| 质粒名称: | pALOD4 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 111026 |
| 存储实验室: | Arun Radhakrishnan |
|
载体骨架基本信息 |
|
| 载体名称: | pRSET-B |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 2887 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Domain 4 of Anthrolysin O,ALOD4 |
| 物种来源: | 炭疽芽孢杆菌 |
| 插入大小: | 429 bp |
| 基因突变: | Mutations: S404C, C472A |
| GenBank编号 | AAT_32463.1 |
| 启动子 | T7 |
| 标签/融合蛋白: | His6标签（插入件上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamH1（如果已销毁，则未知） |
| 3' 克隆位点: | EcoR1（如果已销毁，则未知） |
| 5' 测序引物 | TAATACGACTCACTATAGGG |
| 3' 测序引物 | GCTAGTTATTGCTCAGCGG |
|
文献引用方法 |
|
| 材料和方法部分: | pALOD4 was a gift from Arun Radhakrishnan (Addgene plasmid # 111026 ; http://n2t.net/addgene:111026 ; RRID:Addgene_111026) |
| 参考文献部分: | Switch-like responses of two cholesterol sensors do not require protein oligomerization in membranes. Gay A, Rye D, Radhakrishnan A. Biophys J. 2015 Mar 24;108(6):1459-69. doi: 10.1016/j.bpj.2015.02.008. 10.1016/j.bpj.2015.02.008 PubMed 25809258 |
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
载体简介
载体序列
LOCUS pALOD4 3190 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses domain 4 of anthrolysin O cholesterol sensor, has
N-terminal His6-tag and two point mutations, S404C and C472A.
ACCESSION .
VERSION .
KEYWORDS pALOD4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3190)
AUTHORS Gay A, Rye D, Radhakrishnan A
TITLE Switch-like responses of two cholesterol sensors do not require
protein oligomerization in membranes.
JOURNAL Biophys J. 2015 Mar 24;108(6):1459-69. doi:
10.1016/j.bpj.2015.02.008.
PUBMED 25809258
REFERENCE 2 (bases 1 to 3190)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3190
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 20..39
/label=T7
/note="T7 promoter, forward primer"
promoter 20..38
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 70..92
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 112..129
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 133..165
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag (gene 10 leader)
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS 169..192
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/label=Xpress(TM) tag
/translation="DLYDDDDK"
primer_bind complement(591..609)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 605..652
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 749..1204
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(836..855)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1046..1067
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 1230..1334
/gene="bla"
/label=AmpR promoter
CDS 1335..2195
/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(1553..1572)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2366..2954
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2855..2874
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3108..3125
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
181 gatgacgata aggatccgtg cgctaaaatg accctggatc actatggcgc atacgtcgct
241 cagtttgatg ttagttggga cgaatttacc ttcgatcaaa acggcaagga agttctgacc
301 cataaaacgt gggaaggcag tggtaaagat aagaccgcgc actattccac ggttatcccg
361 ctgccgccga actcaaaaaa tattaagatc gtcgcacgtg aagcgaccgg tctggcatgg
421 gaatggtggc gtacgattat caacgaacag aatgtgccgc tgaccaatga aatcaaggtc
481 tctattggcg gcaccacgct gtatccgacg gcaaccatct cacactaaga attcgaagct
541 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
601 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
661 aggaactata tccggatctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
721 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
781 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
841 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
901 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
961 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
1021 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
1081 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
1141 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
1201 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
1261 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
1321 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
1381 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
1441 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
1501 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
1561 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
1621 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
1681 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
1741 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
1801 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
1861 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
1921 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
1981 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
2041 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
2101 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
2161 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
2221 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
2281 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
2341 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
2401 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
2461 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
2521 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
2581 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
2641 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
2701 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
2761 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
2821 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
2881 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
2941 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
3001 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
3061 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
3121 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
3181 attaatgcag
//
