| 质粒名称: | pcDNA3.1 mycBioID |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 35700 |
| 存储实验室: | Kyle Roux |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1(-) |
| 载体出品公司: | InVitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5400 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | BirA |
| 物种来源: | E. coli |
| 插入大小: | 1116 bp |
| 基因突变: | R118G (highly promiscuous form) |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | Myc（插入N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI(未破坏) |
| 3' 克隆位点: | PmeI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3.1 mycBioID was a gift from Kyle Roux (Addgene plasmid # 35700 ; http://n2t.net/addgene:35700 ; RRID:Addgene_35700) |
| 参考文献部分: | A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells. Roux KJ, Kim DI, Raida M, Burke B. J Cell Biol. 2012 Mar 12. 10.1083/jcb.201112098 PubMed 22412018 |
买家导航
pcDNA3.1mycBioID载体基本信息
pcDNA3.1mycBioID载体图谱质粒图谱和多克隆位点信息
pcDNA3.1mycBioID载体简介
pcDNA3.1mycBioID载体序列
LOCUS pcDNA3.1 mycBioID 6410 bp ds-DNA circular SYN 15-NOV-2018
DEFINITION To fuse your protein of interest to the C-terminus of BirA(R118G)
and use in proximity-dependent biotin identification (BioID); myc
tag.
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1 mycBioID
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6410)
AUTHORS Roux KJ, Kim DI, Raida M, Burke B
TITLE A promiscuous biotin ligase fusion protein identifies proximal and
interacting proteins in mammalian cells.
JOURNAL J Cell Biol. 2012 Mar 12.
PUBMED 22412018
REFERENCE 2 (bases 1 to 6410)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 15, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6410
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(46..65)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 237..616
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 617..820
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 771..791
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 865..884
/label=T7
/note="T7 promoter, forward primer"
promoter 865..883
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 901..910
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 910..939
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 943..1902
/codon_start=1
/product="promiscuous R118G mutant of the E. coli biotin
protein ligase (Cronan, 2005)"
/label=BirA(R118G)
/note="also known as BirA* or BioID"
/translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
primer_bind complement(2003..2020)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2009..2233
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2279..2707
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2366..2385)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2576..2597
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2716..2736)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2721..3050
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2901..3036
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2963..2982
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3117..3911
/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(3171..3190)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3781..3800
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4085..4206
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4122..4141)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4176..4195
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4255..4271)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4255..4271)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4268..4290)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4279..4295
/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(4303..4333)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4348..4369
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4486..4503)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4657..5245)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4737..4756)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5416..6276)
/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 6039..6058
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6277..6381)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
901 gccaccatgg aacaaaaact catctcagaa gaggatctcg acaaggacaa caccgtgccc
961 ctgaagctga tcgccctgct ggccaacggc gagttccact ctggcgagca gctgggagag
1021 accctgggaa tgagcagagc cgccatcaac aagcacatcc agacactgag agactgggga
1081 gtggacgtgt tcaccgtgcc tggcaagggc tacagcctgc ctgagcctat ccagctgctg
1141 aacgccaagc agatcctggg acagctggat ggcggaagcg tggccgtgct gcctgtgatc
1201 gactccacca atcagtacct gctggacaga atcggagagc tgaagtccgg cgacgcctgc
1261 atcgccgagt accagcaggc tggcagagga ggcagaggac ggaagtggtt cagcccattc
1321 ggagccaacc tgtacctgtc catgttctgg agactggagc agggacctgc tgctgccatc
1381 ggactgagtc tggtgatcgg aatcgtgatg gccgaggtgc tgagaaagct gggagccgac
1441 aaggtgagag tgaagtggcc taatgacctg tacctccagg accgcaagct ggctggcatc
1501 ctggtggagc tgacaggcaa gacaggcgat gccgctcaga tcgtgatcgg agccggaatc
1561 aacatggcca tgagaagagt ggaggagagc gtggtgaacc agggctggat caccctgcag
1621 gaggctggca tcaacctgga ccggaacacc ctggccgcca tgctgatcag agagctgaga
1681 gccgctctgg agctgttcga gcaggaggga ctggctcctt acctgagcag atgggagaag
1741 ctggacaact tcatcaacag acctgtgaag ctgatcatcg gcgacaagga aatcttcggc
1801 atctccagag gaatcgacaa gcagggagct ctgctgctgg agcaggacgg aatcatcaag
1861 ccctggatgg gcggagaaat ctccctgaga agcgcagaga agctcgagcg gccgccactg
1921 tgctggatat ctgcagaatt ccaccacact ggactagtgg atccgagctc ggtaccaagc
1981 ttaagtttaa accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
2041 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
2101 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
2161 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
2221 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
2281 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
2341 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
2401 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
2461 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
2521 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
2581 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
2641 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
2701 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
2761 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
2821 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
2881 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
2941 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
3001 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
3061 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
3121 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
3181 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
3241 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
3301 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
3361 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
3421 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
3481 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
3541 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
3601 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
3661 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
3721 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
3781 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
3841 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
3901 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
3961 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
4021 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
4081 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
4141 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
4201 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
4261 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
4321 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
4381 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
4441 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
4501 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
4561 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
4621 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
4681 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
4741 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
4801 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
4861 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
4921 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
4981 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
5041 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
5101 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
5161 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
5221 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
5281 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
5341 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
5401 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
5461 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
5521 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
5581 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
5641 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
5701 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
5761 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
5821 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
5881 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
5941 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
6001 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
6061 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
6121 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
6181 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
6241 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
6301 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
6361 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
//
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