质粒名称: | pEYK3.1 |
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出品公司: | 爱迪基因 |
目录编号: | 17366 |
存储实验室: | George Daley |
载体骨架基本信息 |
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载体名称: | pEYK3.1 |
载体出品公司: | -- |
载体抗性: | 博来霉素 |
空载体大小: | 3422 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 逆转录病毒 |
筛选标记: | 博来霉素 |
高拷贝/低拷贝: | -- |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | GFP |
物种来源: | -- |
插入大小: | 719 bp |
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 | -- |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pEYK3.1 was a gift from George Daley (Addgene plasmid # 17366 ; http://n2t.net/addgene:17366 ; RRID:Addgene_17366) |
参考文献部分: | Novel retroviral vectors to facilitate expression screens in mammalian cells. Koh EY, Chen T, Daley GQ. Nucleic Acids Res. 2002 Dec 15. 30(24):e142. 10.1093/nar/gnf142 PubMed 12490733 |
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载体基本信息
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载体序列
LOCUS pEYK3.1 4139 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pEYK3.1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4139) AUTHORS Koh EY, Chen T, Daley GQ TITLE Novel retroviral vectors to facilitate expression screens in mammalian cells. JOURNAL Nucleic Acids Res. 2002 Dec 15. 30(24):e142. PUBMED 12490733 REFERENCE 2 (bases 1 to 4139) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4139 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..470 /note="long terminal repeat from Moloney murine leukemia virus" protein_bind complement(579..612) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." rep_origin complement(858..1446) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(938..957) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1479..1853) /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" promoter complement(1872..1919) /label=EM7 promoter /note="synthetic bacterial promoter " CDS complement(2088..2807) /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2133..2154) /label=EGFP-C /note="EGFP, forward primer" primer_bind 2482..2501 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2741..2762 /label=EGFP-N /note="EGFP, reverse primer" regulatory 2804..2813 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" misc_feature 2853..3227 /label=pol region /note="Moloney murine leukemia virus (MMLV) pol region containing the splice acceptor site" primer_bind complement(2899..2915) /label=pBMN 5' /note="MMLV sequence, for inserts in pBMN retroviral vector" CDS complement(3237..3653) /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind complement(3253..3269) /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind complement(3285..3307) /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" misc_feature 3718..4075 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" ORIGIN 1 tgaaagaccc ccgctgacgg gtagtcaatc actcagagga gaccctccca aggaacagcg 61 agaccacaag tcggatgcaa ctgcaagagg gtttattgga tacacgggta cccgggcgac 121 tcagtcaatc ggaggactgg cgccccgagt gaggggttgt gggctctttt attgagctcg 181 gggagcagaa gcgcgcgaac agaagcgaga agcgaactga ttggttagtt caaataaggc 241 acagggtcat ttcaggtcct tggggcaccc tggaaacatc tgatggttct ctagaaactg 301 ctgagggctg gaccgcatct ggggaccatc tgttcttggc cctgagccgg ggcaggaact 361 gcttaccaca gatatcctgt ttggcccata ttcagctgtt ccatctgttc ttggccctga 421 gccggggcag gaactgctta ccacagatat cctgtttggc ccatattcag ctgttccatc 481 tgttcctgac cttgatctga acttctctat tctcagttat gtatttttcc atgccttgca 541 aaatggcgtt acttaagcta gaggcgcgcc ttaattaaat aacttcgtat aatgtatgct 601 atacgaagtt atgcggccgc gctagcttgc caaacctaca ggtggggtct ttcattcccc 661 cctttttctg gagactaaat aaaatctttt attttatcta tggctcgtac tctataggct 721 tcagctggtg atattgttga gtcaaaacta gagcctggac cactgatatc ctgtctttaa 781 caaattggac taatcgatat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 841 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 901 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 961 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 1021 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 1081 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 1141 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 1201 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 1261 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 1321 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 1381 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 1441 tctcaagaag atcctttgat cttttctacg ggacgcgttc agtcctgctc ctcggccacg 1501 aagtgcacgc agttgccggc cgggtcgcgc agggcgaact cccgccccca cggctgctcg 1561 ccgatctcgg tcatggccgg cccggaggcg tcccggaagt tcgtggacac gacctccgac 1621 cactcggcgt acagctcgtc caggccgcgc acccacaccc aggccagggt gttgtccggc 1681 accacctggt cctggaccgc gctgatgaac agagtcacgt cgtcccggac cacaccggcg 1741 aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt cggtccagaa ctcgaccgct 1801 ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg tcaacttggc catggtttag 1861 ttcctcacct tgtcgtatta tactatgccg atatactatg ccgatgatta attgtcaaca 1921 gtcgaccact gtgctggcgg ccggccgctc gagtttaaat agtacataat ggatttcctt 1981 acgcgaaata cgggcagaca tggcctgccc ggttattatt atttttgaca ccagaccaac 2041 tggtaatggt agcgaccggc gctcagctgg aattcttaat taattcatta cttgtacagc 2101 tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg 2161 cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc 2221 agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg 2281 ctgccgtcct cgatgttgtg gcggatcttg aagttcgcct tgatgccgtt cttctgcttg 2341 tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg 2401 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc 2461 tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc 2521 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg 2581 gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc 2641 acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg 2701 ccctcgccct cgccagacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc 2761 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgttaatt 2821 aagaattccc gtaccaccac actgggatcc ggcagtctag aggatggtcc acccccgggg 2881 tcggcagcct tcacgtgggc ggcgtgtatc caagctgcga tgccgtctac tttgagggcg 2941 gtgggggtgg tcagcaggac tgtgtaaggt cctttccagc gaggttctag gttcttagtc 3001 tggtgtcggc ggacccacac tgtgtcgccg actcggtaag ggtgaggtac caccggtcgg 3061 tccagttgtt cttggtaggc tgccgccaga ggtctccaga cttcgtgctg gactaagtag 3121 agagcctgta agtgagcttg gagagagggg ctgttagtaa ctcttgtctt gtcagggtca 3181 gggaagttta caaggggcgg gggtgcccca tataagatct catatggcca tatgggggcg 3241 cctagagaag gagtgagggc tggataaagg gaggatcgag gcggggtcga acgaggaggt 3301 tcaaggggga gagacggggc ggatggagga agaggaggcg gaggcttagg gtgtacaaag 3361 ggcttgaccc agggaggggg gtcaaaagcc aaggcttccc aggtcacgat gtaggggacc 3421 tggtctgggt gtccatgcgg gccaggtgaa aagaccttga tcttaacctg ggtgatgagg 3481 tctcggttaa aggtgccgtc tcgcggccat ccgacgttaa aggttggcca ttctgcagag 3541 cagaaggtaa cccaacgtct cttcttgaca tctaccgact ggttgtgagc gatccgctcg 3601 acatctttcc agtgacctaa ggtcaaactt aagggagtgg taacagtctg gcccgggccc 3661 ttattttcag acaaatacag aaacacagtc agacagagac aacacagaac gatgctgcag 3721 cagacaagac gcgcggcgcg gcttcggtcc caaaccgaaa gcaaaaattc agacggaggc 3781 gggaactgtt ttaggttctc gtctcctacc agaaccacat atccctcctc taaggggggt 3841 gcaccaaaga gtccaaaacg atcgggattt ttggactcag gtcgggccac aaaaacggcc 3901 cccgaagtcc ctgggacgtc tcccagggtt gcggccgggt gttccgaact cgtcagttcc 3961 accacgggtc cgccagatac agagctagtt agctaactag taccgacgca ggcgcataaa 4021 atcagtcata gacactagac aatcggacag acacagataa gttgctggcc agcttacctc 4081 ccggtggtgg gtcggtggtc cctgggcagg ggtctcccga tcccggacga gcccccaaa //
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