质粒名称: | pcDNA-GalNAc-T2-GFP-ESCargo(FTV) |
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出品公司: | Addgene |
目录编号: | 140163 |
存储实验室: | Benjamin Glick |
载体骨架基本信息 |
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载体名称: | pcDNA5/FRT/TO |
载体出品公司: | -- |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | -- |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | -- |
插入基因信息 |
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插入基因名称: | GalNAc-T2-msGFP2 and piGH-FTVNTT-DsRed-Express2-FKBP(LV)(C22V) |
物种来源: | H. sapiens (human), Synthetic |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子: | EF-1a |
标签/融合蛋白: | ER信号序列(MGWSCIILFLVATATGAHS)(主干上的N端)(插入上的N端子) |
克隆方法: | Gibson克隆 |
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: | pcDNA-GalNAc-T2-GFP-ESCargo(FTV) was a gift from Benjamin Glick (Addgene plasmid # 140163 ; http://n2t.net/addgene:140163 ; RRID:Addgene_140163) |
参考文献部分: | ESCargo: a regulatable fluorescent secretory cargo for diverse model organisms. Casler JC, Zajac AL, Valbuena FM, Sparvoli D, Jeyifous O, Turkewitz AP, Horne-Badovinac S, Green WN, Glick BS. Mol Biol Cell. 2020 Oct 28:mbcE20090591. doi: 10.1091/mbc.E20-09-0591. 10.1091/mbc.E20-09-0591 PubMed 33112725 |
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载体基本信息
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载体简介
载体序列
LOCUS pcDNA-GalNAc-T2-GFP-ESCargo(FTV) 10051 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses a regulatable secretory cargo for mammalian cells and a Golgi marker. ACCESSION . VERSION . KEYWORDS pcDNA-GalNAc-T2-GFP-ESCargo(FTV) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10051) AUTHORS Casler JC, Zajac AL, Valbuena FM, Sparvoli D, Jeyifous O, Turkewitz AP, Horne-Badovinac S, Green WN, Glick BS TITLE ESCargo: a regulatable fluorescent secretory cargo for diverse model organisms. JOURNAL Mol Biol Cell. 2020 Oct 28:mbcE20090591. doi: 10.1091/mbc.E20-09-0591. PUBMED 33112725 REFERENCE 2 (bases 1 to 10051) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10051 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 51..1213 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 282..1204 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1161..1181 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 2964..3671 /codon_start=1 /product="photostable monomeric superfolder GFP (Valbuena et al., 2020)" /label=msGFP2 /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE FVTAAGITGGSGGS" primer_bind complement(3006..3027) /label=EGFP-N /note="EGFP, reverse primer" primer_bind 3614..3635 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3701..3925 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" enhancer 4019..4398 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4399..4602 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4553..4573 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4612..4636 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 4727..4736 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" sig_peptide join(4733..4777,4860..4871) /product="modified signal sequence from the mouse immunoglobulin heavy chain variable region" /label=IgH signal sequence primer_bind complement(5067..5087) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 5285..5304 /label=mCherry-F /note="mCherry, forward primer" primer_bind 5479..5502 /label=DsRed1-C /note="DsRed1, forward primer" CDS 5574..5894 /codon_start=1 /product="destabilization domain that can be stabilized by Shield1 in the ProteoTuner(TM) system" /label=DD /note="L106P mutant of FKBP12" /translation="GVQVETISPGDGRTFPKRGQTVVVHYTGMLEDGKKLDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGVIPPHATLVFDVELL KLE" polyA_signal 6028..6252 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(6385..6404) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 6536..6583 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 6591..7613 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 7743..7864 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7780..7799) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7834..7853 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7913..7929) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7913..7929) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7926..7948) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7937..7953 /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(7961..7991) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8006..8027 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8144..8161) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8315..8903) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8395..8414) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9074..9934) /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 9697..9716 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9935..10039) /gene="bla" /label=AmpR promoter ORIGIN 1 aaaagtgcca cctgacgtcg acggatcggg agatccttca agcttcgtga ggctccggtg 61 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg 121 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt 181 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg 241 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt 301 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct 361 ggctccagta cgtgattctt gatcccgagc tggagccagg ggcgggcctt gcgctttagg 421 agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg 481 aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa 541 tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca 601 ggatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc 661 ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg 721 gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc 781 gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct 841 tcccggccct gctccagggg gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg 901 tgagtcaccc acacaaagga aaggggcctt tccgtcctca gccgtcgctt catgtgactc 961 cacggagtac cgggcgccgt ccaggcacct cgattagttc tggagctttt ggagtacgtc 1021 gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga 1081 gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga 1141 gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt 1201 tcaggtgtcg tgaacacgtg gccaccgcca ccatgaggcg gcgctcgagg atgctgctct 1261 gcttcgcctt cctgtgggtg ctgggaatcg cctactacat gtactctgga ggaggatctg 1321 cgctggccgg aggagcggga ggcggagccg gaaggaagga ggactggaat gaaattgacc 1381 ccattaaaaa gaaagacctt catcacagca atggagaaga gaaagcacaa agcatggaga 1441 ccctccctcc agggaaagta cggtggccag actttaacca ggaagcttat gttggaggga 1501 cgatggtccg ctccgggcag gacccttacg cccgcaacaa gttcaaccag gtggagagtg 1561 ataagcttcg aatggacaga gccatccctg acacccggca tgaccagtgt cagcggaagc 1621 agtggcgggt ggatctgccg gccaccagcg tggtgatcac gtttcacaat gaagccaggt 1681 cggccctact caggaccgtg gtcagcgtgc ttaagaaaag cccgccccat ctcataaaag 1741 aaatcatctt ggtggatgac tacagcaatg atcctgagga cggggctctc ttggggaaaa 1801 ttgagaaagt gcgagttctt agaaatgatc gacgagaagg cctcatgcgc tcacgggttc 1861 ggggggccga tgctgcccaa gccaaggtcc tgaccttcct ggacagtcac tgcgagtgta 1921 atgagcactg gctggagccc ctcctggaaa gggtggcgga ggacaggact cgggttgtgt 1981 cacccatcat cgatgtcatt aatatggaca actttcagta tgtgggggca tctgctgact 2041 tgaagggcgg ttttgattgg aacttggtat tcaagtggga ttacatgacg cctgagcaga 2101 gaaggtcccg gcaggggaac ccagtcgccc ctataaaaac ccccatgatt gctggtgggc 2161 tgtttgtgat ggataagttc tattttgaag aactggggaa gtacgacatg atgatggatg 2221 tgtggggagg agagaaccta gagatctcgt tccgcgtgtg gcagtgtggt ggcagcctgg 2281 agatcatccc gtgcagccgt gtgggacacg tgttccggaa gcagcacccc tacacgttcc 2341 cgggtggcag tggcactgtc tttgcccgaa acacccgccg ggcagcagag gtctggatgg 2401 atgaatacaa aaatttctat tatgcagcag tgccttctgc tagaaacgtt ccttatggaa 2461 atattcagag cagattggag cttaggaaga aactcagctg caagcctttc aaatggtacc 2521 ttgaaaatgt ctatccagag ttaagggttc cagaccatca ggatatagct tttggggcct 2581 tgcagcaggg aactaactgc ctcgacactt tgggacactt tgctgatggt gtggttggag 2641 tttatgaatg tcacaatgct gggggaaacc aggaatgggc cttgacgaag gagaagtcgg 2701 tgaagcacat ggatttgtgc cttactgtgg tggaccgggc accgggctct cttataaagc 2761 tgcagggctg ccgagaaaat gacagcagac agaaatggga acagatcgag ggcaactcca 2821 agctgaggca cgtgggcagc aacctgtgcc tggacagtcg cacggccaag agcgggggcc 2881 taagcgtgga ggtgtgtggc ccggcccttt cgcagcagtg gaagttcacg ctcaacctgc 2941 agcagggagg aagcggagga agcatggata gcactgagag cctgttcacc ggggtggtgc 3001 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg cgcggcgagg 3061 gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc 3121 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc 3181 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg 3241 tccaggagcg caccatcacc ttcaaggacg acggcaccta caagacccgc gccgaggtga 3301 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg 3361 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca 3421 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacgtggagg 3481 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg 3541 tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg 3601 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actggaggca 3661 gtggcggttc ttagtctaga agctcgctga tcagcctcga ctgtgccttc tagttgccag 3721 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 3781 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 3841 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 3901 gctggggatg gcccgggctc tatggcttct gaggcggaaa gaacgaattg catgaagaat 3961 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga 4021 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca 4081 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac 4141 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact 4201 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa 4261 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg 4321 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta 4381 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg 4441 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg 4501 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg 4561 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag cttcgtcgac gagctcgttt 4621 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca 4681 ccgggaccga tccagcctcc ggactctagc gtttaaactt aagcttgcca ccatgggatg 4741 gagctgtatc atcctcttct tggtagcaac agctacaggt aaggggttaa cagtagcagg 4801 cttgaggtct ggacatatat atgggtgaca atgacatcca ctttgccttt ctctccacag 4861 gcgcgcactc ctttaccgtg aacaccaccg atagcactga gaacgtcatc aagcccttca 4921 tgcgcttcaa ggtgcacatg gagggctcag taaatggtca tgagttcgag atcgagggcg 4981 agggcgaggg caagccctac gagggcaccc agaccgccaa gctgcaggtg accaagggcg 5041 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt 5101 acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca 5161 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct 5221 ccctgcagga cggcaccttc atctaccacg tgaagttcat cggcgtgaac ttcccctccg 5281 acggccccgt aatgcagaag aagactctgg gctgggagcc ctccaccgag cgcctgtacc 5341 cccgcgacgg cgtgctgaag ggcgagatcc acaaggcgct gaagctgaag ggcggcggcc 5401 actacctggt ggagttcaag tcaatctaca tggccaagaa gcccgtgaag ctgcccggct 5461 actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac accgtggtgg 5521 agcagtacga gcgcgccgag gcccgccacc acctgttcca gggaggatcc agaggagtgc 5581 aggtggaaac catctcccca ggagacgggc gcaccttccc caagcgcggc cagaccgttg 5641 tggtgcacta caccgggatg cttgaagatg gaaagaaact agattcctcc cgggacagaa 5701 acaagccctt taagtttatg ctaggcaagc aggaggtgat ccgaggctgg gaagaagggg 5761 ttgcccagat gagtgtgggt cagagagcca aactgactat atctccagat tatgcctatg 5821 gtgccactgg gcacccaggc gttatcccac cacatgccac tctcgtcttc gatgtggagc 5881 ttctaaaact ggaatagtct agaagctcgc tgatcaggta ccgagctcgg atccactagt 5941 ccagtgtggt ggaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc 6001 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 6061 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 6121 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 6181 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 6241 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg 6301 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 6361 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 6421 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 6481 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt acctagaagt 6541 tcctattccg aagttcctat tctctagaaa gtataggaac ttccttggcc aaaaagcctg 6601 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc 6661 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg 6721 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc 6781 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg 6841 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg 6901 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg 6961 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca 7021 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg 7081 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg 7141 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc 7201 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt 7261 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc 7321 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt 7381 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg 7441 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg 7501 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac 7561 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagcacgtac 7621 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 7681 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 7741 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 7801 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 7861 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 7921 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 7981 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 8041 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 8101 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 8161 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 8221 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 8281 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 8341 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 8401 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 8461 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 8521 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 8581 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 8641 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 8701 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 8761 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 8821 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 8881 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 8941 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 9001 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 9061 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 9121 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 9181 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 9241 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 9301 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 9361 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 9421 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 9481 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 9541 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 9601 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 9661 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 9721 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 9781 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 9841 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 9901 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 9961 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 10021 aacaaatagg ggttccgcgc acatttcccc g //
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