OGS511

PSF-CMV-ILUMENA - MAMMALIAN SECRETED LUCIFERASE PLASMID

plasmid vector for molecular cloning

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

Properties

• form: buffered aqueous solution
• mol wt: size 4919 bp
• bacteria selection: ampicillin
• Origin of replication: pUC (500 copies)
• Peptide cleavage: no cleavage
• Promoter: Promoter name: CMV; Promoter activity: constitutive; Promoter type: mammalian
• reporter gene: iLumena luciferase
• shipped in: ambient
• storage temp.: −20°C

Description

General description

A plasmid designed for the expression of secreted luciferase in mammalian cells. The iLumena reporter gene in this vector has been specifically engineered for expression in mammalian cells and yields high levels of luminsceence in the supernatant. The substrate for the luciferase reporter is coelenterazine.

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. The ubiquitin promoter driving reporter gene expression is the strongest endogenous human promoter that we provide for most cell types.

Application

Cloning in a gene: This plasmid contains a gene within the main multiple cloning site (NotI-ClaI). Any plasmid that we sell where the gene is this configuration will be located in the exact same position in relation to the start and stop codon of the gene. The only exceptions to this rule are fusions proteins where the fusion gene may be positioned at the front or end of the MCS to allow gene fusion.

By positioning all of our genes in the same location it allows them to be transferred between plasmids using the same cloning method and restriction sites regardless of the plasmid being used from our product range. Inserting a new gene into this plasmid should be easily possible using a range of standard restriction enzyme sites that flank the gene currently in the vector.

Multiple cloning site notes: In the multiple cloning site there are two important restriction sites called BsgI and BseRI sites. These sites both cut the DNA at the same position and cleave the stop codon of the gene in the multiple cloning site in this plasmid thereby producing a TA overhang. This overhang is compatible with any of our peptide or reporter fusion tag plasmids also cut with either of these enzymes. This allows seamless C-terminal fusions to be made with the gene in this multiple cloning site using a single cloning step from our C-terminal peptide and reporter tag product range. Normally the easiest method is to clone the C-terminal tag from our other plasmid products into this plasmid using BsgI or BseRI and the downstream ClaI restriction site.

BseRI and BsgI sites are non-palindromic and cleave a defined number of bases away from their binding sites. This allows them to cut the upstream stop codon in the gene in this plasmid regardless of the gene sequence.

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 Code: 12 - Non Combustible Liquids
• Flash Point(C): Not applicable