质粒名称: | pH-PABE-7-esgRNA |
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出品公司: | 爱迪基因 |
目录编号: | 115620 |
存储实验室: | Caixia Gao |
载体骨架基本信息 |
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载体名称: | pCAMBIA1300 |
载体出品公司: | -- |
载体抗性: | 卡那霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 植物表达,CRISPR |
筛选标记: | -- |
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: | wtTadA-TadA7.10-nCas9-3*NLS |
物种来源: | -- |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | -- |
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: | pH-PABE-7-esgRNA was a gift from Caixia Gao (Addgene plasmid # 115620 ; http://n2t.net/addgene:115620 ; RRID:Addgene_115620) |
参考文献部分: | Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Li C, Zong Y, Wang Y, Jin S, Zhang D, Song Q, Zhang R, Gao C. Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. 10.1186/s13059-018-1443-z PubMed 29807545 |
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 pH-PABE-7-esgRNA 18869 bp ds-DNA circular SYN 11-JAN-2019 DEFINITION Targeted A to G in rice. ACCESSION . VERSION . KEYWORDS pH-PABE-7-esgRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 18869) AUTHORS Li C, Zong Y, Wang Y, Jin S, Zhang D, Song Q, Zhang R, Gao C TITLE Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. JOURNAL Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. PUBMED 29807545 REFERENCE 2 (bases 1 to 18869) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 11, 2019 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..18869 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 261..285 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" primer_bind 473..495 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 487..504 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 488..504 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 571..950 /label=OsU3 promoter /note="Oryza sativa (rice) snRNA U3 promoter" terminator 1058..1077 /label=SUP4 terminator /note="transcription terminator for the S. cerevisiae SUP4 tRNA gene" promoter 1090..3080 /label=Ubi promoter /note="maize polyubiquitin gene promoter" CDS 4288..8388 /codon_start=1 /product="nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9(D10A) /note="generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" RBS 8301..8309 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" CDS 8401..8421 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 8434..8454 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 8467..8487 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(9146..9162) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9146..9162) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(9159..9181) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 9170..9186 /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(9194..9224) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9239..9260 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 9451..10127 /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" primer_bind 10064..10084 /label=35S promoter /note="CaMV 35S promoter, forward primer" CDS 10194..11219 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK K" polyA_signal 11260..11434 /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" misc_feature 11512..11536 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS 11961..12755 /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" rep_origin 14180..14768 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 14669..14688 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 14922..14939 /label=L4440 /note="L4440 vector, forward primer" misc_feature 14954..15094 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(15020..15039) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 15180..15202 /label=pGEX 3' /note="pGEX vectors, reverse primer" rep_origin 15438..15632 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(15698..16771) /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI LVMRYRNLIEGEASAGS" CDS complement(17200..17829) /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" ORIGIN 1 gcgacaagaa aacgccagga aaagggcagg gcggcagcct gtcgcgtaac ttaggacttg 61 tgcgacatgt cgttttcaga agacggctgc actgaacgtc agaagccgac tgcactatag 121 cagcggaggg gttggatcaa agtactttga tcccgagggg aaccctgtgg ttggcatgca 181 catacaaatg gacgaacgga taaacctttt cacgcccttt taaatatccg attattctaa 241 taaacgctct tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta 301 aactgaaggc gggaaacgac aatctgatcc aagctcaagc tgctctagca ttcgccattc 361 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg 421 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca 481 cgacgttgta aaacgacggc cagtgccaag cttagtaatt catccaggtc accaagttct 541 aggattttca gaactgcaac ttattttatc aaggaatctt taaacatacg aacagatcac 601 ttaaagttct tctgaagcaa cttaaagtta tcaggcatgc atggatcttg gaggaatcag 661 atgtgcagtc agggaccata gcacaagaca ggcgtcttct actggtgcta ccagcaaatg 721 ctggaagccg ggaacactgg gtacgttgga aaccacgtga tgtgaagaag taagataaac 781 tgtaggagaa aagcatttcg tagtgggcca tgaagccttt caggacatgt attgcagtat 841 gggccggccc attacgcaat tggacgacaa caaagactag tattagtacc acctcggcta 901 tccacataga tcaaagctga tttaaaagag ttgtgcagat gatccgtggc gtgagaccct 961 gcagggtctc tgtttaagag ctatgctgga aacagcatag caagtttaaa taaggctagt 1021 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttgttttt tatgtctaag 1081 cttgcatgcc tgcagtgcag cgtgacccgg tcgtgcccct ctctagagat aatgagcatt 1141 gcatgtctaa gttataaaaa attaccacat attttttttg tcacacttgt ttgaagtgca 1201 gtttatctat ctttatacat atatttaaac tttactctac gaataatata atctatagta 1261 ctacaataat atcagtgttt tagagaatca tataaatgaa cagttagaca tggtctaaag 1321 gacaattgag tattttgaca acaggactct acagttttat ctttttagtg tgcatgtgtt 1381 ctcctttttt tttgcaaata gcttcaccta tataatactt catccatttt attagtacat 1441 ccatttaggg tttagggtta atggttttta tagactaatt tttttagtac atctatttta 1501 ttctatttta gcctctaaat taagaaaact aaaactctat tttagttttt ttatttaata 1561 atttagatat aaaatagaat aaaataaagt gactaaaaat taaacaaata ccctttaaga 1621 aattaaaaaa actaaggaaa catttttctt gtttcgagta gataatgcca gcctgttaaa 1681 cgccgtcgac gagtctaacg gacaccaacc agcgaaccag cagcgtcgcg tcgggccaag 1741 cgaagcagac ggcacggcat ctctgtcgct gcctctggac ccctctcgag agttccgctc 1801 caccgttgga cttgctccgc tgtcggcatc cagaaattgc gtggcggagc ggcagacgtg 1861 agccggcacg gcaggcggcc tcctcctcct ctcacggcac cggcagctac gggggattcc 1921 tttcccaccg ctccttcgct ttcccttcct cgcccgccgt aataaataga caccccctcc 1981 acaccctctt tccccaacct cgtgttgttc ggagcgcaca cacacacaac cagatctccc 2041 ccaaatccac ccgtcggcac ctccgcttca aggtacgccg ctcgtcctcc ccccccccct 2101 ctctaccttc tctagatcgg cgttccggtc catggttagg gcccggtagt tctacttctg 2161 ttcatgtttg tgttagatcc gtgtttgtgt tagatccgtg ctgctagcgt tcgtacacgg 2221 atgcgacctg tacgtcagac acgttctgat tgctaacttg ccagtgtttc tctttgggga 2281 atcctgggat ggctctagcc gttccgcaga cgggatcgat ttcatgattt tttttgtttc 2341 gttgcatagg gtttggtttg cccttttcct ttatttcaat atatgccgtg cacttgtttg 2401 tcgggtcatc ttttcatgct tttttttgtc ttggttgtga tgatgtggtc tggttgggcg 2461 gtcgttctag atcggagtag aattctgttt caaactacct ggtggattta ttaattttgg 2521 atctgtatgt gtgtgccata catattcata gttacgaatt gaagatgatg gatggaaata 2581 tcgatctagg ataggtatac atgttgatgc gggttttact gatgcatata cagagatgct 2641 ttttgttcgc ttggttgtga tgatgtggtg tggttgggcg gtcgttcatt cgttctagat 2701 cggagtagaa tactgtttca aactacctgg tgtatttatt aattttggaa ctgtatgtgt 2761 gtgtcataca tcttcatagt tacgagttta agatggatgg aaatatcgat ctaggatagg 2821 tatacatgtt gatgtgggtt ttactgatgc atatacatga tggcatatgc agcatctatt 2881 catatgctct aaccttgagt acctatctat tataataaac aagtatgttt tataattatt 2941 ttgatcttga tatacttgga tgatggcata tgcagcagct atatgtggat ttttttagcc 3001 ctgccttcat acgctattta tttgcttggt actgtttctt ttgtcgatgc tcaccctgtt 3061 gtttggtgtt acttctgcag ccctaggcct atgtccgagg tggagttctc tcacgagtat 3121 tggatgaggc acgctcttac acttgctaag agagcttggg acgaaagaga agtgccagtt 3181 ggcgccgttc ttgtgcataa taatagggtg atcggcgagg gttggaatag accaattgga 3241 aggcatgatc caacagctca cgcagagatt atggctctca gacaaggcgg cctcgttatg 3301 cagaactaca ggctcattga cgctacactc tacgtgacac tcgaaccttg cgttatgtgc 3361 gccggagcta tgattcattc taggattggc agggtcgtgt ttggagctag ggacgctaaa 3421 acaggagccg ccggatctct tatggacgtg ttgcatcatc caggcatgaa ccatagggtg 3481 gagattacag agggcattct tgcagacgag tgcgctgctc ttctttccga tttcttcagg 3541 atgagaaggc aggagattaa ggcccagaag aaggctcagt cttctacaga tagcggagga 3601 tcttccggag gatctagcgg ctccgagaca ccaggaacat ccgaaagcgc tacaccagaa 3661 tctagcggag gctcttccgg aggatcttct gaagtggagt tctcccacga gtattggatg 3721 aggcacgctc ttacacttgc taaaagggct agggacgaaa gggaagttcc agttggagct 3781 gttctcgtgc tcaataacag ggtgattggc gagggttgga atagagccat tggactccat 3841 gatccaacag ctcacgcaga gattatggct cttagacaag gcggcctcgt tatgcagaat 3901 tacagactca tcgacgccac actctacgtt accttcgaac cttgcgttat gtgcgccgga 3961 gctatgatcc attctaggat tggcagggtc gtgttcggcg ttagaaacgc taagacagga 4021 gctgcaggct ctcttatgga cgttcttcat tacccaggca tgaatcatag agtggagatc 4081 acagaaggca ttcttgcaga cgagtgcgca gctctccttt gctatttctt caggatgccg 4141 aggcaagttt tcaacgctca gaagaaggcc cagtcttcta cagattccgg cggatcttct 4201 ggaggatcta gcggctccga gacaccagga acatccgaat ccgctacacc agagtcttct 4261 ggaggatcta gcggaggatc tcttaaggac aagaagtact cgatcggcct cgccattggg 4321 actaactctg ttggctgggc cgtgatcacc gacgagtaca aggtgccctc aaagaagttc 4381 aaggtcctgg gcaacaccga tcggcattcc atcaagaaga atctcattgg cgctctcctg 4441 ttcgacagcg gcgagacggc tgaggctacg cggctcaagc gcaccgcccg caggcggtac 4501 acgcgcagga agaatcgcat ctgctacctg caggagattt tctccaacga gatggcgaag 4561 gttgacgatt ctttcttcca caggctggag gagtcattcc tcgtggagga ggataagaag 4621 cacgagcggc atccaatctt cggcaacatt gtcgacgagg ttgcctacca cgagaagtac 4681 cctacgatct accatctgcg gaagaagctc gtggactcca cagataaggc ggacctccgc 4741 ctgatctacc tcgctctggc ccacatgatt aagttcaggg gccatttcct gatcgagggg 4801 gatctcaacc cggacaatag cgatgttgac aagctgttca tccagctcgt gcagacgtac 4861 aaccagctct tcgaggagaa ccccattaat gcgtcaggcg tcgacgcgaa ggctatcctg 4921 tccgctaggc tctcgaagtc tcggcgcctc gagaacctga tcgcccagct gccgggcgag 4981 aagaagaacg gcctgttcgg gaatctcatt gcgctcagcc tggggctcac gcccaacttc 5041 aagtcgaatt tcgatctcgc tgaggacgcc aagctgcagc tctccaagga cacatacgac 5101 gatgacctgg ataacctcct ggcccagatc ggcgatcagt acgcggacct gttcctcgct 5161 gccaagaatc tgtcggacgc catcctcctg tctgatattc tcagggtgaa caccgagatt 5221 acgaaggctc cgctctcagc ctccatgatc aagcgctacg acgagcacca tcaggatctg 5281 accctcctga aggcgctggt caggcagcag ctccccgaga agtacaagga gatcttcttc 5341 gatcagtcga agaacggcta cgctgggtac attgacggcg gggcctctca ggaggagttc 5401 tacaagttca tcaagccgat tctggagaag atggacggca cggaggagct gctggtgaag 5461 ctcaatcgcg aggacctcct gaggaagcag cggacattcg ataacggcag catcccacac 5521 cagattcatc tcggggagct gcacgctatc ctgaggaggc aggaggactt ctaccctttc 5581 ctcaaggata accgcgagaa gatcgagaag attctgactt tcaggatccc gtactacgtc 5641 ggcccactcg ctaggggcaa ctcccgcttc gcttggatga cccgcaagtc agaggagacg 5701 atcacgccgt ggaacttcga ggaggtggtc gacaagggcg ctagcgctca gtcgttcatc 5761 gagaggatga cgaatttcga caagaacctg ccaaatgaga aggtgctccc taagcactcg 5821 ctcctgtacg agtacttcac agtctacaac gagctgacta aggtgaagta tgtgaccgag 5881 ggcatgagga agccggcttt cctgtctggg gagcagaaga aggccatcgt ggacctcctg 5941 ttcaagacca accggaaggt cacggttaag cagctcaagg aggactactt caagaagatt 6001 gagtgcttcg attcggtcga gatctctggc gttgaggacc gcttcaacgc ctccctgggg 6061 acctaccacg atctcctgaa gatcattaag gataaggact tcctggacaa cgaggagaat 6121 gaggatatcc tcgaggacat tgtgctgaca ctcactctgt tcgaggaccg ggagatgatc 6181 gaggagcgcc tgaagactta cgcccatctc ttcgatgaca aggtcatgaa gcagctcaag 6241 aggaggaggt acaccggctg ggggaggctg agcaggaagc tcatcaacgg cattcgggac 6301 aagcagtccg ggaagacgat cctcgacttc ctgaagagcg atggcttcgc gaaccgcaat 6361 ttcatgcagc tgattcacga tgacagcctc acattcaagg aggatatcca gaaggctcag 6421 gtgagcggcc agggggactc gctgcacgag catatcgcga acctcgctgg ctcgccagct 6481 atcaagaagg ggattctgca gaccgtgaag gttgtggacg agctggtgaa ggtcatgggc 6541 aggcacaagc ctgagaacat cgtcattgag atggcccggg agaatcagac cacgcagaag 6601 ggccagaaga actcacgcga gaggatgaag aggatcgagg agggcattaa ggagctgggg 6661 tcccagatcc tcaaggagca cccggtggag aacacgcagc tgcagaatga gaagctctac 6721 ctgtactacc tccagaatgg ccgcgatatg tatgtggacc aggagctgga tattaacagg 6781 ctcagcgatt acgacgtcga tcatatcgtt ccacagtcat tcctgaagga tgactccatt 6841 gacaacaagg tcctcaccag gtcggacaag aaccggggca agtctgataa tgttccttca 6901 gaggaggtcg ttaagaagat gaagaactac tggcgccagc tcctgaatgc caagctgatc 6961 acgcagcgga agttcgataa cctcacaaag gctgagaggg gcgggctctc tgagctggac 7021 aaggcgggct tcatcaagag gcagctggtc gagacacggc agatcactaa gcacgttgcg 7081 cagattctcg actcacggat gaacactaag tacgatgaga atgacaagct gatccgcgag 7141 gtgaaggtca tcaccctgaa gtcaaagctc gtctccgact tcaggaagga tttccagttc 7201 tacaaggttc gggagatcaa caattaccac catgcccatg acgcgtacct gaacgcggtg 7261 gtcggcacag ctctgatcaa gaagtaccca aagctcgaga gcgagttcgt gtacggggac 7321 tacaaggttt acgatgtgag gaagatgatc gccaagtcgg agcaggagat tggcaaggct 7381 accgccaagt acttcttcta ctctaacatt atgaatttct tcaagacaga gatcactctg 7441 gccaatggcg agatccggaa gcgccccctc atcgagacga acggcgagac gggggagatc 7501 gtgtgggaca agggcaggga tttcgcgacc gtcaggaagg ttctctccat gccacaagtg 7561 aatatcgtca agaagacaga ggtccagact ggcgggttct ctaaggagtc aattctgcct 7621 aagcggaaca gcgacaagct catcgcccgc aagaaggact gggatccgaa gaagtacggc 7681 gggttcgaca gccccactgt ggcctactcg gtcctggttg tggcgaaggt tgagaagggc 7741 aagtccaaga agctcaagag cgtgaaggag ctgctgggga tcacgattat ggagcgctcc 7801 agcttcgaga agaacccgat cgatttcctg gaggcgaagg gctacaagga ggtgaagaag 7861 gacctgatca ttaagctccc caagtactca ctcttcgagc tggagaacgg caggaagcgg 7921 atgctggctt ccgctggcga gctgcagaag gggaacgagc tggctctgcc gtccaagtat 7981 gtgaacttcc tctacctggc ctcccactac gagaagctca agggcagccc cgaggacaac 8041 gagcagaagc agctgttcgt cgagcagcac aagcattacc tcgacgagat cattgagcag 8101 atttccgagt tctccaagcg cgtgatcctg gccgacgcga atctggataa ggtcctctcc 8161 gcgtacaaca agcaccgcga caagccaatc agggagcagg ctgagaatat cattcatctc 8221 ttcaccctga cgaacctcgg cgcccctgct gctttcaagt acttcgacac aactatcgat 8281 cgcaagaggt acacaagcac taaggaggtc ctggacgcga ccctcatcca ccagtcgatt 8341 accggcctct acgagacgcg catcgacctg tctcagctcg ggggcgactc cggcggcagc 8401 ccaaagaaga agcggaaggt gtctggaggt tctcctaaga aaaagagaaa agtgtccggc 8461 ggctccccga agaagaagcg caaggtgtag gagctcagag ctttcgttcg tatcatcggt 8521 ttcgacaacg ttcgtcaagt tcaatgcatc agtttcattg cgcacacacc agaatcctac 8581 tgagtttgag tattatggca ttgggaaaac tgtttttctt gtaccatttg ttgtgcttgt 8641 aatttactgt gttttttatt cggttttcgc tatcgaactg tgaaatggaa atggatggag 8701 aagagttaat gaatgatatg gtccttttgt tcattctcaa attaatatta tttgtttttt 8761 ctcttatttg ttgtgtgttg aatttgaaat tataagagat atgcaaacat tttgttttga 8821 gtaaaaatgt gtcaaatcgt ggcctctaat gaccgaagtt aatatgagga gtaaaacact 8881 tgtagttgta ccattatgct tattcactag gcaacaaata tattttcaga cctagaaaag 8941 ctgcaaatgt tactgaatac aagtatgtcc tcttgtgttt tagacattta tgaactttcc 9001 tttatgtaat tttccagaat ccttgtcaga ttctaatcat tgctttataa ttatagttat 9061 actcatggat ttgtagttga gtatgaaaat attttttaat gcattttatg acttgccaat 9121 tgattgacaa cgaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc 9181 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 9241 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 9301 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 9361 gctagagcag cttgccaaca tggtggagca cgacactctc gtctactcca agaatatcaa 9421 agatacagtc tcagaagacc aaagggctat tgagactttt caacaaaggg taatatcggg 9481 aaacctcctc ggattccatt gcccagctat ctgtcacttc atcaaaagga cagtagaaaa 9541 ggaaggtggc acctacaaat gccatcattg cgataaagga aaggctatcg ttcaagatgc 9601 ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga 9661 agacgttcca accacgtctt caaagcaagt ggattgatgt gaacatggtg gagcacgaca 9721 ctctcgtcta ctccaagaat atcaaagata cagtctcaga agaccaaagg gctattgaga 9781 cttttcaaca aagggtaata tcgggaaacc tcctcggatt ccattgccca gctatctgtc 9841 acttcatcaa aaggacagta gaaaaggaag gtggcaccta caaatgccat cattgcgata 9901 aaggaaaggc tatcgttcaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac 9961 ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt 10021 gatgtgatat ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc 10081 cttcctctat ataaggaagt tcatttcatt tggagaggac acgctgaaat caccagtctc 10141 tctctacaaa tctatctctc tcgagctttc gcagatccgg ggggcaatga gatatgaaaa 10201 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct 10261 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag 10321 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg 10381 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggagt 10441 ttagcgagag cctgacctat tgcatctccc gccgttcaca gggtgtcacg ttgcaagacc 10501 tgcctgaaac cgaactgccc gctgttctac aaccggtcgc ggaggctatg gatgcgatcg 10561 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc 10621 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc 10681 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc 10741 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca 10801 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg 10861 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg 10921 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccacgactcc 10981 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt 11041 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga 11101 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag 11161 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aagaaataga 11221 gtagatgccg accgggatct gtcgatcgac aagctcgagt ttctccataa taatgtgtga 11281 gtagttccca gataagggaa ttagggttcc tatagggttt cgctcatgtg ttgagcatat 11341 aagaaaccct tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt 11401 cctaaaacca aaatccagta ctaaaatcca gatcccccga attaattcgg cgttaattca 11461 gtacattaaa aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc 11521 acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc 11581 accactcgat acaggcagcc catcagtccg ggacggcgtc agcgggagag ccgttgtaag 11641 gcggcagact ttgctcatgt taccgatgct attcggaaga acggcaacta agctgccggg 11701 tttgaaacac ggatgatctc gcggagggta gcatgttgat tgtaacgatg acagagcgtt 11761 gctgcctgtg atcaccgcgg tttcaaaatc ggctccgtcg atactatgtt atacgccaac 11821 tttgaaaaca actttgaaaa agctgttttc tggtatttaa ggttttagaa tgcaaggaac 11881 agtgaattgg agttcgtctt gttataatta gcttcttggg gtatctttaa atactgtaga 11941 aaagaggaag gaaataataa atggctaaaa tgagaatatc accggaattg aaaaaactga 12001 tcgaaaaata ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag gtatataagc 12061 tggtgggaga aaatgaaaac ctatatttaa aaatgacgga cagccggtat aaagggacca 12121 cctatgatgt ggaacgggaa aaggacatga tgctatggct ggaaggaaag ctgcctgttc 12181 caaaggtcct gcactttgaa cggcatgatg gctggagcaa tctgctcatg agtgaggccg 12241 atggcgtcct ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag attatcgagc 12301 tgtatgcgga gtgcatcagg ctctttcact ccatcgacat atcggattgt ccctatacga 12361 atagcttaga cagccgctta gccgaattgg attacttact gaataacgat ctggccgatg 12421 tggattgcga aaactgggaa gaagacactc catttaaaga tccgcgcgag ctgtatgatt 12481 ttttaaagac ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac ctgggagaca 12541 gcaacatctt tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg agaagcggca 12601 gggcggacaa gtggtatgac attgccttct gcgtccggtc gatcagggag gatatcgggg 12661 aagaacagta tgtcgagcta ttttttgact tactggggat caagcctgat tgggagaaaa 12721 taaaatatta tattttactg gatgaattgt tttagtacct agaatgcatg accaaaatcc 12781 cttaacgctg atgaatcccc taatgatttt ggtaaaaatc attaagttaa ggtggataca 12841 catcttgtca tatgatcaaa tggtttcgcg aaaaatcaat aatcagacaa caagatgtgc 12901 gaactcgata ttttacacga ctctctttac caattctgcc ccgaattaca cttaaaacga 12961 ctcaacagct taacgttggc ttgccacgca ttacttgact gtaaaactct cactcttacc 13021 gaacttggcc gtaacctgcc aaccaaagcg agaacaaaac ataacatcaa acgaatcgac 13081 cgattgttag gtaatcgtca cctccacaaa gagcgactcg ctgtataccg ttggcatgct 13141 agctttatct gttcgggcaa tacgatgccc attgtacttg ttgactggtc tgatattcgt 13201 gagcaaaaac gacttatggt attgcgagct tcagtcgcac tacacggtcg ttctgttact 13261 ctttatgaga aagcgttccc gctttcagag caatgttcaa agaaagctca tgaccaattt 13321 ctagccgacc ttgcgagcat tctaccgagt aacaccacac cgctcattgt cagtgatgct 13381 ggctttaaag tgccatggta taaatccgtt gagaagctgg gttggtactg gttaagtcga 13441 gtaagaggaa aagtacaata tgcagaccta ggagcggaaa actggaaacc tatcagcaac 13501 ttacatgata tgtcatctag tcactcaaag actttaggct ataagaggct gactaaaagc 13561 aatccaatct catgccaaat tctattgtat aaatctcgct ctaaaggccg aaaaaatcag 13621 cgctcgacac ggactcattg tcaccacccg tcacctaaaa tctactcagc gtcggcaaag 13681 gagccatggg ttctagcaac taacttacct gttgaaattc gaacacccaa acaacttgtt 13741 aatatctatt cgaagcgaat gcagattgaa gaaaccttcc gagacttgaa aagtcctgcc 13801 tacggactag gcctacgcca tagccgaacg agcagctcag agcgttttga tatcatgctg 13861 ctaatcgccc tgatgcttca actaacatgt tggcttgcgg gcgttcatgc tcagaaacaa 13921 ggttgggaca agcacttcca ggctaacaca gtcagaaatc gaaacgtact ctcaacagtt 13981 cgcttaggca tggaagtttt gcggcattct ggctacacaa taacaaggga agacttactc 14041 gtggctgcaa ccctactagc tcaaaattta ttcacacatg gttacgcttt ggggaaatta 14101 tgaggggatc tctcagccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 14161 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 14221 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 14281 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc 14341 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 14401 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 14461 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 14521 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 14581 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 14641 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 14701 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 14761 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 14821 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg 14881 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg 14941 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc 15001 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac 15061 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga 15121 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc 15181 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagggtgc 15241 cttgatgtgg gcgccggcgg tcgagtggcg acggcgcggc ttgtccgcgc cctggtagat 15301 tgcctggccg taggccagcc atttttgagc ggccagcggc cgcgataggc cgacgcgaag 15361 cggcggggcg tagggagcgc agcgaccgaa gggtaggcgc tttttgcagc tcttcggctg 15421 tgcgctggcc agacagttat gcacaggcca ggcgggtttt aagagtttta ataagtttta 15481 aagagtttta ggcggaaaaa tcgccttttt tctcttttat atcagtcact tacatgtgtg 15541 accggttccc aatgtacggc tttgggttcc caatgtacgg gttccggttc ccaatgtacg 15601 gctttgggtt cccaatgtac gtgctatcca caggaaacag accttttcga cctttttccc 15661 ctgctagggc aatttgccct agcatctgct ccgtacatta ggaaccggcg gatgcttcgc 15721 cctcgatcag gttgcggtag cgcatgacta ggatcgggcc agcctgcccc gcctcctcct 15781 tcaaatcgta ctccggcagg tcatttgacc cgatcagctt gcgcacggtg aaacagaact 15841 tcttgaactc tccggcgctg ccactgcgtt cgtagatcgt cttgaacaac catctggctt 15901 ctgccttgcc tgcggcgcgg cgtgccaggc ggtagagaaa acggccgatg ccgggatcga 15961 tcaaaaagta atcggggtga accgtcagca cgtccgggtt cttgccttct gtgatctcgc 16021 ggtacatcca atcagctagc tcgatctcga tgtactccgg ccgcccggtt tcgctcttta 16081 cgatcttgta gcggctaatc aaggcttcac cctcggatac cgtcaccagg cggccgttct 16141 tggccttctt cgtacgctgc atggcaacgt gcgtggtgtt taaccgaatg caggtttcta 16201 ccaggtcgtc tttctgcttt ccgccatcgg ctcgccggca gaacttgagt acgtccgcaa 16261 cgtgtggacg gaacacgcgg ccgggcttgt ctcccttccc ttcccggtat cggttcatgg 16321 attcggttag atgggaaacc gccatcagta ccaggtcgta atcccacaca ctggccatgc 16381 cggccggccc tgcggaaacc tctacgtgcc cgtctggaag ctcgtagcgg atcacctcgc 16441 cagctcgtcg gtcacgcttc gacagacgga aaacggccac gtccatgatg ctgcgactat 16501 cgcgggtgcc cacgtcatag agcatcggaa cgaaaaaatc tggttgctcg tcgcccttgg 16561 gcggcttcct aatcgacggc gcaccggctg ccggcggttg ccgggattct ttgcggattc 16621 gatcagcggc cgcttgccac gattcaccgg ggcgtgcttc tgcctcgatg cgttgccgct 16681 gggcggcctg cgcggccttc aacttctcca ccaggtcatc acccagcgcc gcgccgattt 16741 gtaccgggcc ggatggtttg cgaccgctca cgccgattcc tcgggcttgg gggttccagt 16801 gccattgcag ggccggcagg caacccagcc gcttacgcct ggccaaccgc ccgttcctcc 16861 acacatgggg cattccacgg cgtcggtgcc tggttgttct tgattttcca tgccgcctcc 16921 tttagccgct aaaattcatc tactcattta ttcatttgct catttactct ggtagctgcg 16981 cgatgtattc agatagcagc tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca 17041 gcttggtgtg atcctccgcc ggcaactgaa agttgacccg cttcatggct ggcgtgtctg 17101 ccaggctggc caacgttgca gccttgctgc tgcgtgcgct cggacggccg gcacttagcg 17161 tgtttgtgct tttgctcatt ttctctttac ctcattaact caaatgagtt ttgatttaat 17221 ttcagcggcc agcgcctgga cctcgcgggc agcgtcgccc tcgggttctg attcaagaac 17281 ggttgtgccg gcggcggcag tgcctgggta gctcacgcgc tgcgtgatac gggactcaag 17341 aatgggcagc tcgtacccgg ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt 17401 gatcgcccgc gacacgacaa aggccgcttg tagccttcca tccgtgacct caatgcgctg 17461 cttaaccagc tccaccaggt cggcggtggc ccatatgtcg taagggcttg gctgcaccgg 17521 aatcagcacg aagtcggctg ccttgatcgc ggacacagcc aagtccgccg cctggggcgc 17581 tccgtcgatc actacgaagt cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg 17641 gcggtcgatg ccgacaacgg ttagcggttg atcttcccgc acggccgccc aatcgcgggc 17701 actgccctgg ggatcggaat cgactaacag aacatcggcc ccggcgagtt gcagggcgcg 17761 ggctagatgg gttgcgatgg tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat 17821 aaccttcatg cgttcccctt gcgtatttgt ttatttactc atcgcatcat atacgcagcg 17881 accgcatgac gcaagctgtt ttactcaaat acacatcacc tttttagacg gcggcgctcg 17941 gtttcttcag cggccaagct ggccggccag gccgccagct tggcatcaga caaaccggcc 18001 aggatttcat gcagccgcac ggttgagacg tgcgcgggcg gctcgaacac gtacccggcc 18061 gcgatcatct ccgcctcgat ctcttcggta atgaaaaacg gttcgtcctg gccgtcctgg 18121 tgcggtttca tgcttgttcc tcttggcgtt cattctcggc ggccgccagg gcgtcggcct 18181 cggtcaatgc gtcctcacgg aaggcaccgc gccgcctggc ctcggtgggc gtcacttcct 18241 cgctgcgctc aagtgcgcgg tacagggtcg agcgatgcac gccaagcagt gcagccgcct 18301 ctttcacggt gcggccttcc tggtcgatca gctcgcgggc gtgcgcgatc tgtgccgggg 18361 tgagggtagg gcgggggcca aacttcacgc ctcgggcctt ggcggcctcg cgcccgctcc 18421 gggtgcggtc gatgattagg gaacgctcga actcggcaat gccggcgaac acggtcaaca 18481 ccatgcggcc ggccggcgtg gtggtgtcgg cccacggctc tgccaggcta cgcaggcccg 18541 cgccggcctc ctggatgcgc tcggcaatgt ccagtaggtc gcgggtgctg cgggccaggc 18601 ggtctagcct ggtcactgtc acaacgtcgc cagggcgtag gtggtcaagc atcctggcca 18661 gctccgggcg gtcgcgcctg gtgccggtga tcttctcgga aaacagcttg gtgcagccgg 18721 ccgcgtgcag ttcggcccgt tggttggtca agtcctggtc gtcggtgctg acgcgggcat 18781 agcccagcag gccagcggcg gcgctcttgt tcatggcgta atgtctccgg ttctagtcgc 18841 aagtattcta ctttatgcga ctaaaacac //
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