| 质粒名称: | pMax_BFP_A191G |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 177823 |
| 存储实验室: | Zachary Nagel, Leona Samson |
|
载体骨架基本信息 |
|
| 载体名称: | pMax |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 3503 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: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | agBFP_A191G |
| 物种来源: | Synthetic |
| 插入大小: | 702bp |
| 基因突变: | A191G |
| 启动子: | CMV |
| 克隆方法: | Ligation Independent Cloning |
| 5’测序引物 | CGCAAATGGGCGGTAGGCGTG |
| 3’测序引物 |
GCAATAGCATCACAAATTTCACA
(Common Sequencing Primers)
|
|
文献引用方法 |
|
| 材料和方法部分: |
pMax_BFP_A191G was a gift from Zachary Nagel & Leona Samson (Addgene plasmid # 177823 ; http://n2t.net/addgene:177823 ; RRID:Addgene_177823)
|
| 参考文献部分: | Multiplexed DNA repair assays for multiple lesions and multiple doses via transcription inhibition and transcriptional mutagenesis. Nagel ZD, Margulies CM, Chaim IA, McRee SK, Mazzucato P, Ahmad A, Abo RP, Butty VL, Forget AL, Samson LD. Proc Natl Acad Sci U S A. 2014 May 6;111(18):E1823-32. doi: 10.1073/pnas.1401182111. Epub 2014 Apr 22. 10.1073/pnas.1401182111 PubMed 24757057 |
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pMax_BFP_A191G载体质粒基本信息
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pMax_BFP_A191G质粒载体简介
pMax_BFP_A191G质粒序列载体序列
LOCUS pMax_BFP_A191G 3503 bp DNA circular SYN 14-APR-2025
DEFINITION Site-specific mutagenesis of tagBFP for the purposes of creating a
site-specific U:A mispair for the purposes of measuring UDG activity
via Host Cell Reactivation (HCR)..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3503)
AUTHORS Nagel ZD, Margulies CM, Chaim IA, McRee SK, Mazzucato P, Ahmad A,
Abo RP, Butty VL, Forget AL, Samson LD
TITLE Multiplexed DNA repair assays for multiple lesions and multiple
doses via transcription inhibition and transcriptional mutagenesis.
JOURNAL Proc Natl Acad Sci U S A. 2014 May 6;111(18):E1823-32. doi:
10.1073/pnas.1401182111. Epub 2014 Apr 22.
PUBMED 24757057
REFERENCE 2 (bases 1 to 3503)
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"; doi:
"10.1073/pnas.1401182111"; journalName: "Proc Natl Acad Sci U S A";
date: "2014-05-6- 6"; volume: "111"; issue: "18"; pages: "E1823-32"
COMMENT Sequence Label: pMax_BFP_A191G
FEATURES Location/Qualifiers
source 1..3503
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..204
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 155..175
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
intron 296..428
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
CDS 451..1152
/codon_start=1
/product="monomeric blue fluorescent protein"
/label=TagBFP
/note="mammalian codon-optimized"
/translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
VEGGPLPFAFDILATSFLCGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
DLPSKLGHKLN"
primer_bind complement(1159..1178)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
polyA_signal 1188..1269
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind 1213..1232
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin complement(1467..2055)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1547..1566)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2136..2930)
/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 complement(2136..2930)
/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(2247..2266)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 2857..2876
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
enhancer 3124..3503
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
181 tgggaggtct atataagcag aggtcgttta gtgaaccgtc agatcactag tagctttatt
241 gcggtagttt atcacagtta aattgctaac gcagtcagtg ctcgactgat cacaggtaag
301 tatcaaggtt acaagacagg tttaaggagg ccaatagaaa ctgggcttgt cgagacagag
361 aagattcttg cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct
421 ctccacaggg gtaccgctag cgctgccacc atgagcgagc tgattaagga gaacatgcac
481 atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc
541 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct
601 ctccccttcg ccttcgacat cctggctact agcttcctct gcggcagcaa gaccttcatc
661 aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg
721 gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc
781 caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc
841 cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct
901 gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg
961 atcgcaaaca tcaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct
1021 ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac
1081 gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac
1141 aagcttaatt gagagctcga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
1201 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
1261 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
1321 gaggtttttt aaagcaagta aaacctctac aaatgtggta cttaagaggg ggagaccaaa
1381 gggcgagacg ttaaggcctc acgtgacatg tgagcaaaag gccagcaaaa ggccaggaac
1441 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
1501 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
1561 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct tacgggatac
1621 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
1681 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
1741 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
1801 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
1861 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
1921 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
1981 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
2041 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
2101 gaaaactcac gttaagggat tttggtcatg ccgtctcaga agaactcgtc aagaaggcga
2161 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
2221 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
2281 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
2341 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
2401 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
2461 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctctctcgat gcgatgtttc
2521 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
2581 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
2641 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag tacagctgcg
2701 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
2761 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
2821 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
2881 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat aatattattg
2941 aagcatttat cagggttcgt ctcgtcccgg tctcctccca tgcatgtcaa tattggccat
3001 tagccatatt attcattggt tatatagcat aaatcaatat tggctattgg ccattgcata
3061 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgccat
3121 gttggcattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
3181 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
3241 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
3301 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
3361 atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg
3421 cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg
3481 tattagtcat cgctattacc atg
//
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