半岛bd体育手机客户端 说明
一般描述
The purpose of this expression vector is the expression of genes inSaccharomyces cerevisiaeusing selective removal of histidine from the media for cell selection. The plasmid contains a gene that is an essential part of the histidine synthesis pathway. By transforming the plasmid into yeast cells that are deficient in this gene and growing the cells on media that does not contain histidine cells can be selected that contain the plasmid. This is one of a range of metabolic selection genes that are available for yeast. Others include tryptophan leucine and uracil. We also provide plasmids using more standard selection methods such as puromycin and hygromycin.
Promoter Expression Level:This snapfast vector contains the yeast translation elongation factor 1 promoter. It is the strongest promoter that we provide for expression inSaccharomyces cerevisiae.
应用
Cloning in a gene:PSF-TEFI-HIS3 - HISTIDINE YEAST SELECTION 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
基本信息
NACRES | NA.85 |
半岛bd体育手机客户端 性质
形式 | buffered aqueous solution |
分子量 | size 6800 bp |
复制起点 | 2Micron 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℃ |
安全信息
储存分类代码 | 12 - Non Combustible Liquids |
闪点(F) | Not applicable |
闪点(C) | Not applicable |
Sigma-Aldrich