tcaagattcg cttggatgac cagaaagtct 9121 gaggaaacca tcaccccttg gaacttcgaa gaggtggtgg ataagggtgc tagtgctcag 9181 tctttcatcg agaggatgac caacttcgat aagaaccttc caaacgagaa ggtgctccct 9241 aagcactctt tgctctacga gtacttcacc gtgtacaacg agttgaccaa ggttaagtac 9301 gtgaccgagg gaatgaggaa gcctgctttt ttgtcaggtg agcaaaagaa ggctatcgtt 9361 gatctcttgt tcaagaccaa cagaaaggtg accgtgaagc agctcaaaga ggattacttc 9421 aagaaaatcg agtgcttcga ttcagttgag atttctggtg ttgaggatag gttcaacgca 9481 tctctcggaa cctaccacga tctcctcaag atcattaagg ataaggattt cttggataac 9541 gaggaaaacg aggatatctt ggaggatatc gttcttaccc tcaccctctt tgaagataga 9601 gagatgattg aagaaaggct caagacctac gctcatctct tcgatgataa ggtgatgaag 9661 cagttgaaga gaagaagata cactggttgg ggaaggctct caagaaagct cattaacgga 9721 atcagggata agcagtctgg aaagacaatc cttgatttcc tcaagtctga tggattcgct 9781 aacagaaact tcatgcagct catccacgat gattctctca cctttaaaga ggatatccag 9841 aaggctcagg tttcaggaca gggtgatagt ctccatgagc atatcgctaa cctcgctgga 9901 tctcctgcaa tcaagaaggg aatcctccag actgtgaagg ttgtggatga gttggtgaag 9961 gtgatgggaa ggcataagcc tgagaacatc gtgatcgaaa tggctagaga gaaccagacc 10021 actcagaagg gacagaagaa ctctagggaa aggatgaaga ggatcgagga aggtatcaaa 10081 gagcttggat ctcagatcct caaagagcac cctgttgaga acactcagct ccagaatgag 10141 aagctctacc tctactacct ccagaacgga agggatatgt atgtggatca agagttggat 10201 atcaacaggc tctctgatta cgatgttgat catatcgtgc cacagtcatt cttgaaggat 10261 gattctatcg ataacaaggt gctcaccagg tctgataaga acaggggtaa gagtgataac 10321 gtgccaagtg aagaggttgt gaagaaaatg aagaactatt ggaggcagct cctcaacgct 10381 aagctcatca ctcagagaaa gttcgataac ttgactaagg ctgagagggg aggactctct 10441 gaattggata aggcaggatt catcaagagg cagcttgtgg aaaccaggca gatcactaag 10501 cacgttgcac agatcctcga ttctaggatg aacaccaagt acgatgagaa cgataagttg 10561 atcagggaag tgaaggttat caccctcaag tcaaagctcg tgtctgattt cagaaaggat 10621 ttccaattct acaaggtgag ggaaatcaac aactaccacc acgctcacga tgcttacctt 10681 aacgctgttg ttggaaccgc tctcatcaag aagtatccta agctcgagtc agagttcgtg 10741 tacggtgatt acaaggtgta cgatgtgagg aagatgatcg ctaagtctga gcaagagatc 10801 ggaaaggcta ccgctaagta tttcttctac tctaacatca tgaatttctt caagaccgag 10861 attaccctcg ctaacggtga gatcagaaag aggccactca tcgagacaaa cggtgaaaca 10921 ggtgagatcg tgtgggataa gggaagggat ttcgctaccg ttagaaaggt gctctctatg 10981 ccacaggtga acatcgttaa gaaaaccgag gtgcagaccg gtggattctc taaagagtct 11041 atcctcccta agaggaactc tgataagctc attgctagga agaaggattg ggaccctaag 11101 aaatacggtg gtttcgattc tcctaccgtg gcttactctg ttctcgttgt ggctaaggtt 11161 gagaagggaa agagtaagaa gctcaagtct gttaaggaac ttctcggaat cactatcatg 11221 gaaaggtcat ctttcgagaa gaacccaatc gatttcctcg aggctaaggg atacaaagag 11281 gttaagaagg atctcatcat caagctccca aagtactcac tcttcgaact cgagaacggt 11341 agaaagagga tgctcgcttc tgctggtgag cttcaaaagg gaaacgagct tgctctccca 11401 tctaagtacg ttaactttct ttacctcgct tctcactacg agaagttgaa gggatctcca 11461 gaagataacg agcagaagca acttttcgtt gagcagcaca agcactactt ggatgagatc 11521 atcgagcaga tctctgagtt ctctaaaagg gtgatcctcg ctgatgcaaa cctcgataag 11581 gtgttgtctg cttacaacaa gcacagagat aagcctatca gggaacaggc agagaacatc 11641 atccatctct tcacccttac caacctcggt gctcctgctg ctttcaagta cttcgataca 11701 accatcgata ggaagagata cacctctacc aaagaagtgc tcgatgctac cctcatccat 11761 cagtctatca ctggactcta cgagactagg atcgatctct cacagctcgg tggtgattca 11821 agggctgatc ctaagaagaa gaggaaggtt tgaggcgcgc cgagctccag gcctcccagc 11881 tttcgtccgt atcatcggtt tcgacaacgt tcgtcaagtt caatgcatca gtttcattgc 11941 ccacacacca gaatcctact aagtttgagt attatggcat tggaaaagct gttttcttct 12001 atcatttgtt ctgcttgtaa tttactgtgt tctttcagtt tttgttttcg gacatcaaaa 12061 tgcaaatgga tggataagag ttaataaatg atatggtcct tttgttcatt ctcaaattat 12121 tattatctgt tgtttttact ttaatgggtt gaatttaagt aagaaaggaa ctaacagtgt 12181 gatattaagg tgcaatgtta gacatataaa acagtctttc acctctcttt ggttatgtct 12241 tgaattggtt tgtttcttca cttatctgtg taatcaagtt tactatgagt ctatgatcaa 12301 gtaattatgc aatcaagtta agtacagtat aggcttgagc tccctaggcc cgggcctgag 12361 gacgcgtcca tggttaatta agacgtccgg accgactagt ggatcctcta gagtcgacct 12421 gcaggcatgc aagcttcttc gtcaacatgg tggagcacga cacgcttgtc tactccaaaa 12481 atatcaaaga tacagtctca gaagaccaaa gggcaattga gacttttcaa caaagggtaa 12541 tatccggaaa cctcctcgga ttccattgcc cagctatctg tcactttatt gtgaagatag 12601 tggaaaagga aggtggctcc tacaaatgcc atcattgcga taaaggaaag gccatcgttg 12661 aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg 12721 aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat atctccactg 12781 acgtaaggga tgacgcacaa tcaatcccac tatccttcgc aagacccttt taagggggaa 12841 gttcatttca tttggagagg acacgctgaa atcaccagtc tctctgtaca aatcnatctc 12901 tctctataat attgtgtaag tagttcccag ataagggaat tagggttctt atagggtttc 12961 gctcagctgt tgagcatata agaaaccctt agtcgataga tctgttgggg atctaccatg 13021 agcccagaac gacgcccggc cgacatccgc cgtgccaccg aggcggacat gccggcggtc 13081 tgcaccatcg tcaaccacta catcgagaca agcacggtca acttccgtac cgagccgcag 13141 gaaccgcagg agtggacgga cgacctcgtc cgtctgcggg agcgctatcc ctggctcgtc 13201 gccgaggtgg acggcgaggt cgccggcatc gcctacgcgg gcccctggaa ggcacgcaac 13261 gcctacgact ggacggccga gtcgaccgtg tacgtctccc cccgccacca gcggacggga 13321 ctgggctcca cgctctacac ccacctgctg aagtccctgg aggcacaggg cttcaagagc 13381 gtggtcgctg tcatcgggct gcccaacgac ccgagcgtgc gcatgcacga ggcgctcgga 13441 tatgcccccc gcggcatgct gcgggcggcc ggcttcaagc acgggaactg gcatgacgtg 13501 ggtttctggc agctggactt cagcctgccg gtaccgcccc gtccggtcct gcccgtcacc 13561 gagatctgat gacccaactt agtatgtatt tgtatttgta aaatacttct atcaataaaa 13621 tttctaattc ctaaaaccaa aatccagggg taccgaacaa gcttggcact ggccgtcgtt 13681 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat 13741 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag 13801 ttgcgcagcc tgaatggcga atgagcttga gcttggatca gattgtcgtt tcccgccttc 13861 agtttaaact atcagtgttt gacaggatat attggcgggt aaacctaaga gaaaagagcg 13921 tttattagaa taacggatat ttaaaagggc gtgaaaaggt ttatccgttc gtccatttgt 13981 atgtgcatgc caaccacagg gttcccctcg ggatcaa //
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