| 质粒名称: | pEGFP-N1-mt-ro1GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 82407 |
| 存储实验室: | S. James Remington |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | BD Biosciences Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4700 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: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
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插入基因信息 |
|
| 插入基因名称: | ro1GFP |
| 物种来源: | jellyfish |
| 插入大小: | -- |
| 基因突变: | C48S/F64L/T65S/S147C/Q204C |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | 丙酮酸脱氢酶E1α亚基线粒体靶向先导序列（N端插入） |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
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文献引用方法 |
|
| 材料和方法部分: | pEGFP-N1-mt-ro1GFP was a gift from S. James Remington (Addgene plasmid # 82407 ; http://n2t.net/addgene:82407 ; RRID:Addgene_82407) |
| 参考文献部分: | Investigating mitochondrial redox potential with redox-sensitive green fluorescent protein indicators. Hanson GT, Aggeler R, Oglesbee D, Cannon M, Capaldi RA, Tsien RY, Remington SJ. J Biol Chem. 2004 Mar 26;279(13):13044-53. Epub 2004 Jan 13. 10.1074/jbc.M312846200 PubMed 14722062 |
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LOCUS pEGFP-N1-mt-ro1GFP 4916 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Expression of redox sensitive GFP with a mitochondrial targeting sequence in mammalian cells. ACCESSION . VERSION . KEYWORDS pEGFP-N1-mt-ro1GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4916) AUTHORS Hanson GT, Aggeler R, Oglesbee D, Cannon M, Capaldi RA, Tsien RY, Remington SJ TITLE Investigating mitochondrial redox potential with redox-sensitive green fluorescent protein indicators. JOURNAL J Biol Chem. 2004 Mar 26;279(13):13044-53. Epub 2004 Jan 13. PUBMED 14722062 REFERENCE 2 (bases 1 to 4916) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4916 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 138..441 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 442..645 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 596..616 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 933..942 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 939..1658 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFISTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(984..1005) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1245..1264) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1592..1613 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1781..1902 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1818..1837) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1872..1891 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1909..2364) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2046..2067) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2258..2277 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2391..2495 /gene="bla" /label=AmpR promoter promoter 2497..2854 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2520..2540) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2705..2840 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2767..2786 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2889..3683 /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(2943..2962) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3553..3572 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3871..3890) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3915..3962 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4291..4879 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4780..4799 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc 61 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc 121 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc 181 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg 241 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat 301 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc 361 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta 421 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag 481 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt 541 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa 601 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt 661 cagatccgct agcgctaccg gactcagatc tcgagctcga attcgggctg aaggagactt 721 gggggcaccc gcgtcgtgcc tcctgggttg tgaggagtcg ccgctgccgc cactgcctgt 781 gcttcatgag gaagatgctc gccgccgtct cccgcgtgct gtctggcgct tctcagaagc 841 cggcaagcag agtgctggta gcatcccgta attttgcaaa tgatgctaca tttctgcagt 901 cgacggtacc gcgggcccgg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc 961 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt 1021 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca 1081 tctccaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgtcctacg 1141 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg 1201 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca 1261 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg 1321 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaact 1381 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga 1441 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc 1501 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc tgctccgccc 1561 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg 1621 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat 1681 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc 1741 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta 1801 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 1861 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg 1921 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 1981 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg 2041 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 2101 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt 2161 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag 2221 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg 2281 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc 2341 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc 2401 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 2461 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg 2521 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 2581 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 2641 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 2701 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 2761 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg 2821 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat 2881 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 2941 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 3001 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 3061 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 3121 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 3181 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 3241 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 3301 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 3361 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca 3421 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 3481 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 3541 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 3601 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 3661 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 3721 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 3781 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 3841 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg 3901 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt 3961 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca 4021 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga 4081 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc 4141 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 4201 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 4261 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 4321 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 4381 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 4441 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 4501 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 4561 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 4621 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 4681 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 4741 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 4801 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 4861 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggcct //
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