| 质粒名称: | C1(1-29)-TurboID-V5_pCDNA3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 107173 |
| 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: | -- |
| 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: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 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: | -- |
| 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: | C1(1-29)-TurboID-V5_pCDNA3 was a gift from Alice Ting (Addgene plasmid # 107173 ; http://n2t.net/addgene:107173 ; RRID:Addgene_107173) |
| 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
-TurboID-V5_pCDNA3载体图谱.png)
载体简介
载体序列
LOCUS C1(1-29)-TurboID-V5_pCDNA3 6564 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION expresses V5-tagged TurboID on the mammalian ER membrane.
ACCESSION .
VERSION .
KEYWORDS C1(1-29)-TurboID-V5_pCDNA3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6564)
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 6564)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6564
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 115..494
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 495..698
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 649..669
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 743..762
/label=T7
/note="T7 promoter, forward primer"
promoter 743..761
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 780..789
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 945..1901
/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 1908..1949
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
promoter complement(1997..2015)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1998..2015)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2035..2052)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2041..2265
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2311..2739
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2398..2417)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2608..2629
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2748..2768)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2753..3082
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2933..3068
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2995..3014
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3149..3943
/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(3203..3222)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3813..3832
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4117..4238
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4154..4173)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4208..4227
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4287..4303)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4287..4303)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4300..4322)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4311..4327
/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(4335..4365)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4380..4401
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4518..4535)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4689..5277)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4769..4788)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5448..6308)
/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 6071..6090
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6309..6413)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6488..6507)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
61 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
121 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
181 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
241 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
301 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
361 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
421 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
481 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
541 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
601 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
661 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
721 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgggccg
781 ccaccatgga ccctgtggtc gtgctggggc tgtgcctgag ctgtctgctg ctgctgagcc
841 tgtggaagca gtcctacgga ggcgggggat caggaagcgg cggccgcgga tcaggaagcg
901 gatcaggaag cggatcagga agcggatcag gaagcggatc aggaaaagac aatactgtgc
961 ctctgaagct gatcgctctc ctggctaatg gcgagttcca tagtggcgaa cagctgggag
1021 aaaccctggg catgtccagg gccgctatca acaagcacat tcagactctg cgcgactggg
1081 gcgtggacgt gttcaccgtg cccggaaagg gctactctct gcccgagcct atcccgctgc
1141 tgaacgctaa acagattctg ggacagctgg acggcgggag cgtggcagtc ctgcctgtgg
1201 tcgactccac caatcagtac ctgctggatc gaatcggcga gctgaagagt ggggatgctt
1261 gcattgcaga atatcagcag gcagggagag gaagcagagg gaggaaatgg ttctctcctt
1321 ttggagctaa cctgtacctg agtatgtttt ggcgcctgaa gcggggacca gcagcaatcg
1381 gcctgggccc ggtcatcgga attgtcatgg cagaagcgct gcgaaagctg ggagcagaca
1441 aggtgcgagt caaatggccc aatgacctgt atctgcagga tagaaagctg gcaggcatcc
1501 tggtggagct ggccggaata acaggcgatg ctgcacagat cgtcattggc gccgggatta
1561 acgtggctat gaggcgcgtg gaggaaagcg tggtcaatca gggctggatc acactgcagg
1621 aagcagggat taacctggac aggaatactc tggccgctac gctgatccga gagctgcggg
1681 cagccctgga actgttcgag caggaaggcc tggctccata tctgccacgg tgggagaagc
1741 tggataactt catcaataga cccgtgaagc tgatcattgg ggacaaagag attttcggga
1801 ttagccgggg gattgataaa cagggagccc tgctgctgga acaggacgga gttatcaaac
1861 cctggatggg cggagaaatc agtctgcggt ctgccgaaaa ggctagcggc aagcccatcc
1921 ccaaccccct gctgggcctg gacagcacct aatgaggatc cggcaagccc atccccaacc
1981 ccctgctggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
2041 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
2101 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
2161 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
2221 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
2281 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
2341 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
2401 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
2461 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
2521 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
2581 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
2641 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
2701 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
2761 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
2821 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
2881 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
2941 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
3001 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
3061 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
3121 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
3181 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
3241 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
3301 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
3361 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
3421 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
3481 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
3541 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
3601 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
3661 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
3721 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
3781 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
3841 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
3901 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
3961 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
4021 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
4081 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
4141 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
4201 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
4261 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
4321 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
4381 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
4441 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
4501 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
4561 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
4621 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
4681 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
4741 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
4801 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
4861 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
4921 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
4981 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
5041 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
5101 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
5161 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
5221 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
5281 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
5341 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
5401 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
5461 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
5521 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
5581 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
5641 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
5701 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
5761 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
5821 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
5881 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
5941 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
6001 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
6061 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
6121 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
6181 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
6241 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
6301 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
6361 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
6421 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
6481 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
6541 tgttggaggt cgctgagtag tgcg
//
