Description_x000D_ General description_x000D_ PSF-CMV-CMV-SBFI - dual CMV expression plasmid is a versatile expression vector for the production of two proteins under the control of two separate CMV promoter expression cassettes in mammalian cells. This vector also contains a Kanamycin resistance cassette for growth and maintenance in E. coli._x000D_ Promoter Expression Level: This plasmid vector 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._x000D_ Application_x000D_ First multiple cloning site notes: There is start codon in the NcoI site that can be removed by digestion with KpnI if required. The first MCS for gene insertions extends from NotI to XbaI`But the Shine-Dalgarno sequences and Kozak sequences are aligned with the start codon in the NcoI restriction site. The ClaI to NheI sIt`s have other functions such as adding peptide tags or IRES elements. The BsgI and BseRI restriction sIt`s cleave within the stop codon in the XbaI site and allow the retrospective fusion of coding sequences. These sIt`s are normally only used on genes that we sell in the main multiple cloning site._x000D_ Second multiple cloning site notes: The second MCS in PSF-CMV-CMV-SBFI - dual CMV expression plasmid extends from PspOMI to SpeI. This MCS contains a Kozak sequence immediately upstream of the start codon in the PciI restriction site. We insert the start codons of our genes into here where possible._x000D_ This MCS has been designed to be compatible with genes that are within our main MCS by using enzymes sIt`s in the same order that produce compatible cohesive ends that can be ligated together. This allows genes to be transferred from the main MCS (NotI to NheI) into the second MCS (PspOMI to SpeI) is required. Enzymes that are compatible between these two MCSs include:_x000D_ • PspOMI compatible with NotI_x000D_ • ScaI compatible with EcoRV (Blunt)_x000D_ • SalI compatible with XhoI_x000D_ • PciI compatible with NcoI and BspHI_x000D_ • AclI compatible with ClaI_x000D_ • SpeI compatible with XbaI and NheI Avr2_x000D_ Sequence_x000D_ Quick-reference Plasmid Map_x000D_ Please select the file type you require. For reference most cloning programs will import a .gb (Genbank) file and will show all of the plasmids features automatically when downloaded and imported._x000D_ Genebank Vector Sequence File_x000D_ FASTA Vector Sequence File_x000D_ Full Plasmid Map_x000D_ Analysis Note_x000D_ To view the Certificate of Analysis for this product, please visit www.oxfordgenetics.com._x000D_ Other Notes_x000D_ Looking for more vector options to move your experiments forward faster? Consider a custom cloning vector designed and built by Oxford Genetics™. Find out more at Oxford Genetics - Sigma's partner for cloning and expression vectors for molecular biology and synthetic biology applications._x000D_ Legal Information_x000D_ Oxford Genetics is a trademark of Oxford Genetics Ltd