| 质粒名称: | DsRed-Rab5 WT |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 13050 |
| 存储实验室： | Richard Pagano |
|
载体骨架基本信息 |
|
| 载体名称: | pDsRed2-C1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 4700 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 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 (Canis familiaris) |
| 物种来源: | 光合细菌 Rhodopseudomonas palustris |
| 插入大小: | 699bp |
| 基因突变： | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | RAB5A (Canis familiaris)DsRed-Rab5 WT was a gift from Richard Pagano (Addgene plasmid # 13050 ; http://n2t.net/addgene:13050 ; RRID:Addgene_13050) |
| 参考文献部分: | Glycosphingolipids internalized via caveolar-related endocytosis rapidly merge with the clathrin pathway in early endosomes and form microdomains for recycling. Sharma DK, Choudhury A, Singh RD, Wheatley CL, Marks DL, Pagano RE. J Biol Chem. 2003 Feb 28. 278(9):7564-72. 10.1074/jbc.M210457200 PubMed 12482757 |
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 DsRed-Rab5 WT 5369 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS DsRed-Rab5 WT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5369)
AUTHORS Sharma DK, Choudhury A, Singh RD, Wheatley CL, Marks DL, Pagano RE
TITLE Glycosphingolipids internalized via caveolar-related endocytosis
rapidly merge with the clathrin pathway in early endosomes and form
microdomains for recycling.
JOURNAL J Biol Chem. 2003 Feb 28. 278(9):7564-72.
PUBMED 12482757
REFERENCE 2 (bases 1 to 5369)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5369
/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..1242
/codon_start=1
/product="improved tetrameric variant of DsRed fluorescent
protein"
/label=DsRed2
/note="mammalian codon-optimized"
/translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
LFL"
primer_bind complement(748..768)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 1160..1183
/label=DsRed1-C
/note="DsRed1, forward primer"
regulatory 1249..1258
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1261..1287
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
misc_feature 1936..1988
/label=MCS
/note="multiple cloning site"
polyA_signal 2112..2233
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2149..2168)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2203..2222
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2240..2695)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2377..2398)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2589..2608
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2722..2826
/gene="bla"
/label=AmpR promoter
promoter 2828..3185
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2851..2871)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3036..3171
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3098..3117
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3220..4014
/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(3274..3293)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3884..3903
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4202..4221)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4246..4293
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4622..5210
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5111..5130
/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 gcctcctccg agaacgtcat caccgagttc
601 atgcgcttca aggtgcgcat ggagggcacc gtgaacggcc acgagttcga gatcgagggc
661 gagggcgagg gccgccccta cgagggccac aacaccgtga agctgaaggt gaccaagggc
721 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg
781 tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc
841 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg cgaccgtgac ccaggactcc
901 tccctgcagg acggctgctt catctacaag gtgaagttca tcggcgtgaa cttcccctcc
961 gacggccccg tgatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac
1021 ccccgcgacg gcgtgctgaa gggcgagacc cacaaggccc tgaagctgaa ggacggcggc
1081 cactacctgg tggagttcaa gtccatctac atggccaaga agcccgtgca gctgcccggc
1141 tactactacg tggacgccaa gctggacatc acctcccaca acgaggacta caccatcgtg
1201 gagcagtacg agcgcaccga gggccgccac cacctgttcc tgagatctgc cgccatggca
1261 tatccatatg acgtcccaga ctatgctggc gctagtcgag gcgcaacaag acccaacggg
1321 ccaaatactg gaaataaaat atgccagttc aaactagtac ttctgggaga gtctgctgtt
1381 ggcaaatcaa gcctagtgct tcgttttgtg aagggccaat ttcatgaatt tcaagagagt
1441 accatagggg ctgcttttct aacccaaact gtgtgtcttg atgatacaac agtaaagttt
1501 gaaatatggg atacagctgg tcaagaacga taccatagct tagcaccaat gtactacaga
1561 ggagcacaag cagccatagt tgtatatgat atcacaaatg aggagtcctt tgccagagcc
1621 aaaaactggg ttaaagaact tcagaggcaa gccagtccta acattgtaat agctttatca
1681 ggaaacaagg ctgatcttgc aaataaaaga gctgtcgatt tccaggaagc acagtcctat
1741 gcagatgaca acagtttatt attcatggag acatcagcta aaacatcgat gaatgtaaat
1801 gaaatattca tggcaatagc taaaaagttg ccaaagaacg aaccacagaa tccaggagca
1861 aattctgcca gaggaagagg agtagacctt actgaaccca cgcagccaac caggaatcag
1921 tgttgtagta actagctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc
1981 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
2041 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
2101 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
2161 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
2221 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
2281 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
2341 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
2401 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
2461 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
2521 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
2581 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
2641 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
2701 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
2761 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
2821 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
2881 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
2941 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
3001 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
3061 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
3121 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
3181 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
3241 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
3301 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
3361 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
3421 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
3481 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
3541 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
3601 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
3661 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
3721 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
3781 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
3841 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
3901 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
3961 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
4021 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
4081 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
4141 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
4201 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
4261 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
4321 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
4381 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
4441 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
4501 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
4561 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
4621 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
4681 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
4741 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
4801 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
4861 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
4921 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
4981 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
5041 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
5101 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
5161 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
5221 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
5281 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
5341 tcaattacgg ggtcattagt tcatagccc
//
