pSF-CMV-pGem (OG109) CMV pGEM MCS Plasmid

Product Code: OG109R1
Product Code: OG109C1
Product Code: OG109G1

Product Description

pGem multiple cloning site CMV expression vector. This vector is modified to contain the restriction enzyme pattern from the cloning plasmid pGEM.

Plasmid Information

Product Name: pSF-CMV-pGem

Product Code: OG109

Size (bp): 4354 bp

Bacterial Antibiotic Selection: AmpR

Origin and Compatibility: pUC high copy derived from pBR322

Bacterial Copy Number: 500-700 per cell

Promoter: Cytomegalovirus (CMV) immediate early promoter

Plasmid Purpose:

A versatile cloning vector for the expression of genes in mammalian cells. This 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 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 this vector 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.

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Transcription Termination:

This plasmid contains three alternative transcription terminators for mammalian bacterial and bacteriophage (T7) expression. This means that only the promoter needs to be changed to alter the expression system you are using. We sell multiple promoters that can be used in each of these systems. The presence of each terminator does not reduce expression in the alternative systems.

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.

Intellectual Property Status

This product is part of our SnapFastâ„¢ plasmid range, for more information on the intellectual property status of this plasmid please click here. For more information on the terms of our licences please click here.