| 质粒名称: | pMax_mOrange_A215C |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 177828 |
| 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: | 3518bp |
| 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: | mOrange_A215C |
| 物种来源: | Synthetic |
| 插入大小: | 711bp |
| 基因突变: | A215C |
| 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_mOrange_A215C was a gift from Zachary Nagel & Leona Samson (Addgene plasmid # 177828 ; http://n2t.net/addgene:177828 ; RRID:Addgene_177828) |
| 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_mOrange_A215C载体质粒基本信息
pMax_mOrange_A215C质粒图谱载体图谱和pMax_mOrange_A215C载体序列质粒序列多克隆位点信息
pMax_mOrange_A215C质粒载体简介
pMax_mOrange_A215C质粒序列载体序列
LOCUS pMax_mOrange_A215C 3518 bp DNA circular SYN 14-APR-2025
DEFINITION Site-specific mutagenesis of mOrange for the purposes of creating a
site-specific 8-oxo-G:C lesion for the purposes of measuring OGG1
activity via Host Cell Reactivation (HCR)..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3518)
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 3518)
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_mOrange_A215C
FEATURES Location/Qualifiers
source 1..3518
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 77..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
intron 676..808
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
regulatory 825..834
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 831..1541
/codon_start=1
/product="orange monomeric derivative of DsRed fluorescent
protein (Shanet et al., 2004)"
/label=mOrange
/note="mammalian codon-optimized"
/translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
FQTAKLKVTKGGPLPFAWDILSPQFTSGSKAYVKHPADIPDYFKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind 1244..1263
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind complement(1554..1573)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
polyA_signal 1583..1664
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind 1608..1627
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin complement(1862..2450)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1942..1961)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2531..3325)
/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(2531..3325)
/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(2642..2661)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 3252..3271
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
ORIGIN
1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cctgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag aggtcgttta gtgaaccgtc
601 agatcactag tagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
661 ctcgactgat cacaggtaag tatcaaggtt acaagacagg tttaaggagg ccaatagaaa
721 ctgggcttgt cgagacagag aagattcttg cgtttctgat aggcacctat tggtcttact
781 gacatccact ttgcctttct ctccacaggg gtaccgctag cgctgccacc atggtgagca
841 agggcgagga gaataacatg gccatcatca aggagttcat gcgcttcaag gtgcgcatgg
901 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
961 agggctttca gaccgctaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
1021 acatcctgtc ccctcagttc acctccggct ccaaggccta cgtgaagcac cccgccgaca
1081 tccccgacta cttcaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
1141 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
1201 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
1261 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
1321 gcgagatcaa gatgaggctg aagctgaagg acggcggcca ctacacctcc gaggtcaaga
1381 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc ggcatcaagt
1441 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
1501 gccgccactc caccggcggc atggacgagc tgtacaagta gaagcttgag ctcgatgagt
1561 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
1621 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
1681 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
1741 tctacaaatg tggtacttaa gagggggaga ccaaagggcg agacgttaag gcctcacgtg
1801 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
1861 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
1921 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
1981 gctctcctgt tccgaccctg ccgcttacgg gatacctgtc cgcctttctc ccttcgggaa
2041 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
2101 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
2161 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
2221 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
2281 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
2341 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
2401 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
2461 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
2521 tcatgccgtc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
2581 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
2641 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
2701 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
2761 catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt
2821 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
2881 ccatccgagt acgtgctctc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
2941 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
3001 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
3061 ttcccgcttc agtgacaacg tcgagtacag ctgcgcaagg aacgcccgtc gtggccagcc
3121 acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga
3181 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
3241 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
3301 cgtgcaatcc atcttgttca atcataatat tattgaagca tttatcaggg ttcgtctcgt
3361 cccggtctcc tcccatgcat gtcaatattg gccattagcc atattattca ttggttatat
3421 agcataaatc aatattggct attggccatt gcatacgttg tatctatatc ataatatgta
3481 catttatatt ggctcatgtc caatatgacc gccatgtt
//
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