质粒名称: | pST1374-GCN4-D10A |
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出品公司: | 爱迪基因 |
目录编号: | 113022 |
存储实验室: | Xingxu Huang |
载体骨架基本信息 |
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载体名称: | pST1374 |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 4968 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
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: | GCN4-D10A,2xNLS-10xGCN4-NLS-D10A-NLS |
物种来源: | S. cerevisiae (budding yeast); S. pyogenes |
插入大小: | 5634 bp |
基因突变: | D10A |
GenBank编号 | -- |
启动子 | CMV |
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 | CMV-fwd |
3' 测序引物 | BGH-rev |
文献引用方法 |
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材料和方法部分: | pST1374-GCN4-D10A was a gift from Xingxu Huang (Addgene plasmid # 113022 ; http://n2t.net/addgene:113022 ; RRID:Addgene_113022) |
参考文献部分: | BE-PLUS: a new base editing tool with broadened editing window and enhanced fidelity. Jiang W, Feng S, Huang S, Yu W, Li G, Yang G, Liu Y, Zhang Y, Zhang L, Hou Y, Chen J, Chen J, Huang X. Cell Res. 2018 Jun 6. pii: 10.1038/s41422-018-0052-4. doi: 10.1038/s41422-018-0052-4. 10.1038/s41422-018-0052-4 [pii] PubMed 29875396 |
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载体基本信息
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载体序列
LOCUS pST1374-GCN4-D10A 10601 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Expresses GCN4-D10A in mammalian cells. ACCESSION . VERSION . KEYWORDS pST1374-GCN4-D10A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10601) AUTHORS Jiang W, Feng S, Huang S, Yu W, Li G, Yang G, Liu Y, Zhang Y, Zhang L, Hou Y, Chen J, Chen J, Huang X TITLE BE-PLUS: a new base editing tool with broadened editing window and enhanced fidelity. JOURNAL Cell Res. 2018 Jun 6. pii: 10.1038/s41422-018-0052-4. doi: 10.1038/s41422-018-0052-4. PUBMED 29875396 REFERENCE 2 (bases 1 to 10601) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10601 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 676..696 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 703..723 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 760..816 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 898..954 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 1036..1092 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 1174..1230 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 1312..1368 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 1450..1506 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 1588..1644 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 1726..1782 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 1864..1920 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 2002..2058 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 2098..2118 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2125..6225 /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" CDS 6256..6276 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 6283..6300 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(6323..6340) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6329..6553 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 6599..7027 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6686..6705) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6896..6917 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7036..7056) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 7041..7370 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 7221..7356 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7283..7302 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 7418..7465 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 7484..7882 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" polyA_signal 8040..8161 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8077..8096) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8131..8150 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(8210..8226) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8210..8226) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8223..8245) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8234..8250 /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(8258..8288) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8303..8324 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8441..8458) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8612..9200) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8692..8711) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9371..10231) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 9994..10013 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10232..10336) /gene="bla" /label=AmpR promoter primer_bind complement(10411..10430) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg 661 gctagcacca tgggacccaa gaaaaaacgc aaggtggaag atcctaagaa aaagcggaaa 721 gtggacggca ttggtagtgg gagcaacggc agcagcggag aggaactgct gagcaagaac 781 taccacctgg aaaacgaggt ggccagactg aagaaaggct ctggctctgg cggaagcggt 841 tctggcggat caggatctgg aagtggcggc tctggaagcg gaggttctgg atctggcgaa 901 gaactgctgt ctaagaatta tcacctcgag aacgaagtgg ctcggctcaa gaaaggcagt 961 ggcagcggag gaagtggatc cggcggtagt ggtagtggaa gcggcggatc cggctcaggc 1021 ggatctggtt caggggaaga actcctctcc aaaaactacc atctcgagaa tgaggtcgcc 1081 cgcctgaaaa aaggatcagg ttccggtggt tctggcagcg gtggttcagg ctcaggcagc 1141 ggcggtagcg gtagcggtgg aagcggaagc ggagaagaac ttctcagcaa aaattaccac 1201 ttggagaatg aagttgcaag actcaaaaaa ggttccggca gtggcggcag cggcagcggc 1261 ggatctggta gtggatctgg tggcagtggt tcaggcggaa gtggtagcgg agaggaattg 1321 ctctcaaaga actatcattt ggagaacgag gttgcacgct tgaagaaagg cagcggatca 1381 ggcggatctg gcagcggtgg atctggttct ggatccggcg gctccggtag tggtggaagt 1441 ggctctgggg aagaattgct tagcaagaat tatcatcttg aaaatgaggt tgccaggctt 1501 aaaaaaggca gtggctccgg cggatccgga agcggaggca gcggatctgg atctggtggt 1561 tcaggatctg gcggttctgg tagcggggaa gaactgttga gtaaaaacta tcaccttgag 1621 aacgaggtcg caaggttgaa aaaaggatcc ggctctggcg gctccggaag tggcggatct 1681 ggctccggta gcggaggatc aggatccggc ggaagcggat caggcgagga actgctttcc 1741 aaaaattacc accttgaaaa cgaagtcgcc cgcctcaaga aaggttctgg cagcggaggc 1801 tctggcagtg gtggtagcgg aagtggaagt ggtggcagtg gtagcggtgg atctggaagc 1861 ggcgaggaac tcctgtcaaa gaattaccat ctcgaaaacg aggtcgcaag gctcaagaaa 1921 ggctcaggat caggcggctc tggatccggc ggttctggtt ccggctcagg tggaagtgga 1981 tctggcggct caggttccgg cgaagaattg ctttccaaga actaccattt ggaaaatgaa 2041 gtcgctcgtt tgaagaaagg ttcaggctcc ggccagcggc cgcaaggtgg aggtggaccc 2101 aagaagaagc gcaaggtgtc tagagacaag aaatactcta ttggactggc tatcgggaca 2161 aactccgttg gctgggccgt cataaccgac gagtataagg tgccaagcaa gaaattcaag 2221 gtgctgggta atactgaccg ccattcaatc aagaagaacc tgatcggagc actcctcttc 2281 gactccggtg aaaccgctga agctactcgg ctgaagcgga ccgcaaggcg gagatacacc 2341 cgccgcaaga atcggatatg ttatctgcaa gagatcttta gcaacgaaat ggctaaggtg 2401 gacgactcct tctttcaccg cctggaagag agctttctgg tggaggagga taagaaacac 2461 gagaggcacc ctatattcgg aaatatcgtg gatgaggtgg cttaccatga aaagtatcct 2521 acaatctacc atctgaggaa gaagctggtg gacagcaccg ataaagcaga cctgaggctc 2581 atctatctgg ccctggctca tatgataaag tttagaggac actttctgat cgagggcgac 2641 ctgaatcccg ataattccga tgtggataaa ctcttcattc aactggtgca gacatataac 2701 caactgttcg aggagaatcc cataaacgct tctggtgtgg atgccaaggc tattctgtcc 2761 gctcggctgt ccaagtcacg cagactggag aatctgattg cccaactgcc aggagaaaag 2821 aagaacggcc tgtttgggaa cctcatcgcc ctgagcctgg gcctgacacc taacttcaag 2881 tccaattttg atctggccga agatgctaaa ctccagctct ccaaggacac ctatgacgat 2941 gatctggaca acctgctcgc acagataggc gaccagtacg ccgatctctt tctggctgct 3001 aagaatctct ccgacgccat tctgctgagc gacatactcc gggtcaacac tgagatcacc 3061 aaagcacctc tgagcgcctc catgataaaa cgctatgatg aacaccatca agacctgact 3121 ctgctcaaag ccctcgtgag gcaacagctg ccagagaagt acaaagagat attcttcgac 3181 cagagcaaga atggatatgc cggatacatc gatggcggag catcacagga agaattttac 3241 aagttcatca aaccaatcct cgagaagatg gacggtactg aagagctgct ggtgaagctg 3301 aacagggagg acctgctgag gaagcagagg acctttgata atggctccat tccacatcag 3361 atacacctgg gagagctgca tgcaatcctc cgcaggcagg aggatttcta tcctttcctg 3421 aaggataacc gggagaagat agagaagatc ctgaccttca ggatccctta ttacgtcggc 3481 cctctggcta gaggcaactc ccgcttcgct tggatgacca ggaaatctga ggagacaatt 3541 actccttgga acttcgaaga ggtcgtggat aagggcgcaa gcgcccagtc attcatcgaa 3601 cggatgacca atttcgataa gaacctgccc aacgagaagg tcctgcccaa acattcactc 3661 ctgtacgagt atttcaccgt ctataacgag ctgactaaag tgaagtacgt gaccgagggc 3721 atgaggaagc ctgccttcct gtccggagag cagaagaagg ctatcgttga tctgctcttc 3781 aagactaata gaaaggtgac agtgaagcag ctcaaggagg attactttaa gaagatcgaa 3841 tgctttgact cagtggaaat ctctggcgtg gaggaccgct ttaatgccag cctgggcact 3901 taccatgatc tgctgaagat aatcaaagac aaagatttcc tcgataatga ggagaacgag 3961 gacatcctgg aagatatcgt gctgaccctg actctgttcg aggatagaga gatgatcgaa 4021 gagcgcctga agacctatgc ccatctgttt gacgataaag tcatgaaaca gctcaagcgg 4081 cggcgctaca ctgggtgggg tagactctcc aggaaactca taaacggcat ccgcgacaaa 4141 cagagcggaa agaccatcct ggatttcctg aaatccgacg gattcgctaa caggaacttc 4201 atgcaactga ttcacgatga ctctctgaca tttaaagagg acatccagaa ggcacaggtg 4261 agcggtcaag gcgacagcct gcacgagcac atcgccaacc tcgctggatc acccgccata 4321 aagaagggaa tactgcagac agtcaaggtc gtggacgaac tcgtcaaagt gatgggtcgg 4381 cacaagccag agaatatcgt tatcgaaatg gcaagggaga accaaaccac ccagaagggc 4441 cagaagaact ctcgggaacg gatgaaaaga atcgaagagg gaattaagga gctgggatct 4501 cagatactga aggagcaccc tgtggagaat acacagctcc agaacgagaa actctacctg 4561 tactacctcc agaacgggcg ggacatgtac gttgaccagg aactcgacat caaccggctg 4621 tccgattatg acgtggacca tattgttcca cagtccttcc tcaaagatga ctccattgac 4681 aacaaggtgc tgaccagatc cgataagaat cgcggtaagt ctgacaatgt tccatcagaa 4741 gaggtggtca agaagatgaa gaattactgg cggcagctcc tcaacgccaa actgatcacc 4801 cagcggaagt ttgacaatct gactaaggca gaaagaggag gtctgagcga actcgacaag 4861 gccggcttta ttaagaggca actggtcgaa acacgccaga ttaccaaaca cgtggcacaa 4921 atcctcgact ctaggatgaa cactaagtac gatgagaacg ataagctgat cagggaagtg 4981 aaagtgataa ctctgaagag caagctggtg tctgacttcc ggaaggactt tcaattctac 5041 aaagttcgcg aaataaacaa ttaccatcat gctcacgatg cctatctcaa tgctgtcgtt 5101 ggcaccgccc tgatcaagaa ataccctaaa ctggagtctg agttcgtgta cggtgactat 5161 aaagtctacg atgtgaggaa gatgatagca aagtctgagc aagagattgg caaagccacc 5221 gccaagtact tcttctactc taatatcatg aatttcttta agactgagat aaccctggct 5281 aacggcgaaa tccggaagcg cccactgatc gaaacaaacg gagaaacagg agaaatcgtg 5341 tgggataaag gcagggactt cgcaactgtg cggaaggtgc tgtccatgcc acaagtcaat 5401 atcgtgaaga agaccgaagt gcagaccggc ggattctcaa aggagagcat cctgccaaag 5461 cggaactctg acaagctgat cgccaggaag aaagattggg acccaaagaa gtatggcggt 5521 ttcgattccc ctacagtggc ttattccgtt ctggtcgtgg caaaagtgga gaaaggcaag 5581 tccaagaaac tcaagtctgt taaggagctg ctcggaatta ctattatgga gagatccagc 5641 ttcgagaaga atccaatcga tttcctggaa gctaagggct ataaagaagt gaagaaagat 5701 ctcatcatca aactgcccaa gtactctctc tttgagctgg agaatggtag gaagcggatg 5761 ctggcctccg ccggagagct gcagaaagga aacgagctgg ctctgccctc caaatacgtg 5821 aacttcctgt atctggcctc ccactacgag aaactcaaag gtagccctga agacaatgag 5881 cagaagcaac tctttgttga gcaacataaa cactacctgg acgaaatcat tgaacagatt 5941 agcgagttca gcaagcgggt tattctggcc gatgcaaacc tcgataaagt gctgagcgca 6001 tataataagc acagggacaa gccaattcgc gaacaagcag agaatattat ccacctcttt 6061 actctgacta atctgggcgc tcctgctgcc ttcaagtatt tcgatacaac tattgacagg 6121 aagcggtaca cctctaccaa agaagttctc gatgccaccc tgatacacca gtcaattacc 6181 ggactgtacg agactcgcat cgacctgtct cagctcggcg gcgacggttc tgaattcagc 6241 ctgggcagcg gctcccccaa gaagaagcgc aaggtgaccg gtcatcatca ccatcaccat 6301 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt 6361 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 6421 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 6481 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 6541 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac 6601 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 6661 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 6721 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 6781 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 6841 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 6901 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 6961 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 7021 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 7081 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 7141 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 7201 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 7261 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 7321 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 7381 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata 7441 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agcctttgtc 7501 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc 7561 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt 7621 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc 7681 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat 7741 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc 7801 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc 7861 tggttatgtg tgggagggct aagcacttcg tggccgagga gcaggactga cacgtgctac 7921 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 7981 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 8041 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 8101 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 8161 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 8221 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 8281 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 8341 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 8401 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 8461 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 8521 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 8581 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 8641 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 8701 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 8761 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 8821 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 8881 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 8941 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 9001 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 9061 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 9121 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 9181 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 9241 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 9301 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 9361 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 9421 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 9481 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 9541 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 9601 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 9661 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 9721 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 9781 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 9841 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 9901 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 9961 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 10021 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 10081 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 10141 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 10201 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 10261 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 10321 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 10381 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag 10441 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta 10501 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg 10561 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t //
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