| 质粒名称: | pEGFP-VSP4-E228Q |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 80351 |
| 存储实验室: | Wesley Sundquist |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4038 bp |
| 完整质粒大小: | 6024 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | VPS4-E228Q |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1353 bp |
| 启动子: | CMV |
| 标签/融合蛋白: | GFP (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI (unknown if destroyed) |
| 3’克隆位点: | BamHI (unknown if destroyed) |
| 5’测序引物 | CMV-F |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP-VSP4-E228Q was a gift from Wesley Sundquist (Addgene plasmid # 80351 ; http://n2t.net/addgene:80351 ; RRID:Addgene_80351) |
| 参考文献部分: | Designed proteins induce the formation of nanocage-containing extracellular vesicles. Votteler J, Ogohara C, Yi S, Hsia Y, Nattermann U, Belnap DM, King NP, Sundquist WI. Nature. 2016 Dec 8;540(7632):292-295. doi: 10.1038/nature20607. Epub 2016 Nov 30. 10.1038/nature20607 PubMed 27919066 |
买家导航
pEGFP-VSP4-E228Q载体质粒基本信息
pEGFP-VSP4-E228Q质粒图谱载体图谱和pEGFP-VSP4-E228Q载体序列质粒序列多克隆位点信息
pEGFP-VSP4-E228Q质粒载体简介
pEGFP-VSP4-E228Q质粒序列载体序列
LOCUS pEGFP-VSP4-E228Q 6024 bp DNA circular SYN 14-APR-2025
DEFINITION Expression of GFP-tagged, dominant negative VPS4 mutant.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6024)
AUTHORS Votteler J, Ogohara C, Yi S, Hsia Y, Nattermann U, Belnap DM, King
NP, Sundquist WI
TITLE Designed proteins induce the formation of nanocage-containing
extracellular vesicles.
JOURNAL Nature. 2016 Dec 8;540(7632):292-295. doi: 10.1038/nature20607. Epub
2016 Nov 30.
PUBMED 27919066
REFERENCE 2 (bases 1 to 6024)
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:
"10.1038/nature20607"; journalName: "Nature"; date: "2016-12-8- 8";
volume: "540"; issue: "7632"; pages: "292-295"
COMMENT Sequence Label: pEGFP-VSP4-E228Q
FEATURES Location/Qualifiers
source 1..6024
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(join(825..1535,1536..1538,1539..1541))
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS complement(join(825..1535,1536..1538,1539..1541))
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(867..888)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 1216..1235
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1475..1496
/label=EGFP-N
/note="EGFP, reverse primer"
regulatory 1538..1547
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter complement(1586..1789)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(1615..1635)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 1790..2093
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
rep_origin complement(2268..2856)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2348..2367)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
polyA_signal 3185..3232
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 3257..3276
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(3464..4258)
/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(3464..4258)
/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(3575..3594)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 4185..4204
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(4293..4650)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4307..4442
/label=SV40 ori
/note="SV40 origin of replication"
enhancer 4459..4650
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind 4607..4627
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter complement(4652..4756)
/gene="bla"
/label=AmpR promoter
rep_origin 4783..5238
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4870..4889)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5080..5101
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 5245..5366
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5256..5275)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5310..5329
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
ORIGIN
1 ttgttattcc ccaccccctg catctggacc aagaactccg ttttgatcct ccgggcggcc
61 tcactctcat tttcatttcg ggacccgcag agggaatcca cctggtcgat gaagatgatg
121 gagggcttgt gctgcctggc cagctcaaac aggttcttga ccagcttttc actctccccc
181 agccacttgg acatcagatc tgaggaggac acagagaaga aggtggagtt gttggcctct
241 gttgccacgg ctttggccag gtaggatttc cctgtgccag ggggtccgaa cagcagaatc
301 ccccgccagg gggtgcgctt gcctgtgaac aagtgtggga atttgattgg caaaatgaca
361 gcttctttga gggcctcctt ggccccctcc agcccggcca cgtcgttcca ccgtatgttg
421 ggcttctcca tcacgacggc acccatcagc tgttcttgca gtttcttttt ctccggatta
481 tccccttcac tgtcactgtc actgcccttg ccctcactct ggttctcttt gactggcttc
541 ttgccgtgtt tctctttgct tcgtaaataa tccttcagct tctcggcccg gtctaggtac
601 tgcacgcact tggctcgaat gctctccttg gccttgtcgc tgtgggcctc atacttgata
661 gcgtggagga agtactccac cgcatgctgg tacagccgca gcgcctcctc gtagttcttg
721 gctttgtcct cctctgtggc tttcgtcacc agatcaatgg ctttctggag ggttgacgtt
781 gtcatcgcag aattcgaagc ttgagctcga gatctgagtc cggacttgta cagctcgtcc
841 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
901 tcgttggggt ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc
961 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
1021 tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc
1081 atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg
1141 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
1201 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
1261 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
1321 cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc
1381 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg
1441 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
1501 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtagcgct
1561 agcggatctg acggttcact aaaccagctc tgcttatata gacctcccac cgtacacgcc
1621 taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat
1681 tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg
1741 agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag
1801 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
1861 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
1921 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
1981 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
2041 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
2101 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctaatgcatg
2161 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
2221 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
2281 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
2341 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
2401 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
2461 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
2521 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
2581 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
2641 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
2701 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
2761 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
2821 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
2881 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
2941 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
3001 atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
3061 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
3121 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
3181 ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg
3241 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg
3301 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
3361 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
3421 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
3481 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
3541 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
3601 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
3661 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
3721 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
3781 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
3841 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
3901 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
3961 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
4021 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
4081 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
4141 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
4201 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
4261 gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag
4321 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
4381 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
4441 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
4501 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
4561 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
4621 acattccaca gctggttctt tccgcctcag gactcttcct ttttcaatat tattgaagca
4681 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
4741 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
4801 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
4861 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
4921 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
4981 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
5041 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
5101 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
5161 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
5221 tattaacgct tacaatttac gcgttaagat acattgatga gtttggacaa accacaacta
5281 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
5341 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
5401 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
5461 ctgattatga tcagttatct agatccggtg gatccttaac tctcttgccc aaagtcctct
5521 gagaatttct tcactttcag gaggtcgtct gcattcaccg tgggccgggt ggtggccaga
5581 gaccgcagca tgtccgacat gcaaaccaca ggctctaaga gtttgtcccc agggacatcc
5641 atccaagtca tctccatggc tcctgggtcc cctggtgagc atggagtcag gaggtcatca
5701 atcatcatgc tggggttggt gcgagagggg ccacagacct ttttgaagtg tgtggccgac
5761 tgcaccttcc tcacgggctg catgagagag tcccgcacga tgatgctgat gtccgcgccc
5821 gagtagcctt ccgtcttccg ggccagctcg tggatgtttg catccgtgag gttgtgggga
5881 gtgctcccga gatgcaaccg gaacatctgg gcgcgggcag cttcctccgg caaggggata
5941 taaattcgct tttcaaacct cctcctgatg gccgaatcca acacccatgg gatgtttgtg
6001 gctccaagaa ccagagtccc atca
//
