| 质粒名称: | EGFP-Vimentin-7 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 56439 |
| 存储实验室: | Michael Davidson |
|
载体骨架基本信息 |
|
| 载体名称: | EGFP |
| 载体出品公司: | Clontech |
| 空载体大小: | 4750 bp |
| 完整质粒大小: | 6110 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 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: | Vimentin / NM_003380.3 |
| 物种来源: | H. sapiens (human) |
| 启动子: | CMV |
| 标签/融合蛋白: | EGFP (C terminal on backbone) |
| 克隆方法: | 未知 |
| 5’测序引物 | CMV-F |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | EGFP-Vimentin-7 was a gift from Michael Davidson (Addgene plasmid # 56439 ; http://n2t.net/addgene:56439 ; RRID:Addgene_56439) |
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 EGFP-Vimentin-7 6110 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Localization: Intermediate Filaments, Excitation: 488, Emission:
507.
ACCESSION .
VERSION .
KEYWORDS EGFP-Vimentin-7
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6110)
TITLE Davidson Green fluorescent proteins
REFERENCE 2 (bases 1 to 6110)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6110
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 68..371
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 372..575
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 526..546
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 2057..2066
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2063..2782
/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 2063..2779
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2108..2129)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2369..2388)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2716..2737
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2905..3026
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2942..2961)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2996..3015
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3033..3488)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3170..3191)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3382..3401
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3515..3619
/gene="bla"
/label=AmpR promoter
promoter 3621..3978
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3644..3664)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3829..3964
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3891..3910
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4013..4807
/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(4067..4086)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4677..4696
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4995..5014)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5039..5086
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5415..6003
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5904..5923
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt cccatgtcca ccaggtccgt
661 gtcctcgtcc tcctaccgca ggatgttcgg cggcccgggc accgcgagcc ggccgagctc
721 cagccggagc tacgtgacta cgtccacccg cacctacagc ctgggcagcg cgctgcgccc
781 cagcaccagc cgcagcctct acgcctcgtc cccgggcggc gtgtatgcca cgcgctcctc
841 tgccgtgcgc ctgcggagca gcgtgcccgg ggtgcggctc ctgcaggact cggtggactt
901 ctcgctggcc gacgccatca acaccgagtt caagaacacc cgcaccaacg agaaggtgga
961 gctgcaggag ctgaatgacc gcttcgccaa ctacatcgac aaggtgcgct tcctggagca
1021 gcagaataag atcctgctgg ccgagctcga gcagctcaag ggccaaggca agtcgcgcct
1081 gggggacctc tacgaggagg agatgcggga gctgcgccgg caggtggacc agctaaccaa
1141 cgacaaagcc cgcgtcgagg tggagcgcga caacctggcc gaggacatca tgcgcctccg
1201 ggagaaattg caggaggaga tgcttcagag agaggaagcc gaaaacaccc tgcaatcttt
1261 cagacaggat gttgacaatg cgtctctggc acgtcttgac cttgaacgca aagtggaatc
1321 tttgcaagaa gagattgcct ttttgaagaa actccacgaa gaggaaatcc aggagctgca
1381 ggctcagatt caggaacagc atgtccaaat cgatgtggat gtttccaagc ctgacctcac
1441 ggctgccctg cgtgacgtac gtcagcaata tgaaagtgtg gctgccaaga acctgcagga
1501 ggcagaagaa tggtacaaat ccaagtttgc tgacctctct gaggctgcca accggaacaa
1561 tgacgccctg cgccaggcaa agcaggagtc cactgagtac cggagacagg tgcagtccct
1621 cacctgtgaa gtggatgccc ttaaaggaac caatgagtcc ctggaacgcc agatgcgtga
1681 aatggaagag aactttgccg ttgaagctgc taactaccaa gacactattg gccgcctgca
1741 ggatgagatt cagaatatga aggaggaaat ggctcgtcac cttcgtgaat accaagacct
1801 gctcaatgtt aagatggccc ttgacattga gattgccacc tacaggaagc tgctggaagg
1861 cgaggagagc aggatttctc tgcctcttcc aaacttttcc tccctgaacc tgagggaaac
1921 taatctggat tcactccctc tggttgatac ccactcaaaa aggacacttc tgattaagac
1981 ggttgaaact agagatggac aggttatcaa cgaaacttct cagcatcacg atgaccttga
2041 aggggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
2101 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
2161 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
2221 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
2281 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
2341 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
2401 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
2461 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
2521 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
2581 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
2641 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
2701 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
2761 catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca
2821 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
2881 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
2941 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
3001 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
3061 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
3121 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
3181 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
3241 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
3301 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
3361 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
3421 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
3481 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
3541 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
3601 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
3661 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
3721 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
3781 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
3841 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
3901 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
3961 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
4021 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
4081 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
4141 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
4201 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
4261 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
4321 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
4381 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
4441 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
4501 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
4561 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
4621 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
4681 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
4741 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
4801 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
4861 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
4921 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
4981 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
5041 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
5101 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
5161 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
5221 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
5281 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
5341 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
5401 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
5461 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
5521 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
5581 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
5641 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
5701 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
5761 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
5821 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
5881 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
5941 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
6001 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
6061 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
//
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