| 质粒名称: | pET-Sac-Abeta(M1-42) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 71875 |
| 存储实验室: | Dominic Walsh |
|
载体骨架基本信息 |
|
| 载体名称: | pET vector |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | -- |
| 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 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Abeta(M1-42),amyloid-beta 42，abeta 42 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 129 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | T7 |
| 标签/融合蛋白: | -- |
| 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'- GTGGGTTCTAACAAGGGTGCTATCATCGGTCTGATGGTTGGTGGCGTTGTG -3' |
|
文献引用方法 |
|
| 材料和方法部分: | pET-Sac-Abeta(M1-42) was a gift from Dominic Walsh (Addgene plasmid # 71875 ; http://n2t.net/addgene:71875 ; RRID:Addgene_71875) |
| 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 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
质粒图谱.png)
载体简介
载体序列
LOCUS pET-Sac-Abeta(M1-42) 4741 bp ds-DNA circular SYN 05-JUN-2019
DEFINITION Expresses the amyloid-beta protein (42 aa), containing an exogenous
methionine (Abeta(M1-42)), when expressed in BL21* DE3 p LysS E.
coli cells.
ACCESSION .
VERSION .
KEYWORDS pET-Sac-Abeta(M1-42)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4741)
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 4741)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 5, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4741
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 375..394
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 628..645
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 660..799
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(725..744)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 885..907
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(901..1092)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind 2550..2569
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 2653..2672
/label=T7
/note="T7 promoter, forward primer"
promoter 2653..2671
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 2703..2725
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
primer_bind complement(2923..2941)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2937..2984
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(3350..3378)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(3405..3423)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3491..3595
/gene="bla"
/label=AmpR promoter
CDS 3596..4456
/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(3814..3833)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4627..474
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
61 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
121 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
181 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
241 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
301 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
361 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
421 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
481 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
541 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
601 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
661 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc
721 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
781 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
841 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
901 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
961 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
1021 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
1081 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
1141 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
1201 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
1261 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
1321 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
1381 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
1441 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
1501 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
1561 ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg cggacgcgat
1621 ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga attgattggc
1681 tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag
1741 gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg
1801 cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat cgccgtgacg
1861 atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc ttgaagctgt
1921 ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg catcccgatg
1981 ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc
2041 agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg cttctcgccg
2101 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
2161 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
2221 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
2281 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
2341 aagggcatcg gtcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
2401 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
2461 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
2521 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
2581 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
2641 atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata
2701 attttgttta actttaagaa ggagatatac atatggacgc tgaattccgt cacgactctg
2761 gttacgaagt tcaccaccag aagctggtgt tcttcgctga agacgtgggt tctaacaagg
2821 gtgctatcat cggtctgatg gttggtggcg ttgtgatcgc ttaataggag ctcgatccgg
2881 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
2941 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
3001 tatccggata tccacaggac gggtgtggtc gccatgatcg cgtagtcgat agtggctcca
3061 agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg
3121 ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata gtgactggcg
3181 atgctgtcgg aatggacgat atcccgcaag aggcccggca gtaccggcat aaccaagcct
3241 atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc
3301 atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta
3361 tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg tgatacgcct
3421 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
3481 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
3541 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
3601 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
3661 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
3721 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
3781 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
3841 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
3901 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
3961 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
4021 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
4081 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
4141 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
4201 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
4261 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
4321 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
4381 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
4441 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
4501 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
4561 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
4621 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
4681 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
4741 t
//
