质粒名称: | GST-HA-GFP11-N1 |
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出品公司: | 爱迪基因 |
目录编号: | 91953 |
存储实验室: | Rong Li |
载体骨架基本信息 |
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载体名称: | pAcGFP1-N1 |
载体出品公司: | Clontech |
载体抗性: | 卡那霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | Kan/Neo |
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: | GST-HA-GFP11 |
物种来源: | Synthetic |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | CMV |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | HindIII(销毁后未知) |
3' 克隆位点: | NotI(销毁后未知) |
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: | GST-HA-GFP11-N1 was a gift from Rong Li (Addgene plasmid # 91953 ; http://n2t.net/addgene:91953 ; RRID:Addgene_91953) |
参考文献部分: | Cytosolic proteostasis through importing of misfolded proteins into mitochondria. Ruan L, Zhou C, Jin E, Kucharavy A, Zhang Y, Wen Z, Florens L, Li R. Nature. 2017 Mar 16;543(7645):443-446. doi: 10.1038/nature21695. Epub 2017 Mar 1. 10.1038/nature21695 PubMed 28241148 |
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LOCUS GST-HA-GFP11-N1 4759 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Expresses GST-HA-GFP11 in mammalian cells. ACCESSION . VERSION . KEYWORDS GST-HA-GFP11-N1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4759) AUTHORS Ruan L, Zhou C, Jin E, Kucharavy A, Zhang Y, Wen Z, Florens L, Li R TITLE Cytosolic proteostasis through importing of misfolded proteins into mitochondria. JOURNAL Nature. 2017 Mar 16;543(7645):443-446. doi: 10.1038/nature21695. Epub 2017 Mar 1. PUBMED 28241148 REFERENCE 2 (bases 1 to 4759) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4759 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 124..427 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 428..631 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 582..602 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 691..1344 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" primer_bind 1302..1324 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" CDS 1372..1398 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1438..1485 /codon_start=1 /product="11th beta-strand of superfolder GFP" /label=GFP11 /note="associates with GFP(1-10), consisting of the first 10 beta-strands, to reconstitute fluorescent GFP (Cabantous et al., 2005; Kamiyama et al., 2016)" /translation="RDHMVLHEYVNAAGIT" polyA_signal 1610..1731 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1647..1666) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1701..1720 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1738..2193) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1875..1896) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2087..2106 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2220..2324 /gene="bla" /label=AmpR promoter promoter 2326..2683 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2349..2369) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2534..2669 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2596..2615 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2718..3512 /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(2772..2791) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3382..3401 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3700..3719) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3744..3791 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4120..4708 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4609..4628 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg 61 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt 121 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc 181 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg 241 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat 301 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca 361 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat 421 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg 481 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca 541 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg 601 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg 661 ctaccggact cagatctcga gctcaagctt atgtccccta tactaggtta ttggaaaatt 721 aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag 781 catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg 841 gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc 901 atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca 961 gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca 1021 tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg 1081 aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat 1141 cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg 1201 gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat 1261 aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt 1321 ggtggtggcg accatcctcc aaaaggatcc ggtgacggtc cggtcgccac ctacccatac 1381 gatgttccag attacgctgg tcgacggatc ggtggtggtt ctggaggcgg ttctatgaga 1441 gatcatatgg ttttgcatga atatgttaat gctgctggta ttacttaagc ggccgcgact 1501 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc 1561 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat 1621 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 1681 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa 1741 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt 1801 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag 1861 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg 1921 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat 1981 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc 2041 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga 2101 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac 2161 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc 2221 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 2281 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa 2341 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 2401 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 2461 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 2521 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 2581 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca 2641 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag 2701 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt 2761 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg 2821 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg 2881 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg 2941 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg 3001 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca 3061 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc 3121 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc 3181 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca 3241 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga 3301 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg 3361 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg 3421 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg 3481 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga 3541 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag 3601 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct 3661 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat 3721 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg 3781 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc 3841 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag 3901 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc 3961 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 4021 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 4081 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 4141 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 4201 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 4261 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 4321 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 4381 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 4441 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 4501 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 4561 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 4621 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 4681 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 4741 ggccttttgc tggcctttt //
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