质粒名称: | pLenti CMV GFP Blast (659-1) |
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出品公司: | 爱迪基因 |
目录编号: | 17445 |
存储实验室: | Eric Campeau, Paul Kaufman |
载体骨架基本信息 |
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载体名称: | p156RRL-sinPPT-CMV-GFP-PRE/Nhe I |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 7603 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 |
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: | GFP,enhanced GFP |
物种来源: | -- |
插入大小: | 719 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | BamHI(未破坏) |
3' 克隆位点: | Sal I(未破坏) |
5' 测序引物 | CMVforw |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pLenti CMV GFP Blast (659-1) was a gift from Eric Campeau & Paul Kaufman (Addgene plasmid # 17445 ; http://n2t.net/addgene:17445 ; RRID:Addgene_17445) |
参考文献部分: | A versatile viral system for expression and depletion of proteins in mammalian cells. Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD.. PLoS One. 2009 Aug 6;4(8):e6529. 10.1371/journal.pone.0006529 PubMed 19657394 |
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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 pLenti CMV GFP Blast (659-1) 8311 bp ds-DNA circular SYN 03-MAY-2019 DEFINITION 3rd gen lentiviral eGFP expression vector, CMV promoter, Blast. ACCESSION . VERSION . KEYWORDS pLenti CMV GFP Blast (659-1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8311) AUTHORS Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD. TITLE A versatile viral system for expression and depletion of proteins in mammalian cells. JOURNAL PLoS One. 2009 Aug 6;4(8):e6529. PUBMED 19657394 REFERENCE 2 (bases 1 to 8311) AUTHORS . TITLE Direct Submission JOURNAL Exported May 3, 2019 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8311 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 26..130 /gene="bla" /label=AmpR promoter CDS 131..991 /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(349..368) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1162..1750 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1651..1670 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1904..1921 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2038..2059 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2074..2104 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2112..2128 /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)." primer_bind 2117..2139 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2136..2152 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2136..2152 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2171..2191 /label=T3 /note="T3 promoter, forward primer" promoter 2173..2191 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 2219..2445 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 2446..2626 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 2673..2798 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 3291..3524 /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 4020..4136 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 4159..4462 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4463..4666 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4617..4637 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4663..4687 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 4756..4765 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4762..5481 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4807..4828) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5068..5087) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5415..5436 /label=EGFP-C /note="EGFP, forward primer" misc_feature 5497..6085 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5550..5570) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5968..5979) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(6136..6156) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6141..6470 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6321..6456 /label=SV40 ori /note="SV40 origin of replication" promoter 6518..6565 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 6584..6982 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" LTR 7069..7302 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 7374..7495 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7411..7430) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7465..7484 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 7514..7649 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(7669..7688) /label=T7 /note="T7 promoter, forward primer" promoter complement(7670..7688) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7698..7715) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7698..7714) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7707..7729) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 7856..8311 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7943..7962) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8153..8174 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca 61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa 121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt 181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca 241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag 301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc 361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca 421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt 481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct 541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt 601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga 661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact 721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc 781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga 841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt 901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga 961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact 1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga 1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta 1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa 2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat 2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta 2161 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg 2221 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc 2281 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt 2341 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg 2401 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta 2461 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa 2521 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac 2581 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca 2641 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg 2701 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta 2761 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta 2821 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac 2881 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa 2941 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag 3001 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag 3061 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga 3121 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat 3181 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc 3241 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg 3301 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg 3361 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct 3421 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca 3481 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc 3541 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct 3601 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac 3661 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa 3721 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg 3781 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt 3841 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag 3901 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga 3961 gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact 4021 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata 4081 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaattcaa aattttatcg 4141 ataagcttgg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc 4201 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg 4261 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca 4321 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc 4381 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt 4441 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata 4501 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt 4561 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca 4621 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg 4681 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgactcta 4741 gaggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg 4801 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 4861 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 4921 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 4981 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 5041 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 5101 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 5161 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 5221 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 5281 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 5341 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 5401 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 5461 atggacgagc tgtacaagta aagcggccgc gtcgacaatc aacctctgga ttacaaaatt 5521 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5581 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5641 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5701 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5761 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5821 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 5881 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg 5941 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 6001 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 6061 atctcccttt gggccgcctc cccgcctgga attctgcaga tatccggtta gtaatgagtt 6121 tggaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 6181 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 6241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 6301 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 6361 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 6421 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 6481 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata 6541 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agcctttgtc 6601 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc 6661 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt 6721 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc 6781 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat 6841 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc 6901 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc 6961 tggttatgtg tgggagggct aagcacaatt cgagctcggt acctttaaga ccaatgactt 7021 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctag 7081 ctcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc 7141 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa 7201 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga 7261 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc 7321 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt 7381 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 7441 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 7501 tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 7561 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca 7621 gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag 7681 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc 7741 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa 7801 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg 7861 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 7921 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 7981 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 8041 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 8101 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 8161 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 8221 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 8281 aattttaaca aaatattaac gcttacaatt t //