| 质粒名称: | pMax_GFP |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 177825 |
| 存储实验室: | Zachary Nagel, Leona Samson |
|
载体骨架基本信息 |
|
| 载体名称: | pMax |
| 完整质粒大小: | 3527 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: | GFP |
| 物种来源: | Synthetic |
| 插入大小: | 720 bp |
| 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 |
|
| 材料和方法部分: | pMax_GFP was a gift from Zachary Nagel & Leona Samson (Addgene plasmid # 177825 ; http://n2t.net/addgene:177825 ; RRID:Addgene_177825) |
| 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
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pMax_GFP 3527 bp ds-DNA circular SYN 30-NOV-2021
DEFINITION Host Cell Reactivation (HCR) control GFP plasmid (pMax backbone).
ACCESSION .
VERSION .
KEYWORDS pMax_GFP
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 Nov 30, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3527
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..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
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 831..1547
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(876..897)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1137..1156)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1484..1505
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1563..1582)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
polyA_signal 1592..1673
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind 1617..1636
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin complement(1871..2459)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1951..1970)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2540..3334)
/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(2651..2670)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 3261..3280
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
ORIGIN
1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc 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 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
901 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
961 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
1021 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
1081 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
1141 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
1201 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
1261 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
1321 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
1381 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
1441 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
1501 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtga aagcttgagc
1561 tcgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
1621 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
1681 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
1741 agtaaaacct ctacaaatgt ggtacttaag agggggagac caaagggcga gacgttaagg
1801 cctcacgtga catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
1861 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
1921 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
1981 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttacggg atacctgtcc gcctttctcc
2041 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
2101 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
2161 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
2221 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
2281 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
2341 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
2401 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
2461 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
2521 ggattttggt catgccgtct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
2581 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
2641 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
2701 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
2761 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgctcgcc ttgagcctgg
2821 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
2881 gaccggcttc catccgagta cgtgctctct cgatgcgatg tttcgcttgg tggtcgaatg
2941 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
3001 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
3061 gccagtccct tcccgcttca gtgacaacgt cgagtacagc tgcgcaagga acgcccgtcg
3121 tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt
3181 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
3241 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
3301 gagaacctgc gtgcaatcca tcttgttcaa tcataatatt attgaagcat ttatcagggt
3361 tcgtctcgtc ccggtctcct cccatgcatg tcaatattgg ccattagcca tattattcat
3421 tggttatata gcataaatca atattggcta ttggccattg catacgttgt atctatatca
3481 taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgtt
//
