OGS114

Sigma-Aldrich

PSF-CMV-SV40 ORI SBFI - SV40 ORIGIN PLASMID

plasmid vector for molecular cloning

别名:
expression vector, molecular cloning vector, vector, snapfast vector, plasmid vector, cloning vector, plasmid
NACRES:
NA.85

形式

buffered aqueous solution

分子量

size 4436 bp

复制起点

pUC (500 copies)

肽切割

no cleavage

启动子

Promoter name: CMV
Promoter activity: constitutive
Promoter type: mammalian

菌种筛选

kanamycin

报告基因

none

运输

ambient

储存温度

−20°C

一般描述

A versatile cloning vector for the expression of genes in mammalian cells. This vector also contains the origin of replication from Simian Virus 40 (SV40) which enables it to be replicated in cells that express the SV40 large T antigen (such as 293T cells). The SV40 origin can be used to increase the level of transcription in cells that are transfected with the plasmid because the plasmid is replicated. It is also reported that by using selectable markers in conjunction with the SV40 origin stable cell lines can be produced. However we have not validated it for this purpose. We currently sell a range of antibiotic selection genes that can be used for this purpose in a range of positions within our vector system. In this vector the SV40 origin is inserted within the SbfI site and is flanked by two SbfI sites.

Promoter Expression Level: This plasmid contains the mammalian CMV promoter to drive gene expression. We have tested all of our mammalian promoters in a range of cell types and CMV is consistently the strongest in those we have studied. However there are many reports of the CMV promoter demonstrating silencing by methylation in long-term culture.

应用

Cloning in a gene: This 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...

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