| 质粒名称: | pGFP-Rab8A[T22N] |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 24899 |
| 存储实验室: | Maxence Nachury |
|
载体骨架基本信息 |
|
| 载体名称: | pDEST53 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6100 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: | -- |
| 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[T22N] |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 624 bp |
| 基因突变: | 苏氨酸22改为天冬酰胺 |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | GFP（主干网上的N个终端） |
| 克隆方法: | Gateway 克隆 |
| 5' 克隆位点: | Gateway重组(未破坏) |
| 3' 克隆位点: | Gateway重组(未破坏) |
| 5' 测序引物 | n/a |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pGFP-Rab8A[T22N] was a gift from Maxence Nachury (Addgene plasmid # 24899 ; http://n2t.net/addgene:24899 ; RRID:Addgene_24899) |
| 参考文献部分: | A core complex of BBS proteins cooperates with the GTPase Rab8 to promote ciliary membrane biogenesis. Nachury MV, Loktev AV, Zhang Q, Westlake CJ, Peranen J, Merdes A, Slusarski DC, Scheller RH, Bazan JF, Sheffield VC, Jackson PK. Cell. 2007 Jun 15. 129(6):1201-13. 10.1016/j.cell.2007.03.053 PubMed 17574030 |
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
![pGFP-Rab8A[T22N]质粒图谱](/uploads/载体图谱/pGFP-Rab8A[T22N]质粒图谱.png)
载体简介
载体序列
LOCUS pGFP-Rab8A[T22N] 6760 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pGFP-Rab8A[T22N]
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6760)
AUTHORS Nachury MV, Loktev AV, Zhang Q, Westlake CJ, Peranen J, Merdes A,
Slusarski DC, Scheller RH, Bazan JF, Sheffield VC, Jackson PK
TITLE A core complex of BBS proteins cooperates with the GTPase Rab8 to
promote ciliary membrane biogenesis.
JOURNAL Cell. 2007 Jun 15. 129(6):1201-13.
PUBMED 17574030
REFERENCE 2 (bases 1 to 6760)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6760
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 671..1387
/codon_start=1
/product="Cycle 3 GFP (Crameri et al., 1996)"
/label=Cycle 3 GFP
/note="mammalian codon-optimized"
/translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
primer_bind complement(704..732)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 1327..1347
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
protein_bind 1409..1433
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(2061..2085)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
primer_bind complement(2117..2134)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2123..2347
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2393..2821
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2480..2499)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2690..2711
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2830..2850)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2835..3164
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3015..3150
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3077..3096
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3231..4025
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3285..3304)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3895..3914
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4199..4320
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4236..4255)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4290..4309
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4369..4385)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4369..4385)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4382..4404)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4393..4409
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4417..4447)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4462..4483
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4600..4617)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4771..5359)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4851..4870)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5530..6390)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6153..6172
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6391..6495)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6570..6589)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
661 gctagacacc atggccagca aaggagaaga acttttcact ggagttgtcc caattcttgt
721 tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga
781 tgctacatac ggaaagctta cccttaaatt tatttgcact actggaaaac tacctgttcc
841 atggccaaca cttgtcacta ctttctctta tggtgttcaa tgcttttccc gttatccgga
901 tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaacg
961 cactatatct ttcaaagatg acgggaacta caagacgcgt gctgaagtca agtttgaagg
1021 tgataccctt gttaatcgta tcgagttaaa aggtattgat tttaaagaag atggaaacat
1081 tctcggacac aaactcgagt acaactataa ctcacacaat gtatacatca cggcagacaa
1141 acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacattgaag atggatccgt
1201 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
1261 agacaaccat tacctgtcga cacaatctgc cctttcgaaa gatcccaacg aaaagcgtga
1321 ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca tggatgagct
1381 ctacaaaagc ggttccggac cggatcaaac aagtttgtac aaaaaagcag gcaccatggc
1441 gaagacctac gattacctgt tcaagctgct gctgatcggg gactcggggg tggggaagaa
1501 ctgtgtcctg ttccgcttct ccgaggacgc cttcaactcc acttttatct ccaccatagg
1561 aattgacttt aaaattagga ccatagagct cgatggcaag agaattaaac tgcagatatg
1621 ggacacagcc ggtcaggaac ggtttcggac gatcacaacg gcctactaca ggggtgcaat
1681 gggcatcatg ctggtctacg acatcaccaa cgagaagtcc ttcgacaaca tccggaactg
1741 gattcgcaac attgaggagc acgcctctgc agacgtcgaa aagatgatac tcgggaacaa
1801 gtgtgatgtg aatgacaaga gacaagtttc caaggaacgg ggagaaaagc tggccctcga
1861 ctatggaatc aagttcatgg agaccagcgc gaaggccaac atcaatgtgg aaaatgcatt
1921 tttcactctc gccagagata tcaaagcaaa aatggacaaa aaattggaag gcaacagccc
1981 ccaggggagc aaccagggag tcaaaatcac accggaccag cagaagagga gcagcttttt
2041 ccgatgtgtt cttctgtaga acccagcttt cttgtacaaa gtggtgataa ttaattaaga
2101 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
2161 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
2221 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
2281 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
2341 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
2401 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
2461 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
2521 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
2581 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
2641 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
2701 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
2761 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
2821 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
2881 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
2941 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
3001 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
3061 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
3121 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
3181 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
3241 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
3301 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
3361 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
3421 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
3481 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
3541 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
3601 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
3661 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
3721 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
3781 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
3841 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
3901 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
3961 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
4021 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
4081 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
4141 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
4201 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
4261 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
4321 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
4381 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
4441 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
4501 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
4561 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
4621 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
4681 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
4741 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
4801 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
4861 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
4921 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
4981 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
5041 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
5101 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
5161 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
5221 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
5281 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
5341 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
5401 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
5461 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
5521 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
5581 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
5641 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
5701 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
5761 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
5821 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
5881 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
5941 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
6001 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
6061 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
6121 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
6181 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
6241 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
6301 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
6361 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
6421 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
6481 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
6541 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
6601 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
6661 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
6721 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
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