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pHcRed1-C1

pHcRed1-C1
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  • 产品名称:pHcRed1-C1
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简单介绍
pHcRed1-C1质粒是一个哺乳细胞红色荧光蛋白表达载体,CMV IE启动子驱动HcRed1红色荧光蛋白基因和目的基因融合表达。pHcRed1-C1
产品描述

pHcRed1-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
XY1393 pHcRed1-C1

100μl菌种冻干粉

¥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 IE promoter
复制子: pUC ori,F1 ori,SV40 ori
终止子: SV40 early polyA signals
质粒分类: 哺乳细胞质粒;哺乳荧光质粒;哺乳红色质粒
质粒大小: 4698bp
质粒标签: N-HcRed1
原核抗性: 卡那霉素Kan(50μg/μl)
筛选标记: 新霉素Neo/G418
克隆菌株: DH5α等大肠杆菌
培养条件: 37℃,有氧,LB
表达宿主: 293T等哺乳细胞
培养条件: 37℃,5%CO2
诱导方式: 无需诱导,组成型表达
5'测序引物: CMV-F:CGCAAATGGGCGGTAGGCGTG
3'测序引物: Sv40-polyA-R:GAAATTTGTGATGCTATTGC
备注: 高拷贝

质粒简介

      pHcRed1-C1质粒是一个哺乳细胞红色荧光蛋白表达载体,CMV IE启动子驱动HcRed1红色荧光蛋白基因和目的基因融合表达。
      pHcRed1-C1 is a mammalian expression vector designed to express a protein of interest fused to the C-terminus of the far-red fluorescent protein HcRed1. pHcRed1-C1 can be used to monitor gene expression and protein localization in vivo or as a cotransfection marker; the unmodified vector will express HcRed1 in mammalian cells. HcRed1 was generated by mutagenesis of a non-fluorescent chromoprotein from the reef coral Heteractis crispa . The coding sequence for HcRed1 is human codon-optimized for higher expression in mammalian cells . Genes cloned into the multiple cloning site (MCS) downstream of the HcRed1 coding sequence are expressed as fusions to the C-terminus of HcRed1. The MCS in pHcRed1-C1 is positioned between the HcRed1 coding sequence and the SV40 polyadenylation signal (SV40 poly A). Genes cloned into the MCS will be expressed as fusions to the C-terminus of HcRed1 if they are in the same reading frame as HcRed1 and there are no intervening stop codons. A Kozak consensus sequence upstream of HcRed1 increases the translation efficiency in eukaryotic cells . SV40 poly A signals downstream of the MCS direct proper processing of the 3' end of mRNA transcripts. The vector also contains an SV40 origin for replication in mammalian cells expressing 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 (Neor)—consisting of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene—allows stably transfected eukaryotic cells to be selected using G418. A bacterial promoter upstream of this cassette expresses kanamycin resistance in E. coli. pHcRed1-C1 can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 .

质粒图谱



质粒序列

LOCUS       Exported                4698 bp ds-DNA     circular SYN 22-AUG-2017
DEFINITION  synthetic circular DNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4698)
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 2017 from MiaoLingPlasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..4698
                     /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"
     CDS             613..1296
                     /codon_start=1
                     /product="far-red fluorescent variant of Heteractis crispa 
                     chromoprotein"
                     /label=HcRed1
                     /note="mammalian codon-optimized"
                     /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH
                     VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT
                     AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL
                     KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD
                     LPEKAN"
     misc_feature    1297..1362
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1486..1607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1614..2069)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2096..2200
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2202..2559
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2410..2545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2594..3388
                     /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    3620..3667
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3996..4584
                     /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 ccggtcgcca ccatggtgag cggcctgctg aaggagagta tgcgcatcaa gatgtacatg
      661 gagggcaccg tgaacggcca ctacttcaag tgcgagggcg agggcgacgg caaccccttc
      721 gccggcaccc agagcatgag aatccacgtg accgagggcg cccccctgcc cttcgccttc
      781 gacatcctgg ccccctgctg cgagtacggc agcaggacct tcgtgcacca caccgccgag
      841 atccccgact tcttcaagca gagcttcccc gagggcttca cctgggagag aaccaccacc
      901 tacgaggacg gcggcatcct gaccgcccac caggacacca gcctggaggg caactgcctg
      961 atctacaagg tgaaggtgca cggcaccaac ttccccgccg acggccccgt gatgaagaac
     1021 aagagcggcg gctgggagcc cagcaccgag gtggtgtacc ccgagaacgg cgtgctgtgc
     1081 ggccggaacg tgatggccct gaaggtgggc gaccggcacc tgatctgcca ccactacacc
     1141 agctaccgga gcaagaaggc cgtgcgcgcc ctgaccatgc ccggcttcca cttcaccgac
     1201 atccggctcc agatgctgcg gaagaagaag gacgagtact tcgagctgta cgaggccagc
     1261 gtggcccggt acagcgacct gcccgagaag gccaactccg gactcagatc tcgagctcaa
     1321 gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg
     1381 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     1441 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     1501 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1561 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     1621 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     1681 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     1741 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     1801 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     1861 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     1921 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     1981 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2041 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2101 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2161 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     2221 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2281 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2341 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2401 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2461 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2521 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     2581 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2641 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2701 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2761 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2821 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2881 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     2941 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3001 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3061 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3121 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3181 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3241 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3301 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3361 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3421 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3481 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3541 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     3601 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     3661 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3721 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     3781 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     3841 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3901 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3961 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4021 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4081 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4141 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4201 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4261 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4321 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4381 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4441 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4501 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4561 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4621 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4681 cgtattaccg ccatgcat
//
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