| 质粒名称: | pLenti X2 Neo/pTER shLUC (w181-1) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 17483 |
| 存储实验室: | Eric Campeau, Paul Kaufman |
|
载体骨架基本信息 |
|
| 载体名称: | p156RRL-sinPPT-CMV-GFP-PRE/Nhe I |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 7919 bp |
| 完整质粒大小: | 7986 bp |
| 载体类型: | Lentiviral, RNAi |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Firefly Luciferase shRNA |
| 插入大小: | 67 bp |
| 基因又名: | shLUC |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Bgl II(未破坏) |
| 3' 克隆位点: | Hind III(未破坏) |
| 5' 测序引物 | n/a |
|
文献引用方法 |
|
| 材料和方法部分: | pLenti X2 Neo/pTER shLUC (w181-1) was a gift from Eric Campeau & Paul Kaufman (爱迪基因 plasmid # 17483) |
| 参考文献部分: | A versatile viral system for expression and depletion of proteins in mammalian cells. Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD.. PLoS One. 2009 Aug 6;4(8):e6529. 10.1371/journal.pone.0006529 PubMed 19657394 |
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 pLenti X2 Neo/pTER shLUC (w181-1) 7986 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION 3rd gen lentiviral Luciferase shRNA (H1/TO promoter), 3'LTR
insertion, Neo .
ACCESSION .
VERSION .
KEYWORDS pLenti X2 Neo/pTER shLUC (w181-1)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7986)
AUTHORS Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO,
Campisi J, Yaswen P, Cooper PK, Kaufman PD.
TITLE A versatile viral system for expression and depletion of proteins in
mammalian cells.
JOURNAL PLoS One. 2009 Aug 6;4(8):e6529.
PUBMED 19657394
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pLenti X2 Neo/pTER shLUC (w181-1)|
FEATURES Location/Qualifiers
source 1..7986
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 27..131
/gene="bla"
/label=AmpR promoter
CDS 132..992
/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(350..369)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1163..1751
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1652..1671
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1905..1922
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2039..2060
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2075..2105
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2113..2129
/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 2118..2140
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2137..2153
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2137..2153
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 2172..2192
/label=T3
/note="T3 promoter, forward primer"
promoter 2174..2192
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
LTR 2448..2628
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 2672..2797
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 3290..3523
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 4012..4129
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
misc_feature 4183..4771
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4236..4256)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4654..4665)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
promoter 4814..5313
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(4849..4872)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 5246..5265
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 5327..6130
/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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(5390..5409)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6000..6019
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
protein_bind 6264..6288
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind 6421..6439
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind complement(6485..6504)
/label=H1
/note="Human H1 promoter, forward primer"
protein_bind complement(6681..6705)
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
LTR 6780..6960
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 7032..7153
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7069..7088)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7123..7142
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 7172..7307
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 7234..7253
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(7339..7358)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(7340..7358)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(7368..7385)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(7368..7384)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7377..7399)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 7529..7984
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7616..7635)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7826..7847
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
2221 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
2281 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
2341 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
2401 gcattgcaga gatattgtat ttaagtgcct agctcgatac aataaacggg tctctctggt
2461 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
2521 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
2581 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
2641 cagggacctg aaagcgaaag ggaaaccaga gctctctcga cgcaggactc ggcttgctga
2701 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
2761 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
2821 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
2881 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
2941 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
3001 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
3061 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
3121 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
3181 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
3241 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
3301 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcctcaatg acgctgacgg
3361 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
3421 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
3481 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
3541 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
3601 tggaacagat tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
3661 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
3721 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
3781 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
3841 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
3901 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
3961 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggttaa cttttaaaag
4021 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
4081 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta ttccagtgtg
4141 gtggaattct gcagatatcc agcacagtgg cggccgctcg acaatcaacc tctggattac
4201 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
4261 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
4321 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
4381 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
4441 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
4501 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
4561 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
4621 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
4681 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
4741 agtcggatct ccctttgggc cgcctccccg cctggaattc tgcagatatc cggactagtg
4801 atctaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta
4861 gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca
4921 tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact
4981 cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa
5041 atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag
5101 cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag
5161 aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc
5221 gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg
5281 ctgttctcct cttcctcatc tccgggcctt tcgacctgca gccaatatgg gatcggccat
5341 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
5401 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
5461 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
5521 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
5581 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
5641 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
5701 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
5761 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
5821 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
5881 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
5941 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
6001 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
6061 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
6121 gttcttctga ggggatccgt cgagggcccg gtacctttaa gaccaatgac ttacaaggca
6181 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct agactcgagc
6241 ggccgccact gtgctggata tcaaccactt tgtacaagaa agctgggtct agatatctcg
6301 agtgcggccg cgaattccac cacactggac tagtggatcc gagctcggta ccaagctttt
6361 ccaaaaactt acgctgagta cttcgaggac agcacactcg aagtactcag cgtaagggga
6421 tctctatcac tgatagggaa cttataagat tcccaaatcc aaagacattt cacgtttatg
6481 gtgatttccc agaacacata gcgacatgca aatattgcag ggcgccactc ccctgtccct
6541 cacagccatc ttcctgccag ggcgcacgcg cgctgggtgt tcccgcctag tgacactggg
6601 ccgcgattcc ttggagcggg ttgatgacgt cagcgttcga attcggtacc ggatccagtc
6661 gactgaattg gttcctttaa agcctgcttt tttgtacaaa cttgttgata tctgcagaat
6721 tccaccacac tggactagct cactcccaac gaagacaaga tctgcttttt gcttgtactg
6781 ggtctctctg gttagaccag atctgagcct gggagctctc tggctgccta gggaacccac
6841 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
6901 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
6961 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
7021 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
7081 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
7141 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc
7201 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
7261 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcgt
7321 cgagacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
7381 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
7441 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
7501 ttgcgcagcc tgaatggcga atggcgcgac gcgccctgta gcggcgcatt aagcgcggcg
7561 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
7621 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
7681 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
7741 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
7801 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
7861 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
7921 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
7981 atttcc
//
