| 质粒名称: | pVH21-Tier1-PhEF1a |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 169561 |
| 存储实验室: | Martin Fussenegger |
|
载体骨架基本信息 |
|
| 载体名称: | pMM1 (pcDNA3.1-based) |
| 空载体大小: | 4435 bp |
| 完整质粒大小: | 5793 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 |
|
文献引用方法 |
|
| 材料和方法部分: | pVH21-Tier1-PhEF1a was a gift from Martin Fussenegger (Addgene plasmid # 169561 ; http://n2t.net/addgene:169561 ; RRID:Addgene_169561) |
| 参考文献部分: | A versatile plasmid architecture for mammalian synthetic biology (VAMSyB). Haellman V, Strittmatter T, Bertschi A, Stucheli P, Fussenegger M. Metab Eng. 2021 Jul;66:41-50. doi: 10.1016/j.ymben.2021.04.003. Epub 2021 Apr 18. 10.1016/j.ymben.2021.04.003 PubMed 33857582 |
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 pVH21-Tier1-PhEF1a 5793 bp ds-DNA circular SYN 22-JUN-2022
DEFINITION Tier-1 PhEF1a-driven expression vector (PhEF1a-MCS-pA). .
ACCESSION .
VERSION .
KEYWORDS pVH21-Tier1-PhEF1a
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5793)
TITLE Mammalian Synthetic biology vectors designed to provide a platform
for easy building of gene circuits as well as exchange and
recombination of single modules.
REFERENCE 2 (bases 1 to 5793)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 22, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5793
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 133..1311
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 364..1302
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1259..1279
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
primer_bind complement(1399..1416)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1405..1629
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1675..2103
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1762..1781)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1972..1993
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2112..2132)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2117..2446
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2297..2432
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2359..2378
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2513..3307
/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(2567..2586)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3177..3196
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3481..3602
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3518..3537)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3572..3591
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3651..3667)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3651..3667)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3664..3686)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3675..3691
/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(3699..3729)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3744..3765
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3882..3899)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4053..4641)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4133..4152)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4812..5672)
/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 5435..5454
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5673..5777)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtgccacctg acgtcgacgg atcgggagat ctccacgcgt ggtaccctcg agtgcaaaga
61 tggataaagt tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa
121 ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
181 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
241 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
301 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
361 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc
421 gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt
481 ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga
541 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
601 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
661 ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg
721 gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
781 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
841 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
901 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc
961 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
1021 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
1081 aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg
1141 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg
1201 cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc
1261 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg aggtaccttc
1321 gaagcggaat tcaccatgac tagtgctagc ggatccaccg gtgtctagaa agctttgagg
1381 ccggcctgca gggatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
1441 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
1501 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
1561 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
1621 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
1681 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
1741 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
1801 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
1861 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
1921 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
1981 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
2041 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
2101 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
2161 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
2221 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
2281 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
2341 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
2401 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
2461 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
2521 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
2581 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
2641 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
2701 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
2761 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
2821 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
2881 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
2941 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
3001 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
3061 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
3121 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
3181 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
3241 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
3301 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
3361 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
3421 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
3481 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
3541 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
3601 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
3661 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
3721 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
3781 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
3841 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
3901 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
3961 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
4021 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
4081 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
4141 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
4201 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
4261 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
4321 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
4381 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
4441 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
4501 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
4561 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
4621 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
4681 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
4741 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
4801 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
4861 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
4921 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
4981 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
5041 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
5101 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
5161 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
5221 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
5281 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
5341 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
5401 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
5461 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
5521 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
5581 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
5641 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
5701 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
5761 caaatagggg ttccgcgcac atttccccga aaa
//
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