质粒名称: | pENTR/pSM2(CMV/TO) (w55-1) |
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出品公司: | Addgene |
目录编号: | 17388 |
存储实验室: | Eric Campeau, Paul Kaufman |
载体骨架基本信息 |
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载体名称: | pENTR1A |
载体出品公司: | -- |
载体抗性: | 卡那霉素 |
空载体大小: | 3088 dp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | RNAi ; Entry vector |
筛选标记: | -- |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
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 | -- |
标签/融合蛋白: | -- |
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 | ENTRforw |
3' 测序引物 | ENTRrev |
文献引用方法 |
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材料和方法部分: | pENTR/pSM2(CMV/TO) (w55-1) was a gift from Eric Campeau & Paul Kaufman (Addgene plasmid # 17388 ; http://n2t.net/addgene:17388 ; RRID:Addgene_17388) |
参考文献部分: | A versatile viral system for expression and depletion of proteins in mammalian cells. Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD.. PLoS One. 2009 Aug 6;4(8):e6529. 10.1371/journal.pone.0006529 PubMed 19657394 |
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载体基本信息
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LOCUS pENTR/pSM2(CMV/TO) (w55-1) 3088 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Gateway entry vector for miRNA expression under CMV/TO promoter. ACCESSION . VERSION . KEYWORDS pENTR/pSM2(CMV/TO) (w55-1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3088) AUTHORS Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD. TITLE A versatile viral system for expression and depletion of proteins in mammalian cells. JOURNAL PLoS One. 2009 Aug 6;4(8):e6529. PUBMED 19657394 REFERENCE 2 (bases 1 to 3088) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3088 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 103..189 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 281..308 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind 316..339 /label=pENTR-F /note="pENTR vectors, forward primer" protein_bind 358..457 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" enhancer 480..783 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 784..987 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 938..958 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 989..1007 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 1010..1028 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" ncRNA 1066..1159 /label=5' miR-30a /note="sequence upstream of the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" ncRNA 1173..1285 /label=3' miR-30a /note="sequence downstream of the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" protein_bind complement(1317..1416) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1528..1547) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1539..2348 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1606..1625) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2438..3026 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2927..2946 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg 61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct 121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt 181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg 241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat 301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa 361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc 421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaacctg ggagttccgc 481 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg 541 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa 601 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca 661 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac 721 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc 781 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga 841 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg 901 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta 961 cggtgggagg tctatataag cagagctctc cctatcagtg atagagatct ccctatcagt 1021 gatagagatc gtcgactagt cgactaggga taacagggta attgtttgaa tgaggcttca 1081 gtactttaca gaatcgttgc ctgcacatct tggaaacact tgctgggatt acttcttcag 1141 gttaacccaa cagaaggctc gagcaaccag aattcaaggg gctactttag gagcaattat 1201 cttgtttact aaaactgaat accttgctat ctctttgata catttttaca aagctgaatt 1261 aaaatggtat aaattaaatc acttttttca attggaagac taatgcgttt atctagaccc 1321 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga 1381 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgcag ctctggcccg 1441 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa 1501 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa 1561 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc 1621 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc 1681 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg 1741 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta 1801 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat 1861 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg tccctgcgcc 1921 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg 1981 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc 2041 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac 2101 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga 2161 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg 2221 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa 2281 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt 2341 ttttctaatc agaattggtt aattggttgt aacattattc agattgggcc ccgttccact 2401 gagcgtcaga cccggtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 2461 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 2521 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 2581 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 2641 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 2701 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 2761 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 2821 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 2881 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 2941 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 3001 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 3061 ccttttgctg gccttttgct cacatgtt //