| 质粒名称: | pLenti-puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 39481 |
| 存储实验室: | Ie-Ming Shih |
|
载体骨架基本信息 |
|
| 载体名称: | pLenti4/TO/V5-DEST |
| 载体出品公司: | Invitrogen |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 移除2个EcoRV位点之间的网关元件，并用嘌呤霉素抗性基因替换Zeocin抗性基因 |
| 载体类型: | 哺乳动物表达,慢病毒 |
| 筛选标记: | 嘌呤霉素 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: | V5和6XHis（主干上的C终端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | CMV-fwd |
| 3’测序引物 | V5-rev and BGH-Rev |
|
文献引用方法 |
|
| 材料和方法部分: | pLenti-puro was a gift from Ie-Ming Shih (Addgene plasmid # 39481 ; http://n2t.net/addgene:39481 ; RRID:Addgene_39481) |
| 参考文献部分: | ARID1A, a factor that promotes formation of SWI/SNF-mediated chromatin remodeling, is a tumor suppressor in gynecologic cancers. Guan B, Wang TL, Shih IeM. Cancer Res. 2011 Nov 1;71(21):6718-27. Epub 2011 Sep 7. 10.1158/0008-5472.CAN-11-1562 PubMed 21900401 |
买家导航
pLenti-puro载体 pLentipuro质粒基本信息
pLenti-puro质粒图谱 pLentipuro载体图谱和pLenti-puro载体序列 pLentipuro质粒序列多克隆位点信息
pLenti-puro质粒 pLentipuro载体简介
pLenti-puro质粒序列 pLentipuro载体序列
LOCUS pLenti-puro 7066 bp ds-DNA circular SYN 26-MAY-2021
DEFINITION Tetracycline inducible 3rd generation lentiviral vector.
ACCESSION .
VERSION .
KEYWORDS pLenti-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7066)
AUTHORS Guan B, Wang TL, Shih IeM
TITLE ARID1A, a factor that promotes formation of SWI/SNF-mediated
chromatin remodeling, is a tumor suppressor in gynecologic cancers.
JOURNAL Cancer Res. 2011 Nov 1;71(21):6718-27. Epub 2011 Sep 7.
PUBMED 21900401
REFERENCE 2 (bases 1 to 7066)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 26, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7066
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 94..111
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 228..249
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 264..294
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 302..318
/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 307..329
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 326..342
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 326..342
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 361..381
/label=T3
/note="T3 promoter, forward primer"
promoter 363..381
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 409..635
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 636..816
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 863..988
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1481..1714
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1899..1943
/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"
promoter 2525..2728
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2679..2699
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 2730..2748
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2751..2769
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
CDS 2858..2899
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 2909..2926
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2949..2966)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
primer_bind complement(2978..2998)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2983..3312
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3163..3298
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3322..3921
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3322..3341)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3818..3838
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 3993..4226
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4298..4419
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4335..4354)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4389..4408
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4459..4594
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(4614..4633)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4615..4633)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4643..4660)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4643..4659)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4652..4674)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4801..5256
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4888..4907)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5098..5119
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5282..5386
/gene="bla"
/label=AmpR promoter
CDS 5387..6247
/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(5605..5624)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6418..7006
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6907..6926
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
361 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg
421 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
481 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
541 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
601 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct
661 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
721 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
781 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
841 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
901 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
961 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
1021 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
1081 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
1141 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
1201 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
1261 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
1321 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
1381 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
1441 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
1501 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
1561 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
1621 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
1681 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
1741 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
1801 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
1861 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
1921 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
1981 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
2041 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
2101 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
2161 gagacagatc cattcgatta gtgaacggat ctcgacggta tcgataagct tgggagttcc
2221 gcttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
2281 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
2341 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
2401 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
2461 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
2521 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
2581 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
2641 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
2701 acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc tccctatcag
2761 tgatagagat cgtcgactag tggatccagt gtggtggaat tctgcagata tccagcacag
2821 tggcggccgc tcgagtctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct
2881 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacctg
2941 ctgatcagcc tcgactgtgc cttctagtcg accctgtgga atgtgtgtca gttagggtgt
3001 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
3061 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
3121 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
3181 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
3241 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
3301 ggcttttgca aaaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc
3361 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
3421 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
3481 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
3541 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
3601 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
3661 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
3721 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
3781 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
3841 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
3901 acccgcaagc ccggtgcctg aggtaccttt aagaccaatg acttacaagg cagctgtaga
3961 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
4021 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
4081 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
4141 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
4201 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
4261 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
4321 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
4381 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
4441 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
4501 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
4561 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
4621 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
4681 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
4741 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
4801 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
4861 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
4921 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
4981 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
5041 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
5101 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
5161 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
5221 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
5281 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
5341 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
5401 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
5461 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
5521 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
5581 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
5641 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
5701 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
5761 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
5821 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
5881 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
5941 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
6001 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
6061 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
6121 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
6181 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
6241 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
6301 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
6361 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
6421 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
6481 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
6541 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
6601 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
6661 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
6721 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
6781 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
6841 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
6901 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
6961 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
7021 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatg
//
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pLenti-puro质粒 pLentipuro载体应用举例
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