| 质粒名称: | p40PX-EGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 19010 |
| 存储实验室: | Michael Yaffe |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N3 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 5170 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | 新霉素（G418筛选） |
| 载体抗性: | 卡那霉素，50μg/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: | p40phox |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 444 bp |
| 基因突变: | aa1-148 (PX domains) |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: | EGFP（主干上的C末端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BglII(未破坏) |
| 3’克隆位点: | HindIII (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | EGFP-N |
|
文献引用方法 |
|
| 材料和方法部分: | p40PX-EGFP was a gift from Michael Yaffe (Addgene plasmid # 19010 ; http://n2t.net/addgene:19010 ; RRID:Addgene_19010) |
| 参考文献部分: | The PX domains of p47phox and p40phox bind to lipid products of PI(3)K. Kanai F, Liu H, Field SJ, Akbary H, Matsuo T, Brown GE, Cantley LC, Yaffe MB. Nat Cell Biol. 2001 Jul . 3(7):675-8. 10.1038/35083070 PubMed 11433300 |
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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 p40PX-EGFP 5170 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS p40PX-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5170)
AUTHORS Kanai F, Liu H, Field SJ, Akbary H, Matsuo T, Brown GE, Cantley LC,
Yaffe MB
TITLE The PX domains of p47phox and p40phox bind to lipid products of
PI(3)K.
JOURNAL Nat Cell Biol. 2001 Jul . 3(7):675-8.
PUBMED 11433300
REFERENCE 2 (bases 1 to 5170)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5170
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 1110..1119
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1116..1835
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1116..1832
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1161..1182)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1422..1441)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1769..1790
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1958..2079
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1995..2014)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2049..2068
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2086..2541)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2223..2244)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2435..2454
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2568..2672
/gene="bla"
/label=AmpR promoter
promoter 2674..3031
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2697..2717)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2882..3017
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2944..2963
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3066..3860
/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(3120..3139)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3730..3749
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4048..4067)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4092..4139
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4468..5056
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4957..4976
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atctaccatg gctgtggccc agcagctgcg ggccgagagt gactttgaac
661 agcttccgga tgatgttgcc atctcggcca acattgctga catcgaggag aagagaggct
721 tcaccagcca ctttgttttc gtcatcgagg tgaagacaaa aggaggatcc aagtacctca
781 tctaccgccg ctaccgccag ttccatgctt tgcagagcaa gctggaggag cgcttcgggc
841 cagacagcaa gagcagtgcc ctggcctgta ccctgcccac actcccagcc aaagtctacg
901 tgggtgtgaa acaggagatc gccgagatgc ggatacctgc cctcaacgcc tacatgaaga
961 gcctgctcag cctgccggtc tgggtgctga tggatgagga cgtccggatc ttcttttacc
1021 agtcgcccta tgactcagag caggtgcccc aggcactccg ccaagcttcg aattctgcag
1081 tcgacggtac cgcgggcccg ggatccatcg ccaccatggt gagcaagggc gaggagctgt
1141 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
1201 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
1261 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
1321 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
1381 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
1441 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
1501 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
1561 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
1621 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
1681 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
1741 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
1801 ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat
1861 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
1921 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
1981 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
2041 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
2101 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
2161 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
2221 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
2281 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
2341 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
2401 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
2461 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
2521 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
2581 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
2641 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
2701 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
2761 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
2821 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
2881 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
2941 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
3001 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
3061 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
3121 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
3181 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
3241 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
3301 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
3361 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
3421 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
3481 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
3541 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
3601 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
3661 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
3721 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
3781 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
3841 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
3901 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
3961 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
4021 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
4081 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
4141 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
4201 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
4261 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
4321 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
4381 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
4441 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
4501 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
4561 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
4621 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
4681 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
4741 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
4801 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
4861 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
4921 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
4981 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
5041 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
5101 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
5161 cgccatgcat
//
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