| 质粒名称: | pLKO.1 shSV40 neo |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 158646 |
| 存储实验室: | Kevin Janes |
|
载体骨架基本信息 |
|
| 载体名称: | pLKO.1 neo |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 7623bp |
| 载体类型: | Lentiviral |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | SV40Tag |
| 物种来源: | Simian virus 40 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pLKO.1 shSV40 neo was a gift from Kevin Janes (Addgene plasmid # 158646 ; http://n2t.net/addgene:158646 ; RRID:Addgene_158646) |
| 参考文献部分: | Modeling the complete kinetics of coxsackievirus B3 reveals human determinants of host-cell feedback. Lopacinski AB, Sweatt AJ, Smolko CM, Gray-Gaillard E, Borgman CA, Shah M, Janes KA. Cell Syst. 2021 Apr 21;12(4):304-323.e13. doi: 10.1016/j.cels.2021.02.004. Epub 2021 Mar 18. 10.1016/j.cels.2021.02.004 PubMed 33740397 |
买家导航
pLKO.1 shSV40 neo载体质粒基本信息
pLKO.1 shSV40 neo质粒图谱载体图谱和pLKO.1 shSV40 neo载体序列质粒序列多克隆位点信息
pLKO.1 shSV40 neo质粒载体简介
pLKO.1 shSV40 neo质粒序列载体序列
LOCUS pLKO.1 shSV40 neo 7623 bp DNA circular SYN 14-APR-2025
DEFINITION Lentiviral shRNA vector targeting simian virus 40 T antigen
(SV40Tag).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7623)
AUTHORS Lopacinski AB, Sweatt AJ, Smolko CM, Gray-Gaillard E, Borgman CA,
Shah M, Janes KA
TITLE Modeling the complete kinetics of coxsackievirus B3 reveals human
determinants of host-cell feedback.
JOURNAL Cell Syst. 2021 Apr 21;12(4):304-323.e13. doi:
10.1016/j.cels.2021.02.004. Epub 2021 Mar 18.
PUBMED 33740397
REFERENCE 2 (bases 1 to 7623)
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"; doi:
"10.1016/j.cels.2021.02.004"; journalName: "Cell Syst"; date:
"2021-04-21- 21"; volume: "12"; issue: "4"; pages: "304-323.e13"
COMMENT Sequence Label: pLKO.1 shSV40 neo
FEATURES Location/Qualifiers
source 1..7623
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 115..363
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 115..135
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 286..305
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_feature 460..577
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 626..1136
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 1055..1073
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 1509..2303
/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(1563..1582)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2173..2192
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(2325..2341)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2326..2342)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 2424..2657
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 2729..2850
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2766..2785)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2820..2839
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 2890..3025
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(3045..3064)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3046..3064)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3074..3091)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3074..3090)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3083..3105)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3232..3687
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3319..3338)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3529..3550
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3713..3817
/gene="bla"
/label=AmpR promoter
CDS join(3818..3886,3887..4678)
/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 complement(4036..4055)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4849..5437
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5338..5357
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5591..5608
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5725..5746
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter join(5761..5766,5767..5784,5785..5791)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5799..5815
/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)."
primer_bind 5804..5826
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5823..5839
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5823..5839
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 5858..5878
/label=T3
/note="T3 promoter, forward primer"
promoter 5860..5878
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 5906..6132
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 6133..6313
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6360..6485
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6978..7211
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7396..7440
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
ORIGIN
1 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
61 ggatctcgac ggtatcgatc acgagactag cctcgagcgg ccgccccctt caccgagggc
121 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt
181 ggaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa
241 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt
301 accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac
361 accgggcata gagtgtctgc tattaactgc agttaatagc agacactcta tgctttttga
421 attctcgacc tcgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
481 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
541 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
601 agatccactt tggccgcggc tcgagggggt tggggttgcg ccttttccaa ggcagccctg
661 ggtttgcgca gggacgcggc tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc
721 ctgggtctcg cacattcttc acgtccgttc gcagcgtcac ccggatcttc gccgctaccc
781 ttgtgggccc cccggcgacg cttcctgctc cgcccctaag tcgggaaggt tccttgcggt
841 tcgcggcgtg ccggacgtga caaacggaag ccgcacgtct cactagtacc ctcgcagacg
901 gacagcgcca gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct
961 gctcagcagg gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg
1021 gggcggtagt gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg
1081 agcgcacgtc ggcagtcggc tccctcgttg accgaatcac cgacctctct ccccaggggg
1141 atccaccgga tcaagcttca cgctgccgca agcactcagg gcgcaagggc tgctaaagga
1201 agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct
1261 actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg
1321 ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc
1381 cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct
1441 tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca ggatgaggat
1501 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
1561 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
1621 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
1681 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
1741 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
1801 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
1861 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
1921 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
1981 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
2041 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
2101 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
2161 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
2221 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
2281 gccttcttga cgagttcttc tgagcgggac tctggggttc gcatcgatac cgtcgacctc
2341 gagggggggc ccggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
2401 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
2461 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
2521 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
2581 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
2641 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
2701 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
2761 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
2821 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
2881 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
2941 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
3001 tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt
3061 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
3121 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
3181 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
3241 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
3301 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
3361 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
3421 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
3481 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
3541 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
3601 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
3661 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
3721 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
3781 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
3841 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
3901 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
3961 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
4021 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
4081 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
4141 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
4201 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
4261 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
4321 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
4381 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
4441 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
4501 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
4561 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
4621 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
4681 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
4741 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
4801 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
4861 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
4921 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
4981 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
5041 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
5101 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
5161 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
5221 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
5281 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
5341 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
5401 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
5461 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
5521 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
5581 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
5641 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
5701 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
5761 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
5821 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
5881 acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac
5941 atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga
6001 ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca
6061 tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct
6121 cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
6181 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
6241 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
6301 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
6361 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
6421 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
6481 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
6541 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
6601 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
6661 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
6721 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
6781 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
6841 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
6901 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
6961 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
7021 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
7081 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
7141 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
7201 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
7261 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
7321 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
7381 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
7441 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
7501 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
7561 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
7621 ccc
//
