- 商品货号:FH1309
- 规格:10μl质粒
- 启动子:U6 promoter
- 复制子:pUC ori,F1 ori,SV40 ori
- 终止子:cPPT/CTS , 3
- 质粒分类:RNA文库质粒;RNA人源质粒;shRNA人源质粒
- 质粒大小:7084bp
- 原核抗性:Amp(100μg/ml)
- 筛选标记:Puro
- 克隆菌株:Stbl3
- 表达宿主:293T等哺乳细胞
- 诱导方式:无需诱导
- 5测序引物:U6-F(ATGGACTATCATATGCTTACCGTA)
- 3测序引物:pLKO-R(CTGCACTGTACCCCCCAATC)
- 备注:YAP1序列:NM_001130145 YAP1shRNA序列:CCGGGCCACCAAGCTAGATAAAGAACTCGAG TTCTTTATCTAGCTTGGTGGCTTTTT
- 质粒简介: pLKO.1-YAP1shRNA质粒是在pLKO.1-Puro载体上克隆了一段shRNA序列得到的一个哺乳细胞慢病毒RNA干扰质粒。U6启动YAP1shRNA的转录,可特异性的干扰YAP1蛋白的表达。该质粒可作为普通过表达质粒直接转染哺乳细胞,也可包装成慢病毒侵染细胞。质粒本身带有Puro筛选标记,可用嘌呤霉素筛选转染后的细胞得到阳性克隆。
- 质粒图谱1: png/1-1FF914541D46.png
质粒序列: 点击查看序列LOCUS Exported 7084 bp ds-DNA circular SYN 09-七月-2017DEFINITION .ACCESSION .VERSION .KEYWORDS pLKO.1-YAP1shRNASOURCE synthetic DNA constructORGANISM synthetic DNA constructREFERENCE 1 (bases 1 to 7084)AUTHORS MiaoLingBioTITLE Direct SubmissionJOURNAL Exported 2017年7月9日 from MiaoLingPlasmidhttp://www.snapgene.comFEATURES Location/Qualifierssource 1..7084/organism="synthetic DNA construct"/mol_type="other DNA"CDS 529..1128/codon_start=1/gene="pac from Streptomyces alboniger"/product="puromycin N-acetyltransferase"/note="Puro"/note="confers resistance to puromycin"/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"LTR 1256..1489/note="3' LTR (Delta-U3)"/note="self-inactivating 3' long terminal repeat (LTR) fromHIV-1"polyA_signal 1561..1682/note="SV40 poly(A) signal"/note="SV40 polyadenylation signal"rep_origin 1722..1857/note="SV40 ori"/note="SV40 origin of replication"promoter complement(1878..1896)/note="T7 promoter"/note="promoter for bacteriophage T7 RNA polymerase"primer_bind complement(1906..1922)/note="M13 fwd"/note="common sequencing primer, one of multiple similarvariants"rep_origin 2064..2519/direction=RIGHT/note="f1 ori"/note="f1 bacteriophage origin of replication; arrowindicates direction of (+) strand synthesis"promoter 2545..2649/gene="bla"/note="AmpR promoter"CDS 2650..3510/codon_start=1/gene="bla"/product="beta-lactamase"/note="AmpR"/note="confers resistance to ampicillin, carbenicillin, andrelated antibiotics"/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"rep_origin 3681..4269/direction=RIGHT/note="ori"/note="high-copy-number ColE1/pMB1/pBR322/pUC origin ofreplication"protein_bind 4557..4578/bound_moiety="E. coli catabolite activator protein"/note="CAP binding site"/note="CAP binding activates transcription in the presenceof cAMP."promoter 4593..4623/note="lac promoter"/note="promoter for the E. coli lac operon"protein_bind 4631..4647/bound_moiety="lac repressor encoded by lacI"/note="lac operator"/note="The lac repressor binds to the lac operator toinhibit transcription in E. coli. This inhibition can berelieved by adding lactose orisopropyl-beta-D-thiogalactopyranoside (IPTG)."primer_bind 4655..4671/note="M13 rev"/note="common sequencing primer, one of multiple similarvariants"promoter 4692..4710/note="T3 promoter"/note="promoter for bacteriophage T3 RNA polymerase"promoter 4736..4964/note="RSV promoter"/note="Rous sarcoma virus enhancer/promoter"LTR 4965..5145/note="5' LTR (truncated)"/note="truncated 5' long terminal repeat (LTR) from HIV-1"misc_feature 5192..5317/note="HIV-1 Psi"/note="packaging signal of human immunodeficiency virustype 1"misc_feature 5810..6043/note="RRE"/note="The Rev response element (RRE) of HIV-1 allows forRev-dependent mRNA export from the nucleus to thecytoplasm."promoter 6570..6810/label="U6 Promoter"/note="U6 promoter"/note="RNA polymerase III promoter for human U6 snRNA"misc_feature 6817..6873/note="YAP1shRNA"/note="NM_001130145"misc_feature 6915..7032/note="cPPT/CTS"/note="central polypurine tract and central terminationsequence of HIV-1"promoter 7081..507/note="hPGK promoter"/note="human phosphoglycerate kinase 1 promoter"ORIGIN1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt481 gaccgaatca ccgacctctc tccccagggg gatccaccgg agcttaccat gaccgagtac541 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc601 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag661 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg721 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg781 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc841 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc901 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg961 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg1021 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg1081 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac1141 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcggtacc tttaagacca1201 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa1261 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg1321 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct1381 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt1441 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca1501 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg1561 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca1621 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct1681 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc1741 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg1801 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag1861 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt1921 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc&nbs
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