质粒名称: | GFP-Ub |
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出品公司: | Addgene |
目录编号: | 11928 |
存储实验室: | Nico Dantuma |
载体骨架基本信息 |
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载体名称: | EGFP-C1 |
载体出品公司: | Clontech |
载体抗性: | 卡那霉素 |
空载体大小: | 4731 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: | Ubiquitin,Ub,ubiquitin C,UBC |
物种来源: | H. sapiens (human) |
插入大小: | 245 bp |
基因突变: | -- |
GenBank编号 | NM_021009 |
启动子 | -- |
标签/融合蛋白: | GFP(主干网上的N个终端) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | HindIII(未破坏) |
3' 克隆位点: | KpnI(未破坏) |
5' 测序引物 | EGFP-C (CATGGTCCTGCTGGAGTTCGTG) |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | GFP-Ub was a gift from Nico Dantuma (Addgene plasmid # 11928 ; http://n2t.net/addgene:11928 ; RRID:Addgene_11928) |
参考文献部分: | A dynamic ubiquitin equilibrium couples proteasomal activity to chromatin remodeling. Dantuma NP, Groothuis TA, Salomons FA, Neefjes J. J Cell Biol. 2006 Apr 10. 173(1):19-26. 10.1083/jcb.200510071 PubMed 16606690 |
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GFPUb载体基本信息
GFPUb载体图谱质粒图谱和多克隆位点信息
GFPUb载体简介
GFPUb载体序列
LOCUS GFP-Ub 4953 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Used for in vivo monitoring of ubiquitin localization.. ACCESSION . VERSION . KEYWORDS GFP-Ub SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4953) AUTHORS Dantuma NP, Groothuis TA, Salomons FA, Neefjes J TITLE A dynamic ubiquitin equilibrium couples proteasomal activity to chromatin remodeling. JOURNAL J Cell Biol. 2006 Apr 10. 173(1):19-26. PUBMED 16606690 REFERENCE 2 (bases 1 to 4953) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4953 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 119..422 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 423..626 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 577..597 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 665..674 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 671..1387 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(716..737) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(977..996) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1324..1345 /label=EGFP-C /note="EGFP, forward primer" CDS 1427..1651 /codon_start=1 /product="human ubiquitin" /label=ubiquitin /translation="QIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFA GKQLEDGRTLSDYNIQKESTLHLVLRLRGG" polyA_signal 1799..1920 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1836..1855) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1890..1909 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1927..2382) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2064..2085) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2276..2295 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2409..2513 /gene="bla" /label=AmpR promoter promoter 2515..2872 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2538..2558) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2723..2858 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2785..2804 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2907..3701 /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(2961..2980) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3571..3590 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3889..3908) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3933..3980 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4309..4897 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4798..4817 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta 61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc 661 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt 721 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga 781 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc 841 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga 901 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg 961 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg 1021 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat 1081 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa 1141 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt 1201 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc 1261 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga 1321 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct 1381 gtacaagtcc ggactcagat ctcgagctca agcttcgggc cgcctgcaga tcttcgtgaa 1441 gactctgact ggtaagacca tcaccctcga ggttgagccc agtgacacca ttgagaatgt 1501 caaggcaaag atccaagata aggaaggcat ccctcctgac cagcagaggc tgatctttgc 1561 tggaaaacag ctggaagatg ggcgcaccct gtctgactac aacatccaga aagagtccac 1621 cctgcacctg gtactccgtc tcagaggtgg gtgaggtacc gcgggcccgg gatccaccgg 1681 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa 1741 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa 1801 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 1861 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 1921 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca 1981 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga 2041 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg 2101 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat 2161 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag 2221 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga 2281 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa 2341 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg 2401 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 2461 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg 2521 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc 2581 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc 2641 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat 2701 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc 2761 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga 2821 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc 2881 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 2941 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 3001 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 3061 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca 3121 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 3181 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 3241 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 3301 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 3361 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 3421 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 3481 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 3541 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 3601 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 3661 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga 3721 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt 3781 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg 3841 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa 3901 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca 3961 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga 4021 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca 4081 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc 4141 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa 4201 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 4261 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 4321 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 4381 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 4441 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 4501 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 4561 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 4621 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 4681 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 4741 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 4801 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 4861 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 4921 acggttcctg gccttttgct ggccttttgc tca //
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