OGS112

PSF-CMV-SV40E SBFI - SV40 ENHANCER PLASMID

plasmid vector for molecular cloning

• Synonym(s): cloning vector, expression vector, molecular cloning vector, plasmid, plasmid vector, snapfast vector, vector
• UNSPSC Code: 12352200
• NACRES: NA.85

Properties

• form: buffered aqueous solution
• mol wt: size 4503 bp
• peptide cleavage: no cleavage
• reporter gene: none
• shipped in: ambient
• storage temp.: −20°C

Description

General description

A versatile cloning vector for the expression of genes in mammalian cells. This vector also contains the enhancer from Simian Virus 40 (SV40) inserted into the SbfI site. The enhancer is downstream of the terminators in the SnapFast vector but it is upstream of the 3 prime insulator sequence. In this vector the presence of the enhancer does not significantly enhance expression however this is likely to be due to the high level of expression mediated by the CMV promoter which already contains an enhancer region. The SV40 enhancer is provided in this vector backbone to enable it to be transferred into any vectors with weaker promoters to increase expression. If you would like the SV40 enhancer transferring into any of our other vectors please contact us for more information.

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.

Application

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.

Sequence

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

Analysis Note

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

Safety Information

• Storage Class: 12 - Non Combustible Liquids
• flash_point_f: Not applicable
• flash_point_c: Not applicable