Analysis Note

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

Application

Cloning in a gene: PSF-EF1-PURO - ELONGATION FACTOR 1 ALPHA PROMOTER PUROMYCIN 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.

General description

Here the puromycin resistance gene is under transcription regulation of the human Elongation Factor 1 alpha promoter giving long term expression that can be useful for selection of transfected mammalian cells to make stable cell linesPromoter Expression Level: Molecular cloning vector contains the human Elongation factor 1 promoter to drive protein expression. The EF1 alpha promoter contains a large intron that helps to maintain expression of the promoter in long term culture. Although it shows signficantly lower expression in most cell types compared to the CMV promoter by transient transfection the promoter can maintain activity for longer in many cell types when making stable cell lines.

Sequence

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