| 质粒名称: | EGFP-Rab8a-TN |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 86077 |
| 存储实验室: | Lei Lu |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C2 |
| 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: | 5331 bp |
| 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: | hRab8A,ras-related protein Rab-8A |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | 把苏氨酸22改成天冬氨酸 |
| GenBank编号 | NM_005370.4 |
| 启动子 | CMV |
| 标签/融合蛋白: | GFP (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | BamHI(未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | EGFP-Rab8a-TN was a gift from Lei Lu (Addgene plasmid # 86077 ; http://n2t.net/addgene:86077 ; RRID:Addgene_86077) |
| 参考文献部分: | A ternary complex comprising transportin1, Rab8 and the ciliary targeting signal directs proteins to ciliary membranes. Madugula V, Lu L. J Cell Sci. 2016 Oct 15;129(20):3922-3934. Epub 2016 Sep 15. 10.1242/jcs.194019 PubMed 27633000 |
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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
载体简介
载体序列
LOCUS EGFP-Rab8a-TN 5331 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION A mammalian expression plasmid encoding GDP locked Rab8a
(Rab8a-T22N) with N-terminus EGFP..
ACCESSION .
VERSION .
KEYWORDS EGFP-Rab8a-TN
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5331)
AUTHORS Madugula V, Lu L
TITLE A ternary complex comprising transportin1, Rab8 and the ciliary
targeting signal directs proteins to ciliary membranes.
JOURNAL J Cell Sci. 2016 Oct 15;129(20):3922-3934. Epub 2016 Sep 15.
PUBMED 27633000
REFERENCE 2 (bases 1 to 5331)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5331
/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 2119..2240
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2156..2175)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2210..2229
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2247..2702)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2384..2405)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2596..2615
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2729..2833
/gene="bla"
/label=AmpR promoter
promoter 2835..3192
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2858..2878)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3043..3178
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3105..3124
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3227..4021
/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(3281..3300)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3891..3910
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4209..4228)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4253..4300
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4629..5217
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5118..5137
/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 ccggccggac tcagatctcg agctcaagct tcgaattcgc gaagacctac
1381 gattacctgt tcaagctgct gctgatcggg gactcggggg tggggaagaa ctgtgtcctg
1441 ttccgcttct ccgaggacgc cttcaactcc acttttatct ccaccatagg aattgacttt
1501 aaaattagga ccatagagct cgatggcaag agaattaaac tgcagatatg ggacacagcc
1561 ggtcaggaac ggtttcggac gatcacaacg gcctactaca ggggtgcaat gggcatcatg
1621 ctggtctacg acatcaccaa cgagaagtcc ttcgacaaca tccggaactg gattcgcaac
1681 attgaggagc acgcctctgc agacgtcgaa aagatgatac tcgggaacaa gtgtgatgtg
1741 aatgacaaga gacaagtttc caaggaacgg ggagaaaagc tggccctcga ctatggaatc
1801 aagttcatgg agaccagcgc gaaggccaac atcaatgtgg aaaatgcatt tttcactctc
1861 gccagagata tcaaagcaaa aatggacaaa aaattggaag gcaacagccc ccaggggagc
1921 aaccagggag tcaaaatcac accggaccag cagaagagga gcagcttttt ccgatgtgtt
1981 cttctgtagg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
2041 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
2101 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
2161 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
2221 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
2281 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
2341 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
2401 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
2461 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
2521 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
2581 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
2641 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
2701 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
2761 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
2821 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
2881 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
2941 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
3001 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
3061 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
3121 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
3181 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
3241 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
3301 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
3361 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
3421 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
3481 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
3541 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
3601 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
3661 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
3721 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
3781 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
3841 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
3901 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
3961 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
4021 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
4081 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
4141 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
4201 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
4261 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
4321 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
4381 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
4441 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
4501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
4561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
4621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
4681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
4741 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
4801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
4861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
4921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
4981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
5041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
5101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
5161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
5221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
5281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
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