| 质粒名称: | mCherry-Rab5CA(Q79L) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 35138 |
| 存储实验室: | Sergio Grinstein |
|
载体骨架基本信息 |
|
| 载体名称: | pmCherry-C1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4722 bp |
| 完整质粒大小: | 5340 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | 新霉素（G418筛选） |
| 载体抗性: | 卡那霉素，50μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5α |
| 拷贝数: | 高拷贝 |
|
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: | mCherry（主干上的N终端） |
| 克隆方法: | 限制酶 |
| 5’克隆位点: | XhoI (unknown if destroyed) |
| 3’克隆位点: | BamHI (unknown if destroyed) |
| 5’测序引物 | mCherry-F |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | mCherry-Rab5CA(Q79L) was a gift from Sergio Grinstein (Addgene plasmid # 35138 ; http://n2t.net/addgene:35138 ; RRID:Addgene_35138) |
| 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 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS mCherry-Rab5CA(Q79L) 5340 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS mCherry-Rab5CA(Q79L)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5340)
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 5340)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5340
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 16..319
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 320..523
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 474..494
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 562..571
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 568..1275
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(713..731)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 981..1000
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 2083..2204
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2120..2139)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2174..2193
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2211..2666)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2348..2369)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2560..2579
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2693..2797
/gene="bla"
/label=AmpR promoter
promoter 2799..3156
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2822..2842)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3007..3142
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3069..3088
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3191..3985
/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(3245..3264)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3855..3874
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4173..4192)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4217..4264
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4593..5181
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5082..5101
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
541 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc
601 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
661 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
721 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
781 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
841 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
901 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
961 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
1021 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
1081 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
1141 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
1201 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
1261 gacgagctgt acaagtccgg actcagatct cgagctcaag cttctatggc tagtcgaggc
1321 gcaacaagac ccaacgggcc aaatactgga aataaaatat gccagttcaa actagtactt
1381 ctgggagagt ccgctgttgg caaatcaagc ctagtgcttc gttttgtgaa aggccaattt
1441 catgaatttc aagagagtac cattggggct gcttttctaa cccaaactgt atgtcttgat
1501 gacactacag taaagtttga aatatgggat acagctggtc tcgaacgata ccatagccta
1561 gcaccaatgt actacagagg agcacaagca gccatagttg tatatgatat cacaaatgag
1621 gagtcctttg caagagcaaa aaattgggtt aaagaacttc agaggcaagc aagtcctaac
1681 attgtaatag ctttatcggg aaacaaggcc gacctagcaa ataaaagagc agtagatttc
1741 caggaagcac agtcctatgc agatgacaat agtttattat tcatggagac atccgctaaa
1801 acatcaatga atgtaaatga aatattcatg gcaatagcta aaaaattgcc aaagaatgaa
1861 ccacaaaatc caggagcaaa ttctgccaga ggaagaggag tagaccttac cgaacccaca
1921 caaccaacca ggaatcagtg ttgtagtaac tgaggatcca ccggatctag ataactgatc
1981 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
2041 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
2101 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
2161 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
2221 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
2281 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
2341 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
2401 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
2461 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
2521 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
2581 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
2641 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
2701 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
2761 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
2821 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
2881 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
2941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
3001 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
3061 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
3121 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
3181 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
3241 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
3301 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
3361 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
3421 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
3481 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
3541 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
3601 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
3661 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
3721 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
3781 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
3841 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
3901 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
3961 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
4021 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
4081 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
4141 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
4201 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
4261 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
4321 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
4381 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
4441 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
4501 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
4561 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
4621 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
4681 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
4741 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
4801 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
4861 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
4921 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
4981 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
5041 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
5101 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
5161 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
5221 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
5281 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
//
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