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pDsRed2-Peroxi

pDsRed2-Peroxi
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  • 产品名称：pDsRed2-Peroxi
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简单介绍
pDsRed2-Peroxi是一种哺乳动物表达载体，其编码Discosoma sp红色荧光蛋白（DsRed2）和过氧化物酶体靶向信号1（PTS1）.PTS1序列与DsRed2的3'末端融合，DsRed变体设计用于更快的成熟和���低的非特异性聚集.PTS1序列编码三肽SKL，pDsRed2-Peroxi其中将DsRed2-PTS1融合蛋白靶向过氧化物酶体基质。
产品描述

pDsRed2-Peroxi

  • 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
XY0154 pDsRed2-Peroxi

5μg

¥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: pUC ori,f1 ori
终止子: SV40 poly(A) signal
质粒分类: 哺乳系列质粒；哺乳荧光质粒；哺乳红色质粒
质粒大小: 4644bp
质粒标签: N-DsRed2，C-peroxi
原核抗性: 卡纳霉素Kan（50μg/ml）
筛选标记: 新霉素Neo/G418
克隆菌株: DH5α等大肠杆菌
培养条件: 37℃，LB，有氧
表达宿主: 293T等哺乳细胞
培养条件: 37℃，5%CO2
诱导方式: 无须诱导，瞬时表达
5'测序引物: CMV-F（CGCAAATGGGCGGTAGGCGTG）
3'测序引物: Sv40-polyA-R（GAAATTTGTGATGCTATTGC）

质粒简介

        pDsRed2-Peroxi是一种哺乳动物表达载体，其编码Discosoma sp红色荧光蛋白（DsRed2）和过氧化物酶体靶向信号1（PTS1）.PTS1序列与DsRed2的3'末端融合，DsRed变体设计用于更快的成熟和较低的非特异性聚集.PTS1序列编码三肽SKL，其中将DsRed2-PTS1融合蛋白靶向过氧化物酶体基质。
        pDsRed2-Peroxi is a mammalian expression vector that encodes a fusion of Discosoma sp. Red fluorescent protein (DsRed2; 1, 2) and the peroxisomal targeting signal 1 (PTS1). The PTS1 sequence is fused to the 3'-end of DsRed2, a DsRed variant engineered for faster maturation and lower non-specific aggregation. The PTS1 sequence encodes the tripeptide SKL, which targets the DsRed2-PTS1 fusion protein to the matrix of peroxisomes (3–6).
        To drive expression of DsRed2-PTS1, this vector contains the immediate early promoter of cytomegalovirus (PCMV IE). SV40 polyadenylation signals downstream of the DsRed2 gene direct proper processing of the 3'-end of the DsRed2-PTS1 mRNA transcript. Because it encodes DsRed2, a gene variant that uses human-preferred codons (7), the DsRed2-PTS1 transcript is suited for efficient translation in mammalian cells. To further increase the translational efficiency of DsRed2-PTS1, upstream sequences have been converted to a Kozak consensus translation initiation site (8). The vector also contains an SV40 origin for replication in any mammalian cell line that expresses the SV40 T-antigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production. A neomycin resistance cassette—consisting of the SV40 early promoter (PSV40e), the neomycin/kanamycin resistance gene of Tn5 (Neor/Kanr), and polyadenylation signals from the herpes simplex virus thymidine kinase (HSV TK polyA) gene—allow stably transfected eukaryotic cells to be selected using G418 (9). A bacterial promoter (P) upstream of this cassette drives expression of the gene encoding kanamycin resistance in E. coli.
        pDsRed2-Peroxi is designed for fluorescent labeling of peroxisomes. The vector can be introduced into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 (9). The DsRed2-PTS1 fusion (excitation/emission maxima: 558 nm/583 nm) can be detected by fluorescence microscopy and by flow cytometry. Filter sets optimized for detecting DsRed by microscopy are available from Chroma Technology Corporation and Omega Optical Inc. Please see their websites (www.chroma.com and www.omegafilters.com) and the Living Colors® Vol. II User Manual, provided with this vector, for more information. To detect DsRed2-PTS1-expressing cells by flow cytometry, use the instrument’s argon-ion laser to excite the fluorophore at 488 nm and the FL-2 channel to detect the fluorophore’s emission at 583 nm.
        •    Suitable host strains: DH5α, HB101 and other general purpose strains. Single-stranded DNA production requires a host containing an F plasmid such as JM101 or XL1-Blue.
        •    Selectable marker: plasmid confers resistance to kanamycin (50 µg/ml) to E. coli hosts.
        •    E. coli replication origin: pUC
        •    Copy number: ~500
        •    Plasmid incompatibility group: pMB1/ColE1

质粒图谱



质粒序列

LOCUS       Exported                4644 bp ds-DNA    circular SYN 22-10-2015
KEYWORDS    Untitled 7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4644)
  AUTHORS   admin
  TITLE     Direct Submission
  JOURNAL   Exported 2015-10-22  from MiaoLingPlasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..4644
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             613..1287
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /note="DsRed2"
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     polyA_signal    1432..1553
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1560..2015)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2042..2146
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        2148..2505
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      2356..2491
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             2540..3334
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3566..3613
                     /note="HSV TK poly(A) signal"
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      3942..4530
                     /direction=RIGHT
                     /note="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 ccggtcgcca ccatggcctc ctccgagaac gtcatcaccg agttcatgcg cttcaaggtg
      661 cgcatggagg gcaccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      721 ccctacgagg gccacaacac cgtgaagctg aaggtgacca agggcggccc cctgcccttc
      781 gcctgggaca tcctgtcccc ccagttccag tacggctcca aggtgtacgt gaagcacccc
      841 gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
      901 atgaacttcg aggacggcgg cgtggcgacc gtgacccagg actcctccct gcaggacggc
      961 tgcttcatct acaaggtgaa gttcatcggc gtgaacttcc cctccgacgg ccccgtgatg
     1021 cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg
     1081 ctgaagggcg agacccacaa ggccctgaag ctgaaggacg gcggccacta cctggtggag
     1141 ttcaagtcca tctacatggc caagaagccc gtgcagctgc ccggctacta ctacgtggac
     1201 gccaagctgg acatcacctc ccacaacgag gactacacca tcgtggagca gtacgagcgc
     1261 accgagggcc gccaccacct gttcctgaga tcgtacaagt ccaagctgta gcggccgcga
     1321 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     1381 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     1441 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1501 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     1561 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     1621 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     1681 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     1741 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     1801 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     1861 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     1921 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     1981 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     2041 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2101 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     2161 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2221 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2281 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2341 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2401 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2461 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     2521 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     2581 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     2641 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     2701 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     2761 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     2821 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     2881 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     2941 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3001 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3061 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3121 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3181 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3241 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3301 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3361 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     3421 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     3481 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     3541 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     3601 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     3661 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     3721 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     3781 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3841 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3901 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3961 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4021 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4081 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4141 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4201 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4261 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4321 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4381 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4441 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4501 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4561 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4621 gataaccgta ttaccgccat gcat
//
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