| 质粒名称: | pGEX6p-1-PafA-XIAP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 113402 |
| 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: | 7763bp |
| 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: | PafA-XIAP |
| 物种来源: | 未知 |
| 插入大小: | 1785bp |
| 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-PafA-XIAP was a gift from Min Zhuang (Addgene plasmid # 113402 ; http://n2t.net/addgene:113402 ; RRID:Addgene_113402) |
| 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 |
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 pGEX6p-1-PafA-XIAP 7763 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses PafA-XIAP in bacteria.
ACCESSION .
VERSION .
KEYWORDS pGEX6p-1-PafA-XIAP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7763)
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 7763)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7763
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 7..30
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
primer_bind complement(2902..2924)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3069..3087)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3155..3259
/gene="bla"
/label=AmpR promoter
CDS 3260..4120
/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(3478..3497)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4291..4879
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4780..4799
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5033..5050
/label=L4440
/note="L4440 vector, forward primer"
promoter 5123..5200
/gene="lacI"
/label=lacI promoter
CDS 5201..6283
/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(5220..5239)
/label=LacI-R
/note="LacI, reverse primer"
promoter 6332..6362
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6370..6386
/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 6394..6410
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6394..6410
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 6406..6933
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
QPSEAVVWLCRS"
primer_bind complement(6426..6443)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6426..6442)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6435..6457)
/label=M13/pUC Forward
/note="In lacZ gene"
promoter 7035..7063
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 7071..7087
/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)."
CDS 7110..7763
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
primer_bind 7721..7743
/label=pGEX 5'
/note="pGEX vectors, Glutathione-S-transferase, forward
primer"
ORIGIN
1 tcggatctgg aagttctgtt ccaggggccc ctgggatccc cggaattccc gggtcgacga
61 ctcgagatga gtaccgtgga atccgcattg acccgcagga tcatgggcat tgaaacggag
121 tatggcctca cctttgttga tggtgattcc aaaaagcttc gcccagatga gatagctcga
181 aggatgtttc gtcccatcgt ggagaaatat tccagctcta atatcttcat acccaatggt
241 tcccggttgt atcttgatgt gggttcccac ccggagtacg ccaccgccga gtgtgataat
301 ttgacccagc tgatcaattt tgaaaaagct ggcgatgtta ttgcagatcg catggctgta
361 gatgccgaag agtcgctggc gaaagaagac attgctgggc aggtgtacct gtttaaaaac
421 aatgtcgatt ccgtgggcaa ttcttatggc tgccacgaaa actaccttgt gggtcgctcc
481 atgccgttga aggcgttggg taaaaggctg atgccgtttc tgattacccg ccagctcatc
541 tgcggcgccg gcaggatcca tcaccccaat cctttggata aaggcgaatc cttccccttg
601 ggctactgca tatcccagcg ctctgaccac gtgtgggagg gcgtatcaag tgccaccact
661 agatcacgcc ccattatcaa cacccgtgat gagccacatg cggattccca ttcttaccgc
721 aggctgcacg tgattgtggg tgatgccaac atggcagagc ccagcatcgc gttgaaggtc
781 ggctccacgt tgctggttct ggaaatgatt gaggcagatt tcggtttgcc cagcttagag
841 cttgccaatg atattgcctc aattagggaa atctcccgcg atgcaacagg atccacactg
901 ttgtccctga aagatggcac caccatgact gccttgcaga tccagcaggt ggtctttgag
961 catgcctcga agtggttgga gcagcgcccc gaaccagaat tttctggcac ctccaacaca
1021 gagatggccc gcgtgctgga tctgtggggt cgcatgttga aagcgattga gtccggtgat
1081 ttcagcgaag tggatacaga aattgactgg gtgatcaaaa agaagctcat tgatcgtttc
1141 attcagcgcg gcaaccttgg gttggatgat ccaaaacttg cccaagtgga cttgacttat
1201 cacgatatta ggccaggtag aggcctattt agcgtgctgc aaagccgcgg catgatcaaa
1261 cggtggacta ctgatgaggc gattttagct gcggtggata ccgctcctga tacaacacgt
1321 gctcatttgc gcgggcgaat ccttaaagcg gcggatactc tgggagtacc tgtgactgtc
1381 gattggatgc gtcacaaggt caaccgaccg gagccacaat cggtggaatt gggggatcct
1441 ttttccgcag tgaattccga ggttgatcag ttgattgagt atatgaccgt tcatgcggag
1501 agctatcgca gtggcggttc tggtggcggt tctggtatga cttttaacag ttttgaagga
1561 tctaaaactt gtgtacctgc agacatcaat aaggaagaag aatttgtaga agagtttaat
1621 agattaaaaa cttttgctaa ttttccaagt ggtagtcctg tttcagcatc aacactggca
1681 cgagcagggt ttctttatac tggtgaagga gataccgtgc ggtgctttag ttgtcatgca
1741 gctgtagata gatggcaata tggagactca gcagttggaa gacacaggaa agtatcccca
1801 aattgcagat ttatcaacgg cttttatctt gaaaatagtg ccacgcagtc tacaaattct
1861 ggtatccaga atggtcagta caaagttgaa aactatctgg gaagcagaga tcattttgcc
1921 ttagacaggc catctgagac acatgcagac tatcttttga gaactgggca ggttgtagat
1981 atatcagaca ccatataccc gaggaaccct gccatgtata gtgaagaagc tagattaaag
2041 tcctttcaga actggccaga ctatgctcac ctaaccccaa gagagttagc aagtgctgga
2101 ctctactaca caggtattgg tgaccaagtg cagtgctttt gttgtggtgg aaaactgaaa
2161 aattgggaac cttgtgatcg tgcctggtca gaacacaggc gacactttcc taattgcttc
2221 tttgttttgg gccggaatct taatattcga agtgaatctg atgctgtgag ttctgatagg
2281 aatttcccaa attcaacaaa tcttccaaga aatccatcca tggcagatta tgaagcacgg
2341 atctttactt ttgggacatg gatatactca gttaacaagg agcagcttgc aagagctgga
2401 ttttatgctt taggtgaagg tgataaagta aagtgctttc actgtggagg agggctaact
2461 gattggaagc ccagtgaaga cccttgggaa caacatgcta aatggtatcc agggtgcaaa
2521 tatctgttag aacagaaggg acaagaatat ataaacaata ttcatttaac tcattcactt
2581 gaggagtgtc tggtaagaac tactgagaaa acaccatcac taactagaag aattgatgat
2641 accatcttcc aaaatcctat ggtacaagaa gctatacgaa tggggttcag tttcaaggac
2701 attaagaaaa taatggagga aaaaattcag atatctggga gcaactataa atcacttgag
2761 gttctggttg cagatctagt gaatgctcag aaagacagta tgcaagatga gtcaagtcag
2821 acttcattac agaaagagta agcggccgca tcgtgactga ctgacgatct gcctcgcgcg
2881 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
2941 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
3001 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata attcttgaag
3061 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
3121 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
3181 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
3241 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
3301 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
3361 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
3421 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
3481 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
3541 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
3601 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
3661 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
3721 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
3781 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
3841 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
3901 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
3961 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
4021 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
4081 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
4141 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
4201 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
4261 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
4321 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
4381 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
4441 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
4501 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
4561 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
4621 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
4681 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
4741 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
4801 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
4861 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
4921 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
4981 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
5041 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
5101 tatttcacac cgcataaatt ccgacaccat cgaatggcgc aaaacctttc gcggtatggc
5161 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
5221 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
5281 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
5341 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
5401 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
5461 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
5521 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
5581 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
5641 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
5701 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
5761 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
5821 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
5881 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
5941 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
6001 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
6061 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
6121 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
6181 aaccacgcta gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
6241 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
6301 taagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
6361 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
6421 gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
6481 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
6541 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg
6601 gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc
6661 gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtgacc
6721 tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg
6781 ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat
6841 ggcgttggaa ttagcttatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca
6901 tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag
6961 gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat
7021 tctgaaatga gctgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg
7081 ataacaattt cacacaggaa acagtattca tgtcccctat actaggttat tggaaaatta
7141 agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa tatgaagagc
7201 atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg
7261 agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag tctatggcca
7321 tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag
7381 agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg agaattgcat
7441 atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct gaaatgctga
7501 aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat gtaacccatc
7561 ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca atgtgcctgg
7621 atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca caaattgata
7681 agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa gccacgtttg
7741 gtggtggcga ccatcctcca aaa
//
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