| 质粒名称: | GFP-Rab5CA (Q79L) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 35140 |
| 存储实验室: | Sergio Grinstein |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C1 |
| 载体出品公司: | 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: | 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: | Rab5a |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | Constitutively Active- Q79L |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | EGFP（主干上的N端） |
| 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: | BamHI（销毁后未知） |
| 5' 测序引物 | EGFP-C |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | GFP-Rab5CA (Q79L) was a gift from Sergio Grinstein (Addgene plasmid # 35140 ; http://n2t.net/addgene:35140 ; RRID:Addgene_35140) |
| 参考文献部分: | Recruitment of OCRL and Inpp5B to phagosomes by Rab5 and APPL1 depletes phosphoinositides and attenuates Akt signaling. Bohdanowicz M, Balkin DM, De Camilli P, Grinstein S. Mol Biol Cell. 2012 Jan;23(1):176-87. Epub 2011 Nov 9. 10.1091/mbc.E11-06-0489 PubMed 22072788 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
质粒图谱.png)
载体简介
载体序列
LOCUS GFP-Rab5CA (Q79L) 5349 bp ds-DNA circular SYN 02-MAY-2019
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS GFP-Rab5CA (Q79L)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5349)
AUTHORS Bohdanowicz M, Balkin DM, De Camilli P, Grinstein S
TITLE Recruitment of OCRL and Inpp5B to phagosomes by Rab5 and APPL1
depletes phosphoinositides and attenuates Akt signaling.
JOURNAL Mol Biol Cell. 2012 Jan;23(1):176-87. Epub 2011 Nov 9.
PUBMED 22072788
REFERENCE 2 (bases 1 to 5349)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 2, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5349
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 607..616
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 613..1329
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(658..679)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(919..938)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1266..1287
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2137..2258
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2174..2193)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2228..2247
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2265..2720)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2402..2423)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2614..2633
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2747..2851
/gene="bla"
/label=AmpR promoter
promoter 2853..3210
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2876..2896)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3061..3196
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3123..3142
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3245..4039
/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(3299..3318)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3909..3928
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4227..4246)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4271..4318
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4647..5235
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5136..5155
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
1321 ctgtacaagt ccggactcag atctcgagct caagcttcta tggctagtcg aggcgcaaca
1381 agacccaacg ggccaaatac tggaaataaa atatgccagt tcaaactagt acttctggga
1441 gagtccgctg ttggcaaatc aagcctagtg cttcgttttg tgaaaggcca atttcatgaa
1501 tttcaagaga gtaccattgg ggctgctttt ctaacccaaa ctgtatgtct tgatgacact
1561 acagtaaagt ttgaaatatg ggatacagct ggtctcgaac gataccatag cctagcacca
1621 atgtactaca gaggagcaca agcagccata gttgtatatg atatcacaaa tgaggagtcc
1681 tttgcaagag caaaaaattg ggttaaagaa cttcagaggc aagcaagtcc taacattgta
1741 atagctttat cgggaaacaa ggccgaccta gcaaataaaa gagcagtaga tttccaggaa
1801 gcacagtcct atgcagatga caatagttta ttattcatgg agacatccgc taaaacatca
1861 atgaatgtaa atgaaatatt catggcaata gctaaaaaat tgccaaagaa tgaaccacaa
1921 aatccaggag caaattctgc cagaggaaga ggagtagacc ttaccgaacc cacacaacca
1981 accaggaatc agtgttgtag taactgagga tccaccggat ctagataact gatcataatc
2041 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
2101 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
2161 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
2221 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
2281 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
2341 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
2401 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
2461 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
2521 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
2581 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
2641 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
2701 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
2761 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
2821 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
2881 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
2941 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
3001 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
3061 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
3121 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
3181 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
3241 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
3301 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
3361 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
3421 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
3481 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
3541 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
3601 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
3661 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
3721 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
3781 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
3841 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
3901 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
3961 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
4021 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
4081 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
4141 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
4201 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
4261 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
4321 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
4381 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
4441 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
4501 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
4561 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
4621 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
4681 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
4741 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
4801 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
4861 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
4921 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
4981 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
5041 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
5101 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
5161 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
5221 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
5281 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
5341 gccatgcat
//
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