质粒名称: | dmBACCS2-IRES-dOrai-IRES-GFP |
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出品公司: | Addgene |
目录编号: | 72893 |
存储实验室: | Takao Nakata |
载体骨架基本信息 |
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载体名称: | pWPRE/C1 (modified pEGFP-C1) |
载体出品公司: | Modified from Clontech |
空载体大小: | 4581 bp |
完整质粒大小: | 9127 bp |
载体修饰: | 将土拨鼠肝炎病毒转录后调节元件(WPRE)插入pEGFP-C1的BamHI位点,并去除EGFP。 |
载体类型: | 哺乳细胞表达 |
筛选标记: | 新霉素(G418) |
载体抗性: | 卡那霉素50 μg/mL |
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: | dmBACCS2-IRES-dOrai-IRES-GFP |
物种来源: | D. melanogaster (fly); Avena sativa (oat) |
插入大小: | 4546 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子: | CMV |
标签/融合蛋白: | IRES-GFP(主干上的C终端) |
克隆方法: | 限制性内切酶 |
5’克隆位点: | XhoI(未破坏) |
3’克隆位点: | SalI(未破坏) |
5’测序引物 | CMV Forward |
3’测序引物 | WPRE-R |
文献引用方法 |
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材料和方法部分: | dmBACCS2-IRES-dOrai-IRES-GFP was a gift from Takao Nakata (Addgene plasmid # 72893 ; http://n2t.net/addgene:72893 ; RRID:Addgene_72893) |
参考文献部分: | Light generation of intracellular Ca(2+) signals by a genetically encoded protein BACCS. Ishii T, Sato K, Kakumoto T, Miura S, Touhara K, Takeuchi S, Nakata T. Nat Commun. 2015 Aug 18;6:8021. doi: 10.1038/ncomms9021. 10.1038/ncomms9021 PubMed 26282514 |
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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 dmBACCS2-IRES-dOrai-IRES-GFP 9127 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of dmBACCS2, a Drosophila melanogaster blue light-activated Ca2+ channel switch, along with dOrai and GFP. ACCESSION . VERSION . KEYWORDS dmBACCS2-IRES-dOrai-IRES-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9127) AUTHORS Ishii T, Sato K, Kakumoto T, Miura S, Touhara K, Takeuchi S, Nakata T TITLE Light generation of intracellular Ca(2+) signals by a genetically encoded protein BACCS. JOURNAL Nat Commun. 2015 Aug 18;6:8021. doi: 10.1038/ncomms9021. PUBMED 26282514 REFERENCE 2 (bases 1 to 9127) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9127 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 603..612 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" misc_feature 2186..2759 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 2209..2761 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" regulatory 3281..3290 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" misc_feature 3843..4416 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 3866..4418 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 4418..5137 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 4418..5134 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4463..4484) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4724..4743) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5071..5092 /label=EGFP-C /note="EGFP, forward primer" misc_feature 5182..5770 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5235..5255) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5653..5664) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 5915..6036 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5952..5971) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6006..6025 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(6043..6498) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6180..6201) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6392..6411 /label=F1ori-R /note="F1 origin, reverse primer" promoter 6525..6629 /gene="bla" /label=AmpR promoter promoter 6631..6988 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(6654..6674) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6839..6974 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6901..6920 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 7023..7817 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(7077..7096) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7687..7706 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(8005..8024) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 8049..8096 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 8425..9013 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8914..8933 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcctcg 601 aggccaccat gggaccggtc ggaggaagcg gtggatcttt ggctactaca cttgaacgta 661 ttgagaagaa ctttgtcatt actgacccaa gattgccaga taatcccatt atattcgcgt 721 ccgatagttt cttgcagttg acagaatata gccgtgaaga aattttggga agaaactgca 781 ggtttctaca aggtcctgaa actgatcgcg cgacagtgag aaaaattaga gatgccatag 841 ataaccaaac agaggtcact gttcagctga ttaattatac aaagagtggt aaaaagttct 901 ggaacctctt tcacttgcag cctatgcgag atcagaaggg agatgtccag tactttattg 961 gggttcagtt ggatggaact gagcatgtcc gagatgctgc cgagagagag ggagtcatgc 1021 tgattaagaa aactgcagaa aatctgcaat catggcttca atacacatat gaactagaaa 1081 gtaagaatca tcagaagaag cgcacgtcgg ctgagaagca gctacagtcg gccagagagg 1141 cttgtgagaa attgcgtaag aaacggtcaa gtttggtggg tgcgttcgtt tccacgcacg 1201 gaaagagtat tgatgatgtg gatcggtcga ttgttgaggc acggaatgcc ctcggagatg 1261 taacaaacga gctgcaagaa cgactgcatc gctggaagca aatcgagacg tgccttggct 1321 taaacattgt gaacaacaat ggtctgcccg gaggatccgg aggaagtgga ggaggtatcc 1381 tgcagtctcg gggtggcagt ggtggttcag gcggttctgg atctttggct actacacttg 1441 aacgtattga gaagaacttt gtcattactg acccaagatt gccagataat cccattatat 1501 tcgcgtccga tagtttcttg cagttgacag aatatagccg tgaagaaatt ttgggaagaa 1561 actgcaggtt tctacaaggt cctgaaactg atcgcgcgac agtgagaaaa attagagatg 1621 ccatagataa ccaaacagag gtcactgttc agctgattaa ttatacaaag agtggtaaaa 1681 agttctggaa cctctttcac ttgcagccta tgcgagatca gaagggagat gtccagtact 1741 ttattggggt tcagttggat ggaactgagc atgtccgaga tgctgccgag agagagggag 1801 tcatgctgat taagaaaact gcagaaaatc tgcaatcatg gcttcaatac acatatgaac 1861 tagaaagtaa gaatcatcag aagaagcgca cgtcggctga gaagcagcta cagtcggcca 1921 gagaggcttg tgagaaattg cgtaagaaac ggtcaagttt ggtgggtgcg ttcgtttcca 1981 cgcacggaaa gagtattgat gatgtggatc ggtcgattgt tgaggcacgg aatgccctcg 2041 gagatgtaac aaacgagctg caagaacgac tgcatcgctg gaagcaaatc gagacgtgcc 2101 ttggcttaaa cattgtgaac aacaatggtc tgcccggcgg ctctggaggt agcggactag 2161 tttgagtcga gccgcggcaa ttccgcccct ctccctcccc cccccctaac gttactggcc 2221 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc 2281 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta 2341 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag 2401 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga 2461 accccccacc tggcaacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg 2521 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat 2581 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta 2641 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa 2701 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat 2761 atgtctgtgt ggaccacggc caacaattcg ggcctagaga cgccaacaaa gtcgccgatc 2821 acgtcgtcgg ttccgcgggc atcgagaagt tctgcggtga tcaccactgg aaatcaccag 2881 cagcaccact tccagcacgt tgtggccgcc gccgttgcag ccgccacttc agtggccacc 2941 ggtcatcagt tccagcagca gttcccgctg cacgcacatc cgcatccgca tccacagcac 3001 cacagcaaca gtcccacggg tagcggcagc aacagcaaca acagcgcagg tttccagcgc 3061 accagcatca gcaactcgct gctgcagttc ccgccgcccc cgcccccttc ctcacagaac 3121 caggcgaagc cgcgcggaca ccatcgcacc gccagcagca gcatgtcgca atccggcgag 3181 gatctccact cgcccaccta cctgtcctgg cggaagctcc agcttagtcg ggccaagcta 3241 aaggcatcca gcaagacgtc cgccctgctc tccggattcg ccatggtggc gatggtggag 3301 gtgcaactgg atcacgatac gaatgtacca ccgggcatgc tgatagcctt cgccatctgc 3361 accaccctcc tggtggcggt gcacatgctg gccctgatga tcagcacctg tatcctgcca 3421 aatattgaga cggtgtgtaa tttgcatagt atttccctgg tccacgaatc gccacacgag 3481 cgtctccatt ggtacattga aacggcgtgg gcattctcca cgctgctggg actcatactc 3541 tttctactgg agatagccat cctgtgctgg gtgaaattct atgacctaag tccgccggcg 3601 gcttggtcag cgtgtgtggt cctcataccg gtgatgatca tctttatggc cttcgccatt 3661 cacttctatc gctccctggt gtcgcacaaa tatgaggtga cagtctcggg catccgtgag 3721 ctggagatgc tcaaggagca gatggagcag gatcatttgg agcatcacaa taacatacga 3781 aataatggca tgaactatgg cgcatccggg gacattgtct aggtcgagcc gcggcaattc 3841 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg 3901 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc 3961 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa 4021 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga 4081 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg caacaggtgc 4141 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc 4201 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac 4261 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg 4321 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac 4381 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gtgagcaagg gcgaggagct 4441 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt 4501 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat 4561 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg 4621 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc 4681 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa 4741 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg 4801 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag 4861 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat 4921 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc 4981 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct 5041 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc 5101 cgggatcact ctcggcatgg acgagctgta caagtaaccg cggtcgacgg taccgcgggc 5161 ccgggatcta agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact 5221 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 5281 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 5341 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 5401 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 5461 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 5521 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca 5581 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 5641 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 5701 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 5761 gcctccccgc atcgataccg tggatcgatc caccggatct agataactga tcataatcag 5821 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa 5881 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg 5941 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 6001 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata 6061 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg 6121 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc 6181 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa 6241 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt 6301 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac 6361 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta 6421 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg 6481 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 6541 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 6601 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt 6661 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 6721 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 6781 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 6841 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 6901 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg 6961 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc 7021 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 7081 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 7141 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 7201 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 7261 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 7321 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 7381 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 7441 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 7501 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg 7561 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 7621 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 7681 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 7741 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 7801 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 7861 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 7921 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 7981 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg 8041 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa 8101 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg 8161 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc 8221 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat 8281 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 8341 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 8401 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 8461 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 8521 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt 8581 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 8641 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 8701 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 8761 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 8821 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 8881 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 8941 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 9001 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 9061 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 9121 catgcat //