质粒名称: | pRGE32 |
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出品公司: | 爱迪基因 |
目录编号: | 63159 |
存储实验室: | Yinong Yang |
载体骨架基本信息 |
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载体名称: | pRGE31 |
载体出品公司: | Kabin Xie |
载体抗性: | 氨苄青霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | 用水稻UBI启动子+5'-UTR替换35S启动子。 |
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: | -- |
物种来源: | -- |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | Rice snoRNA U3 promoter for gRNA/PTG expression and UBI promoter for Cas9 expression |
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 | M13R(-48) |
3' 测序引物 | M13F(-20) |
文献引用方法 |
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材料和方法部分: | pRGE32 was a gift from Yinong Yang (Addgene plasmid # 63159 ; http://n2t.net/addgene:63159 ; RRID:Addgene_63159) |
参考文献部分: | Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. Xie K, Minkenberg B, Yang Y. Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. 10.1073/pnas.1420294112 PubMed 25733849 |
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 pRGE32 10092 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Express sgRNA/PTG with rice snoRNA U3 promoter and Cas9 with rice ubiquitin promoter for transient expression in protoplast. ACCESSION . VERSION . KEYWORDS pRGE32 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10092) AUTHORS Xie K, Minkenberg B, Yang Y TITLE Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. JOURNAL Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. PUBMED 25733849 REFERENCE 2 (bases 1 to 10092) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10092 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 26..49 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" terminator 71..323 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" primer_bind complement(332..349) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(332..348) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(341..363) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 561..1016 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(648..667) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 858..879 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1033..1052) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1152..1174 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1212..1230) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1298..1402 /gene="bla" /label=AmpR promoter CDS 1403..2263 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGTRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1621..1640) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2434..3022 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2923..2942 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3176..3193 /label=L4440 /note="L4440 vector, forward primer" protein_bind 3310..3331 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3346..3376 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3384..3400 /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 3389..3411 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 3408..3424 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 3408..3424 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 3443..3823 /label=OsU3 promoter /note="Oryza sativa (rice) snRNA U3 promoter" misc_RNA 3840..3915 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter 3996..5705 /gene="RUBQ2" /label=Ubi promoter /note="promoter for the rice (Oryza sativa) polyubiquitin gene RUBQ2" intron 4744..5705 /note="intron upstream of the coding sequence for the rice polyubiquitin gene RUBQ2" protein_bind 5716..5740 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 5785..5850 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 5857..5877 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5902..10002 /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 10003..10050 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" protein_bind complement(10083..15) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" ORIGIN 1 cttgtacaaa gtggttgata acagcgacta caaggatgac gatgacaagg cttagagctc 61 gaatttcccc gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc 121 cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg taataattaa 181 catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata 241 catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc 301 ggtgtcatct atgttactag atcgggaatt cactggccgt cgttttacaa cgtcgtgact 361 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct 421 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg 481 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca 541 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 601 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 661 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 721 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg 781 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 841 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 901 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 961 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg 1021 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc 1081 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc 1141 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc 1201 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt 1261 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 1321 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 1381 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt 1441 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga 1501 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa 1561 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct 1621 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat 1681 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga 1741 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc 1801 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat 1861 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa 1921 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac 1981 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa 2041 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc 2101 tggagccggt gagcgtggca ctcgcggtat cattgcagca ctggggccag atggtaagcc 2161 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag 2221 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta 2281 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 2341 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 2401 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 2461 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 2521 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 2581 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 2641 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 2701 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 2761 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 2821 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 2881 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 2941 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 3001 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 3061 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 3121 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga 3181 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg 3241 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg 3301 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct 3361 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta 3421 tgaccatgat tacgccaagc ttaaggaatc tttaaacata cgaacagatc acttaaagtt 3481 cttctgaagc aacttaaagt tatcaggcat gcatggatct tggaggaatc agatgtgcag 3541 tcagggacca tagcacaaga caggcgtctt ctactggtgc taccagcaaa tgctggaagc 3601 cgggaacact gggtacgttg gaaaccacgt gatgtgaaga agtaagataa actgtaggag 3661 aaaagcattt cgtagtgggc catgaagcct ttcaggacat gtattgcagt atgggccggc 3721 ccattacgca attggacgac aacaaagact agtattagta ccacctcggc tatccacata 3781 gatcaaagct gatttaaaag agttgtgcag atgatccgtg gcaggagacc gaggtctcgg 3841 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg 3901 gcaccgagtc ggtgcttttt tgttttagag ctagaaatag caagttaaaa taaggctagt 3961 ccgtttttag cgcgtgcatg cctgcaggtc cacaaattcg ggtcaaggcg gaagccagcg 4021 cgccacccca cgtcagcaaa tacggaggcg cggggttgac ggcgtcaccc ggtcctaacg 4081 gcgaccaaca aaccagccag aagaaattac agtaaaaaaa aagtaaattg cactttgatc 4141 caccttttat tacctaagtc tcaatttgga tcacccttaa acctatcttt tcaatttggg 4201 ccgggttgtg gtttggacta ccatgaacaa cttttcgtca tgtctaactt ccctttcagc 4261 aaacatatga accatatata gaggagatcg gccgtatact agagctgatg tgtttaaggt 4321 cgttgattgc acgagaaaaa aaaatccaaa tcgcaacaat agcaaattta tctggttcaa 4381 agtgaaaaga tatgtttaaa ggtagtccaa agtaaaactt atagataata aaatgtggtc 4441 caaagcgtaa ttcactcaaa aaaaatcaac gagacgtgta ccaaacggag acaaacggca 4501 tcttctcgaa atttcccaac cgctcgctcg cccgcctcgt cttcccggaa accgcggtgg 4561 tttcagcgtg gcggattctc caagcagacg gagacgtcac ggcacgggac tcctcccacc 4621 acccaaccgc cataaatacc agccccctca tctcctctcc tcgcatcagc tccacccccg 4681 aaaaatttct ccccaatctc gcgaggctct cgtcgtcgaa tcgaatcctc tcgcgtcctc 4741 aaggtacgct gcttctcctc tcctcgcttc gtttcgattc gatttcggac gggtgaggtt 4801 gttttgttgc tagatccgat tggtggttag ggttgtcgat gtgattatcg tgagatgttt 4861 aggggttgta gatctgatgg ttgtgatttg ggcacggttg gttcgatagg tggaatcgtg 4921 gttaggtttt gggattggat gttggttctg atgattgggg ggaattttta cggttagatg 4981 aattgttgga tgattcgatt ggggaaatcg gtgtagatct gttggggaat tgtggaacta 5041 gtcatgcctg agtgattggt gcgatttgta gcgtgttcca tcttgtaggc cttgttgcga 5101 gcatgttcag atctactgtt ccgctcttga ttgagttatt ggtgccatgg gttggtgcaa 5161 acacaggctt taatatgtta tatctgtttt gtgtttgatg tagatctgta gggtagttct 5221 tcttagacat ggttcaatta tgtagcttgt gcgtttcgat ttgatttcat atgttcacag 5281 attagataat gatgaactct tttaattaat tgtcaatggt aaataggaag tcttgtcgct 5341 atatctgtca taatgatctc atgttactat ctgccagtaa tttatgctaa gaactatatt 5401 agaatatcat gttacaatct gtagtaatat catgttacaa tctgtagttc atctatataa 5461 tctattgtgg taatttcttt ttactatctg tgtgaagatt attgccacta gttcattcta 5521 cttatttctg aagttcagga tacgtgtgct gttactacct atctgaatac atgtgtgatg 5581 tgcctgttac tatctttttg aatacatgta tgttctgttg gaatatgttt gctgtttgat 5641 ccgttgttgt gtccttaatc ttgtgctagt tcttacccta tctgtttggt gattatttct 5701 tgcagatagt tatcaacaag tttgtacaaa aaagcaggct tcgaaggaga tagaaccaat 5761 tctctaagga aatacttaac catggactat aaggaccacg acggagacta caaggatcat 5821 gatattgatt acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt 5881 atccacggag tcccagcagc cgacaagaag tacagcatcg gcctggacat cggcaccaac 5941 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg 6001 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac 6061 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga 6121 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac 6181 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag 6241 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc 6301 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc 6361 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg 6421 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag 6481 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc 6541 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag 6601 aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc 6661 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac 6721 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag 6781 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag 6841 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg 6901 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag 6961 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag 7021 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac 7081 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc 7141 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag 7201 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct 7261 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc 7321 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg 7381 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg 7441 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg 7501 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag 7561 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc 7621 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac 7681 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac 7741 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa 7801 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg 7861 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag 7921 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg 7981 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc 8041 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag 8101 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac 8161 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag 8221 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag 8281 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac 8341 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc 8401 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac 8461 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag 8521 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag 8581 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc 8641 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc 8701 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa 8761 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa 8821 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga 8881 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag 8941 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc 9001 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac 9061 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg 9121 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc 9181 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg 9241 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc 9301 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc 9361 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc 9421 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg 9481 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg 9541 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac 9601 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag 9661 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc 9721 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac 9781 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc 9841 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag 9901 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc 9961 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg 10021 aaaaaggccg gccaggcaaa aaagaaaaag taagaattcg cggccgcact cgagatatct 10081 agacccagct tt //
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