| 质粒名称: | pMax_mOrange_G299C |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 177829 |
| 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_G299C |
| 物种来源: | Synthetic |
| 插入大小: | 711 bp |
| 基因突变: | G299C |
| 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_G299C was a gift from Zachary Nagel & Leona Samson (Addgene plasmid # 177829 ; http://n2t.net/addgene:177829 ; RRID:Addgene_177829) |
| 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 |
买家导航
pMax_mOrange_G299C载体质粒基本信息
pMax_mOrange_G299C质粒图谱载体图谱和pMax_mOrange_G299C载体序列质粒序列多克隆位点信息
pMax_mOrange_G299C质粒载体简介
pMax_mOrange_G299C质粒序列载体序列
LOCUS pMax_mOrange_G299C 3518 bp DNA circular SYN 14-APR-2025
DEFINITION Site-specific mutagenesis of mOrange for the purposes of creating a
site-specific G:G mispair for the purposes of measuring MMR 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_G299C
FEATURES Location/Qualifiers
source 1..3518
/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"
regulatory 445..454
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 451..1161
/codon_start=1
/product="orange monomeric derivative of DsRed fluorescent
protein (Shanet et al., 2004)"
/label=mOrange
/note="mammalian codon-optimized"
/translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
FQTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYFKLSFPEGFKWEPVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind 864..883
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind complement(1174..1193)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
polyA_signal 1203..1284
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind 1228..1247
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin complement(1482..2070)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1562..1581)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2151..2945)
/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(2151..2945)
/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(2262..2281)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 2872..2891
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
enhancer 3139..3518
/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 atggtgagca agggcgagga gaataacatg
481 gccatcatca aggagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac
541 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggctttca gaccgctaag
601 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
661 acctacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttcaagctg
721 tccttccccg agggcttcaa gtgggagccc gtgatgaact tcgaggacgg cggcgtggtg
781 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
841 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
901 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gatgaggctg
961 aagctgaagg acggcggcca ctacacctcc gaggtcaaga ccacctacaa ggccaagaag
1021 cccgtgcagc tgcccggcgc ctacatcgtc ggcatcaagt tggacatcac ctcccacaac
1081 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
1141 atggacgagc tgtacaagta gaagcttgag ctcgatgagt ttggacaaac cacaactaga
1201 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
1261 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
1321 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtacttaa
1381 gagggggaga ccaaagggcg agacgttaag gcctcacgtg acatgtgagc aaaaggccag
1441 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
1501 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
1561 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
1621 ccgcttacgg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
1681 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
1741 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
1801 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
1861 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
1921 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
1981 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
2041 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
2101 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgccgtc tcagaagaac
2161 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
2221 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
2281 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
2341 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
2401 tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
2461 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctctc
2521 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
2581 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
2641 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
2701 tcgagtacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
2761 tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
2821 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
2881 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
2941 atcataatat tattgaagca tttatcaggg ttcgtctcgt cccggtctcc tcccatgcat
3001 gtcaatattg gccattagcc atattattca ttggttatat agcataaatc aatattggct
3061 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
3121 caatatgacc gccatgttgg cattgattat tgactagtta ttaatagtaa tcaattacgg
3181 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
3241 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
3301 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
3361 cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg
3421 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt
3481 ggcagtacat ctacgtatta gtcatcgcta ttaccatg
//
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