| 质粒名称: | pIRE4-Azi |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 105829 |
| 存储实验室: | Irene Coin |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4038 bp |
| 完整质粒大小: | 6658 bp |
| 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: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称1: | Humanized EAziRS |
| 物种来源: | Synthetic |
| 基因突变: | (Y37L, D182S, F183M, L186A, D265R)TyrRS |
| 启动子: | PGK |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Unknown (unknown if destroyed) |
| 3’克隆位点: | Unknown (unknown if destroyed) |
| 5’测序引物 | PGK-F 5' cattctgcacgcttcaaaag 3' |
| 插入基因名称2: | 4 copies of the U6-BstYam tRNA cassette |
| 物种来源: | Synthetic |
| 启动子: | U6 |
| 克隆方法: | Restriction Enzyme |
| 5‘’克隆位点: | Unknown (unknown if destroyed) |
| 3’克隆位点: | Unknown (unknown if destroyed) |
| 5’测序引物 |
Unknown
(Common Sequencing Primers)
|
|
文献引用方法 |
|
| 材料和方法部分: | |
| 参考文献部分: | |
买家导航
pIRE4-Azi载体质粒基本信息
pIRE4-Azi质粒图谱载体图谱和pIRE4-Azi载体序列质粒序列多克隆位点信息
pIRE4-Azi质粒载体简介
pIRE4-Azi质粒序列载体序列
LOCUS pIRE4-Azi 6658 bp DNA circular SYN 14-APR-2025
DEFINITION Plasmid encoding for the translational pair that enables efficent
incoporation of p-Azido-phenylalanine in response to the amber codon
into proteins in mammmalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6658)
AUTHORS Seidel L, Zarzycka B, Zaidi SA, Katritch V, Coin I
TITLE Structural insight into the activation of a class B
G-protein-coupled receptor by peptide hormones in live human cells.
JOURNAL Elife. 2017 Aug 3;6. doi: 10.7554/eLife.27711.
PUBMED 28771403
REFERENCE 2 (bases 1 to 6658)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife.";
date: "2017-08-3"; volume: "6. doi"; pages: " 10.7554/eLife.27711"
COMMENT Sequence Label: pIRE4-Azi
FEATURES Location/Qualifiers
source 1..6658
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 26..525
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(61..84)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 458..477
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
regulatory 552..561
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
polyA_signal 1981..2102
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2018..2037)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2072..2091
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
promoter 2127..2375
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
promoter 2485..2733
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
promoter 2843..3091
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
promoter 3201..3449
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
rep_origin complement(3552..4007)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3689..3710)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3901..3920
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4034..4138
/gene="bla"
/label=AmpR promoter
promoter 4140..4497
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 4140..4331
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(4163..4183)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4348..4483
/label=SV40 ori
/note="SV40 origin of replication"
CDS 4532..5326
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 4532..5326
/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(4586..4605)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5196..5215
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5514..5533)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5558..5605
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5956..6544
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6445..6464
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 acgcgttagt tattaaaatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga
61 gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc
121 acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg
181 ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc
241 aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct
301 gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct
361 ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg
421 ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc
481 acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg ctagcgttta
541 aacttaagct tgccaccatg gccagcagca acctgatcaa gcagctgcag gaacggggcc
601 tggtggccca agtgacagat gaagaagccc tggccgagag actggcccag ggacctattg
661 ccctgctgtg cggctttgac cccaccgccg attctctgca cctgggacat ctggtgcctc
721 tgctgtgtct gaagcggttc cagcaggccg gccacaaacc tgtggctctc gtgggcggag
781 ccaccggcct gattggcgat cctagcttta aggccgccga gcggaagctg aacaccgaag
841 agacagtgca ggaatgggtg gacaagatcc ggaaacaggt ggcccccttc ctggacttcg
901 actgcggcga gaatagcgcc attgccgcca acaactacga ttggttcggc aatatgaacg
961 tgctgacctt cctgcgggac atcggcaagc acttcagcgt gaaccagatg atcaacaaag
1021 aagccgtgaa gcagcggctg aacagagagg accagggcat cagcttcacc gagttcagct
1081 acaacctgct gcagggctac agcatggcct gcgccaacaa gcagtacggc gtggtgctgc
1141 agatcggcgg ctctgaccag tggggcaata tcaccagcgg catcgacctg accagacggc
1201 tgcaccagaa ccaggtgttc ggcctgaccg tgcccctgat cacaaaggcc gacggcacca
1261 agttcggcaa gaccgaaggc ggagccgtgt ggctggaccc caagaaaacc agcccctaca
1321 agttctacca gttctggatc aacaccgcca gagccgacgt gtaccggttc ctgaagttct
1381 tcaccttcat gagcatcgag gaaatcaacg ccctggaaga agaggacaag aacagcggca
1441 aggcccccag agcccagtac gtgctggcag aacaagtgac caggctggtg cacggcgagg
1501 aaggactgca ggccgccaag agaatcaccg agtgcctgtt cagcggcagc ctgagcgctc
1561 tgagcgaggc cgattttgag cagctggctc aggacggcgt gcccatggtg gaaatggaaa
1621 agggcgccga cctgatgcag gccctggtgg attctgagct gcagcccagc agaggacagg
1681 ccagaaagac aatcgccagc aacgccatca ccatcaacgg cgagaagcag agcgaccccg
1741 agtacttctt caaagaggaa gatcggctgt tcggccggtt caccctgctg cggagaggca
1801 agaagaacta ctgcctgatc tgctggaagt aatgaattct gcagatatcc agcacagtgg
1861 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
1921 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
1981 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
2041 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
2101 taaggtctag actcgagggg caggaagagg gcctatttcc catgattcct tcatatttgc
2161 atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga
2221 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
2281 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
2341 tggctttata tatcttgtgg aaaggacgaa acaccggagg ggtagcgaag tggctaaacg
2401 cggcggactc taaatccgct ccctttgggt tcggcggttc gaatccgtcc ccctccagtc
2461 ctttttttgg tcgaggggca ggaagagggc ctatttccca tgattccttc atatttgcat
2521 atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata
2581 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
2641 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
2701 gctttatata tcttgtggaa aggacgaaac accggagggg tagcgaagtg gctaaacgcg
2761 gcggactcta aatccgctcc ctttgggttc ggcggttcga atccgtcccc ctccagtcct
2821 ttttttggtc gaggggcagg aagagggcct atttcccatg attccttcat atttgcatat
2881 acgatacaag gctgttagag agataattag aattaatttg actgtaaaca caaagatatt
2941 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
3001 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
3061 tttatatatc ttgtggaaag gacgaaacac cggaggggta gcgaagtggc taaacgcggc
3121 ggactctaaa tccgctccct ttgggttcgg cggttcgaat ccgtccccct ccagtccttt
3181 ttttggtcga ggggcaggaa gagggcctat ttcccatgat tccttcatat ttgcatatac
3241 gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag
3301 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
3361 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
3421 tatatatctt gtggaaagga cgaaacaccg gaggggtagc gaagtggcta aacgcggcgg
3481 actctaaatc cgctcccttt gggttcggcg gttcgaatcc gtccccctcc agtccttttt
3541 ttggtcgacg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
3601 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
3661 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
3721 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
3781 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
3841 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
3901 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
3961 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
4021 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
4081 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
4141 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
4201 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
4261 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
4321 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
4381 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
4441 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
4501 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
4561 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
4621 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
4681 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
4741 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
4801 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
4861 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
4921 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
4981 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
5041 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
5101 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
5161 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
5221 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
5281 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
5341 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
5401 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
5461 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
5521 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
5581 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
5641 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
5701 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
5761 cctgccatag cagtactaaa tatttatccc gggctcaggt tactcatata tactttagat
5821 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
5881 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
5941 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
6001 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
6061 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
6121 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
6181 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
6241 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
6301 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
6361 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
6421 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
6481 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
6541 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
6601 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
pIRE4-Azi载体图谱质粒图谱pdf版和pIRE4-Azi载体序列质粒序列等相关资料下载
pIRE4-Azi质粒载体应用举例
- 上一篇：pRPF215
- 下一篇：mCherry-ARPp34-C-14
