| 质粒名称: | mycBioID2-pBABE-puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 80900 |
| 存储实验室: | Kyle Roux |
|
载体骨架基本信息 |
|
| 载体名称: | pBABE-puro |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5949 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，逆转录病毒 |
| 筛选标记: | 嘌呤霉素 |
| 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 | -- |
| 标签/融合蛋白: |
人性化A。带R40G（主干上的N端）的aeolicus BioID2
myc（主干上的N终端）
|
| 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 | pBABE 5' |
| 3' 测序引物 | pBABE 3' |
|
文献引用方法 |
|
| 材料和方法部分: | mycBioID2-pBABE-puro was a gift from Kyle Roux (Addgene plasmid # 80900 ; http://n2t.net/addgene:80900 ; RRID:Addgene_80900) |
| 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-pBABE-puro 5858 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION To fuse your protein of interest to the C-terminus of BioID2 and use
in proximity-dependent biotin identification (BioID); a small MCS is
present downstream of myc tag and BioID2 tag.
ACCESSION .
VERSION .
KEYWORDS mycBioID2-pBABE-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5858)
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 5858)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5858
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 12..481
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 546..745
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 946..1362
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1292..1314
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1330..1346
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
regulatory 1415..1424
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1424..1453
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1457..2152
/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(2192..2212)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2197..2526
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2377..2512
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2439..2458
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2536..3135
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(2536..2555)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3032..3052
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 3356..3825
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(3955..3972)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4126..4714)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4206..4225)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4874..5734)
/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 5497..5516
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5735..5839)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ccacctgcag cctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
61 ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
121 cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca
181 gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
241 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
301 ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg
361 agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
421 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
481 atttgggggc tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg
541 gaggcaagct ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat
601 tttatgcgcc tgcgtcggta ctagttagct aactagctct gtatctggcg gacccgtggt
661 ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg
721 ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga
781 tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg
841 ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg
901 tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc
961 actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag
1021 tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt
1081 aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc
1141 aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg
1201 gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg
1261 cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct
1321 cgatcctccc tttatccagc cctcactcct tctctaggcg ccggccggat cccagtgtgg
1381 tggtacgtag gaattcggag acccaagctg gctagccacc atggaacaaa aactcatctc
1441 agaagaggat ctcgacttca agaacctgat ctggctgaag gaggtggaca gcacccagga
1501 gagactgaag gagtggaacg tgagctacgg caccgccctg gtggccgaca gacagaccaa
1561 gggcagaggc ggcctgggca gaaagtggct gagccaggag ggcggcctgt acttcagctt
1621 cctgctgaac cccaaggagt tcgagaacct gctgcagctg cccctggtgc tgggcctgag
1681 cgtgagcgag gccctggagg agatcaccga gatccccttc agcctgaagt ggcccaacga
1741 cgtgtacttc caggagaaga aggtgagcgg cgtgctgtgc gagctgagca aggacaagct
1801 gatcgtgggc atcggcatca acgtgaacca gagagagatc cccgaggaga tcaaggacag
1861 agccaccacc ctgtacgaga tcaccggcaa ggactgggac agaaaggagg tgctgctgaa
1921 ggtgctgaag agaatcagcg agaacctgaa gaagttcaag gagaagagct tcaaggagtt
1981 caagggcaag atcgagagca agatgctgta cctgggcgag gaggtgaagc tgctgggcga
2041 gggcaagatc accggcaagc tggtgggcct gagcgagaag ggcggcgccc tgatcctgac
2101 cgaggagggc atcaaggaga tcctgagcgg cgagttcagc ctgagaagaa gcctcgagtg
2161 acttaagttt aaaccgctga gtcgaccctg tggaatgtgt gtcagttagg gtgtggaaag
2221 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
2281 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
2341 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
2401 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
2461 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
2521 tgcaaaaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
2581 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
2641 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
2701 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
2761 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
2821 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
2881 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
2941 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
3001 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
3061 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
3121 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
3181 ccccatgcat cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
3241 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
3301 aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa
3361 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
3421 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
3481 tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac
3541 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
3601 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
3661 agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc
3721 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
3781 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcacatgc agcatgtatc
3841 aaaattaatt tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg
3901 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
3961 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
4021 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
4081 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
4141 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
4201 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
4261 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
4321 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
4381 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
4441 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
4501 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
4561 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
4621 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
4681 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
4741 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
4801 aaggatcttc acctagatcc ttttgcggcc gcaaatcaat ctaaagtata tatgagtaaa
4861 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
4921 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
4981 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
5041 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
5101 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
5161 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
5221 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
5281 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
5341 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
5401 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
5461 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
5521 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
5581 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
5641 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
5701 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
5761 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
5821 aacaaatagg ggttccgcgc acatttcccc gaaaagtg
//
