| 质粒名称: | GFP-rab11 WT |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 12674 |
| 存储实验室: | Richard Pagano |
|
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: | RAB11A (Homo sapiens),GFP-rab11 wild type |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 717 bp |
| 基因突变: | -- |
| GenBank编号 | NM_004663 |
| 启动子 | -- |
| 标签/融合蛋白: |
GFP（主干网上的N个终端）
HA（插入N端子）
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BAMHI(未破坏) |
| 3' 克隆位点: | BAMHI(未破坏) |
| 5' 测序引物 | EGFP-C |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | GFP-rab11 WT was a gift from Richard Pagano (Addgene plasmid # 12674 ; http://n2t.net/addgene:12674 ; RRID:Addgene_12674) |
| 参考文献部分: | Rab proteins mediate Golgi transport of caveola-internalized glycosphingolipids and correct lipid trafficking in Niemann-Pick C cells. Choudhury A, Dominguez M, Puri V, Sharma DK, Narita K, Wheatley CL, Marks DL, Pagano RE. J Clin Invest. 2002 Jun . 109(12):1541-50. 10.1172/JCI15420 PubMed 12070301 |
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 GFP-rab11 WT 5442 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS GFP-rab11 WT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5442)
AUTHORS Choudhury A, Dominguez M, Puri V, Sharma DK, Narita K, Wheatley CL,
Marks DL, Pagano RE
TITLE Rab proteins mediate Golgi transport of caveola-internalized
glycosphingolipids and correct lipid trafficking in Niemann-Pick C
cells.
JOURNAL J Clin Invest. 2002 Jun . 109(12):1541-50.
PUBMED 12070301
REFERENCE 2 (bases 1 to 5442)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5442
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 68..371
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 372..575
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 526..546
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 614..623
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 620..1336
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(665..686)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(926..945)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1273..1294
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 1337..1402
/label=MCS
/note="multiple cloning site"
CDS 1406..1432
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1433..1459
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
polyA_signal 2237..2358
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2274..2293)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2328..2347
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2365..2820)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2502..2523)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2714..2733
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2847..2951
/gene="bla"
/label=AmpR promoter
promoter 2953..3310
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2976..2996)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3161..3296
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3223..3242
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3345..4139
/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(3399..3418)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4009..4028
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4327..4346)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4371..4418
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4747..5335
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5236..5255
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
721 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
781 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
901 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
961 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
1021 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
1081 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
1201 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
1321 ggacgagctg tacaagtccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga
1381 cggtaccgcg ggcccgggat ccatgtaccc atacgatgtt ccggattacg cttacccata
1441 cgatgttccg gattacgctg gtacccgcga cgacgagtac gactacctct ttaaagttgt
1501 ccttattgga gattctggtg ttggaaagag taatctcctg tctcgattta ctcgaaatga
1561 gtttaatctg gaaagcaaga gcaccattgg agtagagttt gcaacaagaa gcatccaggt
1621 tgatggaaaa acaataaagg cacagatatg ggacacagca gggcaagagc gatatcgagc
1681 tataacatca gcatattatc gtggagctgt aggtgcctta ttggtttatg acattgctaa
1741 acatctcaca tatgaaaatg tagagcgatg gctgaaagaa ctgagagatc atgctgatag
1801 taacattgtt atcatgcttg tgggcaataa gagtgatcta cgtcatctca gggcagttcc
1861 tacagatgaa gcaagagctt ttgcagaaaa gaatggtttg tcattcattg aaacttcggc
1921 cctagactct acaaatgtag aagctgcttt tcagacaatt ttaacagaga tttaccgcat
1981 tgtttctcag aagcaaatgt cagacagacg cgaaaatgac atgtctccaa gcaacaatgt
2041 ggttcctatt catgttccac caaccactga aaacaagcca aaggtgcagt gctgtcagaa
2101 catctaagga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
2161 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
2221 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
2281 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
2341 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
2401 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
2461 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
2521 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
2581 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
2641 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
2701 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
2761 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
2821 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
2881 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
2941 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
3001 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
3061 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
3121 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
3181 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
3241 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
3301 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
3361 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
3421 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
3481 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
3541 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
3601 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
3661 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
3721 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
3781 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
3841 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
3901 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
3961 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
4021 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
4081 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
4141 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
4201 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
4261 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
4321 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
4381 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
4441 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
4501 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
4561 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
4621 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
4681 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
4741 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
4801 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
4861 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
4921 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
4981 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
5041 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
5101 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
5161 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
5221 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
5281 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
5341 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
5401 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gc
//
