PSF-CMV-PGEM - CMV PGEM MCS PLASMID OGS109-5UG

NACRES:

NA.85

PSF-CMV-PGEM -CMV PGEM MCS plasmid is a versatile cloning vector for the expression of genes in mammalian cells. The molecular cloning vector contains the multiple cloning site (MCS) from the Promega pGEM vector however it has been modified slightly to accommodate some restriction sites in the SnapFast system. These changes are described in the cloning tab. The use of this MCS plasmid instead of the normal SnapFast vector MCS will limit the ability to use some of the inserts that we sell in other plasmid that immediately flank or are inserted within the standard MCS. In CMV PGEM MCS plasmid, these limitations mainly extend to 5 prime tags and signal peptides. Most other inserts should still be compatible with this vector. pGEM is a registered trademark of the Promega Corporation.
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. For this reason we stock a range of other promoters that are compatible with this plasmid and are available on request.
Multiple cloning site notes: This vector contains a different multiple cloning site (MCS) from the standard SnapFast vector. The MCS is derived from pGEM. The order of the restriction sites in this vector are almost the same as in pGEM however the sequence is not. The pGEM MCS has also been modified as follows:

• The normal NotI SbfI and PstI sites have been ablated in this MCS. The NotI site has been moved to the 5 prime of the MCS this allows the use of many of our 5 prime coding sequences and tags.
• An XbaI site has been added to the end of the MCS.
• The BamHI site that is normally found in all SnapFast vectors (in the downstream fusion MCS) has been replaced with a BclI site. BamHI and BclI produce compatible cohesive ends when cleaved although BclI is methylated in this vector and will require growth in a Dam methylase negative bacterial strain (such as JM110 cells) before it can be used.

The ClaI to NheI sites have other functions such as adding peptide tags or IRES elements. The BsgI and BseRI restriction sites cleave within the stop codon in the XbaI site and allow the retrospective fusion of coding sequences. These sites are normally only used on genes that we sell in the main multiple cloning site. This concept is explained in more detail on our website using the drop down box on the homepage and selecting: Fuse a coding sequence to the 3 prime end of a gene already in the SnapFast plasmid.
Sequence
Analysis Note

form	buffered aqueous solution
mol wt	size 4354 bp
Origin of replication	pUC (500 copies)
Peptide cleavage	no cleavage
Promoter	Promoter name: CMV Promoter activity: constitutive Promoter type: mammalian
bacteria selection	ampicillin
reporter gene	none
shipped in	ambient
storage temp.	20°C

RIDADR	NONH for all modes of transport
Flash Point F	Not applicable
Flash Point C	Not applicable