| 质粒名称: | pEGFP-VSVG |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 11912 |
| 存储实验室: | Jennifer Lippincott-Schwartz |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4733bp |
| 完整质粒大小: | 4989 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: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | VSVG、Vesicular stomatitus virus G protein |
| 插入大小: | 1500bp |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BglII (not destroyed) |
| 3’克隆位点: | EcoRI (not destroyed) |
| 5’测序引物 | CMV-F |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP-VSVG was a gift from Jennifer Lippincott-Schwartz (Addgene plasmid # 11912 ; http://n2t.net/addgene:11912 ; RRID:Addgene_11912) |
| 参考文献部分: | ER-to-Golgi transport visualized in living cells. Presley JF, Cole NB, Schroer TA, Hirschberg K, Zaal KJ, Lippincott-Schwartz J. Nature. 1997 Sep 4. 389(6646):81-5. 10.1038/38001 PubMed 9288971 |
买家导航
pEGFP-VSVG载体质粒基本信息
pEGFP-VSVG质粒图谱载体图谱和pEGFP-VSVG载体序列质粒序列多克隆位点信息
pEGFP-VSVG质粒载体简介
pEGFP-VSVG质粒序列载体序列
LOCUS pEGFP-VSVG 6254 bp DNA circular SYN 25-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6254)
AUTHORS Presley JF, Cole NB, Schroer TA, Hirschberg K, Zaal KJ,
Lippincott-Schwartz J
TITLE ER-to-Golgi transport visualized in living cells.
JOURNAL Nature. 1997 Sep 4. 389(6646):81-5.
PUBMED 9288971
REFERENCE 2 (bases 1 to 6254)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature.
1997 Sep 4. 389(6646):81-5."
FEATURES Location/Qualifiers
source 1..6254
/mol_type="other DNA"
/organism="synthetic DNA construct"
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"
CDS 625..2157
/codon_start=1
/product="vesicular stomatitis virus G glycoprotein,
thermosensitive tsO45 mutant"
/label=VSV-G tsO45
/note="thermosensitive variant of VSV-G Orsay"
/translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAAIVQVTPHHVLVDEYTGEWVDSQ
FINGKCSNDICPTVHNSTTWHSDYKVKGLCDSNLISMDITSFSEDGELSSLGKEGTGFR
SNYFAYETGDKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPVFTIINGTLK
YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDETLFFFGDTGLSKNPIEFVEGW
FSSWKSSIASFFFIIGLIIGLFLVLRVGIYLCIKLKHTKKRQIYTDIEMNRLG"
CDS 2200..2919
/codon_start=1
/product="enhanced GFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/label=mEGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2245..2266)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2506..2525)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2853..2874
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 3042..3163
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3079..3098)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3133..3152
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3170..3625)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3307..3328)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3519..3538
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3652..3756
/gene="bla"
/label=AmpR promoter
promoter 3758..4115
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 3758..3949
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(3781..3801)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3966..4101
/label=SV40 ori
/note="SV40 origin of replication"
CDS 4150..4944
/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 4150..4944
/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(4204..4223)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4814..4833
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5132..5151)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5176..5223
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5552..6140
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6041..6060
/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 atctcgccgc caccatgaag tgccttttgt acttagcttt tttattcatc
661 ggggtgaatt gcaagttcac catagttttt ccacacaacc aaaaaggaaa ctggaaaaat
721 gttccttcca attaccatta ttgcccgtca agctcagatt taaattggca taatgactta
781 ataggcacag ccttacaagt caaaatgccc aagagtcaca aggctattca agcagacggt
841 tggatgtgtc atgcttccaa atgggtcact acttgtgatt tccgctggta cggaccgaag
901 tatataacac attccatccg atccttcact ccatctgtag aacaatgcaa ggaaagcatt
961 gaacaaacga aacaaggaac ttggctgaat ccaggcttcc ctcctcaaag ttgtggatat
1021 gcaactgtga cggatgctga agcagcgatt gtccaggtga ctcctcacca tgtgcttgtt
1081 gatgaataca caggagaatg ggttgattca cagttcatca acggaaaatg cagcaatgac
1141 atatgcccca ctgtccataa ctccacaacc tggcattccg actataaggt caaagggcta
1201 tgtgattcta acctcatttc catggacatc acctccttct cagaggacgg agagctatca
1261 tccctaggaa aggagggcac agggttcaga agtaactact ttgcttatga aaccggagac
1321 aaggcctgca aaatgcagta ctgcaagcat tggggagtca gactcccatc aggtgtctgg
1381 ttcgagatgg ctgataagga tctctttgct gcagccagat tccctgaatg cccagaaggg
1441 tcaagtatct ctgctccatc tcagacctca gtggatgtaa gtctcattca ggacgttgag
1501 aggatcttgg attattccct ctgccaagaa acctggagca aaatcagagc gggtcttccc
1561 atctctccag tggatctcag ctatcttgct cctaaaaacc caggaaccgg tcctgtcttt
1621 accataatca atggtaccct aaaatacttt gagaccagat acatcagagt cgatattgct
1681 gctccaatcc tctcaagaat ggtcggaatg atcagtggaa ctaccacaga aagggaactg
1741 tgggatgact gggctccata tgaagacgtg gaaattggac ccaatggagt tctgaggacc
1801 agttcaggat ataagtttcc tttatatatg attggacatg gtatgttgga ctccgatctt
1861 catcttagct caaaggctca ggtgtttgaa catcctcaca ttcaagacgc tgcttcgcag
1921 cttcctgatg atgagacttt attttttttt ggtgatactg ggctatccaa aaatccaatc
1981 gagtttgtag aaggttggtt cagtagttgg aagagctcta ttgcctcttt tttctttatc
2041 atagggttaa tcattggact attcttggtt ctccgagttg gtatttatct ttgcattaaa
2101 ttaaagcaca ccaagaaaag acagatttat acagacatag agatgaaccg acttggaaac
2161 agaattctgc agtcgacggt accgcgggcc cgggatccaa tggtgagcaa gggcgaggag
2221 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
2281 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
2341 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
2401 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
2461 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
2521 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
2581 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
2641 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
2701 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
2761 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
2821 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
2881 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca
2941 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
3001 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
3061 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
3121 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
3181 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
3241 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
3301 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
3361 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
3421 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
3481 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
3541 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
3601 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
3661 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
3721 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
3781 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
3841 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
3901 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
3961 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
4021 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
4081 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
4141 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
4201 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
4261 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
4321 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
4381 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
4441 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
4501 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
4561 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
4621 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
4681 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
4741 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
4801 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
4861 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
4921 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
4981 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
5041 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
5101 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
5161 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
5221 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
5281 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
5341 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
5401 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
5461 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
5521 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
5581 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
5641 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
5701 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
5761 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
5821 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
5881 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
5941 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
6001 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
6061 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
6121 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
6181 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
6241 ttaccgccat gcat
//
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