- 商品货号:FH1036
- 规格:10μl质粒
- 启动子:mouse U6 promoter
- 复制子:pUC ori,F1 ori,SV40 ori
- 终止子:WPRE
- 质粒分类:RNA文库质粒;RNA人源质粒;miRNA人源质粒
- 质粒大小:8994bp
- 原核抗性:Amp(100μg/ml)
- 筛选标记:EGFP,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:mU6-F(CAGCACAAAAGGAAACTCACC)
- 3测序引物:SV40-R(GGACTTTCCACACCTGGTTG)
- 质粒简介: pLL3.7-Hsa-Let-7a-1miRNA质粒是在pLL3.7载体上克隆了一段非编码核苷酸序列得到的一个哺乳细胞慢病毒miRNA(MicroRNA)表达质粒。mU6启动Hsa-Let-7a-1miRNA的转录。该质粒可作为普通过表达质粒直接转染哺乳细胞,其是第三代慢病毒载体,所以也可包装成慢病毒侵染细胞。质粒本身同时带有EGFP荧光标签和Puro抗性标记,可利用EGFP来检测转染效率,或可用流式细胞仪和嘌呤霉素筛选转染后的细胞得到阳性克隆。
- 质粒图谱1: png/1-1FF91643462H.png
质粒序列: 点击查看序列LOCUS Exported 8994 bp ds-DNA circular SYN 09-七月-2017DEFINITION .ACCESSION .VERSION .KEYWORDS pLL3.7-Hsa-Let-7a-1miRNASOURCE synthetic DNA constructORGANISM synthetic DNA constructREFERENCE 1 (bases 1 to 8994)AUTHORS XCTITLE Direct SubmissionJOURNAL Exported 2017年7月9日 from MiaoLingPlasmidhttp://www.snapgene.comFEATURES Location/Qualifierssource 1..8994/organism="synthetic DNA construct"/mol_type="other DNA"enhancer 238..617/note="CMV enhancer"/note="human cytomegalovirus immediate early enhancer"promoter 619..817/note="CMV promoter"/note="human cytomegalovirus (CMV) immediate earlypromoter"LTR 835..1015/note="5' LTR (truncated)"/note="truncated 5' long terminal repeat (LTR) from HIV-1"misc_feature 1062..1187/note="HIV-1 Psi"/note="packaging signal of human immunodeficiency virustype 1"misc_feature 1686..1919/note="RRE"/note="The Rev response element (RRE) of HIV-1 allows forRev-dependent mRNA export from the nucleus to thecytoplasm."misc_feature 2446..2563/note="cPPT/CTS"/note="central polypurine tract and central terminationsequence of HIV-1"promoter 2622..2933/note="mU6 promoter"/note="mouse U6 promoter"misc_feature 2949..2955/note="Xho-SalI拼接点"misc_feature 2956..3304/note="pri-hsa-miR-let-7a-1"misc_feature 3103..3182/note="pre-hsa-let-7a-1"misc_feature 3103..3182/note="pre-hsa-let-7a-1"promoter 3343..3672/note="SV40 promoter"/note="SV40 enhancer and early promoter"rep_origin 3523..3658/note="SV40 ori"/note="SV40 origin of replication"CDS 3688..4287/codon_start=1/gene="pac from Streptomyces alboniger"/product="puromycin N-acetyltransferase"/note="PuroR"/note="confers resistance to puromycin"/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"primer_bind 4296..4312/note="KS primer"/note="common sequencing primer, one of multiple similarvariants"protein_bind 4354..4387/bound_moiety="Cre recombinase"/note="loxP"/note="Cre-mediated recombination occurs in the 8-bp coresequence (GCATACAT)."enhancer 4503..4806/note="CMV enhancer"/note="human cytomegalovirus immediate early enhancer"promoter 4807..5010/note="CMV promoter"/note="human cytomegalovirus (CMV) immediate earlypromoter"CDS 5055..5774/codon_start=1/product="enhanced GFP"/note="EGFP"/note="mammalian codon-optimized"/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"protein_bind 5794..5827/bound_moiety="Cre recombinase"/note="loxP"/note="Cre-mediated recombination occurs in the 8-bp coresequence (GCATACAT)."misc_feature 5883..6471/note="WPRE"/note="woodchuck hepatitis virus posttranscriptionalregulatory element"CDS complement(6354..6365)/codon_start=1/product="Factor Xa recognition and cleavage site"/note="Factor Xa site"/translation="IEGR"primer_bind complement(6474..6490)/note="KS primer"/note="common sequencing primer, one of multiple similarvariants"LTR 7000..7180/note="5' LTR (truncated)"/note="truncated 5' long terminal repeat (LTR) from HIV-1"rep_origin complement(7242..7830)/direction=LEFT/note="ori"/note="high-copy-number ColE1/pMB1/pBR322/pUC origin ofreplication"CDS complement(8001..8861)/codon_start=1/gene="bla"/product="beta-lactamase"/note="AmpR"/note="confers resistance to ampicillin, carbenicillin, andrelated antibiotics"/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"promoter complement(8862..8966)/gene="bla"/note="AmpR promoter"ORIGIN1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa服务时间:24小时在线客服
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