| 质粒名称: | pHaloTag-PS |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 86626 |
| 存储实验室: | Thomas Leonard, Ivan Yudushkin |
|
载体骨架基本信息 |
|
| 载体名称: | pHaloTag-C1 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | HaloTag-PS(RA),PS: Akt pseudosubstrate (VELDPEFEPRARERAYAFGH),HaloTag: Promega HaloTag |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pHaloTag-PS was a gift from Thomas Leonard & Ivan Yudushkin (Addgene plasmid # 86626 ; http://n2t.net/addgene:86626 ; RRID:Addgene_86626) |
| 参考文献部分: | PI(3,4,5)P3 Engagement Restricts Akt Activity to Cellular Membranes. Ebner M, Lucic I, Leonard TA, Yudushkin I. Mol Cell. 2017 Feb 2;65(3):416-431.e6. doi: 10.1016/j.molcel.2016.12.028. 10.1016/j.molcel.2016.12.028 PubMed 28157504 |
买家导航
pHaloTagPS载体基本信息
pHaloTagPS载体图谱质粒图谱和多克隆位点信息
pHaloTagPS载体简介
pHaloTagPS载体序列
LOCUS pHaloTag-PS 4953 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses Akt pseudosubstrate tagged with a HaloTag.
ACCESSION .
VERSION .
KEYWORDS pHaloTag-PS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4953)
AUTHORS Ebner M, Lucic I, Leonard TA, Yudushkin I
TITLE PI(3,4,5)P3 Engagement Restricts Akt Activity to Cellular Membranes.
JOURNAL Mol Cell. 2017 Feb 2;65(3):416-431.e6. doi:
10.1016/j.molcel.2016.12.028.
PUBMED 28157504
REFERENCE 2 (bases 1 to 4953)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4953
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 13..601
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 502..521
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 776..1079
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1080..1283
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1234..1254
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 1315..1324
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1321..2211
/codon_start=1
/product="modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives"
/label=HaloTag(R)
/translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
IGSEIARWLSTLEISG"
polyA_signal 2456..2577
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2493..2512)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2547..2566
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2584..3039)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2721..2742)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2933..2952
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3066..3170
/gene="bla"
/label=AmpR promoter
promoter 3172..3529
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3195..3215)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3380..3515
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3442..3461
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3564..4358
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3618..3637)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4228..4247
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4546..4565)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4590..4637
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
1 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
61 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
121 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
661 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
721 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
781 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
841 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
901 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
961 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
1021 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
1081 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
1141 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
1201 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
1261 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag caaagccacc
1321 atggcagaaa tcggtactgg ctttccattc gacccccatt atgtggaagt cctgggcgag
1381 cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc ctgtgctgtt cctgcacggt
1441 aacccgacct cctcctacgt gtggcgcaac atcatcccgc atgttgcacc gacccatcgc
1501 tgcattgctc cagacctgat cggtatgggc aaatccgaca aaccagacct gggttatttc
1561 ttcgacgacc acgtccgctt catggatgcc ttcatcgaag ccctgggtct ggaagaggtc
1621 gtcctggtca ttcacgactg gggctccgct ctgggtttcc actgggccaa gcgcaatcca
1681 gagcgcgtca aaggtattgc atttatggag ttcatccgcc ctatcccgac ctgggacgaa
1741 tggccagaat ttgcccgcga gaccttccag gccttccgca ccaccgacgt cggccgcaag
1801 ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt cgtccgcccg
1861 ctgactgaag tcgagatgga ccattaccgc gagccgttcc tgaatcctgt tgaccgcgag
1921 ccactgtggc gcttcccaaa cgagctgcca atcgccggtg agccagcgaa catcgtcgcg
1981 ctggtcgaag aatacatgga ctggctgcac cagtcccctg tcccgaagct gctgttctgg
2041 ggcaccccag gcgttctgat cccaccggcc gaagccgctc gcctggccaa aagcctgcct
2101 aactgcaagg ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga caacccggac
2161 ctgatcggca gcgagatcgc gcgctggctg tcgacgctcg agatttccgg ctccggactc
2221 agatctcgag ctgttgaact agatcctgag tttgaacctc gtgccagaga gcgtgcttac
2281 gccttcggac actaagaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc
2341 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
2401 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
2461 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
2521 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
2581 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
2641 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
2701 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
2761 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
2821 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
2881 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
2941 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
3001 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
3061 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
3121 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
3181 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
3241 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
3301 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
3361 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
3421 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
3481 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
3541 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
3601 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
3661 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
3721 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
3781 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
3841 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
3901 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
3961 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
4021 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
4081 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
4141 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
4201 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
4261 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
4321 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
4381 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
4441 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
4501 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
4561 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
4621 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
4681 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
4741 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
4801 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
4861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
4921 gttccactga gcgtcagacc ccgtagaaaa gat
//
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pHaloTagPS载体质粒应用举例
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