| 质粒名称: | V5-TurboID-NES_pCDNA3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 107169 |
| 存储实验室: | Alice Ting |
|
载体骨架基本信息 |
|
| 载体名称: | pCDNA3 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | TurboID (BirA mutant) |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | Q65P, I87V, R118S, E140K, Q141R, S150G, L151P, V160A, T192A, K194I, M209V, M241T, S263P, I305V |
| 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: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | V5-TurboID-NES_pCDNA3 was a gift from Alice Ting (Addgene plasmid # 107169 ; http://n2t.net/addgene:107169 ; RRID:Addgene_107169) |
| 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 V5-TurboID-NES_pCDNA3 6431 bp ds-DNA circular SYN 21-AUG-2019
DEFINITION expresses V5-tagged TurboID in the mammalian cytosol.
ACCESSION .
VERSION .
KEYWORDS V5-TurboID-NES_pCDNA3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6431)
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 6431)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 21, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6431
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 667..676
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 676..717
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 1687..1713
/codon_start=1
/product="nuclear export signal from the HIV Rev protein
(Fischer et al., 1995)"
/label=NES
/translation="LPPLERLTL"
promoter complement(1750..1768)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1751..1768)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1788..1805)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1794..2018
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2064..2492
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2151..2170)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2361..2382
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2501..2521)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2506..2835
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2686..2821
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2748..2767
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2902..3696
/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(2956..2975)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3566..3585
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3870..3991
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3907..3926)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3961..3980
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4040..4056)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4040..4056)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4053..4075)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4064..4080
/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(4088..4118)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4133..4154
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4271..4288)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4442..5030)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4522..4541)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5201..6061)
/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 5824..5843
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6062..6166)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6241..6260)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
661 gcggccgcca ccatgggcaa gcccatcccc aaccccctgc tgggcctgga cagcaccgct
721 agcaaagaca atactgtgcc tctgaagctg atcgctctcc tggctaatgg cgagttccat
781 agtggcgaac agctgggaga aaccctgggc atgtccaggg ccgctatcaa caagcacatt
841 cagactctgc gcgactgggg cgtggacgtg ttcaccgtgc ccggaaaggg ctactctctg
901 cccgagccta tcccgctgct gaacgctaaa cagattctgg gacagctgga cggcgggagc
961 gtggcagtcc tgcctgtggt cgactccacc aatcagtacc tgctggatcg aatcggcgag
1021 ctgaagagtg gggatgcttg cattgcagaa tatcagcagg cagggagagg aagcagaggg
1081 aggaaatggt tctctccttt tggagctaac ctgtacctga gtatgttttg gcgcctgaag
1141 cggggaccag cagcaatcgg cctgggcccg gtcatcggaa ttgtcatggc agaagcgctg
1201 cgaaagctgg gagcagacaa ggtgcgagtc aaatggccca atgacctgta tctgcaggat
1261 agaaagctgg caggcatcct ggtggagctg gccggaataa caggcgatgc tgcacagatc
1321 gtcattggcg ccgggattaa cgtggctatg aggcgcgtgg aggaaagcgt ggtcaatcag
1381 ggctggatca cactgcagga agcagggatt aacctggaca ggaatactct ggccgctacg
1441 ctgatccgag agctgcgggc agccctggaa ctgttcgagc aggaaggcct ggctccatat
1501 ctgccacggt gggagaagct ggataacttc atcaatagac ccgtgaagct gatcattggg
1561 gacaaagaga ttttcgggat tagccggggg attgataaac agggagccct gctgctggaa
1621 caggacggag ttatcaaacc ctggatgggc ggagaaatca gtctgcggtc tgccgaaaag
1681 ctgcagctgc ctcccctgga gcgcctgacc ctggactaat agctcgagca tgcatctaga
1741 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
1801 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
1861 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
1921 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
1981 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
2041 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
2101 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
2161 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
2221 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
2281 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
2341 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
2401 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
2461 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
2521 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
2581 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
2641 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
2701 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
2761 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
2821 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
2881 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
2941 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
3001 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
3061 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
3121 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
3181 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
3241 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
3301 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
3361 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
3421 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
3481 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
3541 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
3601 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
3661 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
3721 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
3781 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
3841 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
3901 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
3961 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
4021 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
4081 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
4141 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
4201 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
4261 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
4321 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
4381 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
4441 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
4501 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
4561 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
4621 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
4681 ccaagctggg ctgtgtgcac aaaccccccg ttcagcccga ccgctgcgcc ttatccggta
4741 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
4801 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
4861 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
4921 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
4981 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
5041 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
5101 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
5161 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
5221 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
5281 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
5341 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
5401 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
5461 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
5521 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
5581 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
5641 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
5701 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
5761 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
5821 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
5881 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
5941 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
6001 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
6061 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
6121 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
6181 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
6241 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
6301 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
6361 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
6421 atatacgcgt t
//
