| 質粒名稱: | pET-Sac-Abeta(M1-40) |
|---|---|
| 出品公司: | 愛迪基因 |
| 目錄編號: | 71876 |
| 存儲實驗室: | Dominic Walsh |
|
載體骨架基本信息 |
|
| 載體名稱: | pET vector |
| 載體抗性: | 氨芐青霉素 |
| 空載體大小: | 4700 bp |
| 載體修飾: | 向量是一個修正的pET向量。Abeta基因的限制性消化位點是Nde1（5’）和Sac1（3’） |
| 載體類型: | 細菌表達 |
| 高拷貝/低拷貝: | 低拷貝 |
| 生長條件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名稱: | Abeta(M1-40),amyloid-beta 40,abeta 40 |
| 物種來源: | H. sapiens (human) |
| 啟動子 | T7 |
| 克隆方法: | 限制性內切酶 |
| 5' 克隆位點: | Nde1(未破壞) |
| 3' 克隆位點: | Sac1 (未破壞) |
| 5' 測序引物 | 5'- ATGGACGCTGAATTCCGTCACGACTCTGGTTACGAAGTTCACCACCAGAAGCTGGTG -3' |
| 3' 測序引物 | 5'- CACAACGCCACCAACCATCAGACCGATGATAGCACCCTTGTTAGAACCCAC -3' |
|
文獻引用方法 |
|
| 材料和方法部分: | pET-Sac-Abeta(M1-40) was a gift from Dominic Walsh (Addgene plasmid # 71876 ; //n2t.net/addgene:71876 ; RRID:Addgene_71876) |
| 參考文獻部分: | 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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pET-Sac-Abeta(M1-40)載體質粒基本信息
pET-Sac-Abeta(M1-40)質粒圖譜載體圖譜和pET-Sac-Abeta(M1-40)載體序列質粒序列多克隆位點信息
質粒圖譜.png)
pET-Sac-Abeta(M1-40)質粒載體簡介
pET-Sac-Abeta(M1-40)質粒序列載體序列
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
//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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