| 质粒名称: | pRC-CMV-Rev1b |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 164443 |
| 存储实验室: | Alejandro Balazs |
|
载体骨架基本信息 |
|
| 载体名称: | pRC |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 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: | HIV-1 Rev |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pRC-CMV-Rev1b was a gift from Alejandro Balazs (Addgene plasmid # 164443 ; http://n2t.net/addgene:164443 ; RRID:Addgene_164443) |
| 参考文献部分: | COVID-19-neutralizing antibodies predict disease severity and survival. Garcia-Beltran WF, Lam EC, Astudillo MG, Yang D, Miller TE, Feldman J, Hauser BM, Caradonna TM, Clayton KL, Nitido AD, Murali MR, Alter G, Charles RC, Dighe A, Branda JA, Lennerz JK, Lingwood D, Schmidt AG, Iafrate AJ, Balazs AB. Cell. 2021 Jan 21;184(2):476-488.e11. doi: 10.1016/j.cell.2020.12.015. Epub 2020 Dec 15. 10.1016/j.cell.2020.12.015 PubMed 33412089 |
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 pRC-CMV-Rev1b 5897 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Encodes HIV-1 Rev accessory protein for pseudovirus production.
ACCESSION .
VERSION .
KEYWORDS pRC-CMV-Rev1b
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5897)
TITLE COVID-19 neutralizing antibodies predict disease severity and
survival
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5897)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5897
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 760..1110
/codon_start=1
/gene="rev"
/product="human immunodeficiency virus 1 (HIV-1) Rev
protein"
/label=Rev
/note="enhances nuclear export of unspliced and
incompletely spliced viral mRNAs"
/translation="MAGRSGDSDEDLLKAVRLIKFLYQSNPPPNPEGTRQARRNRRRRW
RERQRQIHSISERILSTYLGRSAEPVPLQLPPLERLTLDCNEDCGTSGTQGVGSPQILV
ESPTVLESGTKE"
promoter complement(1122..1140)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1123..1140)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1160..1177)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1166..1390
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1436..1949
/direction=RIGHT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1523..1542)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1733..1754
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1972..1992)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 1977..2306
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2157..2292
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2219..2238
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2373..3167
/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(2427..2446)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3037..3056
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3341..3475
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3378..3397)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3432..3451
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3506..3522)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3506..3522)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3519..3541)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3530..3546
/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(3554..3584)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3599..3620
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3737..3754)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3908..4496)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3988..4007)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4667..5527)
/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 5290..5309
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5528..5632)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5707..5726)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattct gcagatatcc
721 atcacactgg cggccgctcg agcatgcatc tagactgcca tggcaggaag aagcggagac
781 agcgacgaag acctcctcaa ggcagtcaga ctcatcaagt ttctctatca aagcaaccca
841 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
901 agacagagac agatccattc gattagtgaa cggatcctta gcacttatct gggacgatct
961 gcggagcctg tgcctcttca gctaccaccg cttgagagac ttactcttga ttgtaacgag
1021 gattgtggaa cttctgggac gcagggggtg ggaagccctc aaatattggt ggaatctcct
1081 acagtattgg agtcaggaac taaagaatag gatccggcta ttctatagtg tcacctaaat
1141 gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
1201 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
1261 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
1321 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
1381 ggctctatgg cttctgaggc ggaaagaacc agctggggct cgagggggga tccccacgcg
1441 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
1501 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
1561 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
1621 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
1681 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
1741 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
1801 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
1861 aattttaaca aaatattaac gcttacaatt taaatatttg cttatacaat cttcctgttt
1921 ttggggcttt tctgattatc aaccggggtg ggtaccgagc tcgaattctg tggaatgtgt
1981 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
2041 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
2101 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
2161 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
2221 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
2281 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttggatatcc attttcggat
2341 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
2401 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
2461 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
2521 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
2581 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
2641 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
2701 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
2761 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
2821 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
2881 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
2941 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
3001 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
3061 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
3121 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
3181 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
3241 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
3301 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
3361 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
3421 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgtcg
3481 acctcgagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
3541 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
3601 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
3661 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
3721 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
3781 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
3841 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
3901 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
3961 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
4021 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
4081 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
4141 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
4201 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
4261 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
4321 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
4381 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
4441 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
4501 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
4561 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
4621 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
4681 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
4741 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
4801 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
4861 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
4921 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
4981 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
5041 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
5101 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
5161 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
5221 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
5281 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
5341 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
5401 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
5461 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
5521 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
5581 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
5641 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
5701 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
5761 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
5821 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
5881 ggccagatat acgcgtt
//
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