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pmirGLO哺乳报告质粒

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基本信息

别名:

pmir-GLO

启动子:

PGK

复制子:

pUC

终止子:

SV40 poly(A) signal

质粒分类:

哺乳细胞,信号通路报告载体,  双萤光素酶报告基因载体

质粒大小:

7350bp

原核抗性:

Amp

真核抗性:

G418

克隆菌株:

DH5a

培养条件:

37度

表达宿主:

哺乳细胞

诱导方式:

无须诱导,瞬时表达

5'测序引物:

PmirGLO-F:GACGAGGTGCCTAAAGGACT

3'测序引物:
根据序列设计引物

备注:

信号报告空载体


质粒属性

质粒宿主:

哺乳细胞

质粒用途:

信号报告

片段类型:

miRNA,UTR

片段物种:

空载体

原核抗性:

Amp

真核抗性:

G418

荧光标记:

Fluc,Rluc


质粒简介

The pmirGLO Dual-Luciferase miRNA Target Expression Vector(a–d) is designed to quantitatively evaluate microRNA (miRNA) activity by the insertion of miRNA target sites 3´ of the firefly luciferase gene (luc2 ). These target sites can be introduced by cloning putative miRNA binding sites alone, or the 3´ untranslated region (UTR) of a gene of interest, to study the influence of these sites on transcript stability and activity. Firefly luciferase is the primary reporter gene; reduced firefly luciferase expression indicates the binding of endogenous or introduced miRNAs to the cloned miRNA target sequence. This vector is based on Promega dual-luciferase technology, with firefly luciferase (luc2 ) used as the primary reporter to monitor mRNA regulation and Renilla luciferase (hRluc-neo) acting as a control reporter for normalization and selection. This vector contains the following features:

  • Human phosphoglycerate kinase (PGK) promoter provides low translational expression, which is advantageous when reduction of signal is the desired response. The PGK promoter is a nonviral universal promoter, which functions across cell lines (yeast, rat, mouse and human).

  • Firefly luciferase reporter gene (luc2 ) inversely reports miRNA activity in mammalian cells.

  • Multiple cloning site (MCS) is located 3´ of the firefly luciferase reporter gene (luc2 ).

  • Humanized Renilla luciferase-neomycin resistance cassette (hRluc-neo) is used as a control reporter for normalization of gene expression and stable cell line selection.

  • Ampr gene allows bacterial selection for vector amplification.

  • SV40 late poly(A) signal sequence is positioned downstream of luc2 to provide efficient transcription termination and mRNA polyadenylation.

  • Synthetic poly(A) signal/transcription stop site.


质粒图谱

image.png

质粒序列

质粒序列请下载:

pmirGLO哺乳报告质粒.txt


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