| 质粒名称: | mEGFP-Lifeact-7 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 54610 |
| 存储实验室: | Michael Davidson |
|
载体骨架基本信息 |
|
| 载体名称: | mEGFP-Lifeact |
| 载体出品公司: | Clontech |
| 空载体大小: | 4800bp |
| 完整质粒大小: | 4729bp |
| 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: | 无 |
| 标签/融合蛋白: | mEGFP-Lifeact (C terminal on backbone) |
|
文献引用方法 |
|
| 材料和方法部分: | mEGFP-Lifeact-7 was a gift from Michael Davidson (Addgene plasmid # 54610 ; http://n2t.net/addgene:54610 ; RRID:Addgene_54610) |
买家导航
mEGFP-Lifeact-7载体质粒基本信息
mEGFP-Lifeact-7质粒图谱载体图谱和mEGFP-Lifeact-7载体序列质粒序列多克隆位点信息
mEGFP-Lifeact-7质粒载体简介
mEGFP-Lifeact-7质粒序列载体序列
LOCUS mEGFP-Lifeact-7 4729 bp DNA circular SYN 14-APR-2025
DEFINITION Localization: Actin, Excitation: 488, Emission: 507.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4729)
AUTHORS Davidson Lab
TITLE Michael Davidson Empty Backbones
JOURNAL unpublished
REFERENCE 2 (bases 1 to 4729)
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"; journalName:
"unpublished"
COMMENT Sequence Label: mEGFP-Lifeact-7
FEATURES Location/Qualifiers
source 1..4729
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 39..342
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 343..546
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 497..517
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 575..584
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 581..631
/codon_start=1
/product="peptide that binds filamentous actin (Riedl et
al., 2008)"
/label=Lifeact
/note="first 17 amino acids of budding yeast Abp140"
/translation="MGVADLIKKFESISKEE"
regulatory 647..656
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS join(653..655,656..658,659..1372)
/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(698..719)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(959..978)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1306..1327
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1495..1616
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1532..1551)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1586..1605
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1623..2078)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1760..1781)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1972..1991
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2105..2209
/gene="bla"
/label=AmpR promoter
promoter 2211..2568
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2211..2402
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2234..2254)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2419..2554
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2603..3397
/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 2603..3397
/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(2657..2676)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3267..3286
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3585..3604)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3629..3676
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4005..4593
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4494..4513
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
61 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
121 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
181 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
241 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
301 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
361 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
421 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
481 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
541 agagctggtt tagtgaaccg tcagatccgc tagcgccacc atgggcgtgg ccgacttgat
601 caagaagttc gagtccatct ccaaggagga gggggatcca ccggtcgcca ccatggtgag
661 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
721 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
781 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
841 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
901 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
961 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
1021 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
1081 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
1141 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
1201 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
1261 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
1321 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg
1381 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
1441 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
1501 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
1561 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
1621 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
1681 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
1741 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
1801 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
1861 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
1921 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
1981 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
2041 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
2101 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
2161 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
2221 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
2281 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
2341 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
2401 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
2461 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
2521 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
2581 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
2641 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
2701 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
2761 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
2821 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
2881 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
2941 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
3001 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
3061 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
3121 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
3181 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
3241 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
3301 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
3361 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
3421 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
3481 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
3541 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
3601 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
3661 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
3721 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
3781 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
3841 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
3901 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
3961 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
4021 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
4081 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
4141 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
4201 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
4261 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
4321 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
4381 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
4441 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
4501 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
4561 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
4621 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
4681 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatt
//
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