pMIR-Report Luciferase

载体基本信息

载体质粒图谱和多克隆位点信息

载体简介

pMIR-REPORT miRNA表达报告基因载体系统由一个实验萤火虫荧光素酶报告载体和相关的β-半乳糖苷酶报告对照质粒组成。通过在多克隆位点插入预测的miRNA靶序列， pMIR-REPORT荧光素酶报告miRNA表达报告载体可以被用来进行准确的，定量的，评估细胞内miRNA的功能。它也可以被用来检测siRNA的靶位点和分析3'端非编码区序列对基因表达的影响。 pMIR-REPORT Luciferase载体包含一个CMV启动子和终止子控制下的萤火虫荧光素酶报告基因。在荧光素酶基因的3'端非编码区包含一个多克隆位点，用于插入的预测miRNA的结合靶序列或其他核苷酸序列。通过将预测的miRNA靶序列克隆插入到pMIR-REPORT载体，荧光素酶报告表达就收到调节。这模仿的miRNA靶序列的作用方式。

pMIR-REPORT β-gal质粒是一个β-半乳糖苷酶报告质粒，其被设计用于细胞转染操作流程的标准化探索。 该对照质粒表达的β-半乳糖苷酶，可以用来标准化由于细胞活力和转染效率差异导致的细胞表达水平的多样性。

作为附加的功能，pMIR-REPORT Luciferase和pMIR-REPORT β-gal分别含有嘌呤霉素和新霉素抗性基因，这可以用于选择转染成功的哺乳动物细胞。研究需要的话，抗生素选择可用于构建pMIR-REPORT载体质粒稳表达细胞系。对于大多数应用，瞬表达是足够的，抗生素的筛选也是没有必要的。

载体序列

LOCUS pMIR-Report\Luciferase 	6470 bp 	DNA	SYNDEFINITION pMIR-reportACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors.COMMENT This file is created by Vector NTI http://www.biofeng.com/COMMENT VNTAUTHORNAME|biofeng.com|FEATURES Location/Qualifiers source 1..6470 /organism="pMIR-report" /mol_type="other DNA" misc_feature complement(29..51) /label="pGEX_3_primer" misc_feature complement(250..393) /label="lacZ_a" misc_feature 364..386 /label="M13_pUC_fwd_primer" promoter 379..395 /label="M13_forward20_primer" CDS complement(542..2269) /label="ORF frame 2" gene complement(560..2200) /label="firefly_Luciferase (variant)" /gene="firefly_Luciferase (variant)" misc_feature 2116..2134 /label="LucNrev_primer" promoter complement(2227..2791) /label="CMV_immearly_promoter" misc_feature complement(2254..2274) /label="CMV_fwd_primer" misc_feature complement(2440..2727) /label="CAG_enhancer" promoter complement(2854..2872) /label="T7_promoter" misc_feature complement(2885..2902) /label="Sp6_primer" misc_feature complement(2935..2957) /label="M13_pUC_rev_primer" promoter complement(2971..3000) /label="lac_promoter" gene complement(4083..4943) /label="Ampicillin" /gene="Ampicillin" CDS complement(4083..4943) /label="ORF frame 1" promoter complement(4985..5013) /label="AmpR_promoter" CDS 5396..6139 /label="ORF frame 2" CDS complement(5447..6046) /label="ORF frame 2" promoter complement(6191..6459) /label="SV40_promoter" misc_feature complement(6211..6230) /label="SV40pro_F_primer" rep_origin complement(6215..6292) /label="SV40_origin" misc_feature 6262..6453 /label="SV40_enhancer"ORIGIN 1 TCGCGCGTTT CGGTGATGAC GGTGAAAACC TCTGACACAT GCAGCTCCCG GAGACGGTCA 61 CAGCTTGTCT GTAAGCGGAT GCCGGGAGCA GACAAGCCCG TCAGGGCGCG TCAGCGGGTG 121 TTGGCGGGTG TCGGGGCTGG CTTAACTATG CGGCATCAGA GCAGATTGTA CTGAGAGTGC 181 ACCATATGCG GTGTGAAATA CCGCACAGAT GCGTAAGGAG AAAATACCGC ATCAGGCGCC 241 ATTCGCCATT CAGGCTGCGC AACTGTTGGG AAGGGCGATC GGTGCGGGCC TCTTCGCTAT 301 TACGCCAGCT GGCGAAAGGG GGATGTGCTG CAAGGCGATT AAGTTGGGTA ACGCCAGGGT 361 TTTCCCAGTC ACGACGTTGT AAAACGACGG CCAGTGCCAA GCTAGCGGCC GCATACAAAA 421 AACCAACACA CAGATCCAAT GAAAATAAAA GATCCTTTAT TAAGCTTAAC TGTTTAAACC 481 TATGATATCA CGCACGCGTC TATGCCGGCC TATGAGCTCC CTCACTAGTG CGCAGCTTTC 541 ATCACTCGAG CAATTTGGAC TTTCCGCCCT TCTTGGCCTT TATGAGGATC TCTCTGATTT 601 TTCTTGCGTC GAGTTTTCCG GTAAGACCTT TCGGTACTTC GTCCACAAAC ACAACTCCTC 661 CGCGCAACTT TTTCGCGGTT GTTACTTGAC TGGCGACGTA ATCCACGATC TCTTTTTCCG 721 TCATCGTCTT TCCGTGCTCC AAAACAACAA CGGCGGCGGG AAGTTCACCG GCGTCATCGT 781 CGGGAAGACC TGCCACGCCC GCGTCGAAGA TGTTGGGGTG TTGTAACAAT ATCGATTCCA 841 ATTCAGCGGG GGCCACCTGA TATCCTTTGT ATTTAATTAA AGACTTCAAG CGGTCAACTA 901 TGAAGAAGTG TTCGTCTTCG TCCCAGTAAG CTATGTCTCC AGAATGTAGC CATCCATCCT 961 TGTCAATCAA GGCGTTGGTC GCTTCCGGAT TGTTTACATA ACCGGACATA ATCATAGGTC 1021 CTCTGACACA TAATTCGCCT CTCTGATTAA CGCCCAGCGT TTTCCCGGTA TCCAGATCCA 1081 CAACCTTCGC TTCAAAAAAT GGAACAACTT TACCGACCGC GCCCGGTTTA TCATCCCCCT 1141 CGGGTGTAAT CAGAATAGCT GATGTAGTCT CAGTGAGCCC ATATCCTTGT CGTATCCCTG 1201 GAAGATGGAA GCGTTTTGCA ACCGCTTCCC CGACTTCTTT CGAAAGAGGT GCGCCCCCAG 1261 AAGCAATTTC GTGTAAATTA GATAAATCGT ATTTGTCAAT CAGAGTGCTT TTGGCGAAGA 1321 ATGAAAATAG GGTTGGTACT AGCAACGCAC TTTGAATTTT GTAATCCTGA AGGGATCGTA 1381 AAAACAGCTC TTCTTCAAAT CTATACATTA AGACGACTCG AAATCCACAT ATCAAATATC 1441 CGAGTGTAGT AAACATTCCA AAACCGTGAT GGAATGGGAC AACACTTAAA ATCGCAGTAT 1501 CCGGAACGAT TTGATTGCCA AAAATAGGAT CTCTGGCATG CGAGAATCTG ACGCAGGCAG 1561 TTCTATGCGG AAGGGCCACA CCCTTAGGTA ACCCAGTAGA TCCAGAGGAA TTCATTATCA 1621 GTGCAATTGT TTTGTCACGA TCAAAGGACT CTGGTACAAA ATCGTATTCA TTAAAACCGG 1681 GAGGTAGATG AGATGTGACG AACGTGTACA TCGACTGAAA TCCCTGGTAA TCCGTTTTAG 1741 AATCCATGAT AATAATTTTC TGGATTATTG GTAATTTTTT TTGCACGTTC AAAATTTTTT 1801 GCAACCCCTT TTTGGAAACA AACACTACGG TAGGCTGCGA AATGTTCATA CTGTTGAGCA 1861 ATTCACGTTC ATTATAAATG TCGTTCGCGG GCGCAACTGC AACTCCGATA AATAACGCGC 1921 CCAACACCGG CATAAAGAAT TGAAGAGAGT TTTCACTGCA TACGACGATT CTGTGATTTG 1981 TATTCAGCCC ATATCGTTTC ATAGCTTCTG CCAACCGAAC GGACATTTCG AAGTATTCCG 2041 CGTACGTGAT GTTCACCTCG ATATGTGCAT CTGTAAAAGC AATTGTTCCA GGAACCAGGG 2101 CGTATCTCTT CATAGCCTTA TGCAGTTGCT CTCCAGCGGT TCCATCCTCT AGAGGATAGA 2161 ATGGCGCCGG GCCTTTCTTT ATGTTTTTGG CGTCTTCCAT GGTGGATCCA CGGTTCACTA 2221 AACCAGCTCT GCTTATATAG ACCTCCCACC GTACACGCCT ACCGCCCATT TGCGTCAATG 2281 GGGCGGAGTT GTTACGACAT TTTGGAAAGT CCCGTTGATT TTGGTGCCAA AACAAACTCC 2341 CATTGACGTC AATGGGGTGG AGACTTGGAA ATCCCCGTGA GTCAAACCGC TATCCACGCC 2401 CATTGATGTA CTGCCAAAAC CGCATCACCA TGGTAATAGC GATGACTAAT ACGTAGATGT 2461 ACTGCCAAGT AGGAAAGTCC CATAAGGTCA TGTACTGGGC ATAATGCCAG GCGGGCCATT 2521 TACCGTCATT GACGTCAATA GGGGGCGTAC TTGGCATATG ATACACTTGA TGTACTGCCA 2581 AGTGGGCAGT TTACCGTAAA TACTCCACCC ATTGACGTCA ATGGAAAGTC CCTATTGGCG 2641 TTACTATGGG AACATACGTC ATTATTGACG TCAATGGGCG GGGGTCGTTG GGCGGTCAGC 2701 CAGGCGGGCC ATTTACCGTA AGTTATGTAA CGCGGAACTC CATATATGGG CTATGAACTA 2761 ATGACCCCGT AATTGATTAC TATTAATAAC TAAGATCTGG TACCTTGAAT TCATGCTTCT 2821 CCTCCCTTTA GTGAGGGTAA TTCTCTCTCT CTCCCTATAG TGAGTCGTAT TAATTCCTTC 2881 TCTTCTATAG TGTCACCTAA ATCGTTGCAA TTCGTAATCA TGTCATAGCT GTTTCCTGTG 2941 TGAAATTGTT ATCCGCTCAC AATTCCACAC AACATACGAG CCGGAAGCAT AAAGTGTAAA 3001 GCCTGGGGTG CCTAATGAGT GAGCTAACTC ACATTAATTG CGTTGCGCTC ACTGCCCGCT 3061 TTCCAGTCGG GAAACCTGTC GTGCCAGCTG CATTAATGAA TCGGCCAACG CGCGGGGAGA 3121 GGCGGTTTGC GTATTGGGCG CTCTTCCGCT TCCTCGCTCA CTGACTCGCT GCGCTCGGTC 3181 GTTCGGCTGC GGCGAGCGGT ATCAGCTCAC TCAAAGGCGG TAATACGGTT ATCCACAGAA 3241 TCAGGGGATA ACGCAGGAAA GAACATGTGA GCAAAAGGCC AGCAAAAGGC CAGGAACCGT 3301 AAAAAGGCCG CGTTGCTGGC GTTTTTCCAT AGGCTCCGCC CCCCTGACGA GCATCACAAA 3361 AATCGACGCT CAAGTCAGAG GTGGCGAAAC CCGACAGGAC TATAAAGATA CCAGGCGTTT 3421 CCCCCTGGAA GCTCCCTCGT GCGCTCTCCT GTTCCGACCC TGCCGCTTAC CGGATACCTG 3481 TCCGCCTTTC TCCCTTCGGG AAGCGTGGCG CTTTCTCATA GCTCACGCTG TAGGTATCTC 3541 AGTTCGGTGT AGGTCGTTCG CTCCAAGCTG GGCTGTGTGC ACGAACCCCC CGTTCAGCCC 3601 GACCGCTGCG CCTTATCCGG TAACTATCGT CTTGAGTCCA ACCCGGTAAG ACACGACTTA 3661 TCGCCACTGG CAGCAGCCAC TGGTAACAGG ATTAGCAGAG CGAGGTATGT AGGCGGTGCT 3721 ACAGAGTTCT TGAAGTGGTG GCCTAACTAC GGCTACACTA GAAGAACAGT ATTTGGTATC 3781 TGCGCTCTGC TGAAGCCAGT TACCTTCGGA AAAAGAGTTG GTAGCTCTTG ATCCGGCAAA 3841 AAAACCACCG CTGGTAGCGG TGGTTTTTTT GTTTGCAAGC AGCAGATTAC GCGCAGAAAA 3901 AAAGGATCTC AAGAAGATCC TTTGATCTTT TCTACGGGGT CTGACGCTCA GTGGAACGAA 3961 AACTCACGTT AAGGGATTTT GGTCATGAGA TTATCAAAAA GGATCTTCAC CTAGATCCTT 4021 TTAAATTAAA AATGAAGTTT TAAATCAATC TAAAGTATAT ATGAGTAAAC TTGGTCTGAC 4081 AGTTACCAAT GCTTAATCAG TGAGGCACCT ATCTCAGCGA TCTGTCTATT TCGTTCATCC 4141 ATAGTTGCCT GACTCCCCGT CGTGTAGATA ACTACGATAC GGGAGGGCTT ACCATCTGGC 4201 CCCAGTGCTG CAATGATACC GCGAGACCCA CGCTCACCGG CTCCAGATTT ATCAGCAATA 4261 AACCAGCCAG CCGGAAGGGC CGAGCGCAGA AGTGGTCCTG CAACTTTATC CGCCTCCATC 4321 CAGTCTATTA ATTGTTGCCG GGAAGCTAGA GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC 4381 AACGTTGTTG CCATTGCTAC AGGCATCGTG GTGTCACGCT CGTCGTTTGG TATGGCTTCA 4441 TTCAGCTCCG GTTCCCAACG ATCAAGGCGA GTTACATGAT CCCCCATGTT GTGCAAAAAA 4501 GCGGTTAGCT CCTTCGGTCC TCCGATCGTT GTCAGAAGTA AGTTGGCCGC AGTGTTATCA 4561 CTCATGGTTA TGGCAGCACT GCATAATTCT CTTACTGTCA TGCCATCCGT AAGATGCTTT 4621 TCTGTGACTG GTGAGTACTC AACCAAGTCA TTCTGAGAAT AGTGTATGCG GCGACCGAGT 4681 TGCTCTTGCC CGGCGTCAAT ACGGGATAAT ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG 4741 CTCATCATTG GAAAACGTTC TTCGGGGCGA AAACTCTCAA GGATCTTACC GCTGTTGAGA 4801 TCCAGTTCGA TGTAACCCAC TCGTGCACCC AACTGATCTT CAGCATCTTT TACTTTCACC 4861 AGCGTTTCTG GGTGAGCAAA AACAGGAAGG CAAAATGCCG CAAAAAAGGG AATAAGGGCG 4921 ACACGGAAAT GTTGAATACT CATACTCTTC CTTTTTCAAT ATTATTGAAG CATTTATCAG 4981 GGTTATTGTC TCATGAGCGG ATACATATTT GAATGTATTT AGAAAAATAA ACAAATAGGG 5041 GTTCCGCGCA CATTTCCCCG AAAAGTGCCA CCTGACGTCT AAGAAACCAT TATTATCATG 5101 ACATTAACCT ATAAAAATAG GCGTATCACG AGATTGCAGT GAAAAAAATG CTTTATTTGT 5161 GAAATTTGTG ATGCTATTGC TTTATTTGTA ACCATTATAA GCTGCAATAA ACAAGTTAAC 5221 AACAACAATT GCATTCATTT TATGTTTCAG GTTCAGGGGG AGGTGTGGGA GGTTTTTTAA 5281 AGCAAGTAAA ACCTCTACAA ATGTGGTATG GCTGATTATG ATCCTCTAGA GTCGGTGGGC 5341 CTCGGGGGCG GGTGCGGGGT CGGCGGGGCC GCCCCGGGTG GCTTCGGTCG GAGCCATGGG 5401 GTCGTGCGCT CCTTTCGGTC GGGCGCTGCG GGTCGTGGGG CGGGCGTCAG GCACCGGGCT 5461 TGCGGGTCAT GCACCAGGTG CGCGGTCCTT CGGGCACCTC GACGTCGGCG GTGACGGTGA 5521 AGCCGAGCCG CTCGTAGAAG GGGAGGTTGC GGGGCGCGGA GGTCTCCAGG AAGGCGGGCA 5581 CCCCGGCGCG CTCGGCCGCC TCCACTCCGG GGAGCACGAC GGCGCTGCCC AGACCCTTGC 5641 CCTGGTGGTC GGGCGAGACG CCGACGGTGG CCAGGAACCA CGCGGGCTCC TTGGGCCGGT 5701 GCGGCGCCAG GAGGCCTTCC ATCTGTTGCT GCGCGGCCAG CCGGGAACCG CTCAACTCGG 5761 CCATGCGCGG GCCGATCTCG GCGAACACCG CCCCCGCTTC GACGCTCTCC GGCGTGGTCC 5821 AGACCGCCAC CGCGGCGCCG TCGTCCGCGA CCCACACCTT GCCGATGTCG AGCCCGACGC 5881 GCGTGAGGAA GAGTTCTTGC AGCTCGGTGA CCCGCTCGAT GTGGCGGTCC GGGTCGACGG 5941 TGTGGCGCGT GGCGGGGTAG TCGGCGAACG CGGCGGCGAG GGTGCGTACG GCCCGGGGGA 6001 CGTCGTCGCG GGTGGCGAGG CGCACCGTGG GCTTGTACTC GGTCATGGAA GGTCGTCTCC 6061 TTGTGAGGGG TCAGGGGCGT GGGTCAGGGG ATGGTGGCGG CACCGGTCGT GGCGGCCGAC 6121 CTGCAGGCAT GCAAGCTAGC TTTTTGCAAA AGCCTAGGCC TCCAAAAAAG CCTCCTCACT 6181 ACTTCTGGAA TAGCTCAGAG GCCGAGGCGG CCTCGGCCTC TGCATAAATA AAAAAAATTA 6241 GTCAGCCATG GGGCGGAGAA TGGGCGGAAC TGGGCGGAGT TAGGGGCGGG ATGGGCGGAG 6301 TTAGGGGCGG GACTATGGTT GCTGACTAAT TGAGATGCAT GCTTTGCATA CTTCTGCCTG 6361 CTGGGGAGCC TGGGGACTTT CCACACCTGG TTGCTGACTA ATTGAGATGC ATGCTTTGCA 6421 TACTTCTGCC TGCTGGGGAG CCTGGGGACT TTCCACACCA ATCTTTCGTC