| 质粒名称: | GFP-Ub |
|---|---|
| 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 |
|
| 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 |
|
| 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 |
|
| 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 |
买家导航
GFP-Ub载体 GFPUb质粒基本信息
GFP-Ub质粒图谱 GFPUb载体图谱和GFP-Ub载体序列 GFPUb质粒序列多克隆位点信息
GFP-Ub质粒 GFPUb载体简介
GFP-Ub质粒序列 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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