OGS504

Sigma-Aldrich

PSF-CAG-AMP - CAG PROMOTER PLASMID

plasmid vector for molecular cloning

别名:
expression vector, molecular cloning vector, vector, snapfast vector, plasmid vector, cloning vector, plasmid

形式

buffered aqueous solution

分子量

size 5431 bp

复制起点

pUC (500 copies)

肽切割

no cleavage

启动子

Promoter name: CAG
Promoter activity: constitutive
Promoter type: mammalian

菌种筛选

ampicillin

报告基因

none

运输

ambient

储存温度

−20°C

一般描述

This mammalian expression vector contains the chimeric CAG promoter which is a composite of the CMV enhancer the chicken beta actin promoter (CBA) and the rabbit beta globin intron. This promoter is often used instead of the CMV promoter because in some cell types the CMV promoter can get silenced. To prevent this the CAG promoter contains the CpG island from the CBA promoter to help prevent promoter methylation. The plasmid can be used to drive protein expression in a range of mammalian cell types and we have consistently found that expression levels are equal to CMV in short term experiments in standard cell types.

Promoter Expression Level: The plasmid vector contains the mammalian CAG promoter which is a synthetic composite of the CMV immediate early enhancer followed by the CBA promoter and the rabbit beta globin intron. The Chicken beta actin contains a CpG island that can help to keep the promoter active for longer in stable culture when compared to the CMV promoter.

应用

Cloning in a gene: PSF-CAG-AMP - CAG PROMOTER PLASMID has been designed to be compatible with a range of cloning techniques. The multiple cloning site contains a range of standard commonly used restriction sites for cloning. Using these sites genes can be inserted using standard cloning methods with DNA ligase. Other methods such as ligase independent cloning (LIC) Gibson Assembly InFusionHD or Seamless GeneArt can also be used and because all of our plasmids are based on the same backbone the same method can be used for cloning into all of our catalogue vectors.

Multiple cloning site notes: There are a few important sites within the MCS. These include the NcoI site the XbaI site and the BsgI and BseRI sites. The NcoI site contains a start codon that is immediately downstream of both a Kozak and Shine-Dalgarno ribosomal binding site. These allow for optimal positioning of genes when the start codon is placed in this location. If this is not required and you wish to use a downstream site for gene cloning you can remove the NcoI site by cleaving the plasmid with KpnI.

The XbaI site contains a stop codon. This stop codon is positioned in a specific position in relation to the BsgI and BseRI sites that are immediately downstream. When either BseRI or BsgI cleave the plasmid they produce a TA overhang from the stop codon in the XbaI site that is compatible with all of our peptide tag plasmids cut with the same sites. BseRI and BsgI sites are non-palindromic and cleave a defined number of bases away from their binding site.

Whenever we clone a gene into our multiple cloning site we always position the start and stop codon in the same positions in the MCS. If the start and ends of the genes are not compatible with NcoI and XbaI we extend the sequence to the nearest external sites but keep the start and stop codons locations consistent.

序列

To view sequence information for this product, please visit the product page

分析说明

To view the Certificate of Analysis for this product, please visit www.oxgene.com

storage_class_code

12 - Non Combustible Liquids

闪点(F)

Not applicable

闪点(C)

Not applicable

分析证书

原产地证书 (CofO)

Jin-Gyoung Jung et al.
PLoS genetics, 10(10), e1004751-e1004751 (2014-10-31)
The Notch3 signaling pathway is thought to play a critical role in cancer development, as evidenced by the Notch3 amplification and rearrangement observed in human cancers. However, the molecular mechanism by which Notch3 signaling contributes to tumorigenesis is largely unknown....
Alexander C Cerny et al.
PLoS genetics, 11(10), e1005578-e1005578 (2015-10-29)
Recycling of signaling proteins is a common phenomenon in diverse signaling pathways. In photoreceptors of Drosophila, light absorption by rhodopsin triggers a phospholipase Cβ-mediated opening of the ion channels transient receptor potential (TRP) and TRP-like (TRPL) and generates the visual...
Diana Romero et al.
Carcinogenesis, 37(1), 18-29 (2015-10-28)
Dickkopf-3 (Dkk-3) is a secreted protein whose expression is downregulated in many types of cancer. Endogenous Dkk-3 is required for formation of acini in 3D cultures of prostate epithelial cells, where it inhibits transforming growth factor (TGF)-β/Smad signaling. Here, we...
Geoffrey M Lynn et al.
Nature biotechnology, 33(11), 1201-1210 (2015-10-27)
The efficacy of vaccine adjuvants such as Toll-like receptor agonists (TLRa) can be improved through formulation and delivery approaches. Here, we attached small molecule TLR-7/8a to polymer scaffolds (polymer-TLR-7/8a) and evaluated how different physicochemical properties of the TLR-7/8a and polymer...
技术文章
Our plasmids are composed of both a core vector backbone and a series of DNA components. Our plasmid platform is all built around the same core backbone, which means that the DNA components within each plasmids can be interchanged into each other.
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