| 質粒名稱: | pcDNA3-FLAG-VSVG |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 80606 |
| 存儲實驗室: | Sven Eyckerman |
|
載體骨架基本信息 |
|
| 載體名稱: | pcDNA3.1 |
| 載體出品公司: | Life Technologies |
| 空載體大小: | 5349bp |
| 完整質粒大小: | 6939bp |
| 載體類型: | 哺乳細胞表達 |
| 篩選標記: | 新霉素（G418） |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | VSV-G、Vesicular Stomatitis Virus Glycoprotein |
| 物種來源: | Vesicular Stomatitis Virus |
| 插入大小: | 1590bp |
| 啟動子: | CMV |
| 標簽/融合蛋白: | FLAG (N terminal on insert) |
| 克隆方法: | 限制性內切酶 |
| 5’克隆位點: | NheI (not destroyed) |
| 3’克隆位點: | XbaI (not destroyed) |
| 5’測序引物 | GAACCCACTGCTTACTGGC |
| 3’測序引物 | GTCGAGGCTGATCAGCGAGC |
|
文獻引用方法 |
|
| 材料和方法部分: | pcDNA3-FLAG-VSVG was a gift from Sven Eyckerman (Addgene plasmid # 80606 ; //n2t.net/addgene:80606 ; RRID:Addgene_80606) |
| 參考文獻部分: | Trapping mammalian protein complexes in viral particles. Eyckerman S, Titeca K, Van Quickelberghe E, Cloots E, Verhee A, Samyn N, De Ceuninck L, Timmerman E, De Sutter D, Lievens S, Van Calenbergh S, Gevaert K, Tavernier J. Nat Commun. 2016 Apr 28;7:11416. doi: 10.1038/ncomms11416. 10.1038/ncomms11416 PubMed 27122307 |
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pcDNA3-FLAG-VSVG載體質粒基本信息
pcDNA3-FLAG-VSVG質粒圖譜載體圖譜和pcDNA3-FLAG-VSVG載體序列質粒序列多克隆位點信息
pcDNA3-FLAG-VSVG質粒載體簡介
pcDNA3-FLAG-VSVG質粒序列載體序列
LOCUS pcDNA3-FLAG-VSVG 6939 bp DNA circular SYN 14-APR-2025
DEFINITION Mammalian expression vector that expresses a VSV-G protein with a
FLAG tag near the N-terminus of the protein.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6939)
AUTHORS Eyckerman S, Titeca K, Van Quickelberghe E, Cloots E, Verhee A,
Samyn N, De Ceuninck L, Timmerman E, De Sutter D, Lievens S, Van
Calenbergh S, Gevaert K, Tavernier J
TITLE Trapping mammalian protein complexes in viral particles.
JOURNAL Nat Commun. 2016 Apr 28;7:11416. doi: 10.1038/ncomms11416.
PUBMED 27122307
REFERENCE 2 (bases 1 to 6939)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun."; date: "2016-04-28"; volume: "7:11416. doi"; pages: "
10.1038/ncomms11416"
COMMENT Sequence Label: pcDNA3-FLAG-VSVG
FEATURES Location/Qualifiers
source 1..6939
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 984..1007
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2454..2486
/codon_start=1
/product="epitope tag from vesicular stomatitis virus G
protein"
/label=VSV-G tag
/translation="YTDIEMNRLGK"
primer_bind complement(2533..2550)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2539..2763
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2809..3237
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2896..2915)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3106..3127
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3246..3266)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3251..3580
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3431..3566
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3493..3512
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3647..4441
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3701..3720)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4311..4330
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4615..4736
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4652..4671)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4706..4725
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4785..4801)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4785..4801)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4798..4820)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4809..4825
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(join(4833..4839,4840..4857,4858..4863))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4878..4899
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5016..5033)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5187..5772)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5267..5286)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(5943..6734,6735..6803))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6566..6585
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6804..6908)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 ccaccatgaa gtgccttttg tacttagcct ttttattcat tggggtgaat tgcaagttca
961 ccatagtttt tccatccgga ggagattaca aggatgacga cgataagggc ggaagcttgg
1021 gacacaacca aaaaggaaac tggaaaaatg ttccttctaa ttaccattat tgcccgtcaa
1081 gctcagattt aaattggcat aatgacttaa taggcacagc cttacaagtc aaaatgccca
1141 agagtcacaa ggctattcaa gcagacggtt ggatgtgtca tgcttccaaa tgggtcacta
1201 cttgtgattt ccgctggtat ggaccgaagt atataacaca ttccatccga tccttcactc
1261 catctgtaga acaatgcaag gaaagcattg aacaaacgaa acaaggaact tggctgaatc
1321 caggcttccc tcctcaaagt tgtggatatg caactgtgac ggatgccgaa gcagtgattg
1381 tccaggtgac tcctcaccat gtgctggttg atgaatacac aggagaatgg gttgattcac
1441 agttcatcaa cggaaaatgc agcaattaca tatgccccac tgtccataac tctacaacct
1501 ggcattctga ctataaggtc aaagggctat gtgattctaa cctcatttcc atggacatca
1561 ccttcttctc agaggacgga gagctatcat ccctgggaaa ggagggcaca gggttcagaa
1621 gtaactactt tgcttatgaa actggaggca aggcctgcaa aatgcaatac tgcaagcatt
1681 ggggagtcag actcccatca ggtgtctggt tcgagatggc tgataaggat ctctttgctg
1741 cagccagatt ccctgaatgc ccagaagggt caagtatctc tgctccatct cagacctcag
1801 tggatgtaag tctaattcag gacgttgaga ggatcttgga ttattccctc tgccaagaaa
1861 cctggagcaa aatcagagcg ggtcttccaa tctctccagt ggatctcagc tatcttgctc
1921 ctaaaaaccc aggaaccggt cctgctttca ccataatcaa tggtacccta aaatactttg
1981 agaccagata catcagagtc gatattgctg ctccaatcct ctcaagaatg gtcggaatga
2041 tcagtggaac taccacagaa agggaactgt gggatgactg ggcaccatat gaagacgtgg
2101 aaattggacc caatggagtt ctgaggacca gttcaggata taagtttcct ttatacatga
2161 ttggacatgg tatgttggac tccgatcttc atcttagctc aaaggctcag gtgttcgaac
2221 atcctcacat tcaagacgct gcttcgcaac ttcctgatga tgagagttta ttttttggtg
2281 atactgggct atccaaaaat ccaatcgagc ttgtagaagg ttggttcagt agttggaaaa
2341 gctctattgc ctcttttttc tttatcatag ggttaatcat tggactattc ttggttctcc
2401 gagttggtat ccatctttgc attaaattaa agcacaccaa gaaaagacag atttatacag
2461 acatagagat gaaccgactt ggaaaggaat tctaactcga gtctagaggg cccgtttaaa
2521 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
2581 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
2641 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
2701 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
2761 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
2821 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
2881 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
2941 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
3001 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
3061 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
3121 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
3181 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
3241 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
3301 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
3361 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
3421 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
3481 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
3541 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
3601 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
3661 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
3721 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
3781 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
3841 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
3901 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
3961 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
4021 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
4081 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
4141 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
4201 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
4261 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
4321 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
4381 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
4441 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
4501 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
4561 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
4621 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
4681 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
4741 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
4801 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
4861 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
4921 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
4981 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
5041 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
5101 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
5161 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
5221 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
5281 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
5341 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
5401 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
5461 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
5521 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
5581 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
5641 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
5701 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
5761 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
5821 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
5881 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
5941 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
6001 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
6061 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
6121 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
6181 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
6241 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
6301 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
6361 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
6421 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
6481 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
6541 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
6601 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
6661 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
6721 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
6781 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
6841 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
6901 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
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