质粒名称: | pMax_mOrange |
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出品公司: | Addgene |
目录编号: | 177827 |
存储实验室: | Zachary Nagel, Leona Samson |
载体骨架基本信息 |
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载体名称: | pMax |
完整质粒大小: | 3518 bp |
载体类型: | 哺乳细胞表达 |
载体抗性: | Kanamycin |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | Orange |
物种来源: | Synthetic |
插入大小: | 711bp |
启动子: | CMV |
克隆方法: | Ligation Independent Cloning |
5’测序引物 | CGCAAATGGGCGGTAGGCGTG |
3’测序引物 | GCAATAGCATCACAAATTTCACA |
文献引用方法 |
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材料和方法部分: | pMax_mOrange was a gift from Zachary Nagel & Leona Samson (Addgene plasmid # 177827 ; http://n2t.net/addgene:177827 ; RRID:Addgene_177827) |
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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LOCUS pMax_mOrange 3518 bp ds-DNA circular SYN 30-NOV-2021 DEFINITION Host Cell Reactivation (HCR) control mOrange plasmid (pMax backbone). ACCESSION . VERSION . KEYWORDS pMax_mOrange 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 Nov 30, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..3518 /organism="synthetic DNA construct" /mol_type="other DNA" 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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 FQTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNF 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 /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 gtgggagcgc 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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