质粒名称: | pJFRC7-RFPnls-PQRv2-mCD8:GFP |
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出品公司: | Addgene |
目录编号: | 73974 |
存储实验室: | Brian Chen |
载体骨架基本信息 |
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载体名称: | pJFRC7 |
载体出品公司: | Janelia Farms |
空载体大小: | 8142bp |
完整质粒大小: | 10413 bp |
载体类型: | Insect Expression |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称1: | sfGFP / folder GFP |
插入大小: | 1202 bp |
克隆方法: | 20XUAS-HS |
标签/融合蛋白: | mCD8 (N terminal on insert) |
克隆方法: | Gibson Cloning |
5’测序引物 | GGCCACCTGATCTGCAAC |
3’测序引物 | CCATTCATCAGTTCCATAGG |
插入基因名称2: | RFP / TagRFP-T |
物种来源: | D. melanogaster (fly) |
插入大小: | 816bp |
克隆方法: | 20XUAS-HS |
标签/融合蛋白: | nls (C terminal on insert) |
克隆方法: | Gibson Cloning |
5’测序引物 | GAGCGCCGGAGTATAAATAGAG |
3’测序引物 | TCTGGAAGAGCCAAGAGCAT |
文献引用方法 |
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材料和方法部分: | pJFRC7-RFPnls-PQRv2-mCD8:GFP was a gift from Brian Chen (Addgene plasmid # 73974 ; http://n2t.net/addgene:73974 ; RRID:Addgene_73974) |
参考文献部分: | Quantification of Protein Levels in Single Living Cells. Lo CA, Kays I, Emran F, Lin TJ, Cvetkovska V, Chen BE. Cell Rep. 2015 Dec 22;13(11):2634-44. doi: 10.1016/j.celrep.2015.11.048. Epub 2015 Dec 10. 10.1016/j.celrep.2015.11.048 PubMed 26686644 |
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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 pJFRC7-RFPnls-PQRv2-mCD8:GFP 10413 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Protein Quantitation Reporter (PQR) to quantify a protein of interest in insect cells.. ACCESSION . VERSION . KEYWORDS pJFRC7-RFPnls-PQRv2-mCD8:GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10413) AUTHORS Lo CA, Kays I, Emran F, Lin TJ, Cvetkovska V, Chen BE TITLE Quantification of Protein Levels in Single Living Cells. JOURNAL Cell Rep. 2015 Dec 22;13(11):2634-44. doi: 10.1016/j.celrep.2015.11.048. Epub 2015 Dec 10. PUBMED 26686644 REFERENCE 2 (bases 1 to 10413) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..10413 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 31..135 /gene="bla" /label=AmpR promoter CDS 136..996 /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 complement(354..373) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1167..1755 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1656..1675 /label=pBR322ori-F /note="pBR322 origin, forward primer" protein_bind 1842..1911 /label=attB /bound_moiety="phage phi-C31 integrase" /note="attB site for the phi-C31 integrase (Groth et al., 2000)" primer_bind complement(2136..2154) /label=pCasper-F /note="Drosophila mini-white gene, reverse primer" CDS join(2606..2677,3079..3352,3427..4081,4143..4458,4662..4793, 4864..5478) /codon_start=1 /gene="white" /product="Drosophila white gene eye color pigment" /label=mini-white /note="This is a modified version of the white gene lacking part of the first intron." /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE" protein_bind 6508..6602 /label=5X UAS /bound_moiety="GAL4" /note="five tandem copies of the ""ScaI site"" 17-mer CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) that efficiently binds yeast Gal4 (Webster et al., 1988; Pfeiffer et al., 2010)" protein_bind 6636..6730 /label=5X UAS /bound_moiety="GAL4" /note="five tandem copies of the ""ScaI site"" 17-mer CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) that efficiently binds yeast Gal4 (Webster et al., 1988; Pfeiffer et al., 2010)" protein_bind 6764..6858 /label=5X UAS /bound_moiety="GAL4" /note="five tandem copies of the ""ScaI site"" 17-mer CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) that efficiently binds yeast Gal4 (Webster et al., 1988; Pfeiffer et al., 2010)" protein_bind 6892..6986 /label=5X UAS /bound_moiety="GAL4" /note="five tandem copies of the ""ScaI site"" 17-mer CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) that efficiently binds yeast Gal4 (Webster et al., 1988; Pfeiffer et al., 2010)" promoter 7011..7249 /label=hsp70 promoter /note="Drosophila melanogaster hsp70Bb promoter" primer_bind 7196..7217 /label=pCasper-hs /note="Drosophila Hsp70 promoter, forward primer" intron 7264..7330 /label=Mhc intron /note="modified Drosophila myosin heavy chain intron 16 (Pfeiffer et al., 2010)" CDS 7339..8070 /codon_start=1 /product="photostable variant of TagRFP (Shaner et al., 2008)" /label=TagRFP-T /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFPSNCPVMQKKTLGWEANTEMLYPADGGLEGRTD MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV ARYCDLPSKLGHKLNGMDELYK" CDS 8080..8100 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 8104..8124 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 8128..8148 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 8170..8223 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 8224..8889 /codon_start=1 /product="mouse lymphocyte antigen CD8 alpha chain" /label=mCD8 /translation="MASPLTRFLSLNLLLLGESIILGSGEAKPQAPELRIFPKKMDAEL GQKVDLVCEVLGSVSQGCSWLFQNSSSKLPQPTFVVYMASSHNKITWDEKLNSSKLFSA MRDTNNKYVLTLNKFSKENEGYYFCSVISNSVMYFSSVVPVLQKVNSTTTKPVLRTPSP VHPTGTSQPQRPEDCRPRGSVKGTGLDFACDIYIWAPLAGICVALLLSLIITLICYHSR " CDS 8896..9609 /codon_start=1 /product="green fluorescent protein" /label=GFP /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(8926..8954) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 9549..9569 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" intron 9696..9761 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 9891..9911 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 10183..10317 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10220..10239) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10274..10293 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(10359..10377) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 gactcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 61 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 121 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 181 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 241 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 301 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 361 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 421 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 481 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 541 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 601 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 661 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 721 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 781 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 841 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 901 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 961 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 1021 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 1081 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 1141 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 1201 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 1261 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 1321 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 1381 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 1441 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 1501 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 1561 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 1621 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 1681 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 1741 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 1801 ccttttgctc acatgttacc gtcgacgatg taggtcacgg tctcgaagcc gcggtgcggg 1861 tgccagggcg tgcccttggg ctccccgggc gcgtactcca cctcacccat ctggtccatc 1921 atgatgaacg ggtcgaggtg gcggtagttg atcccggcga acgcgcggcg caccgggaag 1981 ccctcgccct cgaaaccgct gggcgcggtg gtcacggtga gcacgggacg tgcgacggcg 2041 tcggcgggtg cggatacgcg gggcagcgtc agcgggttct cgacggtcac ggcgggcatg 2101 tcgacaagcc gaacatatgg gcgcgcctag tatgtatgta agttaataaa acccattttt 2161 gcggaaagta gataaaaaaa acattttttt tttttactgc actggatatc attgaactta 2221 tctgatcagt tttaaattta cttcgatcca agggtatttg atgtaccagg ttctttcgat 2281 tacctctcac tcaaaatgac attccactca aagtcagcgc tgtttgcctc cttctctgtc 2341 cacagaaata tcgccgtctc tttcgccgct gcgtccgcta tctctttcgc caccgtttgt 2401 agcgttacgt agcgtcaatg tccgccttca gttgcatttt gtcagcggtt tcgtgacgaa 2461 gctccaagcg gtttacgcca tcaattaaac acaaagtgct gtgccaaaac tcctctcgct 2521 tcttattttt gtttgttttt tgagtgattg gggtggtgat tggttttggg tgggtaagca 2581 ggggaaagtg tgaaaaatcc cggcaatggg ccaagaggat caggagctat taattcgcgg 2641 aggcagcaaa cacccatctg ccgagcatct gaacaatgtg agtagtacat gtgcatacat 2701 cttaagttca cttgatctat aggaactgcg attgcaacat caaattgtat gcggcgtgag 2761 aactgcgacc cacaaaaatc ccaaaccgca attgcacaaa caaatagtga cacgaaacag 2821 attattctgg tagctgttct cgctatataa gacaattttt gagatcatat catgatcaag 2881 acatctaaag gcattcattt tcgactatat tcttttttac aaaaaatata acaaccagat 2941 attttaagct gatcctagat gcacaaaaaa taaataaaag tataaaccta cttcgtagga 3001 tacttcgggg tactttttgt tcggggttag atgagcataa cgctagtagt tgatatttga 3061 gatcccctat cattgcaggg tgacagcgga gcggcttcgc agagctgcat taaccagggc 3121 ttcgggcagg ccaaaaacta cggcacgctc cggccaccca gtccgccgga ggactccggt 3181 tcagggagcg gccaactagc cgagaacctc acctatgcct ggcacaatat ggacatcttt 3241 ggggcggtca atcagccggg ctccggatgg cggcagctgg tcaaccggac acgcggacta 3301 ttctgcaacg agcgacacat accggcgccc aggaaacatt tgctcaagaa cggtgagttt 3361 ctattcgcag tcggctgatc tgtgtgaaat cttaataaag ggtccaatta ccaatttgaa 3421 actcagtttg cggcgtggcc tatccgggcg aacttttggc cgtgatgggc agttccggtg 3481 ccggaaagac gaccctgctg aatgcccttg cctttcgatc gccgcagggc atccaagtat 3541 cgccatccgg gatgcgactg ctcaatggcc aacctgtgga cgccaaggag atgcaggcca 3601 ggtgcgccta tgtccagcag gatgacctct ttatcggctc cctaacggcc agggaacacc 3661 tgattttcca agccatggtg cggatgccac gacatctgac ctatcggcag cgagtggccc 3721 gcgtggatca ggtgatccag gagctttcgc tcagcaaatg tcagcacacg atcatcggtg 3781 tgcccggcag ggtgaaaggt ctgtccggcg gagaaaggaa gcgtctggca ttcgcctccg 3841 aggctctaac cgatccgccg cttctgatct gcgatgagcc cacctccgga ctggactcct 3901 ttaccgccca cagcgtcgtc caggtgctga agaagctgtc gcagaagggc aagaccgtca 3961 tcctgaccat tcatcagccg tcttccgagc tgtttgagct ctttgacaag atccttctga 4021 tggccgaggg cagggtagct ttcttgggca ctcccagcga agccgtcgac ttcttttcct 4081 agtgagttcg atgtgtttat taagggtatc tagtattaca taacatctca actcctatcc 4141 agcgtgggtg cccagtgtcc taccaactac aatccggcgg acttttacgt acaggtgttg 4201 gccgttgtgc ccggacggga gatcgagtcc cgtgatcgga tcgccaagat atgcgacaat 4261 tttgccatta gcaaagtagc ccgggatatg gagcagttgt tggccaccaa aaatctggag 4321 aagccactgg agcagccgga gaatgggtac acctacaagg ccacctggtt catgcagttc 4381 cgggcggtcc tgtggcgatc ctggctgtcg gtgctcaagg aaccactcct cgtaaaagtg 4441 cgacttattc agacaacggt gagtggttcc agtggaaaca aatgatataa cgcttacaat 4501 tcttggaaac aaattcgcta gattttagat agaattgcct gattccacac ccttcttagt 4561 ttttttcaat gagatgtata gtttatagtt ttgcagaaga taaataaatt tcatttaact 4621 cgcgaatatt aatgagatgc gagtaacatt ttaatttgca gatggttgcc atcttgattg 4681 gcctcatctt tttgggccaa caactcacgc aagtgggtgt gatgaatatc aacggagcca 4741 tcttcctctt cctgaccaac atgacctttc aaaacgtctt tgccacgata aatgtaagtc 4801 atgtttagaa tacatttgca tttcaataat ttactaactt tctaatgaat cgattcgatt 4861 taggtgttca cctcagagct gccagttttt atgagggagg cccgaagtcg actttatcgc 4921 tgtgacacat actttctggg caaaacgatt gccgaattgc cgctttttct cacagtgcca 4981 ctggtcttca cggcgattgc ctatccgatg atcggactgc gggccggagt gctgcacttc 5041 ttcaactgcc tggcgctggt cactctggtg gccaatgtgt caacgtcctt cggatatcta 5101 atatcctgcg ccagctcctc gacctcgatg gcgctgtctg tgggtccgcc ggttatcata 5161 ccattcctgc tctttggcgg cttcttcttg aactcgggct cggtgccagt atacctcaaa 5221 tggttgtcgt acctctcatg gttccgttac gccaacgagg gtctgctgat taaccaatgg 5281 gcggacgtgg agccgggcga aattagctgc acatcgtcga acaccacgtg ccccagttcg 5341 ggcaaggtca tcctggagac gcttaacttc tccgccgccg atctgccgct ggactacgtg 5401 ggtctggcca ttctcatcgt gagcttccgg gtgctcgcat atctggctct aagacttcgg 5461 gcccgacgca aggagtagcc gacatatatc cgaaataact gcttgttttt tttttttacc 5521 attattacca tcgtgtttac tgtttattgc cccctcaaaa agctaatgta attatatttg 5581 tgccaataaa aacaagatat gacctataga atacaagtat ttccccttcg aacatcccca 5641 caagtagact ttggatttgt cttctaacca aaagacttac acacctgcat accttacatc 5701 aaaaactcgt ttatcgctac ataaaacacc gggatatatt ttttatatac atacttttca 5761 aatcgcgcgc cctcttcata attcacctcc accacaccac gtttcgtagt tgctctttcg 5821 ctgtctccca cccgctctcc gcaacacatt caccttttgt tcgacgacct tggagcgact 5881 gtcgttagtt ccgcgcgatt cggttcgctc aaatggttcc gagtggttca tttcgtctca 5941 atagaaatta gtaataaata tttgtatgta caatttattt gctccaatat atttgtatat 6001 atttccctca cagctatatt tattctaatt taatattatg actttttaag gtaatttttt 6061 gtgacctgtt cggagtgatt agcgttacaa tttgaactga aagtgacatc cagtgtttgt 6121 tccttgtgta gatgcatctc aaaaaaatgg tgggcataat agtgttgttt atatatatca 6181 aaaataacaa ctataataat aagaatacat ttaatttaga aaatgcttgg atttcactgg 6241 aactagggcg cgcctccgga acataatggt gcagggcgct gacttccgcg tttccagact 6301 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca 6361 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca 6421 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca 6481 gggccgcaag cttgcatgcc tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct 6541 ccgagcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg 6601 agcggagact ctagccctag ggcatgcctg caggtcggag tactgtcctc cgagcggagt 6661 actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact 6721 gtcctccgag cggagactct agcgctagcg catgcctgca ggtcggagta ctgtcctccg 6781 agcggagtac tgtcctccga gcggagtact gtcctccgag cggagtactg tcctccgagc 6841 ggagtactgt cctccgagcg gagactctag cactagtgca tgcctgcagg tcggagtact 6901 gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc 6961 ctccgagcgg agtactgtcc tccgagcgga gactctagcg acgtcgagcg ccggagtata 7021 aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc 7081 gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca atctgcagta 7141 aagtgcaagt taaagtgaat caattaaaag taaccagcaa ccaagtaaat caactgcaac 7201 tactgaaatc tgccaagaag taattattga atacaagaag agaactctga atagatctaa 7261 aaggtaggtt caaccactga tgcctaggca caccgaaacg actaacccta attcttatcc 7321 tttacttcag gcggccgcat ggtgtctaag ggcgaagagc tgattaagga gaacatgcac 7381 atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc 7441 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct 7501 ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcag aaccttcatc 7561 aaccacaccc agggcatccc cgatttcttt aagcagtcct tccctgaggg cttcacatgg 7621 gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc 7681 caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttccc atccaactgc 7741 cctgtgatgc agaagaaaac actcggctgg gaggccaaca ccgagatgct gtaccccgct 7801 gacggcggcc tggaaggcag aaccgacatg gccctgaagc tcgtgggcgg gggccacctg 7861 atctgcaact tcaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc 7921 ggcgtctact atgtggacca cagactggaa agaatcaagg aggccgacaa agagacctac 7981 gtcgagcagc acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac 8041 aaacttaatg gcatggacga gctgtacaag ctcgaggacc caaagaagaa gcggaaagtg 8101 gatccaaaga aaaagcgcaa ggtggaccct aagaagaaac ggaaggtgac cggtatcgat 8161 gggtccgggg aaggtcgtgg tagtctacta acgtgtggtg atgtagaaga aaatcctgga 8221 cctatggcct caccgttgac ccgctttctg tcgctgaacc tgctgctgct gggtgagtcg 8281 attatcctgg ggagtggaga agctaagcca caggcacccg aactccgaat ctttccaaag 8341 aaaatggacg ccgaacttgg tcagaaggtg gacctggtat gtgaagtgtt ggggtccgtt 8401 tcgcaaggat gctcttggct cttccagaac tccagctcca aactccccca gcccaccttc 8461 gttgtctata tggcttcatc ccacaacaag ataacgtggg acgagaagct gaattcgtcg 8521 aaactgtttt ctgccatgag ggacacgaat aataagtacg ttctcaccct gaacaagttc 8581 agcaaggaaa acgaaggcta ctatttctgc tcagtcatca gcaactcggt gatgtacttc 8641 agttctgtcg tgccagtcct tcagaaagtg aactctacta ctaccaagcc agtgctgcga 8701 actccctcac ctgtgcaccc taccgggaca tctcagcccc agagaccaga agattgtcgg 8761 ccccgtggct cagtgaaggg gaccggattg gacttcgcct gtgatattta catctgggca 8821 cccttggccg gaatctgcgt ggcccttctg ctgtccttga tcatcactct catctgctac 8881 cacagccgcg gatccatgag taaaggagaa gaacttttca ctggagttgt cccaattctt 8941 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt 9001 gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt 9061 ccatggccaa cacttgtcac tactttaact tatggtgttc aatgcttttc aagataccca 9121 gatcatatga aacagcatga ctttttcaag agtgccatgc ccgaaggtta tgtccaggaa 9181 agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt caagtttgaa 9241 ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga agatggaaac 9301 attcttggac acaaattgga atacaactat aactcacaca atgtatacat catggcagac 9361 aaacaaaaga atggaatcaa agttaacttc aaaattagac acaacattga agatggaagc 9421 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta 9481 ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga 9541 gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg catggatgaa 9601 ctatacaaat aatctagagg atctttgtga aggaacctta cttctgtggt gtgacataat 9661 tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat 9721 aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc tatggaactg 9781 atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc tcagaagaaa 9841 tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct ccaaaaaaga 9901 agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt ttgagtcatg 9961 ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag gaaaaagctg 10021 cactgctata caagaaaatt atggaaaaat atttgatgta tagtgccttg actagagatc 10081 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc 10141 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct 10201 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 10261 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcgat 10321 ctggccggcc gtttaaacga attcttgaag acgaaagggc ctcgtgatac gcctattttt 10381 ataggttaat gtcatgataa taatggtttc tta //
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