| 质粒名称: | pGEX6p-1-BCCP-PupE |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 73974 |
| 存储实验室: | Min Zhuang |
|
载体骨架基本信息 |
|
| 载体名称: | pGEX6P-1 |
| 完整质粒大小: | 5391 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | BCCP-PupE |
| 物种来源: | 未知 |
| 插入大小: | 417 bp |
| 启动子: | tac |
| 标签/融合蛋白: | GST (N terminal on backbone) |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | self-designed |
|
文献引用方法 |
|
| 材料和方法部分: | pGEX6p-1-BCCP-PupE was a gift from Min Zhuang (Addgene plasmid # 113403 ; http://n2t.net/addgene:113403 ; RRID:Addgene_113403) |
| 参考文献部分: | A proximity-tagging system to identify membrane protein-protein interactions. Liu Q, Zheng J, Sun W, Huo Y, Zhang L, Hao P, Wang H, Zhuang M. Nat Methods. 2018 Aug 13. pii: 10.1038/s41592-018-0100-5. doi: 10.1038/s41592-018-0100-5. 10.1038/s41592-018-0100-5 PubMed 30104635 |
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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 pGEX6p-1-BCCP-PupE 5391 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses BCCP-PupE in bacteria.
ACCESSION .
VERSION .
KEYWORDS pGEX6p-1-BCCP-PupE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5391)
AUTHORS Liu Q, Zheng J, Sun W, Huo Y, Zhang L, Hao P, Wang H, Zhuang M
TITLE A proximity-tagging system to identify membrane protein-protein
interactions.
JOURNAL Nat Methods. 2018 Aug 13. pii: 10.1038/s41592-018-0100-5. doi:
10.1038/s41592-018-0100-5.
PUBMED 30104635
REFERENCE 2 (bases 1 to 5391)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5391
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1..654
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
primer_bind 612..634
/label=pGEX 5'
/note="pGEX vectors, Glutathione-S-transferase, forward
primer"
CDS 661..684
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
primer_bind complement(1184..1206)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1351..1369)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1437..1541
/gene="bla"
/label=AmpR promoter
CDS 1542..2402
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(1760..1779)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2573..3161
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3062..3081
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3315..3332
/label=L4440
/note="L4440 vector, forward primer"
promoter 3405..3482
/gene="lacI"
/label=lacI promoter
CDS 3483..4565
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(3502..3521)
/label=LacI-R
/note="LacI, reverse primer"
promoter 4614..4644
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4652..4668
/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)."
primer_bind 4676..4692
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4676..4692
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 4688..5215
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
QPSEAVVWLCRS"
primer_bind complement(4708..4725)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4708..4724)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4717..4739)
/label=M13/pUC Forward
/note="In lacZ gene"
promoter 5317..5345
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 5353..5369
/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)."
ORIGIN
1 atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt
61 ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa
121 tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat
181 ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac
241 atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg
301 gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt
361 gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa
421 acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat
481 gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa
541 aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca
601 tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat
661 ctggaagttc tgttccaggg gcccctggga tccgccggga aggcaggcga gggagagatc
721 cccgcaccct tggccggcac ggtcagcaaa atcctggtca aggaaggcga caccgtgaag
781 gctggacaga cggtgttggt actggaggcg atgaagatgg agacagagat caatgccccg
841 accgatggga aggtggagaa ggtgttggtt aaggaaaggg acgccgtgca gggcggtcag
901 ggactgatca agatcggcat gaacgcgaaa cagacccaga tcatgggtgg cggtggtcgt
961 gacgaagaca atgcggaaga ctctgctcag gcgtctggtc aggttcagat caataccgaa
1021 ggtgttgact ctctgctgga cgaaatcgac ggcctgctcg aaaacaacgc ggaggaattc
1081 gttcgttctt acgttcagaa aggtggtgaa taagtcgact cgagcggccg catcgtgact
1141 gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
1201 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
1261 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
1321 gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
1381 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
1441 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
1501 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
1561 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
1621 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
1681 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
1741 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
1801 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
1861 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
1921 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
1981 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
2041 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
2101 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
2161 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
2221 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
2281 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
2341 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
2401 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
2461 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
2521 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
2581 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
2641 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
2701 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
2761 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
2821 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
2881 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
2941 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
3001 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
3061 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
3121 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
3181 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
3241 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
3301 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
3361 ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc atcgaatggc
3421 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
3481 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
3541 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
3601 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
3661 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
3721 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
3781 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
3841 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
3901 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
3961 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
4021 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
4081 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
4141 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
4201 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
4261 ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct
4321 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
4381 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
4441 ccgtctcact ggtgaaaaga aaaaccacgc tagcgcccaa tacgcaaacc gcctctcccc
4501 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
4561 agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca ggctttacac
4621 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
4681 aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga
4741 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
4801 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
4861 atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga
4921 tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc
4981 gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga
5041 gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg
5101 ccagacgcga attatttttg atggcgttgg aattagctta tcgactgcac ggtgcaccaa
5161 tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac
5221 tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca
5281 tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca tcggctcgta
5341 taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagtatt c
//
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