质粒名称: | pLenti-GFP1-9 |
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出品公司: | addgene |
目录编号: | 113613 |
存储实验室: | Xiaokun Shu |
载体骨架基本信息 |
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载体名称: | N/A |
完整质粒大小: | 9597 bp |
载体类型: | 哺乳细胞表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 30 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 未知 |
插入基因信息 |
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插入基因名称: | GFP1-9 |
文献引用方法 |
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材料和方法部分: | pLenti-GFP1-9 was a gift from Xiaokun Shu (Addgene plasmid # 113613 ; http://n2t.net/addgene:113613 ; RRID:Addgene_113613) |
参考文献部分: | A rapid fluorogenic GPCR-beta-arrestin interaction assay. Zhang Q, Zheng YW, Coughlin SR, Shu X. Protein Sci. 2018 Apr;27(4):874-879. doi: 10.1002/pro.3385. Epub 2018 Feb 23. 10.1002/pro.3385 PubMed 29411438 |
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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-GFP1-9 9597 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION PPI reporter. ACCESSION . VERSION . KEYWORDS pLenti-GFP1-9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9597) AUTHORS Zhang Q, Zheng YW, Coughlin SR, Shu X TITLE A rapid fluorogenic GPCR-beta-arrestin interaction assay. JOURNAL Protein Sci. 2018 Apr;27(4):874-879. doi: 10.1002/pro.3385. Epub 2018 Feb 23. PUBMED 29411438 REFERENCE 2 (bases 1 to 9597) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..9597 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 487..1075 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(540..560) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(958..969) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(1078..1094) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1079..1095) /label=pBluescriptKS /note="For pBluescript vector" LTR 1185..1418 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" promoter complement(1443..1461) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1444..1461) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1751..1770) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(1984..2003) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2103..2125 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2163..2181) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2249..2353 /gene="bla" /label=AmpR promoter CDS 2354..3214 /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(2572..2591) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3385..3973 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3874..3893 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4127..4144 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(4214..4234) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4219..4548 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4399..4534 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4461..4480 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 5682..5747 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 5877..5897 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 6322..6456 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6359..6378) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6413..6432 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 6624..7257 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 7304..7429 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 7926..8159 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8344..8388 /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 8673..8790 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 8943..9350 /label=SFFV promoter /note="spleen focus-forming virus long terminal repeat (LTR) promoter" ORIGIN 1 gccgtggccg accctggtga ccaccctgac ctatggcgtg cagtgcttta gccgctatcc 61 ggatcatatg aaacgccatg atttttttaa aagcgcgatg ccggaaggct atgtgcagga 121 acgcaccatt tattttaaag atgatggcac ctataaaacc cgcgcggaag tgaaatttga 181 aggcgatacc ctggtgaacc gcattgaact gaaaggcatt gattttaaag aagatggcaa 241 cattctgggc cataaactgg aatataactt taacagccat aaagtgtata ttaccgcgga 301 taaacagaac aacggcatta aagcgaactt taccattcgc cataacgtgg aagatggcag 361 cgtgcagctg gcggatcatt atcagcagaa caccccgatt ggcgatggcc cggttcttct 421 tccttgagcg gccgcgactc tagagtcgac ctgcaggcat gcaagcttga tatcaagctt 481 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 541 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 601 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 661 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 721 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 781 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 841 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg 901 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 961 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 1021 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga 1081 taccgtcgac ctcgagggaa ttaattcgag ctcggtacct ttaagaccaa tgacttacaa 1141 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 1201 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 1261 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 1321 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 1381 cctcagaccc ttttagtcag tgtggaaaat ctctagcagc atctagaatt aattccgtgt 1441 attctatagt gtcacctaaa tcgtatgtgt atgatacata aggttatgta ttaattgtag 1501 ccgcgttcta acgacaatat gtacaagcct aattgtgtag catctggctt actgaagcag 1561 accctatcat ctctctcgta aactgccgtc agagtcggtt tggttggacg aaccttctga 1621 gtttctggta acgccgtccc gcacccggaa atggtcagcg aaccaatcag cagggtcatc 1681 gctagccaga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg 1741 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg 1801 ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg 1861 ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta 1921 ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaat ggtgcactct 1981 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc 2041 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt 2101 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa 2161 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac 2221 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 2281 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 2341 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc 2401 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga 2461 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga 2521 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg 2581 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc 2641 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac 2701 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact 2761 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca 2821 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg 2881 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact 2941 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg 3001 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 3061 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 3121 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc 3181 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 3241 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 3301 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 3361 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 3421 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 3481 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 3541 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 3601 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 3661 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 3721 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 3781 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 3841 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 3901 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 3961 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 4021 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 4081 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag 4141 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca 4201 ttaatgcagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 4261 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 4321 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 4381 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 4441 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta 4501 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttggacac 4561 aagacaggct tgcgagatat gtttgagaat accactttat cccgcgtcag ggagaggcag 4621 tgcgtaaaaa gacgcggact catgtgaaat actggttttt agtgcgccag atctctataa 4681 tctcgcgcaa cctattttcc cctcgaacac tttttaagcc gtagataaac aggctgggac 4741 acttcacatg agcgaaaaat acatcgtcac ctgggacatg ttgcagatcc atgcacgtaa 4801 actcgcaagc cgactgatgc cttctgaaca atggaaaggc attattgccg taagccgtgg 4861 cggtctgtac cgggtgcgtt actggcgcgt gaactgggta ttcgtcatgt cgataccgtt 4921 tgtatttcca gctacgatca cgacaaccag cgcgagctta aagtgctgaa acgcgcagaa 4981 ggcgatggcg aaggcttcat cgttattgat gacctggtgg ataccggtgg tactgcggtt 5041 gcgattcgtg aaatgtatcc aaaagcgcac tttgtcacca tcttcgcaaa accggctggt 5101 cgtccgctgg ttgatgacta tgttgttgat atcccgcaag atacctggat tgaacagccg 5161 tgggatatgg gcgtcgtatt cgtcccgcca atctccggtc gctaatcttt tcaacgcctg 5221 gcactgccgg gcgttgttct ttttaacttc aggcgggtta caatagtttc cagtaagtat 5281 tctggaggct gcatccatga cacaggcaaa cctgagcgaa accctgttca aaccccgctt 5341 taaacatcct gaaacctcga cgctagtccg ccgctttaat cacggcgcac aaccgcctgt 5401 gcagtcggcc cttgatggta aaaccatccc tcactggtat cgcatgatta accgtctgat 5461 gtggatctgg cgcggcattg acccacgcga aatcctcgac gtccaggcac gtattgtgat 5521 gagcgatgcc gaacgtaccg acgatgattt atacgatacg gtgattggct accgtggcgg 5581 caactggatt tatgagtggg ccccggatct ttgtgaagga accttacttc tgtggtgtga 5641 cataattgga caaactacct acagagattt aaagctctaa ggtaaatata aaatttttaa 5701 gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt attttagatt ccaacctatg 5761 gaactgatga atgggagcag tggtggaatg cctttaatga ggaaaacctg ttttgctcag 5821 aagaaatgcc atctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa 5881 aaaagaagag aaaggtagaa gaccccaagg actttccttc agaattgcta agttttttga 5941 gtcatgctgt gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa 6001 aagctgcact gctatacaag aaaattatgg aaaaatattc tgtaaccttt ataagtaggc 6061 ataacagtta taatcataac atactgtttt ttcttactcc acacaggcat agagtgtctg 6121 ctattaataa ctatgctcaa aaattgtgta cctttagctt tttaatttgt aaaggggtta 6181 ataaggaata tttgatgtat agtgccttga ctagagatca taatcagcca taccacattt 6241 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa 6301 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc 6361 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 6421 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa 6481 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat 6541 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa 6601 atatgtacta caaacttagt agttggaagg gctaattcac tcccaaagaa gacaagatat 6661 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc 6721 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga 6781 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag 6841 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct 6901 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg 6961 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac 7021 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt 7081 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 7141 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 7201 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg 7261 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac 7321 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa 7381 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg 7441 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata 7501 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg 7561 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca 7621 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca 7681 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa 7741 caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga 7801 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca 7861 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg 7921 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg 7981 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac 8041 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc 8101 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg 8161 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt 8221 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga 8281 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa 8341 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt 8401 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta 8461 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca 8521 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata 8581 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga 8641 cggtatcgcc aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg 8701 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta 8761 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca 8821 gtttggatcg ataagcttga tatcgaattc ctgcagcccc gataaaataa aagattttat 8881 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctg 8941 cagtaacgcc attttgcaag gcatggaaaa ataccaaacc aagaatagag aagttcagat 9001 caagggcggg tacatgaaaa tagctaacgt tgggccaaac aggatatctg cggtgagcag 9061 tttcggcccc ggcccggggc caagaacaga tggtcaccgc agtttcggcc ccggcccgag 9121 gccaagaaca gatggtcccc agatatggcc caaccctcag cagtttctta agacccatca 9181 gatgtttcca ggctccccca aggacctgaa atgaccctgc gccttatttg aattaaccaa 9241 tcagcctgct tctcgcttct gttcgcgcgc ttctgcttcc cgagctctat aaaagagctc 9301 acaacccctc actcggcgcg ccagtcctcc gacagactga gtcgcccggg ggggatctgg 9361 agctctcgag aattctcacg cgtctgcagg atatcaagct tgcggtaccg cgggcccggg 9421 atccgccgcc accatgcgca aaggcgaaga actgtttacc ggcattgtgc cgattctggt 9481 ggaactggat ggcgatgtga acggccataa attttttgtg cgcggcgaag gcgaaggcga 9541 tgcgaccatt ggcaaactga gcctgaaatt tatttgcacc accggcaaac tgccggt //