| 质粒名称: | myc-BioID2-MCS,pCDNA3.1-Myc-BioID2-MCS |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 74223 |
| 存储实验室: | Kyle Roux |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA 3.1 |
| 载体出品公司: | Thermo Fisher Scientific |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6147 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: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: |
Myc（主干上的N终端）
humanized A. aeolicus BioID2 with R40G（主干上的N端）
|
| 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 | CMV-F |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | myc-BioID2-MCS was a gift from Kyle Roux (Addgene plasmid # 74223 ; http://n2t.net/addgene:74223 ; RRID:Addgene_74223) |
| 参考文献部分: | An improved smaller biotin ligase for BioID proximity labeling. Kim DI, Jensen SC, Noble KA, Kc B, Roux KH, Motamedchaboki K, Roux KJ. Mol Biol Cell. 2016 Feb 24. pii: mbc.E15-12-0844. 10.1091/mbc.E15-12-0844 PubMed 26912792 |
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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 myc-BioID2-MCS 6146 bp ds-DNA circular SYN 19-MAR-2019
DEFINITION To fuse your protein of interest to the C-terminus of BioID2 and use
in proximity-dependent biotin identification (BioID); MCS is present
downstream of myc tag and BioID2 tag.
ACCESSION .
VERSION .
KEYWORDS myc-BioID2-MCS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6146)
AUTHORS Kim DI, Jensen SC, Noble KA, Kc B, Roux KH, Motamedchaboki K, Roux
KJ
TITLE An improved smaller biotin ligase for BioID proximity labeling.
JOURNAL Mol Biol Cell. 2016 Feb 24. pii: mbc.E15-12-0844.
PUBMED 26912792
REFERENCE 2 (bases 1 to 6146)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 19, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6146
/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..1638
/codon_start=1
/product="promiscuous R40G mutant of a biotin protein
ligase from Aquifex aeolicus (Kim et al., 2016)"
/label=BioID2
/translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
LSGEFSLRRS"
primer_bind complement(1739..1756)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1745..1969
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2015..2443
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2102..2121)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2312..2333
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2452..2472)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2457..2786
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2637..2772
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2699..2718
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2853..3647
/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(2907..2926)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3517..3536
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3821..3942
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3858..3877)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3912..3931
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3991..4007)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3991..4007)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4004..4026)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4015..4031
/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(4039..4069)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4084..4105
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4222..4239)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4393..4981)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4473..4492)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5152..6012)
/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 5775..5794
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6013..6117)
/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 acttcaagaa cctgatctgg
961 ctgaaggagg tggacagcac ccaggagaga ctgaaggagt ggaacgtgag ctacggcacc
1021 gccctggtgg ccgacagaca gaccaagggc agaggcggcc tgggcagaaa gtggctgagc
1081 caggagggcg gcctgtactt cagcttcctg ctgaacccca aggagttcga gaacctgctg
1141 cagctgcccc tggtgctggg cctgagcgtg agcgaggccc tggaggagat caccgagatc
1201 cccttcagcc tgaagtggcc caacgacgtg tacttccagg agaagaaggt gagcggcgtg
1261 ctgtgcgagc tgagcaagga caagctgatc gtgggcatcg gcatcaacgt gaaccagaga
1321 gagatccccg aggagatcaa ggacagagcc accaccctgt acgagatcac cggcaaggac
1381 tgggacagaa aggaggtgct gctgaaggtg ctgaagagaa tcagcgagaa cctgaagaag
1441 ttcaaggaga agagcttcaa ggagttcaag ggcaagatcg agagcaagat gctgtacctg
1501 ggcgaggagg tgaagctgct gggcgagggc aagatcaccg gcaagctggt gggcctgagc
1561 gagaagggcg gcgccctgat cctgaccgag gagggcatca aggagatcct gagcggcgag
1621 ttcagcctga gaagaagcct cgagcggccg ccactgtgct ggatatctgc agaattccac
1681 cacactggac tagtggatcc gagctcggta ccaagcttaa gtttaaaccg ctgatcagcc
1741 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
1801 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
1861 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
1921 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
1981 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
2041 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
2101 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
2161 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
2221 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
2281 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
2341 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
2401 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
2461 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
2521 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
2581 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
2641 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
2701 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
2761 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
2821 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
2881 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
2941 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
3001 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
3061 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
3121 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
3181 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
3241 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
3301 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
3361 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
3421 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
3481 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
3541 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
3601 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
3661 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
3721 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
3781 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
3841 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
3901 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
3961 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
4021 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
4081 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
4141 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
4201 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
4261 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
4321 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
4381 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
4441 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
4501 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
4561 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
4621 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
4681 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
4741 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
4801 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
4861 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
4921 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
4981 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
5041 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
5101 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
5161 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
5221 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
5281 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
5341 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
5401 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
5461 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
5521 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
5581 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
5641 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
5701 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
5761 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
5821 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
5881 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
5941 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
6001 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
6061 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
6121 atttccccga aaagtgccac ctgacg
//
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