质粒名称: | pET-Sac-Abeta(M1-40) |
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出品公司: | 爱迪基因 |
目录编号: | 71876 |
存储实验室: | Dominic Walsh |
载体骨架基本信息 |
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载体名称: | pET vector |
载体抗性: | 氨苄青霉素 |
空载体大小: | 4700 bp |
载体修饰: | 向量是一个修正的pET向量。Abeta基因的限制性消化位点是Nde1(5’)和Sac1(3’) |
载体类型: | 细菌表达 |
高拷贝/低拷贝: | 低拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | Abeta(M1-40),amyloid-beta 40,abeta 40 |
物种来源: | H. sapiens (human) |
启动子 | T7 |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | Nde1(未破坏) |
3' 克隆位点: | Sac1 (未破坏) |
5' 测序引物 | 5'- ATGGACGCTGAATTCCGTCACGACTCTGGTTACGAAGTTCACCACCAGAAGCTGGTG -3' |
3' 测序引物 | 5'- CACAACGCCACCAACCATCAGACCGATGATAGCACCCTTGTTAGAACCCAC -3' |
文献引用方法 |
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材料和方法部分: | pET-Sac-Abeta(M1-40) was a gift from Dominic Walsh (Addgene plasmid # 71876 ; http://n2t.net/addgene:71876 ; RRID:Addgene_71876) |
参考文献部分: | A facile method for expression and purification of the Alzheimer's disease-associated amyloid beta-peptide. Walsh DM, Thulin E, Minogue AM, Gustavsson N, Pang E, Teplow DB, Linse S. FEBS J. 2009 Mar;276(5):1266-81. doi: 10.1111/j.1742-4658.2008.06862.x. 10.1111/j.1742-4658.2008.06862.x PubMed 19175671 |
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载体基本信息
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载体简介
载体序列
LOCUS pET-Sac-Abeta(M1-40) 4735 bp ds-DNA circular SYN 20-NOV-2018 DEFINITION Expresses the amyloid-beta protein (40 aa), containing an exogenous methionine (Abeta(M1-40)), when expressed in BL21* DE3 p LysS E. coli cells. ACCESSION . VERSION . KEYWORDS pET-Sac-Abeta(M1-40) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4735) AUTHORS Walsh DM, Thulin E, Minogue AM, Gustavsson N, Pang E, Teplow DB, Linse S TITLE A facile method for expression and purification of the Alzheimer's disease-associated amyloid beta-peptide. JOURNAL FEBS J. 2009 Mar;276(5):1266-81. doi: 10.1111/j.1742-4658.2008.06862.x. PUBMED 19175671 REFERENCE 2 (bases 1 to 4735) AUTHORS . TITLE Direct Submission JOURNAL Exported Nov 20, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4735 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 51..73 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" primer_bind complement(265..283) /label=T7 Term /note="T7 terminator, reverse primer" terminator 279..326 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(692..720) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" primer_bind complement(747..765) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 833..937 /gene="bla" /label=AmpR promoter CDS 938..1798 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1156..1175) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1969..2557 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2458..2477 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2711..2728 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2743..2882 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(2808..2827) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2968..2990 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(2984..3175) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind 4633..4652 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac 61 tttaagaagg agatatacat atggacgctg aattccgtca cgactctggt tacgaagttc 121 accaccagaa gctggtgttc ttcgctgaag acgtgggttc taacaagggt gctatcatcg 181 gtctgatggt tggtggcgtt gtgtaatagg agctcgatcc ggctgctaac aaagcccgaa 241 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct 301 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tatccacagg 361 acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg 421 actgggcggc ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc 481 atcaacgcat atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggacg 541 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg 601 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg 661 cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa 721 acatgagaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt 781 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac 841 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 901 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt 961 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct 1021 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga 1081 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag 1141 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca 1201 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga 1261 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag 1321 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc 1381 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa 1441 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt 1501 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg 1561 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt 1621 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg 1681 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat 1741 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact 1801 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa 1861 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 1921 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 1981 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 2041 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 2101 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt 2161 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 2221 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 2281 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 2341 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 2401 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 2461 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 2521 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 2581 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 2641 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 2701 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct 2761 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct 2821 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg 2881 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat 2941 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt 3001 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt 3061 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg 3121 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg 3181 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga 3241 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta 3301 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc 3361 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga 3421 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga 3481 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc 3541 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta 3601 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc 3661 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag 3721 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg 3781 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc 3841 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa 3901 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat 3961 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc 4021 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag 4081 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag 4141 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg 4201 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc 4261 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg 4321 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat 4381 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgacg 4441 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag 4501 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac 4561 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg 4621 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt 4681 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaat //
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