质粒名称: | pGFP-Rab8A[T22N] |
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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 |
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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 |
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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 |
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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 |
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pGFP-Rab8A[T22N]载体质粒基本信息
pGFP-Rab8A[T22N]质粒图谱载体图谱和pGFP-Rab8A[T22N]载体序列质粒序列多克隆位点信息
pGFP-Rab8A[T22N]质粒载体简介
pGFP-Rab8A[T22N]质粒序列载体序列
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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