质粒名称: | pENTR1A-GFP-N2 (FR1) |
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出品公司: | 爱迪基因 |
目录编号: | 19364 |
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: | Invitrogen |
载体抗性: | 卡那霉素 |
空载体大小: | -- |
完整质粒大小: | 2348 bp |
载体修饰: | 人细胞色素c氧化酶亚基VIII(human cytochrome C oxidase subunit VIII) |
载体类型: | 哺乳细胞表达 |
筛选标记: | 新霉素(G418) |
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: | Enhanced Green Fluorescent Protein |
物种来源: | -- |
插入大小: | 719bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII(N端) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | AgeI(未破坏) |
3' 克隆位点: | NotI (未破坏) |
5' 测序引物 | CMV-F |
3' 测序引物 | SV40pA-R |
文献引用方法 |
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材料和方法部分: | pENTR1A-GFP-N2 (FR1) was a gift from Eric Campeau & Paul Kaufman (Addgene plasmid # 19364 ; http://n2t.net/addgene:19364 ; RRID:Addgene_19364) |
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
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pENTR1A-GFP-N2 (FR1) 3071 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Gateway entry vector for C-term eGFP fusion, frame N2. ACCESSION . VERSION . KEYWORDS pENTR1A-GFP-N2 (FR1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3071) 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 3071) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3071 /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" misc_feature 486..538 /label=MCS /note="multiple cloning site" regulatory 550..559 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 556..1275 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(601..622) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(862..881) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1209..1230 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(1300..1399) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1511..1530) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1522..2331 /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(1589..1608) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2421..3009 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2910..2929 /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 aaggaaccaa ttcagtcgac 481 tggatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac 541 cggccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc 601 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag 661 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc 721 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac 781 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag 841 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc 901 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc 961 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc 1021 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc 1081 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg 1141 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag 1201 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac 1261 gagctgtaca agtaaagcgg ccgcactcga gatatctaga cccagctttc ttgtacaaag 1321 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc 1381 aaaataaaat cattatttgc catccagctg cagctctggc ccgtgtctca aaatctctga 1441 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata 1501 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta 1561 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa 1621 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa 1681 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg 1741 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg 1801 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat 1861 tcaggtgaaa atattgttga tgcgctggca gtgtccctgc gccggttgca ttcgattcct 1921 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga 1981 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt 2041 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact 2101 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt 2161 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc 2221 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat 2281 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg 2341 gttaattggt tgtaacatta ttcagattgg gccccgttcc actgagcgtc agacccggta 2401 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 2461 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 2521 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag 2581 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 2641 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 2701 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 2761 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 2821 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 2881 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 2941 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 3001 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 3061 gctcacatgt t //