| 质粒名称: | pMax_GFP_C289T |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 177826 |
| 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: | 3527bp |
| 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: | GFP_C289T |
| 物种来源: | Synthetic |
| 插入大小: | 720bp |
| 基因突变: | C289T |
| 启动子: | CMV |
| 克隆方法: | Ligation Independent Cloning |
| 5’克隆位点: | AgeI(未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 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_GFP_C289T was a gift from Zachary Nagel & Leona Samson (Addgene plasmid # 177826 ; http://n2t.net/addgene:177826 ; RRID:Addgene_177826) |
| 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_GFP_C289T载体质粒基本信息
pMax_GFP_C289T质粒图谱载体图谱和pMax_GFP_C289T载体序列质粒序列多克隆位点信息
pMax_GFP_C289T质粒载体简介
pMax_GFP_C289T质粒序列载体序列
LOCUS pMax_GFP_C289T 3527 bp DNA circular SYN 14-APR-2025
DEFINITION Site-specific mutagenesis of GFP for the purposes of creating a
site-specific hypoxanthine lesion for the purposes of measuring MPG
activity via Host Cell Reactivation (HCR)..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3527)
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 3527)
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_GFP_C289T
FEATURES Location/Qualifiers
source 1..3527
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 178..197
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 437..458
/label=EGFP-N
/note="EGFP, reverse primer"
regulatory 500..509
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
intron 526..658
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
promoter complement(750..953)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(779..799)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 954..1333
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
CDS 1527..2321
/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 1527..2321
/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(1581..1600)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2191..2210
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
rep_origin 2402..2990
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2891..2910
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
polyA_signal 3188..3269
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3225..3244)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3279..3298
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
CDS complement(join(3311..3527,1..497,498..500,501..503))
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQECTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3356..3377)
/label=EGFP-C
/note="EGFP, forward primer"
ORIGIN
1 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
61 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
121 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
181 ttgtagttgc cgtcgtcctt gaagaagatg gtgcactcct ggacgtagcc ttcgggcatg
241 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
301 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
361 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
421 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
481 agctcctcgc ccttgctcac catggtggca gcgctagcgg tacccctgtg gagagaaagg
541 caaagtggat gtcagtaaga ccaataggtg cctatcagaa acgcaagaat cttctctgtc
601 tcgacaagcc cagtttctat tggcctcctt aaacctgtct tgtaaccttg atacttacct
661 gtgatcagtc gagcactgac tgcgttagca atttaactgt gataaactac cgcaataaag
721 ctactagtga tctgacggtt cactaaacga cctctgctta tatagacctc ccaccgtaca
781 cgcctaccgc ccatttgcgt caacggggcg gggttattac gacattttgg aaagtcccgt
841 tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc
901 cgtgagtcaa accgctatcc acgcccattg gtgtactgcc aaaaccgcat caccatggta
961 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccgtaa ggtcatgtac
1021 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cggacttggc
1081 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
1141 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
1201 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
1261 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactagtc
1321 aataatcaat gccaacatgg cggtcatatt ggacatgagc caatataaat gtacatatta
1381 tgatatagat acaacgtatg caatggccaa tagccaatat tgatttatgc tatataacca
1441 atgaataata tggctaatgg ccaatattga catgcatggg aggagaccgg gacgagacga
1501 accctgataa atgcttcaat aatattatga ttgaacaaga tggattgcac gcaggttctc
1561 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
1621 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
1681 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
1741 cgacgggcgt tccttgcgca gctgtactcg acgttgtcac tgaagcggga agggactggc
1801 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
1861 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
1921 cattcgacca ccaagcgaaa catcgcatcg agagagcacg tactcggatg gaagccggtc
1981 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
2041 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
2101 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
2161 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
2221 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
2281 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agacggcatg accaaaatcc
2341 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
2401 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
2461 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
2521 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
2581 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
2641 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
2701 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
2761 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
2821 ggagaaaggc ggacaggtat cccgtaagcg gcagggtcgg aacaggagag cgcacgaggg
2881 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
2941 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
3001 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcacgtgagg
3061 ccttaacgtc tcgccctttg gtctccccct cttaagtacc acatttgtag aggttttact
3121 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
3181 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
3241 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcga
3301 gctcaagctt tcacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
3361 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggg
3421 tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
3481 ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcgg
//
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