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pDsRed-Express-DR

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  • 产品名称:pDsRed-Express-DR
  • 产品型号:
  • 产品展商:HZbscience
  • 产品文档:无相关文档
  • 发布时间:2018-08-23
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简单介绍
pDsRed-Express-DR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pDsRed-Express-DR后请短暂离心,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。
产品描述

pDsRed-Express-DR载体基本信息

载体名称: pDsRed-Express-DR
质粒类型: 无启动子载体;荧光报告载体
高拷贝/低拷贝: 高拷贝
克隆方法: 限制性内切酶,多克隆位点
启动子: 无启动子
载体大小: 4240 bp
5' 测序引物及序列: 5'd[GTACTGGAACTGGGGGGACAG]
3' 测序引物及序列: --
载体标签: 红色荧光蛋白DsRed-Express-DR
载体抗性: Kanamycin.html' target='_blank'>卡那霉素
筛选标记: 新霉素(Neomycin)
克隆菌株: TOP10, DH5α, HB101
宿主细胞(系): 真核细胞
备注: pDsRed-Express-DR载体不含启动子,将待研究的目的启动子插入MCS区,用于研究启动子及顺式转录元件在细胞内的作用水平;
DsRed-Express-DR与DsRed-Express相比,半衰期短,适用于需要报告基因逆转速度快的研究领域。
产品目录号: 632423
稳定性: 瞬表达 或 稳表达
组成型/诱导型: --
病毒/非病毒: 非病毒

pDsRed-Express-DR载体质粒图谱和多克隆位点信息

pDsRed-Express-DR载体图谱



pDsRed-Express-DR 多克隆位点

pDsRed-Express-DR 载体特征1
pDsRed-Express-DR 载体特征2

pDsRed-Express-DR载体简介

pDsRed-Express-DR载体描述 pDsRed-Express-DR is a promoterless vector that encodes DsRed-Express-DR, a destabilized variant of Discosoma sp. red fluorescent protein (DsRed). In contrast to the original protein, which is extremely stable, DsRed-Express-DR has a short half-life, making it well suited for studies that require rapid reporter turnover. Though its geneology can be traced to wild-type DsRed (1), DsRed-Express-DR is directly derived from DsRed-Express, a DsRed variant engineered for rapid fluorescence development (2, 3). DsRed-Express contains nine amino acid substitutions (listed on page 2), which improve the protein’s solubility and decrease the time from transfection to detection of red fluorescence (2). The substitutions also reduce the level of residual green emission (2). The destabilized variant, DsRed-Express-DR, was constructed by fusing the C-terminus of the protein to amino acid residues 422–461 of mouse ornithine decarboxylase (MODC), one of the most shortlived proteins in mammalian cells (4). This region of MODC contains a PEST sequence that targets the protein for degradation, resulting in rapid protein turnover (4, 5). The DsRed-Express-DR gene is human codon-optimized for high expression in mammalian cells (6), and sequences upstream of the gene have been converted to a Kozak consensus translation initiation site (7) to enhance translation efficiency in eukaryotic cells.
pDsRed-Express-DR can be used to monitor transcription from different promoters and promoter/ enhancer combinations inserted into the multiple cloning site (MCS), located upstream of the DsRed-Express-DR coding sequence. SV40 polyadenylation signals downstream of the DsRed- Express-DR gene direct proper processing of the 3' end of the DsRed-Express-DR 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. A neomycin-resistance cassette (Neor) allows stably transfected eukaryotic cells to be selected using G418. This cassette consists 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. A bacterial promoter upstream of the cassette expresses kanamycin resistance in E. coli.


DsRed-Express-DR can be used as an in vivo reporter of gene expression. Because of its rapid turnover rate, its expression from a promoter of interest provides a more accurate assessment of the promoter's activity over time than does the more stable DsRed-Express. Promoter/enhancer elements should be inserted into the MCS upstream of the DsRed-Express-DR coding sequence. Without the addition of a functional promoter, this vector will not express DsRed-Express-DR. The recombinant pDsRed-Express-DR vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 (8). pDsRed-Express-DR载体含有以下元件: MCS: 12–89
 Destabilized Discosoma sp. Red Fluorescent Protein (DsRed-Express-DR) gene
Kozak consensus translation initiation site: 90–100
Start codon (ATG): 97–99; Stop codon: 904–906
CGC→GCC (Arg-2 to Ala) mutation: 100–102
AAG→GAG (Lys-5 to Glu) mutation: 109–111
AAC→GAC (Asn-6 to Asp) mutation: 112–114
ACC→TCC (Thr-21 to Ser) mutation: 157–159
CAC→ACC (His-41 to Thr) mutation: 217–219
AAC→CAG (Asn-42 to Gln) mutation: 220–222
GTG→GCC (Val-44 to Ala) mutation: 226–228
TGC→TCC (Cys-117 to Ser) mutation: 445–447
ACC→GCC (Thr-217 to Ala) mutation: 745–747
Mouse ornithine decarboxylase PEST sequence: 784–906
 SV40 early mRNA polyadenylation signal
Polyadenylation signals: 1059–1064 & 1088–1093
mRNA 3' ends: 1097 & 1109
 f1 single-strand DNA origin: 1156–1611
(Packages noncoding strand of DsRed-Express-DR.)
 Ampicillin resistance (β-lactamase) promoter
–35 region: 1673–1678; –10 region: 1696–1701
Transcription start point: 1708
 SV40 origin of replication: 1952–2087
 SV40 early promoter
Enhancer (72-bp tandem repeats): 1783–1856 & 1857–1928
21-bp repeats: 1932–1952, 1953–1973 & 1975–1995
Early promoter element: 2008–2014
Major transcription start points: 2004, 2042, 2048 & 2053 Kanamycin/neomycin resistance gene
Neomycin phosphotransferase coding sequences:
Start codon (ATG): 2136–2138; stop codon: 2928–2930
G→A mutation to remove Pst I site: 2318

C→A (Arg→Ser) mutation to remove BssH II site: 2664
 Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signal
Polyadenylation signals: 3166–3171 & 3179–3184
 pUC plasmid replication origin: 3515–4158 Propagation in E. Coli Suitable host strains: DH5α, HB101 and other general purpose strains. Single-stranded DNA production requires
a host containing an F plasmid such as JM109 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/Col E1 Excitation and emission maxima of DsRed-Express Excitation maximum = 557 nm
 Emission maximum = 579 nm 

pDsRed-Express-DR载体序列

hz-8232R FRMD1  FRMD1蛋白抗体
hz-8233R FRMD2/FRMPD1  FRMD2蛋白抗体
hz-8234R FRMD3  FRMD3蛋白抗体
hz-8235R FRMD4A  FRMD4A蛋白抗体
hz-8236R FRMD4B  FRMD4B蛋白抗体
hz-8237R FRMD5  FRMD5蛋白抗体
hz-8238R FRMD6  FRMD6蛋白抗体
hz-8239R FRMD7  FRMD7蛋白抗体
hz-8240R FRMD8  FRMD8蛋白抗体
hz-8241R CTDSPL  CTDSPL蛋白抗体
hz-8242R CTDSPL2  CTDSPL2蛋白抗体
hz-8243R CUEDC1  CUE结构域蛋白1抗体
hz-8244R CA XI/Carbonic Anhydrase XI  碳酸酐酶11抗体
hz-8245R Alpha casein  α酪蛋白抗体
hz-8246R CADM2/IGSF4D/Cell adhesion molecule 2  细胞粘附分子2
hz-8247R COBL/Cordon bleu homolog  耳聋-甲状腺肿综合征相关COBL蛋白抗体
hz-8248R CUTA  乙酰胆碱酯酶相关蛋白抗体
hz-8249R DCAKD  脱磷酸辅酶A结构域蛋白抗体
hz-8250R DCST1  DCST1蛋白抗体
hz-8251R DCUN1D4  DCUN1D4蛋白抗体

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