| 质粒名称: | mycBioID2-13X Linker-MCS |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 92308 |
| 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: | Thermo Fisher Scientific |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6387 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）
具有 R40G 的人源化 A.aeolicus BioID2（骨架上的 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: | mycBioID2-13X Linker-MCS was a gift from Kyle Roux (Addgene plasmid # 92308 ; http://n2t.net/addgene:92308 ; RRID:Addgene_92308) |
| 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 |
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 mycBioID2-13X Linker-MCS 6386 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION To fuse your protein of interest to the C-terminus of BioID2 and use
in proximity-dependent biotin identification (BioID); MCS and a 25nm
GS linker are present downstream of BioID2 and myc tags..
ACCESSION .
VERSION .
KEYWORDS mycBioID2-13X Linker-MCS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6386)
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 6386)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6386
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 6..230
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 276..704
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(363..382)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 573..594
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(713..733)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 718..1047
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 898..1033
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 960..979
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 1114..1908
/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(1168..1187)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1778..1797
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 2082..2203
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2119..2138)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2173..2192
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(2252..2268)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2252..2268)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2265..2287)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2276..2292
/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(2300..2330)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2345..2366
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2483..2500)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2654..3242)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2734..2753)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3413..4273)
/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 4036..4055
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4274..4378)
/gene="bla"
/label=AmpR promoter
primer_bind complement(4453..4472)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 4644..5023
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5024..5227
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 5178..5198
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 5272..5291
/label=T7
/note="T7 promoter, forward primer"
promoter 5272..5290
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 5308..5317
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 5317..5346
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 5350..6045
/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(6386..17)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
ORIGIN
1 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
61 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
121 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
181 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
241 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
301 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
361 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
421 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
481 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
541 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
601 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
661 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
721 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
781 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
841 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
901 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
961 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
1021 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
1081 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
1141 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
1201 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
1261 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
1321 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
1381 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
1441 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
1501 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
1561 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
1621 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
1681 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
1741 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
1801 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
1861 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
1921 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
1981 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
2041 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
2101 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
2161 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
2221 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
2281 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
2341 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
2401 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
2461 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
2521 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
2581 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
2641 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
2701 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
2761 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
2821 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
2881 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
2941 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
3001 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
3061 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
3121 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
3181 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
3241 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
3301 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
3361 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
3421 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
3481 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
3541 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
3601 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
3661 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
3721 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
3781 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
3841 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
3901 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
3961 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
4021 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
4081 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
4141 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
4201 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
4261 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
4321 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
4381 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
4441 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
4501 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
4561 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
4621 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
4681 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
4741 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
4801 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
4861 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
4921 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
4981 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
5041 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
5101 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
5161 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
5221 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
5281 tcactatagg gagacccaag ctggctagcc accatggaac aaaaactcat ctcagaagag
5341 gatctcgact tcaagaacct gatctggctg aaggaggtgg acagcaccca ggagagactg
5401 aaggagtgga acgtgagcta cggcaccgcc ctggtggccg acagacagac caagggcaga
5461 ggcggcctgg gcagaaagtg gctgagccag gagggcggcc tgtacttcag cttcctgctg
5521 aaccccaagg agttcgagaa cctgctgcag ctgcccctgg tgctgggcct gagcgtgagc
5581 gaggccctgg aggagatcac cgagatcccc ttcagcctga agtggcccaa cgacgtgtac
5641 ttccaggaga agaaggtgag cggcgtgctg tgcgagctga gcaaggacaa gctgatcgtg
5701 ggcatcggca tcaacgtgaa ccagagagag atccccgagg agatcaagga cagagccacc
5761 accctgtacg agatcaccgg caaggactgg gacagaaagg aggtgctgct gaaggtgctg
5821 aagagaatca gcgagaacct gaagaagttc aaggagaaga gcttcaagga gttcaagggc
5881 aagatcgaga gcaagatgct gtacctgggc gaggaggtga agctgctggg cgagggcaag
5941 atcaccggca agctggtggg cctgagcgag aagggcggcg ccctgatcct gaccgaggag
6001 ggcatcaagg agatcctgag cggcgagttc agcctgagaa gaagcaccgg tttttcggaa
6061 ttcggatccg gtggaggcgg gtctggaggc gggggtagtg gcgggggtgg aagcgggggt
6121 ggaggcgggt cgggtggcgg aggtagcgga ggcggtggaa gtggaggcgg tggaagtggt
6181 ggaggaggtt ctggtggcgg aggtagcggt ggcggaggta gcggaggcgg tggaagtggt
6241 ggcggaggta gcggaggcgg tggatcggcg cgccgtttca agaacctcga gcggccgcca
6301 ctgtgctgga tatctgcaga attccaccac actggactag tggatccgag ctcggtacca
6361 agcttaagtt taaaccgctg atcagc
//
