质粒名称: | pLenti-puro |
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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 |
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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 |
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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 |
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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
pLentipuro载体基本信息
pLentipuro载体图谱质粒图谱和多克隆位点信息
pLentipuro载体简介
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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