| 质粒名称: | Rab5a-pmCherryC1 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 27679 |
| 存储实验室: | Christien Merrifield |
|
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: | 5358bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | Rab5a |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 650bp |
| 标签/融合蛋白: | mCherry (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BglII(未破坏) |
| 3’克隆位点: | EcoRI(未破坏) |
| 5’测序引物 | mCherry-F 5'-ccccgtaatgcagaagaaga |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | Rab5a-pmCherryC1 was a gift from Christien Merrifield (Addgene plasmid # 27679 ; http://n2t.net/addgene:27679 ; RRID:Addgene_27679) |
| 参考文献部分: | A high precision survey of the molecular dynamics of Mammalian clathrin-mediated endocytosis. Taylor MJ, Perrais D, Merrifield CJ. PLoS Biol. 2011 Mar . 9(3):e1000604. 10.1371/journal.pbio.1000604 PubMed 21445324 |
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 Rab5a-pmCherryC1 5358 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Rab5a-pmCherryC1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5358)
AUTHORS Taylor MJ, Perrais D, Merrifield CJ
TITLE A high precision survey of the molecular dynamics of Mammalian
clathrin-mediated endocytosis.
JOURNAL PLoS Biol. 2011 Mar . 9(3):e1000604.
PUBMED 21445324
REFERENCE 2 (bases 1 to 5358)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5358
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(272..291)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 316..363
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 692..1280
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1181..1200
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 1455..1758
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1759..1962
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1913..1933
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 1994..2701
/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(2139..2157)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2407..2426
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 3540..3661
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3577..3596)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3631..3650
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3668..4123)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3805..3826)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4017..4036
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4150..4254
/gene="bla"
/label=AmpR promoter
promoter 4256..4613
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4279..4299)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4464..4599
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4526..4545
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4648..84
/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(4702..4721)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5312..5331
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
ORIGIN
1 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
61 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
121 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
181 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
241 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
301 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
361 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
421 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
481 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
541 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
601 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
661 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
721 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
781 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
841 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
901 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
961 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
1021 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
1081 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
1141 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
1201 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
1261 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
1321 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
1381 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
1441 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
1501 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
1561 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
1621 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
1681 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
1741 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
1801 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
1861 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
1921 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
1981 atccgctagc accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
2041 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
2101 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
2161 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
2221 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
2281 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
2341 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
2401 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
2461 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
2521 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
2581 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
2641 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
2701 gtccggactc agatctggtt caggatctgg aatggctaat cgaggagcaa caagacccaa
2761 cgggccaaat actggaaata aaatatgcca gttcaaactg gtccttctag gagagtctgc
2821 tgttggcaaa tcaagcctgg ttcttcgctt tgtgaaaggc caatttcatg aatttcaaga
2881 gagtaccatt ggggctgcct ttctaaccca aactgtgtgt cttgatgaca caacagtaaa
2941 atttgaaata tgggatacag ctggtcaaga acggtatcat agcttagcac caatgtacta
3001 ccgaggagca caagcagcca tagttgtgta tgatatcaca aatgaggaat cctttgcgag
3061 agcaaaaaac tgggttaaag aacttcaaag gcaagcaagt cctaatattg tgatagcttt
3121 gtcaggaaac aaagctgact tagcaaataa aagagctgtt gacttccagg aagcacagtc
3181 ctatgcagat gacaacagct tattatttat ggagacatca gctaagacat caatgaatgt
3241 aaatgaaata tttatggcaa tagctaaaaa gctgccaaag aatgaaccac agaatcctgg
3301 tgcaaactca gccagaggac gaggagtaga ccttactgag cctgcacagc cagccagaag
3361 ccagtgttgt agtaactgag aattctgcag tcgacggtac cgcgggcccg ggatccaccg
3421 gatctagata actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
3481 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
3541 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
3601 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
3661 aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
3721 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
3781 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
3841 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
3901 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
3961 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
4021 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
4081 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
4141 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
4201 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
4261 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
4321 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
4381 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
4441 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
4501 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
4561 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
4621 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
4681 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
4741 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
4801 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
4861 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
4921 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
4981 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
5041 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
5101 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
5161 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
5221 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
5281 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
5341 cttggcggcg aatgggct
//
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