| 质粒名称: | 3xHA-TurboID-NLS_pCDNA3 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 107171 |
| 存储实验室: | Alice Ting |
|
载体骨架基本信息 |
|
| 载体名称: | pCDNA3 |
| 完整质粒大小: | 6524bp |
| 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 |
|
| 插入基因名称: | TurboID (BirA mutant) |
| 物种来源: | 光合细菌 Rhodopseudomonas palustris |
| 插入大小: | 951 bp |
| 基因突变: | Q65P, I87V, R118S, E140K, Q141R, S150G, L151P, V160A, T192A, K194I, M209V, M241T, S263P, 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 |
|
| 材料和方法部分: | xHA-TurboID-NLS_pCDNA3 was a gift from Alice Ting (Addgene plasmid # 107171 ; http://n2t.net/addgene:107171 ; RRID:Addgene_107171) |
| 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 |
买家导航
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 3xHA-TurboID-NLS_pCDNA3 6524 bp ds-DNA circular SYN 20-DEC-2021
DEFINITION expresses 3xHA-tagged TurboID in the mammalian nucleus.
ACCESSION .
VERSION .
KEYWORDS 3xHA-TurboID-NLS_pCDNA3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6524)
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 6524)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 20, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6524
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(19..38)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 265..285
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(294..722)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(404..425)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 616..635
/label=F1ori-R
/note="F1 origin, reverse primer"
polyA_signal 768..992
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 981..998
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
promoter 1018..1036
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1018..1035
/label=SP6
/note="SP6 promoter, forward primer"
CDS complement(1067..1087)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(1091..1111)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(1115..1135)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(1154..2110)
/codon_start=1
/product="mutant of the E. coli biotin ligase BioID
engineered for improved catalytic efficiency (Branon et
al., 2018)"
/label=TurboID
/translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
CDS complement(2117..2143)
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS complement(2147..2173)
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS complement(2177..2203)
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind complement(2231..2250)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2232..2250)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(2295..2498)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(2324..2344)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 2499..2878
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 3050..3069
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 3144..3248
/gene="bla"
/label=AmpR promoter
CDS 3249..4109
/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(3467..3486)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4280..4868
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4769..4788
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5022..5039
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5156..5177
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5192..5222
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5230..5246
/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 5235..5257
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5254..5270
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5254..5270
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 5319..5440
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5330..5349)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5384..5403
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(5614..6408)
/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(5725..5744)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 6335..6354
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(6475..280)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6489..100
/label=SV40 ori
/note="SV40 origin of replication"
ORIGIN
1 gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
61 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
121 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
181 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
241 ctggggagcc tggggacttt ccacacccta actgacacac attccacaga attaattcgc
301 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
361 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
421 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
481 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
541 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
601 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
661 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
721 gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca tagagcccac
781 cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac
841 cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta
901 ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca
961 aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcgag ctctagcatt
1021 taggtgacac tatagaatag ggccctctag atgcatgctc gagtcacacc ttcctcttct
1081 tcttggggtc caccttcctc ttcttcttgg ggtccacctt cctcttcttc ttggggtcgg
1141 ccctgctgaa ttccttttcg gcagaccgca gactgatttc tccgcccatc cagggtttga
1201 taactccgtc ctgttccagc agcagggctc cctgtttatc aatcccccgg ctaatcccga
1261 aaatctcttt gtccccaatg atcagcttca cgggtctatt gatgaagtta tccagcttct
1321 cccaccgtgg cagatatgga gccaggcctt cctgctcgaa cagttccagg gctgcccgca
1381 gctctcggat cagcgtagcg gccagagtat tcctgtccag gttaatccct gcttcctgca
1441 gtgtgatcca gccctgattg accacgcttt cctccacgcg cctcatagcc acgttaatcc
1501 cggcgccaat gacgatctgt gcagcatcgc ctgttattcc ggccagctcc accaggatgc
1561 ctgccagctt tctatcctgc agatacaggt cattgggcca tttgactcgc accttgtctg
1621 ctcccagctt tcgcagcgct tctgccatga caattccgat gaccgggccc aggccgattg
1681 ctgctggtcc ccgcttcagg cgccaaaaca tactcaggta caggttagct ccaaaaggag
1741 agaaccattt cctccctctg cttcctctcc ctgcctgctg atattctgca atgcaagcat
1801 ccccactctt cagctcgccg attcgatcca gcaggtactg attggtggag tcgaccacag
1861 gcaggactgc cacgctcccg ccgtccagct gtcccagaat ctgtttagcg ttcagcagcg
1921 ggataggctc gggcagagag tagccctttc cgggcacggt gaacacgtcc acgccccagt
1981 cgcgcagagt ctgaatgtgc ttgttgatag cggccctgga catgcccagg gtttctccca
2041 gctgttcgcc actatggaac tcgccattag ccaggagagc gatcagcttc agaggcacag
2101 tattgtcttt gctagctgcg tagtctggga cgtcataggg gtacccggca tagtcgggca
2161 cgtcgtaggg atagccagcg taatccggaa catcatacgg gtacatggtg gcggccgcaa
2221 gcttgggtct ccctatagtg agtcgtatta atttcgataa gccagtaagc agtgggttct
2281 ctagttagcc agagagctct gcttatatag acctcccacc gtacacgcct accgcccatt
2341 tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa
2401 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
2461 tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
2521 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
2581 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
2641 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
2701 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
2761 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
2821 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcaa
2881 cgcgtatatc tggcccgtac atcgcgaagc agcgcaaaac gcctaaccct aagcagattc
2941 ttcatgcaat tgtcggtcaa gccttgcctt gttgtagctt aaattttgct cgcgcactac
3001 tcagcgacct ccaacacaca agcagggagc agatactggc ttaactatgc ggcatcagag
3061 cagattgtac tgagagtgca ccatagggga tcgggagatc tcccgatccg tcgacgtcag
3121 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
3181 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
3241 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
3301 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
3361 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
3421 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
3481 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
3541 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
3601 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
3661 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
3721 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
3781 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
3841 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
3901 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
3961 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
4021 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
4081 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
4141 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
4201 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
4261 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
4321 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
4381 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
4441 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
4501 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
4561 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
4621 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
4681 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
4741 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
4801 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
4861 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
4921 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
4981 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
5041 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
5101 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
5161 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
5221 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
5281 ccaagctcta gctagaggtc gacggtatac agacatgata agatacattg atgagtttgg
5341 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
5401 tgctttattt gtaaccatta taagctgcaa taaacaagtt ggggtgggcg aagaactcca
5461 gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc
5521 ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg
5581 cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata
5641 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
5701 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
5761 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
5821 gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg
5881 cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
5941 atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc
6001 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
6061 catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac
6121 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
6181 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag
6241 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa
6301 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
6361 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc
6421 tcatcctgtc tcttgatcag atccgaaaat ggatatacaa gctcccggga gctttttgca
6481 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aata
//
