质粒名称: | pT7-7 asyn WT |
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出品公司: | Addgene |
目录编号: | 36046 |
存储实验室: | Hilal Lashuel |
载体骨架基本信息 |
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载体名称: | pT7-7 |
载体出品公司: | Clontech |
空载体大小: | 2438bp |
完整质粒大小: | 2914bp |
载体类型: | 细菌表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | alpha-synuclein |
物种来源: | H. sapiens (human) |
插入大小: | 476bp |
启动子: | T7 |
克隆方法: | 限制性内切酶 |
5’克隆位点: | ANdeI (not destroyed) |
3’克隆位点: | HindIII (not destroyed) |
5’测序引物 | T7 |
3’测序引物 | AmpStop (TCAGGCAACTATGGATGAAC) |
文献引用方法 |
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材料和方法部分: | pT7-7 asyn WT was a gift from Hilal Lashuel (Addgene plasmid # 36046 ; http://n2t.net/addgene:36046 ; RRID:Addgene_36046) |
参考文献部分: | Phosphorylation at Ser-129 but not the phosphomimics S129E/D inhibits the fibrillation of alpha-synuclein. Paleologou KE, Schmid AW, Rospigliosi CC, Kim HY, Lamberto GR, Fredenburg RA, Lansbury PT Jr, Fernandez CO, Eliezer D, Zweckstetter M, Lashuel HA. J Biol Chem. 2008 Jun 13;283(24):16895-905. Epub 2008 Mar 14. 10.1074/jbc.M800747200 PubMed 18343814 |
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载体序列
LOCUS pT7-7 asyn WT 2914 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pT7-7 asyn WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2914) AUTHORS Paleologou KE, Schmid AW, Rospigliosi CC, Kim HY, Lamberto GR, Fredenburg RA, Lansbury PT Jr, Fernandez CO, Eliezer D, Zweckstetter M, Lashuel HA TITLE Phosphorylation at Ser-129 but not the phosphomimics S129E/D inhibits the fibrillation of alpha-synuclein. JOURNAL J Biol Chem. 2008 Jun 13;283(24):16895-905. Epub 2008 Mar 14. PUBMED 18343814 REFERENCE 2 (bases 1 to 2914) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..2914 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(42..902) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 665..684 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(903..1007) /gene="bla" /label=AmpR promoter primer_bind 1075..1093 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" rep_origin 1223..1811 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1712..1731 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1965..1982 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(2073..2092) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2235..2257 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2322..2341 /label=T7 /note="T7 promoter, forward primer" promoter 2322..2340 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 2372..2394 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" ORIGIN 1 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 61 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 121 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 181 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 241 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 301 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 361 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 421 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 481 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 541 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 601 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca 661 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 721 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 781 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 841 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 901 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 961 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 1021 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 1081 cgtatcacga ggccctttcg tcttcaagaa ttaaaaggat ctaggtgaag atcctttttg 1141 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 1201 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 1261 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 1321 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 1381 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 1441 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 1501 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 1561 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag 1621 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 1681 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 1741 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 1801 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 1861 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 1921 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 1981 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 2041 accgcataca gatctgtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 2101 agccagtata tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc 2161 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca 2221 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg 2281 cgcgaggccc agcgattcga acttctgata gacttcgaaa ttaatacgac tcactatagg 2341 gagaccacaa cggtttccct ctagaaataa ttttgtttaa ctttaagaag gagatataca 2401 tatggatgta ttcatgaaag gactttcaaa ggccaaggag ggagttgtgg ctgctgctga 2461 gaaaaccaaa cagggtgtgg cagaagcagc aggaaagaca aaagagggtg ttctctatgt 2521 aggctccaaa accaaggagg gagtggtgca tggtgtggca acagtggctg agaagaccaa 2581 agagcaagtg acaaatgttg gaggagcagt ggtgacgggt gtgacagcag tagcccagaa 2641 gacagtggag ggagcaggga gcattgcagc agccactggc tttgtcaaaa aggaccagtt 2701 gggcaagaat gaagaaggag ccccacagga aggaattctg gaagatatgc ctgtggatcc 2761 tgacaatgag gcttatgaaa tgccttctga ggaagggtat caagactacg aacctgaagc 2821 ctaagaaata tctttgctcc cagtttcttg agatctgctg acagatgttc catccaagct 2881 tatcatcgat gataagctgt caaacatgag aatt //
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