质粒名称: | pLX304-Flag-APEX2-NES |
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出品公司: | Addgene |
目录编号: | 92158 |
存储实验室: | Alice Ting |
载体骨架基本信息 |
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载体名称: | pLX304 |
载体出品公司: | -- |
空载体大小: | 8553 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳动物表达,慢病毒 |
筛选标记: | 杀稻瘟菌素 |
载体抗性: | 氨苄青霉素 |
生长温度: | 30 ℃ |
宿主菌: | NEB Stable |
拷贝数: | -- |
插入基因信息 |
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插入基因名称: | Flag-APEX2-NES |
物种来源: | soybean |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子: | CMV |
标签/融合蛋白: | Flag(插入件上的N端子) |
克隆方法: | Gateway 克隆 |
5’克隆位点: | -- |
3’克隆位点: | -- |
5’测序引物 | CMV-F |
3’测序引物 | WPRE-R |
文献引用方法 |
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材料和方法部分: | pLX304-Flag-APEX2-NES was a gift from Alice Ting (Addgene plasmid # 92158 ; http://n2t.net/addgene:92158 ; RRID:Addgene_92158) |
参考文献部分: | Proteomic mapping of cytosol-facing outer mitochondrial and ER membranes in living human cells by proximity biotinylation. Hung V, Lam SS, Udeshi ND, Svinkina T, Guzman G, Mootha VK, Carr SA, Ting AY. Elife. 2017 Apr 25;6. pii: e24463. doi: 10.7554/eLife.24463. 10.7554/eLife.24463 PubMed 28441135 |
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载体基本信息
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载体序列
LOCUS pLX304-Flag-APEX2-NES 8553 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cytosolic APEX2 construct. ACCESSION . VERSION . KEYWORDS pLX304-Flag-APEX2-NES SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8553) AUTHORS Hung V, Lam SS, Udeshi ND, Svinkina T, Guzman G, Mootha VK, Carr SA, Ting AY TITLE Proteomic mapping of cytosol-facing outer mitochondrial and ER membranes in living human cells by proximity biotinylation. JOURNAL Elife. 2017 Apr 25;6. pii: e24463. doi: 10.7554/eLife.24463. PUBMED 28441135 REFERENCE 2 (bases 1 to 8553) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8553 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(154..742) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(234..253) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(913..1773) /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 1536..1555 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1774..1878) /gene="bla" /label=AmpR promoter rep_origin complement(1904..2359) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2041..2062) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2253..2272 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2486..2508 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2500..2517 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2501..2517 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2527..2546 /label=T7 /note="T7 promoter, forward primer" promoter 2527..2545 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" rep_origin 2566..2701 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(2620..2639) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 2741..2862 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2752..2771) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2806..2825 /label=SV40pA-R /note="SV40 polyA, reverse primer" LTR 2940..3173 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" misc_feature 3245..3833 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS 3351..3362 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind 3760..3780 /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3875..3916) /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" protein_bind 3918..3942 /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS complement(3950..3976) /codon_start=1 /product="nuclear export signal from the HIV Rev protein (Fischer et al., 1995)" /label=NES /translation="LPPLERLTL" CDS complement(3983..4729) /codon_start=1 /product="soybean ascorbate peroxidase, engineered to serve as a monomeric tag for electron microscopy and live-cell proteomics (Lam et al., 2015)" /label=APEX2 /note="more active than the original APEX due to the A134P mutation" /translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK YAADEDAFFADYAEAHQKLSELGFADA" CDS complement(4730..4753) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" protein_bind complement(4774..4798) /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" promoter complement(4821..5024) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(4850..4870) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 5025..5328 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" misc_feature 5445..5562 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" CDS complement(5622..6017) /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" promoter complement(6029..6529) /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind complement(6092..6110) /label=hPGK-F /note="Human PGK promoter, forward primer" CDS complement(6704..6748) /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 6933..7166 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 7659..7784 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" LTR 7831..8011 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" promoter complement(8012..8238) /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" primer_bind complement(8266..8286) /label=T3 /note="T3 promoter, forward primer" promoter complement(8266..8284) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(8305..8321) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8305..8321) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8318..8340) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8329..8345 /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(8353..8383) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8398..8419 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8536..8553) /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 61 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 661 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 841 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 901 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 1021 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 1081 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 1141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 1201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 1261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 1321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 1381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 1441 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 1501 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 1561 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 1621 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 1681 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 1741 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 1801 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 1861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat 1921 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc 1981 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt 2041 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa 2101 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg 2161 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga 2221 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct 2281 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat 2341 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga 2401 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 2461 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag 2521 cgcgcgtaat acgactcact atagggcgaa ttgggtacgt ccctaggcct ccaaaaaagc 2581 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa 2641 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga 2701 tgggcggagt taggggcggg atagctagag ccagacatga taagatacat tgatgagttt 2761 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct 2821 attgctttat ttgtaaccat tataagctgc aataaacaag ttcctctcac tctctgatat 2881 tcatttcttt gcaagttata aatactgaat aataagatga catgaactac tatacgtact 2941 gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca 3001 caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag 3061 tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc 3121 agtacaagca aaaagcagat cttgtcttcg ttgggagtga attagccctt ccagtccccc 3181 cttttctttt aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagt 3241 cgacgcgggg aggcggccca aagggagatc cgactcgtct gagggcgaag gcgaagacgc 3301 ggaagaggcc gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg attgagggcc 3361 gaagggacgt agcagaagga cgtcccgcgc agaatccagg tggcaacaca ggcgagcagc 3421 caaggaaagg acgatgattt ccccgacaac accacggaat tgtcagtgcc caacagccga 3481 gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aatagggagg 3541 gggaaagcga aagtcccgga aaggagctga caggtggtgg caatgcccca accagtgggg 3601 gttgcgtcag caaacacagt gcacaccacg ccacgttgcc tgacaacggg ccacaactcc 3661 tcataaagag acagcaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa 3721 gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca 3781 tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg attgtcgact 3841 taacgcgcca ccggttagcg ctagctcatt actacgtaga atcgagaccg aggagagggt 3901 tagggatagg cttaccaacc actttgtaca agaaagctgg gtcttagtcc agggtcaggc 3961 gctccagggg aggcagctgc agggcatcag caaacccaag ctcggaaagc ttttggtgag 4021 cctcagcgta atcagcaaag aaggcatctt cgtccgctgc atatttgtca acgagagggc 4081 ggaatacagg gtcagacaaa agagccttgt cagaaggtag ctgaaggaga ccttccttct 4141 caccactcaa caactccgtg aagtatgagt tgtcgaaaat aagaggatta gaggtccagg 4201 gaccctcaaa tccagaacgc tccttgtgtg cagctccaat agtgtgaccc ccagatagag 4261 caacgatatc ttggtcagta agccccatag ctttgccaaa cacatctctc aaatggtcag 4321 aacccttagt gggatcgggc aagcgaccct ctggtggtgg ctcaggcttg tcctctcttc 4381 cagggtggaa tggaacctta ggtccacccg tgacctcaac ggcaacaacg ccagccaact 4441 ggtagaaatc ggcgtagctc aaaataggga actccgcctt gagtggctcc aaaagcctaa 4501 cagcgatgtc aagaccgttg ttagcgctgt gagccagttc ggcagggtgc ttgatggttc 4561 cgaagggtcc accggtcttc gtgcccttgt caaaggttcc agcagagtgg aatgccaaac 4621 ggagcattag aggagcgcat ctcttctcag cgatgaagcc tctgagcttc ttcttcgcct 4681 tctcaacggc gtcctggtaa tcagcactca cagttgggta agactttccc ttatcgtcgt 4741 catccttgta gtccatggtt ctatctcctt cgaagcctgc ttttttgtac aaacttgttg 4801 atcccgacag ttagccagag agctctgctt atatagacct cccaccgtac acgcctaccg 4861 cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg 4921 tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca 4981 aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg 5041 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa 5101 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac 5161 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg 5221 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg 5281 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata 5341 tatgggctat gaactaatga ccccgtaatt gattactatt aataaccccg ggccccctcg 5401 agccggcgcc aaagtggatc tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt 5461 gtaatttgtt tttgtaattc tttagtttgt atgtctgttg ctattatgtc tactattctt 5521 tcccctgcac tgtacccccc aatcccccct tttcttttaa aattgtggat gaatactgcc 5581 atttgtctca agatctagaa ttcttactac tgcagctgca ggccctccca cacataacca 5641 gagggcagca attcacgaat cccaactgcc gtcggctgtc catcactgtc cttcactatg 5701 gctttgatcc caggatgcag atcgagaagc acctgtcggc accgtccgca ggggctcaag 5761 atgcccctgt tctcatttcc gatcgcgacg atacaagtca ggttgccagc tgccgcagca 5821 gcagcagtgc ccagcaccac gagttctgca caaggtcccc cagtaaaatg atatacattg 5881 acaccagtga agatgcggcc gtcgctagag agagctgcgc tggcgacgct gtagtcttca 5941 gagatgggga tgctgttgat tgtagccgtt gctctttcaa tgagggtgga ttcttcttga 6001 gacaaaggct tggccatggt ggtaccccct ggggagagag gtcggtgatt cggtcaacga 6061 gggagccgac tgccgacgtg cgctccggag gcttgcagaa tgcggaacac cgcgcgggca 6121 ggaacagggc ccacactacc gccccacacc ccgcctcccg caccgcccct tcccggccgc 6181 tgctctcggc gcgccctgct gagcagccgc tattggccac agcccatcgc ggtcggcgcg 6241 ctgccattgc tccctggcgc tgtccgtctg cgagggtact agtgagacgt gcggcttccg 6301 tttgtcacgt ccggcacgcc gcgaaccgca aggaaccttc ccgacttagg ggcggagcag 6361 gaagcgtcgc cggggggccc acaagggtag cggcgaagat ccgggtgacg ctgcgaacgg 6421 acgtgaagaa tgtgcgagac ccagggtcgg cgccgctgcg tttcccggaa ccacgcccag 6481 agcagccgcg tccctgcgca aacccagggc tgccttggaa aaggcgcaag ggtcccctcg 6541 gggttgggag gtgggtctga aacgataatg gtgaatatcc ctgcctaact ctattcacta 6601 tagaaagtac agcaaaaact attcttaaac ctaccaagcc tcctactatc attatgaata 6661 attttatata ccacagccaa tttgttatgt taaaccaatt ccacaaactt gcccatttat 6721 ctaattccaa taattcttgt tcattctttt cttgctggtt ttgcgattct tcaattaagg 6781 agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca ctccatccag gtcgtgtgat 6841 tccaaatctg ttccagagat ttattactcc aactagcatt ccaaggcaca gcagtggtgc 6901 aaatgagttt tccagagcaa ccccaaatcc ccaggagctg ttgatccttt aggtatcttt 6961 ccacagccag gattcttgcc tggagctgct tgatgcccca gactgtgagt tgcaacagat 7021 gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg ctgcactata ccagacaata 7081 attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt cctgctgctc 7141 ccaagaaccc aaggaacaaa gctcctattc ccactgctct tttttctctc tgcaccactc 7201 ttctctttgc cttggtgggt gctactccta atggttcaat ttttactact ttatatttat 7261 ataattcact tctccaattg tccctcatat ctcctcctcc aggtctgaag atcagcggcc 7321 gcttgctgtg cggtggtctt acttttgttt tgctcttcct ctatcttgtc taaagcttcc 7381 ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac tgtattatat 7441 aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc cagtatttgt 7501 ctacagcctt ctgatgtttc taacaggcca ggattaactg cgaatcgttc tagctccctg 7561 cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct taaccgaatt 7621 ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca cccatctctc 7681 tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg cccctcgcct 7741 cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc tggtttccct 7801 ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact gactaaaagg 7861 gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca 7921 ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc 7981 aggctcagat ctggtctaac cagagagacc cgtttatgta tcgagctagg cacttaaata 8041 caatatctct gcaatgcggc aattcagtgg ttcgtccaat ccatgtcaga cccgtctgtt 8101 gccttcctaa taaggcacga tcgtaccacc ttacttccac caatcggcat gcacggtgct 8161 ttttctctcc ttgtaaggca tgttgctaac tcatcgttac catgttgcaa gactacaaga 8221 gtattgcata agactacatt aagcttgcag ctccagcttt tgttcccttt agtgagggtt 8281 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 8341 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 8401 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 8461 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 8521 gcgctcttcc gcttcctcgc tcactgactc gct //
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