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ptdTomato-C1

ptdTomato-C1
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  • 产品名称:ptdTomato-C1
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  • 产品展商:XYbscience
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简单介绍
ptdTomato-C1质粒是一个哺乳细胞红色荧光过表达质粒,CMV启动子驱动tdTomato红色荧光蛋白和目的基因融合在一起表达。ptdTomato-C1
产品描述

ptdTomato-C1

  • 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
XY1594 ptdTomato-C1

20μl

¥请加QQ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
质粒大小: 5442bp
质粒标签: N-tdTomato
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Stbl3
培养条件: 37℃,有氧 LB
表达宿主: 哺乳细胞293T
诱导方式: 无须诱导,瞬时表达
5'测序引物: 根据序列设计引物
3'测序引物: Sv40-polyA-R:GAAATTTGTGATGCTATTGC

质粒简介

         ptdTomato-C1质粒是一个哺乳细胞红色荧光过表达质粒,CMV启动子驱动tdTomato红色荧光蛋白和目的基因融合在一起表达。
        ptdTomato-C1 is a mammalian expression vector designed to express a protein of interest fused to the C-terminus of tdTomato. td Tomato is a member of the family of fruit fluorescent proteins (1) derived from the Discosoma sp. red fluorescent protein, DsRed (2). Because the Tomato protein has a tendency to dimerize, the vector was designed with two copies of the Tomato coding region linked together to allow intramolecular dimerization. As a result, each tdTomato RNA transcript encodes a tandem dimer of the Tomato protein (excitation and emission maxima equal 554nm and 581nm, respectively). Expression of td Tomato as a tandem dimer prevents the fused protein of interest from being forced into a dimeric complex. Fusions that retain the fluorescence properties of tdTomato can be monitored by flow cytometry and localized by fluorescence microscopy. The multiple cloning site (MCS) in ptdTomato-C1 is positioned downstream of the tdTomato coding sequence. A Kozak consensus sequence, located immediately upstream of the tdTomato coding sequence, enhances translational efficiency in eukaryotic cells (3). SV40 polyadenylation signals downstream of the tdTomato coding sequence and the MCS direct proper processing of the 3' end of the tdTomato (or fusion gene) mRNA. The vector backbone contains an SV40 origin for replication in mammalian cells expressing the SV40 large T antigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production. This vector also has a neomycin resistance cassette (Neor) that allows G418 selection of stably transfected eukaryotic cells. This cassette consists of the SV40 early promoter, the Tn5 neomycin/kanamycin resistance gene, and polyadenylation signals from the herpes simplex virus thymidine kinase (HSV TK) gene. A bacterial promoter upstream of the cassette confers kanamycin resistance in E. Coli.

质粒图谱


质粒序列

LOCUS       Exported                5442 bp ds-DNA     circular SYN 01-SEP-2016
DEFINITION  synthetic circular DNA
KEYWORDS    Untitled 8
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5442)
  TITLE     Direct Submission
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..5442
                     /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..2040
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /note="tdTomato"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    2041..2106
                     /note="MCS"
                     /note="multiple cloning site"
     polyA_signal    2230..2351
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2358..2813)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2840..2944
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        2946..3303
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      3154..3289
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             3338..4132
                     /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    4364..4411
                     /note="HSV TK poly(A) signal"
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4740..5328
                     /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 ccatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag
      661 gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
      721 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
      781 ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac
      841 cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc
      901 gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac
      961 ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta
     1021 atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc
     1081 gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg
     1141 gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg
     1201 gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag
     1261 cgctccgagg gccgccacca cctgttcctg gggcatggca ccggcagcac cggcagcggc
     1321 agctccggca ccgcctcctc cgaggacaac aacatggccg tcatcaaaga gttcatgcgc
     1381 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
     1441 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     1501 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg
     1561 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg
     1621 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg
     1681 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc
     1741 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
     1801 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac
     1861 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
     1921 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1981 tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag
     2041 tccggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg
     2101 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     2161 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2221 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2281 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2341 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2401 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2461 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2521 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2581 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2641 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2701 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2761 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     2821 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2881 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2941 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3001 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3061 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3121 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3181 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3241 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3301 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3361 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3421 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3481 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     3541 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3601 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3661 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3721 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3781 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3841 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     3901 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3961 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4021 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4081 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4141 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4201 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4261 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     4321 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4381 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4441 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     4501 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     4561 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     4621 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4681 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4741 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4801 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4861 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4921 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4981 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5041 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5101 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5161 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5221 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5281 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5341 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5401 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
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