质粒名称: | pRK5-HA-Ubiquitin-WT |
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出品公司: | 爱迪基因 |
目录编号: | 17608 |
存储实验室: | Ted Dawson |
载体骨架基本信息 |
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载体名称: | pRK5-HA |
载体抗性: | 氨苄霉素 |
空载体大小: | 4800 bp |
完整质粒大小: | 5020 bp |
载体类型: | 哺乳细胞表达 |
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: | Ubiquitin C |
物种来源: | H. sapiens (human) |
插入大小: | 220 bp |
基因突变: | Ub |
标签/融合蛋白: | HA (N terminal on backbone) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | SalI(未破坏) |
3' 克隆位点: | NotI (未破坏) |
5' 测序引物 | SP6 |
文献引用方法 |
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材料和方法部分: | pRK5-HA-Ubiquitin-WT was a gift from Ted Dawson (爱迪基因 plasmid # 17608) |
参考文献部分: | Parkin mediates nonclassical, proteasomal-independent ubiquitination of synphilin-1: implications for Lewy body formation. Lim KL, Chew KC, Tan JM, Wang C, Chung KK, Zhang Y, Tanaka Y, Smith W, Engelender S, Ross CA, Dawson VL, Dawson TM. J Neurosci. 2005 Feb 23. 25(8):2002-9. 10.1523/JNEUROSCI.4474-04.2005 PubMed 15728840 |
pRK5-HA-Ubiquitin-WT
价格:1000元
联系方式:I47-825O-882O
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LOCUS pRK5-HA-Ubiquitin-WT 4996 bp ds-DNA circular SYN 10-JAN-2018 DEFINITION Mammalian expression of HA tagged ubiquitin. ACCESSION . VERSION . KEYWORDS pRK5-HA-Ubiquitin-WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4996) AUTHORS Lim KL, Chew KC, Tan JM, Wang C, Chung KK, Zhang Y, Tanaka Y, Smith W, Engelender S, Ross CA, Dawson VL, Dawson TM TITLE Parkin mediates nonclassical, proteasomal-independent ubiquitination of synphilin-1: implications for Lewy body formation. JOURNAL J Neurosci. 2005 Feb 23. 25(8):2002-9. PUBMED 15728840 COMMENT This file is created by Vector NTI http://www.biofeng.com/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|biofeng.com| COMMENT VNTNAME|pRK5-HA-Ubiquitin-WT| FEATURES Location/Qualifiers source 1..4996 /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 581..605 /label=LNCX /note="Human CMV promoter, forward primer" promoter 815..833 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 815..832 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 846..865 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 950..959 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 962..988 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1019..1246 /codon_start=1 /gene="Homo sapiens UBC" /product="human ubiquitin C" /label=ubiquitin /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG" regulatory 1283..1292 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 1296..1430 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1333..1352) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1387..1406 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 1499..1856 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(1522..1542) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 1707..1842 /label=SV40 ori /note="SV40 origin of replication" primer_bind 1769..1788 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(1876..1893) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1876..1892) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1885..1907) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2105..2560 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2192..2211) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2402..2423 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2577..2596) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2696..2718 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2756..2774) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2842..2946 /gene="bla" /label=AmpR promoter CDS 2947..3807 /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 complement(3165..3184) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3978..4566 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4467..4486 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4720..4737 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4854..4875 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4890..4920 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4928..4944 /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)." primer_bind 4933..4955 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4952..4968 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4952..4968 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" 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 agctcgttta gtgaaccgtc 601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat 661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg 721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca 781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat 841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat 901 cgattgaatt cccgaaccga cagtcggtct cttcaccaag gccattcgcg ccaccatggg 961 ctacccctat gatgtgcctg actacgcaga tctcaatggt ggtggtggtg ggtcgaccat 1021 gcagatcttc gtgaagaccc tgactggtaa gaccatcact ctcgaagtgg agccgagtga 1081 caccattgag aatgtcaagg caaagatcca agacaaggaa ggcatccctc ctgaccagca 1141 gaggttgatc tttgctggga aacagctgga agatggacgc accctgtctg actacaacat 1201 ccagaaagag tccaccctgc acctggtcct ccgtctcaga ggtggttgag cggccgctaa 1261 gtaagtaagg atccccagct tggccgccat ggcccaactt gtttattgca gcttataatg 1321 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 1381 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gggaattaat 1441 tcggcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct 1501 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 1561 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 1621 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 1681 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 1741 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 1801 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 1861 tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 1921 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga 1981 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat 2041 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca 2101 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 2161 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 2221 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 2281 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt 2341 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 2401 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttaat 2461 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 2521 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca 2581 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg 2641 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg 2701 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc 2761 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt 2821 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 2881 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 2941 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 3001 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 3061 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 3121 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 3181 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 3241 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 3301 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 3361 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 3421 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 3481 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 3541 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 3601 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 3661 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 3721 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 3781 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 3841 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 3901 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 3961 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 4021 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 4081 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg 4141 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 4201 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 4261 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 4321 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 4381 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 4441 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 4501 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 4561 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 4621 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 4681 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 4741 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc 4801 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg 4861 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg 4921 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat gattacgaat 4981 taattcgagc tcgccc //
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