| 质粒名称: | Ub-G76V-GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 11941 |
| 存储实验室: | Nico Dantuma |
|
载体骨架基本信息 |
|
| 载体名称: | EGFP-N1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 3970 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: | Ub-G76V-GFP,Ubiquitin,Ub |
| 物种来源: | -- |
| 插入大小: | 990 bp |
| 基因突变: | Ubiquitin fused to N-terminus of GFP. Glycine 76 mutated to Valine. |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | EGFP-N (CGTCGCCGTCCAGCTCGACCAG) |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | Ub-G76V-GFP was a gift from Nico Dantuma (Addgene plasmid # 11941 ; http://n2t.net/addgene:11941 ; RRID:Addgene_11941) |
| 参考文献部分: | Short-lived green fluorescent proteins for quantifying ubiquitin/proteasome-dependent proteolysis in living cells. Dantuma NP, Lindsten K, Glas R, Jellne M, Masucci MG. Nat Biotechnol. 2000 May . 18(5):538-43. 10.1038/75406 PubMed 10802622 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS Ub-G76V-GFP 4959 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Ub-G76V-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4959)
AUTHORS Dantuma NP, Lindsten K, Glas R, Jellne M, Masucci MG
TITLE Short-lived green fluorescent proteins for quantifying
ubiquitin/proteasome-dependent proteolysis in living cells.
JOURNAL Nat Biotechnol. 2000 May . 18(5):538-43.
PUBMED 10802622
REFERENCE 2 (bases 1 to 4959)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4959
/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"
CDS 696..920
/codon_start=1
/product="human ubiquitin"
/label=ubiquitin
/translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
AGKQLEDGRTLSDYNIQKESTLHLVLRLRG"
regulatory 957..966
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 963..1682
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1008..1029)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1269..1288)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1616..1637
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1805..1926
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1842..1861)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1896..1915
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1933..2388)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2070..2091)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2282..2301
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2415..2519
/gene="bla"
/label=AmpR promoter
promoter 2521..2878
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2544..2564)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2729..2864
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2791..2810
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2913..3707
/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(2967..2986)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3577..3596
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3895..3914)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3939..3986
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4315..4903
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4804..4823
/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 ggactcagat ctcgagctca agcttcgaat tcaccatgca gatcttcgtg aagactctga
721 ctggtaagac catcaccctc gaggttgagc ccagtgacac cattgagaat gtcaaggcaa
781 agatccaaga taaggaaggc atccctcctg accagcagag gctgatcttt gctggaaaac
841 agctggaaga tgggcgcacc ctgtctgact acaacatcca gaaagagtcc accctgcacc
901 tggtactccg tctcagaggt gtggtgggga agcttggtcg acaggatcca ccggtcgcca
961 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
1021 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
1081 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
1141 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
1201 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
1261 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
1321 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
1381 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
1441 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
1501 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
1561 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
1621 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
1681 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
1741 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
1801 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
1861 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
1921 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
1981 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
2041 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
2101 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
2161 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
2221 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
2281 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
2341 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
2401 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
2461 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
2521 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
2581 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
2641 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
2701 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
2761 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
2821 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
2881 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
2941 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
3001 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
3061 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
3121 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
3181 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
3241 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
3301 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
3361 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
3421 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
3481 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
3541 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
3601 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
3661 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
3721 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
3781 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
3841 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
3901 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
3961 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
4021 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
4081 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
4141 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
4201 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
4261 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
4321 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
4381 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
4441 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
4501 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
4561 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
4621 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
4681 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
4741 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
4801 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
4861 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
4921 ctttttacgg ttcctggcct tttgctggcc ttttgctca
//
