质粒名称: | pY30 |
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出品公司: | 爱迪基因 |
目录编号: | 84745 |
存储实验室: | Feng Zhang |
载体骨架基本信息 |
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载体名称: | pcDNA3.1 |
载体抗性: | 氨苄青霉素 |
空载体大小: | 5620 bp |
完整质粒大小: | 10340 bp |
载体类型: | 哺乳细胞表达 |
筛选标记: | 嘌呤霉素 |
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: | huAsCpf1 |
物种来源: | Synthetic |
启动子 | CMV |
标签/融合蛋白: |
NLS(插接件上的C端子)
3xHA(插入件上的C端子)
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克隆方法: | Gibson克隆 |
5' 测序引物 | cgcaaatgggcggtaggcgtg |
文献引用方法 |
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材料和方法部分: | pY30 was a gift from Feng Zhang (Addgene plasmid # 84745 ; http://n2t.net/addgene:84745 ; RRID:Addgene_84745) |
参考文献部分: | Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array. Zetsche B, Heidenreich M, Mohanraju P, Fedorova I, Kneppers J, DeGennaro EM, Winblad N, Choudhury SR, Abudayyeh OO, Gootenberg JS, Wu WY, Scott DA, Severinov K, van der Oost J, Zhang F. Nat Biotechnol. 2017 Jan;35(1):31-34. doi: 10.1038/nbt.3737. Epub 2016 Dec 5. 10.1038/nbt.3737 PubMed 27918548 |
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 pY30 10340 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Expresses huAsCpf1-P2A-puro and crRNA guide. ACCESSION . VERSION . KEYWORDS pY30 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10340) AUTHORS Zetsche B, Heidenreich M, Mohanraju P, Fedorova I, Kneppers J, DeGennaro EM, Winblad N, Choudhury SR, Abudayyeh OO, Gootenberg JS, Wu WY, Scott DA, Severinov K, van der Oost J, Zhang F TITLE Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array. JOURNAL Nat Biotechnol. 2017 Jan;35(1):31-34. doi: 10.1038/nbt.3737. Epub 2016 Dec 5. PUBMED 27918548 REFERENCE 2 (bases 1 to 10340) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10340 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 170..410 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 170..190 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 341..360 /label=LKO.1 5' /note="Human U6 promoter, forward primer" enhancer 538..917 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 918..1121 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1072..1092 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 1166..1185 /label=T7 /note="T7 promoter, forward primer" promoter 1166..1184 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1232..5152 /codon_start=1 /product="CRISPR-associated protein from Acidaminococcus sp. BV3L6 (Zetsche et al., 2015)" /label=AsCpf1 /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES KDLKLQNGISNQDWLAYIQELRN" regulatory 4872..4881 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5153..5200 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 5210..5266 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 5267..5863 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind 5760..5780 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(5934..5951) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5940..6164 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 6210..6638 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6297..6316) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6507..6528 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6647..6667) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6652..6981 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6832..6967 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6894..6913 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 7048..7842 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(7102..7121) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7712..7731 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 8016..8137 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8053..8072) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8107..8126 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(8186..8202) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8186..8202) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8199..8221) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8210..8226 /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(8234..8264) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8279..8300 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8417..8434) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8588..9173) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8668..8687) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9344..10204) /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 9967..9986 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10205..10309) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga cttaattaag agggcctatt 181 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat 241 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt 301 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt 361 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgt 421 aatttctact cttgtagatg gagacgatat atcgtctctt tttcaattgc atgaagaatc 481 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 541 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 601 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 661 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 721 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 781 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 841 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 901 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 961 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 1021 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 1081 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac 1141 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct 1201 agcgtttaaa cttaagcttg gtaccgccac catgacacag ttcgagggct ttaccaacct 1261 gtatcaggtg agcaagacac tgcggtttga gctgatccca cagggcaaga ccctgaagca 1321 catccaggag cagggcttca tcgaggagga caaggcccgc aatgatcact acaaggagct 1381 gaagcccatc atcgatcgga tctacaagac ctatgccgac cagtgcctgc agctggtgca 1441 gctggattgg gagaacctga gcgccgccat cgactcctat agaaaggaga aaaccgagga 1501 gacaaggaac gccctgatcg aggagcaggc cacatatcgc aatgccatcc acgactactt 1561 catcggccgg acagacaacc tgaccgatgc catcaataag agacacgccg agatctacaa 1621 gggcctgttc aaggccgagc tgtttaatgg caaggtgctg aagcagctgg gcaccgtgac 1681 cacaaccgag cacgagaacg ccctgctgcg gagcttcgac aagtttacaa cctacttctc 1741 cggcttttat gagaacagga agaacgtgtt cagcgccgag gatatcagca cagccatccc 1801 acaccgcatc gtgcaggaca acttccccaa gtttaaggag aattgtcaca tcttcacacg 1861 cctgatcacc gccgtgccca gcctgcggga gcactttgag aacgtgaaga aggccatcgg 1921 catcttcgtg agcacctcca tcgaggaggt gttttccttc cctttttata accagctgct 1981 gacacagacc cagatcgacc tgtataacca gctgctggga ggaatctctc gggaggcagg 2041 caccgagaag atcaagggcc tgaacgaggt gctgaatctg gccatccaga agaatgatga 2101 gacagcccac atcatcgcct ccctgccaca cagattcatc cccctgttta agcagatcct 2161 gtccgatagg aacaccctgt ctttcatcct ggaggagttt aagagcgacg aggaagtgat 2221 ccagtccttc tgcaagtaca agacactgct gagaaacgag aacgtgctgg agacagccga 2281 ggccctgttt aacgagctga acagcatcga cctgacacac atcttcatca gccacaagaa 2341 gctggagaca atcagcagcg ccctgtgcga ccactgggat acactgagga atgccctgta 2401 tgagcggaga atctccgagc tgacaggcaa gatcaccaag tctgccaagg agaaggtgca 2461 gcgcagcctg aagcacgagg atatcaacct gcaggagatc atctctgccg caggcaagga 2521 gctgagcgag gccttcaagc agaaaaccag cgagatcctg tcccacgcac acgccgccct 2581 ggatcagcca ctgcctacaa ccctgaagaa gcaggaggag aaggagatcc tgaagtctca 2641 gctggacagc ctgctgggcc tgtaccacct gctggactgg tttgccgtgg atgagtccaa 2701 cgaggtggac cccgagttct ctgcccggct gaccggcatc aagctggaga tggagccttc 2761 tctgagcttc tacaacaagg ccagaaatta tgccaccaag aagccctact ccgtggagaa 2821 gttcaagctg aactttcaga tgcctacact ggcctctggc tgggacgtga ataaggagaa 2881 gaacaatggc gccatcctgt ttgtgaagaa cggcctgtac tatctgggca tcatgccaaa 2941 gcagaagggc aggtataagg ccctgagctt cgagcccaca gagaaaacca gcgagggctt 3001 tgataagatg tactatgact acttccctga tgccgccaag atgatcccaa agtgcagcac 3061 ccagctgaag gccgtgacag cccactttca gacccacaca acccccatcc tgctgtccaa 3121 caatttcatc gagcctctgg agatcacaaa ggagatctac gacctgaaca atcctgagaa 3181 ggagccaaag aagtttcaga cagcctacgc caagaaaacc ggcgaccaga agggctacag 3241 agaggccctg tgcaagtgga tcgacttcac aagggatttt ctgtccaagt ataccaagac 3301 aacctctatc gatctgtcta gcctgcggcc atcctctcag tataaggacc tgggcgagta 3361 ctatgccgag ctgaatcccc tgctgtacca catcagcttc cagagaatcg ccgagaagga 3421 gatcatggat gccgtggaga caggcaagct gtacctgttc cagatctata acaaggactt 3481 tgccaagggc caccacggca agcctaatct gcacacactg tattggaccg gcctgttttc 3541 tccagagaac ctggccaaga caagcatcaa gctgaatggc caggccgagc tgttctaccg 3601 ccctaagtcc aggatgaaga ggatggcaca ccggctggga gagaagatgc tgaacaagaa 3661 gctgaaggat cagaaaaccc caatccccga caccctgtac caggagctgt acgactatgt 3721 gaatcacaga ctgtcccacg acctgtctga tgaggccagg gccctgctgc ccaacgtgat 3781 caccaaggag gtgtctcacg agatcatcaa ggataggcgc tttaccagcg acaagttctt 3841 tttccacgtg cctatcacac tgaactatca ggccgccaat tccccatcta agttcaacca 3901 gagggtgaat gcctacctga aggagcaccc cgagacacct atcatcggca tcgatcgggg 3961 cgagagaaac ctgatctata tcacagtgat cgactccacc ggcaagatcc tggagcagcg 4021 gagcctgaac accatccagc agtttgatta ccagaagaag ctggacaaca gggagaagga 4081 gagggtggca gcaaggcagg cctggtctgt ggtgggcaca atcaaggatc tgaagcaggg 4141 ctatctgagc caggtcatcc acgagatcgt ggacctgatg atccactacc aggccgtggt 4201 ggtgctggag aacctgaatt tcggctttaa gagcaagagg accggcatcg ccgagaaggc 4261 cgtgtaccag cagttcgaga agatgctgat cgataagctg aattgcctgg tgctgaagga 4321 ctatccagca gagaaagtgg gaggcgtgct gaacccatac cagctgacag accagttcac 4381 ctcctttgcc aagatgggca cccagtctgg cttcctgttt tacgtgcctg ccccatatac 4441 atctaagatc gatcccctga ccggcttcgt ggaccccttc gtgtggaaaa ccatcaagaa 4501 tcacgagagc cgcaagcact tcctggaggg cttcgacttt ctgcactacg acgtgaaaac 4561 cggcgacttc atcctgcact ttaagatgaa cagaaatctg tccttccaga ggggcctgcc 4621 cggctttatg cctgcatggg atatcgtgtt cgagaagaac gagacacagt ttgacgccaa 4681 gggcacccct ttcatcgccg gcaagagaat cgtgccagtg atcgagaatc acagattcac 4741 cggcagatac cgggacctgt atcctgccaa cgagctgatc gccctgctgg aggagaaggg 4801 catcgtgttc agggatggct ccaacatcct gccaaagctg ctggagaatg acgattctca 4861 cgccatcgac accatggtgg ccctgatccg cagcgtgctg cagatgcgga actccaatgc 4921 cgccacaggc gaggactata tcaacagccc cgtgcgcgat ctgaatggcg tgtgcttcga 4981 ctcccggttt cagaacccag agtggcccat ggacgccgat gccaatggcg cctaccacat 5041 cgccctgaag ggccagctgc tgctgaatca cctgaaggag agcaaggatc tgaagctgca 5101 gaacggcatc tccaatcagg actggctggc ctacatccag gagctgcgca acaaaaggcc 5161 ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag ggatccggcg caacaaactt 5221 ctctctgctg aaacaagccg gagatgtcga agagaatcct ggaccgaccg agtacaagcc 5281 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc 5341 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt 5401 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt 5461 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc 5521 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca 5581 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc 5641 caccgtcgga gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc 5701 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg 5761 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga 5821 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc taagaattct gcagatatcc 5881 agcacagtgg cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac 5941 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct 6001 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct 6061 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg 6121 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag 6181 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc 6241 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc 6301 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa 6361 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact 6421 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt 6481 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa 6541 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt 6601 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag 6661 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 6721 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 6781 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 6841 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 6901 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt 6961 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat 7021 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct 7081 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc 7141 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc 7201 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc 7261 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg 7321 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag 7381 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc 7441 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt 7501 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc 7561 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc 7621 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg 7681 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag 7741 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg 7801 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg 7861 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct 7921 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc 7981 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg 8041 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 8101 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct 8161 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 8221 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 8281 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 8341 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 8401 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 8461 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 8521 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 8581 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 8641 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 8701 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 8761 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 8821 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 8881 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 8941 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 9001 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 9061 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 9121 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 9181 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 9241 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 9301 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 9361 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 9421 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 9481 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 9541 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 9601 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 9661 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 9721 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 9781 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 9841 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 9901 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 9961 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 10021 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 10081 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 10141 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 10201 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 10261 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 10321 gaaaagtgcc acctgacgtc //
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