质粒名称: | pEGFP hsp70 |
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出品公司: | 爱迪基因 |
目录编号: | 15215 |
存储实验室: | Lois Greene |
载体骨架基本信息 |
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载体名称: | pEGFP-C3 |
载体出品公司: | Clontech |
载体抗性: | 卡那霉素 |
空载体大小: | 4727 bp |
完整质粒大小: | -- |
载体修饰: | -- |
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: | hsp70 |
物种来源: | H. sapiens (human) |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | M11717 |
启动子 | -- |
标签/融合蛋白: | EGFP(主干上的N端) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | HindIII(未破坏) |
3' 克隆位点: | BamHI (未破坏) |
5' 测序引物 | EGFP-C primer |
3' 测序引物 | SV40pA-R |
文献引用方法 |
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材料和方法部分: | pEGFP hsp70 was a gift from Lois Greene (Addgene plasmid # 15215 ; http://n2t.net/addgene:15215 ; RRID:Addgene_15215) |
参考文献部分: | Hsp70 dynamics in vivo: effect of heat shock and protein aggregation. Zeng XC, Bhasin S, Wu X, Lee JG, Maffi S, Nichols CJ, Lee KJ, Taylor JP, Greene LE, Eisenberg E. J Cell Sci. 2004 Oct 1. 117(Pt 21):4991-5000. 10.1242/jcs.01373 PubMed 15367583 |
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pEGFPhsp70载体基本信息
pEGFPhsp70载体图谱质粒图谱和多克隆位点信息
pEGFPhsp70载体简介
pEGFPhsp70载体序列
LOCUS pEGFP hsp70 6888 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pEGFP hsp70 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6888) AUTHORS Zeng XC, Bhasin S, Wu X, Lee JG, Maffi S, Nichols CJ, Lee KJ, Taylor JP, Greene LE, Eisenberg E TITLE Hsp70 dynamics in vivo: effect of heat shock and protein aggregation. JOURNAL J Cell Sci. 2004 Oct 1. 117(Pt 21):4991-5000. PUBMED 15367583 REFERENCE 2 (bases 1 to 6888) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6888 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 54..357 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 358..561 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 512..532 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 600..609 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 606..1322 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(651..672) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(912..931) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1259..1280 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3669..3790 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3706..3725) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3760..3779 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3797..4252) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3934..3955) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4146..4165 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4279..4383 /gene="bla" /label=AmpR promoter promoter 4385..4742 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4408..4428) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4593..4728 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4655..4674 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4777..5571 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4831..4850) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5441..5460 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5759..5778) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5803..5850 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6179..6767 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6668..6687 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca 61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg 601 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc 661 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca 721 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc 781 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca 841 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca 901 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca 961 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg 1021 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga 1081 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc 1141 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca 1201 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca 1261 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca 1321 agtactcaga tctcgagctc aagcttatgg ccaaagccgc ggcgatcggc atcgacctgg 1381 gcaccaccta ctcctgcgtg ggggtgttcc aacacggcaa ggtggagatc atcgccaacg 1441 accagggcaa ccgcaccacc cccagctacg tggccttcac ggacaccgag cggctcatcg 1501 gggatgcggc caagaaccag gtggcgctga acccgcagaa caccgtgttt gacgcgaagc 1561 ggctgatcgg ccgcaagttc ggcgacccgg tggtgcagtc ggacatgaag cactggcctt 1621 tccaggtgat caacgacgga gacaagccca aggtgcaggt gagctacaag ggggagacca 1681 aggcattcta ccccgaggag atctcgtcca tggtgctgac caagatgaag gagatcgccg 1741 aggcgtacct gggctacccg gtgaccaacg cggtgatcac cgtgccggcc tacttcaacg 1801 actcgcagcg ccaggccacc aaggatgcgg gtgtgatcgc ggggctcaac gtgctgcgga 1861 tcatcaacga gcccacggcc gccgccatcg cctacggcct ggacagaacg ggcaaggggg 1921 agcgcaacgt gctcatcttt gacctgggcg ggggcacctt cgacgtgtcc atcctgacga 1981 tcgacgacgg catcttcgag gtgaaggcca cggccgggga cacccacctg ggtggggagg 2041 actttgacaa caggctggtg aaccacttcg tggaggagtt caagagaaaa cacaagaagg 2101 acatcagcca gaacaagcga gccgtgaggc ggctgcgcac cgcctgcgag agggccaaga 2161 ggaccctgtc gtccagcacc caggccagcc tggagatcga ctccctgttt gagggcatcg 2221 acttctacac gtccatcacc agggcgaggt tcgaggagct gtgctccgac ctgttccgaa 2281 gcaccctgga gcccgtggag aaggctctgc gcgacgccaa gctggacaag gcccagattc 2341 acgacctggt cctggtcggg ggctccaccc gcatccccaa ggtgcagaag ctgctgcagg 2401 acttcttcaa cgggcgcgac ctgaacaaga gcatcaaccc cgacgaggct gtggcctacg 2461 gggcggcggt gcaggcggcc atcctgatgg gggacaagtc cgagaacgtg caggacctgc 2521 tgctgctgga cgtggctccc ctgtcgctgg ggctggagac ggccggaggc gtgatgactg 2581 ccctgatcaa gcgcaactcc accatcccca ccaagcagac gcagatcttc accacctact 2641 ccgacaacca acccggggtg ctgatccagg tgtacgaggg cgagagggcc atgacgaaag 2701 acaacaatct gttggggcgc ttcgagctga gcggcatccc tccggccccc aggggcgtgc 2761 cccagatcga ggtgaccttc gacatcgatg ccaacggcat cctgaacgtc acggccacgg 2821 acaagagcac cggcaaggcc aacaagatca ccatcaccaa cgacaagggc cgcctgagca 2881 aggaggagat cgagcgcatg gtgcaggagg cggagaagta caaagcggag gacgaggtgc 2941 agcgcgagag ggtgtcagcc aagaacgccc tggagtccta cgccttcaac atgaagagcg 3001 ccgtggagga tgaggggctc aagggcaaga tcagcgaggc cgacaagaag aaggtgctgg 3061 acaagtgtca agaggtcatc tcgtggctgg acgccaacac cttggccgag aaggacgagt 3121 ttgagcacaa gaggaaggag ctggagcagg tgtgtaaccc catcatcagc ggactgtacc 3181 agggtgccgg tggtcccggg cctgggggct tcggggctca gggtcccaag ggagggtctg 3241 ggtcaggccc caccattgag gaggtagatt aggggccttt ccaagattgc tgtttttgtt 3301 ttggagcttc aagactttgc atttcctagt atttctgttt gtcagttctc aatttcctgt 3361 gtttgcaatg ttgaaatttt ttggtgaagt cggatccacc ggatctagat aactgatcat 3421 aatcagccat accacatttg tagaggaagg gcgaattcca gcacactggc ggccgttact 3481 agtggatccg agctcggtac caagcttcga attctgcagt cgacggtacc gcgggcccgg 3541 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac 3601 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg 3661 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 3721 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 3781 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt 3841 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca 3901 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta 3961 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta 4021 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg 4081 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga 4141 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg 4201 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc 4261 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 4321 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 4381 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc 4441 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 4501 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 4561 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 4621 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 4681 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 4741 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 4801 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 4861 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 4921 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg 4981 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 5041 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 5101 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 5161 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 5221 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 5281 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat 5341 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 5401 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 5461 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 5521 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 5581 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 5641 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 5701 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg 5761 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga 5821 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc 5881 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt 5941 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg 6001 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt 6061 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 6121 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 6181 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 6241 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 6301 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 6361 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 6421 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 6481 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 6541 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 6601 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 6661 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 6721 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 6781 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 6841 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttat //
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