| 质粒名称: | pEvol-pIRE4-Bpa |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 155342 |
| 存储实验室: | Irene Coin |
|
载体骨架基本信息 |
|
| 载体名称: | EGFP-N1 |
| 载体出品公司: | Clontech, major modifications |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 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: | humanized EBpaRS/BstYam |
| 启动子: | PGK/U6 |
| 标签/融合蛋白: | Synthetic; E.coli |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | 未知(未破坏) |
| 3’克隆位点: | 未知 (未破坏) |
| 5’测序引物 | n/a |
|
文献引用方法 |
|
| 材料和方法部分: | pIRE4-Bpa was a gift from Irene Coin (Addgene plasmid # 155342 ; http://n2t.net/addgene:155342 ; RRID:Addgene_155342) |
| 参考文献部分: | Exploring GPCR-arrestin interfaces with genetically encoded crosslinkers. Bottke T, Ernicke S, Serfling R, Ihling C, Burda E, Gurevich VV, Sinz A, Coin I. EMBO Rep. 2020 Sep 14:e50437. doi: 10.15252/embr.202050437. 10.15252/embr.202050437 PubMed 32929862 |
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 pIRE4-Bpa 6660 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Plasmid encoding the translational pair that enables efficient
incorporation of Benzoyl-phenylalanine in response to the amber
codon into proteins in mammmalian cells..
ACCESSION .
VERSION .
KEYWORDS pIRE4-Bpa
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6660)
AUTHORS Bottke T, Ernicke S, Serfling R, Ihling C, Burda E, Gurevich VV,
Sinz A, Coin I
TITLE Exploring GPCR-arrestin interfaces with genetically encoded
crosslinkers.
JOURNAL EMBO Rep. 2020 Sep 14:e50437. doi: 10.15252/embr.202050437.
PUBMED 32929862
REFERENCE 2 (bases 1 to 6660)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6660
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..500
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(36..59)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 433..452
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
regulatory 527..536
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
polyA_signal 1958..2079
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1995..2014)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2049..2068
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
promoter 2104..2344
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
promoter 2462..2702
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
promoter 2820..3060
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
promoter 3178..3418
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
rep_origin complement(3529..3984)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3666..3687)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3878..3897
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4011..4115
/gene="bla"
/label=AmpR promoter
primer_bind complement(4140..4160)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4256..4466
/label=SV40 promoter
/note="SV40 early promoter"
promoter 4278..4474
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 4325..4460
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4387..4406
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4509..5303
/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
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4563..4582)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5173..5192
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5491..5510)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5535..5582
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5933..6521
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6422..6441
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
61 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
121 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
181 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
241 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
301 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
361 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
421 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
481 cctcatctcc gggcctttcg acctgctagc gtttaaactt aagcttgcca ccatggccag
541 cagcaacctg atcaagcagc tgcaggaacg gggcctggtg gcccaagtga cagatgaaga
601 agccctggcc gagagactgg cccagggacc tattgccctg ggctgcggct ttgaccccac
661 cgccgattct ctgcacctgg gacatctggt gcctctgctg tgtctgaagc ggttccagca
721 ggccggccac aaacctgtgg ctctcgtggg cggagccacc ggcctgattg gcgatcctag
781 ctttaaggcc gccgagcgga agctgaacac cgaagagaca gtgcaggaat gggtggacaa
841 gatccggaaa caggtggccc ccttcctgga cttcgactgc ggcgagaata gcgccattgc
901 cgccaacaac tacgattggt tcggcaatat gaacgtgctg accttcctgc gggacatcgg
961 caagcacttc agcgtgaacc agatgatcaa caaagaagcc gtgaagcagc ggctgaacag
1021 agaggaccag ggcatcagct tcaccgagtt cagctacaac ctgctgcagg gctacggctt
1081 cgcctgcgcc aacaagcagt acggcgtggt gctgcagatc ggcggctctg accagtgggg
1141 caatatcacc agcggcatcg acctgaccag acggctgcac cagaaccagg tgttcggcct
1201 gaccgtgccc ctgatcacaa aggccgacgg caccaagttc ggcaagaccg aaggcggagc
1261 cgtgtggctg gaccccaaga aaaccagccc ctacaagttc taccagttct ggatcaacac
1321 cgccagagcc gacgtgtacc ggttcctgaa gttcttcacc ttcatgagca tcgaggaaat
1381 caacgccctg gaagaagagg acaagaacag cggcaaggcc cccagagccc agtacgtgct
1441 ggcagaacaa gtgaccaggc tggtgcacgg cgaggaagga ctgcaggccg ccaagagaat
1501 caccgagtgc ctgttcagcg gcagcctgag cgctctgagc gaggccgatt ttgagcagct
1561 ggctcaggac ggcgtgccca tggtggaaat ggaaaagggc gccgacctga tgcaggccct
1621 ggtggattct gagctgcagc ccagcagagg acaggccaga aagacaatcg ccagcaacgc
1681 catcaccatc aacggcgaga agcagagcga ccccgagtat ttcttcaaag aggaagatcg
1741 gctgttcggc cggttcaccc tgctgcggag aggcaagaag aactactgcc tgatctgctg
1801 gaagtaataa gaattctgca gatatccagc acagtggcgg ccgcgactct agatcataat
1861 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
1921 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
1981 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
2041 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggtctagact cgaggggcag
2101 gaagagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
2161 gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
2221 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
2281 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
2341 ggacgaaaca ccggaggggt agcgaagtgg ctaaacgcgg cggactctaa atccgctccc
2401 tttgggttcg gcggttcgaa tccgtccccc tccagtcctt tttttggtcg aggggcagga
2461 agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
2521 gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
2581 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
2641 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
2701 acgaaacacc ggaggggtag cgaagtggct aaacgcggcg gactctaaat ccgctccctt
2761 tgggttcggc ggttcgaatc cgtccccctc cagtcctttt tttggtcgag gggcaggaag
2821 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
2881 taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
2941 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
3001 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
3061 gaaacaccgg aggggtagcg aagtggctaa acgcggcgga ctctaaatcc gctccctttg
3121 ggttcggcgg ttcgaatccg tccccctcca gtcctttttt tggtcgaggg gcaggaagag
3181 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
3241 attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
3301 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
3361 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
3421 aacaccggag gggtagcgaa gtggctaaac gcggcggact ctaaatccgc tccctttggg
3481 ttcggcggtt cgaatccgtc cccctccagt cctttttttg gtcgacgtaa attgtaagcg
3541 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
3601 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
3661 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
3721 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
3781 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
3841 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
3901 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
3961 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
4021 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
4081 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
4141 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
4201 aagcatgcat ctcaattagt cagcaaccak rkkyssrmmg tccccaggct ccccagcagg
4261 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
4321 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
4381 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
4441 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
4501 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
4561 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
4621 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
4681 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
4741 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
4801 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
4861 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
4921 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
4981 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
5041 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
5101 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
5161 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
5221 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
5281 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
5341 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
5401 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
5461 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
5521 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
5581 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
5641 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
5701 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag tactaaatat
5761 ttatcccggg ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
5821 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
5881 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
5941 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
6001 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
6061 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
6121 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
6181 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
6241 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
6301 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
6361 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
6421 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
6481 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
6541 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
6601 ctgattctgt ggataaccgt attaccgcca tgcatacgcg ttagttatta aaattctacc
//
