质粒名称: | pMSCV-hygro-STING R232 |
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出品公司: | Addgene |
目录编号: | 102608 |
存储实验室: | Nicolas Manel |
载体骨架基本信息 |
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载体名称: | pMSCV-hygro |
完整质粒大小: | 8085bp |
载体类型: | 哺乳细胞表达, Retroviral |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | STING |
物种来源: | H. sapiens (human) |
基因突变: | R232 |
克隆方法: | 限制性内切酶 |
5’克隆位点: | unknown (unknown if destroyed) |
3’克隆位点: | unknown (unknown if destroyed) |
5’测序引物 | MSCV |
文献引用方法 |
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材料和方法部分: | pMSCV-hygro-STING R232 was a gift from Nicolas Manel (Addgene plasmid # 102608 ; http://n2t.net/addgene:102608 ; RRID:Addgene_102608) |
参考文献部分: | Transmission of innate immune signaling by packaging of cGAMP in viral particles. Gentili M, Kowal J, Tkach M, Satoh T, Lahaye X, Conrad C, Boyron M, Lombard B, Durand S, Kroemer G, Loew D, Dalod M, Thery C, Manel N. Science. 2015 Sep 11;349(6253):1232-6. doi: 10.1126/science.aab3628. Epub 2015 Jul 30. 10.1126/science.aab3628 PubMed 26229115 |
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载体基本信息
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载体序列
LOCUS pMSCV-hygro-STING R232 8085 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Retroviral vector to expression STING R232. ACCESSION . VERSION . KEYWORDS pMSCV-hygro-STING R232 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8085) AUTHORS Gentili M, Kowal J, Tkach M, Satoh T, Lahaye X, Conrad C, Boyron M, Lombard B, Durand S, Kroemer G, Loew D, Dalod M, Thery C, Manel N TITLE Transmission of innate immune signaling by packaging of cGAMP in viral particles. JOURNAL Science. 2015 Sep 11;349(6253):1232-6. doi: 10.1126/science.aab3628. Epub 2015 Jul 30. PUBMED 26229115 REFERENCE 2 (bases 1 to 8085) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..8085 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 94..610 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" misc_feature 674..1015 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 1082..1498 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1428..1450 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1466..1482 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" promoter 2673..3172 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(2708..2731) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 3105..3124 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 3190..4215 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" LTR 4573..5087 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 5249..5265 /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(5273..5303) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5318..5339 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5456..5473) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5627..6215) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5707..5726) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6386..7246) /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 7009..7028 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7247..7351) /gene="bla" /label=AmpR promoter primer_bind 7419..7437 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7475..7497) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7597..7616 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7810..7832 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7960..7979 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 agttttgact caacaatatc accagctgaa gcctatagag tacgagccat agataaaata 61 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg 121 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag 181 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg 241 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc 301 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat 361 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt 421 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga 481 tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa 541 tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc cacctcgggg 601 gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac caccgacccc 661 cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt 721 ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat 781 ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg 841 tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa 901 tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc 961 tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg 1021 ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta 1081 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg 1141 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc 1201 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat 1261 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc 1321 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt 1381 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc 1441 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg 1501 aattagatcc atgccccact ccagcctgca tccatccatc ccgtgtccca ggggtcacgg 1561 ggcccagaag gcagccttgg ttctgctgag tgcctgcctg gtgacccttt gggggctagg 1621 agagccacca gagcacactc tccggtacct ggtgctccac ctagcctccc tgcagctggg 1681 actgctgtta aacggggtct gcagcctggc tgaggagctg cgccacatcc actccaggta 1741 ccggggcagc tactggagga ctgtgcgggc ctgcctgggc tgccccctcc gccgtggggc 1801 cctgttgctg ctgtccatct atttctacta ctccctccca aatgcggtcg gcccgccctt 1861 cacttggatg cttgccctcc tgggcctctc gcaggcactg aacatcctcc tgggcctcaa 1921 gggcctggcc ccagctgaga tctctgcagt gtgtgaaaaa gggaatttca acgtggccca 1981 tgggctggca tggtcatatt acatcggata tctgcggctg atcctgccag agctccaggc 2041 ccggattcga acttacaatc agcattacaa caacctgcta cggggtgcag tgagccagcg 2101 gctgtatatt ctcctcccat tggactgtgg ggtgcctgat aacctgagta tggctgaccc 2161 caacattcgc ttcctggata aactgcccca gcagaccggt gaccgtgctg gcatcaagga 2221 tcgggtttac agcaacagca tctatgagct tctggagaac gggcagcggg cgggcacctg 2281 tgtcctggag tacgccaccc ccttgcagac tttgtttgcc atgtcacaat acagtcaagc 2341 tggctttagc cgggaggata ggcttgagca ggccaaactc ttctgccgga cacttgagga 2401 catcctggca gatgcccctg agtctcagaa caactgccgc ctcattgcct accaggaacc 2461 tgcagatgac agcagcttct cgctgtccca ggaggttctc cggcacctgc ggcaggagga 2521 aaaggaagag gttactgtgg gcagcttgaa gacctcagcg gtgcccagta cctccacgat 2581 gtcccaagag cctgagctcc tcatcagtgg aatggaaaag cccctccctc tccgcacgga 2641 tttctcttga ctcgaggtta acgaattcta ccgggtaggg gaggcgcttt tcccaaggca 2701 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg 2761 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc 2821 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc 2881 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag 2941 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc 3001 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg 3061 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca 3121 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcat 3181 cccgccacca tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa 3241 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc 3301 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc 3361 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg 3421 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt 3481 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag 3541 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga 3601 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc 3661 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct 3721 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg 3781 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg 3841 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg 3901 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca 3961 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc 4021 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc 4081 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg 4141 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg 4201 agggcaaagg aatagagtag atgccgaccg aacaagagct gatttcgaga acgcctcagc 4261 cagcaactcg cgcgagccta gcaaggcaaa tgcgagagaa cggccttacg cttggtggca 4321 cagttctcgt ccacagttcg ctaagctcgc tcggctgggt cgcgggaggg ccggtcgcag 4381 tgattcaggc ccttctggat tgtgttggtc cccagggcac gattgtcatg cccacgcact 4441 cgggtgatct gactgatccc gcagattgga gatcgccgcc cgtgcctgcc gattgggtgc 4501 agatccgtcg acctgcagcc aagcttatcg ataaaataaa agattttatt tagtctccag 4561 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca 4621 ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa 4681 cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg 4741 ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa 4801 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact 4861 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa 4921 gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc 4981 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg 5041 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc 5101 ttgaagttgg agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa 5161 ttagccactg ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc 5221 gcagaaagag ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac 5281 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 5341 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 5401 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 5461 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 5521 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 5581 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 5641 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 5701 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 5761 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 5821 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 5881 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 5941 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 6001 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 6061 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 6121 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 6181 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 6241 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 6301 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 6361 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 6421 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 6481 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 6541 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 6601 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 6661 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 6721 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 6781 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 6841 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 6901 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 6961 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 7021 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 7081 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 7141 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 7201 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 7261 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 7321 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 7381 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct 7441 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga 7501 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag 7561 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga 7621 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca 7681 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt 7741 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc 7801 cagggttttc ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga 7861 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc 7921 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc 7981 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg 8041 attagtccaa tttgttaaag acaggatatc agtggtccag gctct //
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