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pKillerRed-mem

pKillerRed-mem
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  • 产品名称:pKillerRed-mem
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简单介绍
pKillerRed-mem菌株发货前均经过严格的多重验证,如存在质量问题,请在收到产品的三个月内通知我司。pKillerRed-mem收到质粒后请短暂离心,加入20μl ddH2O溶解质粒,取2μl转化���对应感受态中,挑取单克隆重新提取质粒后使用。
产品描述

pKillerRed-mem

  • 产品信息
  • 基本信息
  • 质粒简介
  • 质粒图谱
  • 质粒序列

产品信息

产品货号 产品名称 产品规格 优惠价
XY1385 pKillerRed-mem

5μg质粒

¥请询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证,如存在质量问题,请在收到产品的三个月内通知我司。收到质粒后请短暂离心,加入20μl ddH2O溶解质粒,取2μl转化至对应感受态中,挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

启动子: CMV promoter
复制子: Ori,F1 ori,SV40 ori
终止子: SV40 poly(A) signal
质粒分类: 哺乳细胞质粒;哺乳报告质粒;荧光定位质粒
质粒大小: 4787bp
质粒标签: C-mem,C-KillerRed
原核抗性: 卡那霉素Kan
筛选标记: 新霉素Neo/G418
克隆菌株: 大肠杆菌DH5α
培养条件: 37℃,有氧,LB
表达宿主: 293T等哺乳细胞

质粒简介

          pKillerRed-mem质粒是一个哺乳细胞红色荧光定位于细胞膜的质粒。pKillerRed-mem is a mammalian expression vector encoding membrane-targeted KillerRed. KillerRed localized on cellular membrane can be used for effective light-induced cell killing.KillerRed codon usage is optimized for high expression in mammalian cells (humanized) [Haas et al. 1996]. Membrane localization signal (MLS) of neuromodulin is linked to the KillerRed N-terminus. The MLS (Nterminal 20 amino acid residues of neuromodulin) contains a signal for posttranslational palmitoylation of cysteines 3 and 4 that targets KillerRed to cellular membranes [Skene and Virág 1989]. pKillerRed-mem vector can be used as a source of MLS-KillerRed hybrid sequence. The vector backbone contains unique restriction sites that permit its excision and further insertion into expression vector of choice.The vector backbone contains immediate early promoter of cytomegalovirus (PCMV IE) for protein expression, SV40 origin for replication in mammalian cells expressing SV40 T-antigen, pUC origin of replication for propagation in E. coli, and f1 origin for single-stranded DNA production. SV40 polyadenylation signals (SV40 poly A) direct proper processing of the 3’-end of the reporter mRNA. SV40 early promoter (PSV40) provides neomycin resistance gene (Neor ) expression to select stably transfected eukaryotic cells using G418. Bacterial promoter (P) provides kanamycin resistance gene expression (Kanr ) in E. coli. Kanr /Neor gene is linked with herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signals.pKillerRed-mem vector can be transfected into mammalian cells by any known transfection method. CMV promoter provides strong, constitutive expression of memrane-targeted KillerRed in eukaryotic cells. If required, stable transformants can be selected using G418 [Gorman 1985].Suitable host strains for propagation in E. coli include DH5alpha, HB101, XL1-Blue, and other general purpose strains. Plasmid incompatibility group is pMB1/ColE1. The vector confers resistance to kanamycin (30 µg/ml) to E. coli hosts. Copy number in E. coli is about 500.

质粒图谱

质粒序列

LOCUS       Exported                4787 bp ds-DNA     circular SYN 17-AUG-2017
DEFINITION  synthetic circular DNA
KEYWORDS    pKillerRed-mem.dna
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4787)
FEATURES             Location/Qualifiers
     source          1..4787
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    591..671
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    679..738
                     /label=mem
     misc_feature    739..1452
                     /label=KillerRed
     polyA_signal    1575..1696
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1703..2158)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2185..2289
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2291..2648
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2499..2634
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2683..3477
                     /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"
     polyA_signal    3709..3756
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4085..4673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatccaccgg tcgccaccat gctgtgctgt atgagaagaa ccaaacaggt tgaaaagaat
      721 gatgaggacc aaaagatctc cgagggcggc cccgccctgt tccagagcga catgaccttc
      781 aaaatcttca tcgacggcga ggtgaacggc cagaagttca ccatcgtggc cgacggcagc
      841 agcaagttcc cccacggcga cttcaacgtg cacgccgtgt gcgagaccgg caagctgccc
      901 atgagctgga agcccatctg ccacctgatc cagtacggcg agcccttctt cgcccgctac
      961 cccgacggca tcagccattt cgcccaggag tgcttccccg agggcctgag catcgaccgc
     1021 accgtgcgct tcgagaacga cggcaccatg accagccacc acacctacga gctggacgac
     1081 acctgcgtgg tgagccgcat caccgtgaac tgcgacggct tccagcccga cggccccatc
     1141 atgcgcgacc agctggtgga catcctgccc aacgagaccc acatgttccc ccacggcccc
     1201 aacgccgtgc gccagctggc cttcatcggc ttcaccaccg ccgacggcgg cctgatgatg
     1261 ggccacttcg acagcaagat gaccttcaac ggcagccgcg ccatcgagat ccccggccca
     1321 cacttcgtga ccatcatcac caagcagatg agggacacca gcgacaagcg cgaccacgtg
     1381 tgccagcgcg aggtggccta cgcccacagc gtgccccgca tcaccagcgc catcggtagc
     1441 gacgaggatt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     1501 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     1561 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     1621 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1681 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     1741 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     1801 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     1861 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     1921 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     1981 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2041 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2101 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     2161 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2221 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2281 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     2341 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2401 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2461 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2521 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2581 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2641 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     2701 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     2761 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     2821 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     2881 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     2941 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3001 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3061 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3121 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3181 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     3241 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3301 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3361 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     3421 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     3481 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     3541 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     3601 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     3661 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     3721 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     3781 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     3841 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     3901 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     3961 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4021 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4081 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4141 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4201 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4261 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4321 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4381 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4441 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4501 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4561 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4621 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4681 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4741 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
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