质粒名称: | miniTurbo-His6_pET21a |
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出品公司: | addgene |
目录编号: | 107170 |
存储实验室: | Alice Ting |
载体骨架基本信息 |
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载体名称: | pCDNA3 |
完整质粒大小: | 6245 bp |
载体类型: | 哺乳细胞表达 |
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: | miniTurbo (BirA mutant) |
基因突变: | aa1-63 deleted; Q65P, I87V, R118S, E140K, Q141R, S150G, L151P, V160A, T192A, K194I, M209V, I305V |
克隆方法: | 限制性内切酶 |
5’克隆位点: | 未知 |
3’克隆位点: | 未知 |
文献引用方法 |
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材料和方法部分: | V5-miniTurbo-NES_pCDNA3 was a gift from Alice Ting (Addgene plasmid # 107170 ; http://n2t.net/addgene:107170 ; RRID:Addgene_107170) |
参考文献部分: | Efficient proximity labeling in living cells and organisms with TurboID. Branon TC, Bosch JA, Sanchez AD, Udeshi ND, Svinkina T, Carr SA, Feldman JL, Perrimon N, Ting AY. Nat Biotechnol. 2018 Aug 20. pii: nbt.4201. doi: 10.1038/nbt.4201. 10.1038/nbt.4201 PubMed 30125270 |
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V5-miniTurbo-NES_pCDNA3载体质粒基本信息
V5-miniTurbo-NES_pCDNA3质粒图谱载体图谱和V5-miniTurbo-NES_pCDNA3载体序列质粒序列多克隆位点信息
V5-miniTurbo-NES_pCDNA3质粒载体简介
V5-miniTurbo-NES_pCDNA3质粒序列载体序列
LOCUS V5-miniTurbo-NES_pCDNA3 6245 bp ds-DNA circular SYN 31-MAY-2022 DEFINITION expresses V5-tagged miniTurbo in the mammalian cytosol. ACCESSION . VERSION . KEYWORDS V5-miniTurbo-NES_pCDNA3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6245) AUTHORS Branon TC, Bosch JA, Sanchez AD, Udeshi ND, Svinkina T, Carr SA, Feldman JL, Perrimon N, Ting AY TITLE Efficient proximity labeling in living cells and organisms with TurboID. JOURNAL Nat Biotechnol. 2018 Aug 20. pii: nbt.4201. doi: 10.1038/nbt.4201. PUBMED 30125270 REFERENCE 2 (bases 1 to 6245) AUTHORS . TITLE Direct Submission JOURNAL Exported May 31, 2022 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..6245 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 27..162 /label=SV40 ori /note="SV40 origin of replication" primer_bind 89..108 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 243..1037 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(297..316) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 907..926 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 1211..1332 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1248..1267) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1302..1321 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(1381..1397) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1381..1397) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1394..1416) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1405..1421 /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(1429..1459) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1474..1495 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1612..1629) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1783..2371) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1863..1882) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2542..3402) /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 3165..3184 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3403..3507) /gene="bla" /label=AmpR promoter primer_bind complement(3582..3601) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 3773..4152 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4153..4356 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4307..4327 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4401..4420 /label=T7 /note="T7 promoter, forward primer" promoter 4401..4419 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 4439..4448 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4448..4489 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 4496..5266 /codon_start=1 /product="N-terminally truncated mutant of the E. coli biotin ligase BioID engineered for improved catalytic efficiency (Branon et al., 2018)" /label=miniTurbo /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACI AEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADK VRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQ EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF GISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK" CDS 5273..5299 /codon_start=1 /product="nuclear export signal from the HIV Rev protein (Fischer et al., 1995)" /label=NES /translation="LPPLERLTL" promoter complement(5336..5354) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(5337..5354) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(5374..5391) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5380..5604 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5650..6078 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5737..5756) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5947..5968 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6087..6107) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6092..176 /label=SV40 promoter /note="SV40 enhancer and early promoter" ORIGIN 1 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 61 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 121 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 181 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 241 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 301 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 361 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 421 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 481 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 541 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 601 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 661 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 721 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 781 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 841 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 901 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 961 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 1021 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 1081 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 1141 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 1201 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 1261 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 1321 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 1381 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 1441 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 1501 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 1561 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 1621 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 1681 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 1741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 1801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 1861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 1921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 1981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 2041 caaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 2101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 2161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 2221 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 2281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 2341 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 2401 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 2461 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 2521 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 2581 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 2641 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 2701 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 2761 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 2821 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 2881 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 2941 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 3001 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 3061 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 3121 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 3181 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 3241 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 3301 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 3361 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 3421 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 3481 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga 3541 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc 3601 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg 3661 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt 3721 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga 3781 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg 3841 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 3901 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 3961 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 4021 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 4081 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 4141 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 4201 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 4261 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 4321 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact 4381 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttgcggccgc 4441 caccatgggc aagcccatcc ccaaccccct gctgggcctg gacagcaccg ctagcatccc 4501 gctgctgaac gctaaacaga ttctgggaca gctggacggc gggagcgtgg cagtcctgcc 4561 tgtggtcgac tccaccaatc agtacctgct ggatcgaatc ggcgagctga agagtgggga 4621 tgcttgcatt gcagaatatc agcaggcagg gagaggaagc agagggagga aatggttctc 4681 tccttttgga gctaacctgt acctgagtat gttttggcgc ctgaagcggg gaccagcagc 4741 aatcggcctg ggcccggtca tcggaattgt catggcagaa gcgctgcgaa agctgggagc 4801 agacaaggtg cgagtcaaat ggcccaatga cctgtatctg caggatagaa agctggcagg 4861 catcctggtg gagctggccg gaataacagg cgatgctgca cagatcgtca ttggcgccgg 4921 gattaacgtg gctatgaggc gcgtggagga aagcgtggtc aatcagggct ggatcacact 4981 gcaggaagca gggattaacc tggacaggaa tactctggcc gctatgctga tccgagagct 5041 gcgggcagcc ctggaactgt tcgagcagga aggcctggct ccatatctgt cacggtggga 5101 gaagctggat aacttcatca atagacccgt gaagctgatc attggggaca aagagatttt 5161 cgggattagc cgggggattg ataaacaggg agccctgctg ctggaacagg acggagttat 5221 caaaccctgg atgggcggag aaatcagtct gcggtctgcc gaaaagctgc agctgcctcc 5281 cctggagcgc ctgaccctgg actaatagct cgagcatgca tctagagggc cctattctat 5341 agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc 5401 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg 5461 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc 5521 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg 5581 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg 5641 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 5701 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 5761 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt 5821 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac 5881 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 5941 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt 6001 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac 6061 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc 6121 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg 6181 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 6241 agcaa //
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