OGS552

Sigma-Aldrich

PSF-TEFI-CEN6/ARS4-URA3 - LOW COPY ORIGIN YEAST 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 5892 bp

复制起点

pUC (500 copies)

肽切割

no cleavage

启动子

Promoter name: TEF1
Promoter activity: constitutive
Promoter type: yeast

菌种筛选

kanamycin

报告基因

none

运输

ambient

储存温度

−20°C

酵母筛选

uracil

一般描述

This is a low copy Saccharomyces cerevisiae yeast plasmid that allows for the plasmid to be maintained in cells at an approximate frequency of 1-2 copies per cell. These plasmids are not typically used for protein production because expression is signficantly lower than using the standard 2 micron origin which has a much higher copy number. This plasmid can still be used when large DNA inserts are required and low levels of expression of genes are needed. The plasmid contains the URA3 selection cassette which allows the plasmid to be maintained in cells that are defective for this gene. Cells carrying the plasmid can then be grown in media that is deficient in uracil.

Promoter Expression Level: This expression vector contains the yeast translation elongation factor 1 promoter. It is the strongest promoter that we provide for expression in Saccharomyces cerevisiae.

应用

Cloning in a gene: PSF-TEFI-CEN6/ARS4-URA3 - LOW COPY ORIGIN YEAST 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.oxfordgenetics.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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