PSF-CMV-PURO-NH2-MBP-TEV - N-TERMINAL MBP TAG MAMMALIAN PLASMID OGS3195-5UG

NACRES:

NA.85

This plasmid is designed to express tagged proteins in mammalian cells either by transient transfection or by creating stable cell lines. It contains a puromycin resistance expression cassette using the human Ubiquitin promoter to drive expression and allow for the selection of cells containing the plasmid.
About the Peptide Tag:This plasmid contains an n-terminal Maltose Binding Protein (MBP) affinity tag that can be fused to a gene of interest to allow protein detection and/or purification. The sequence of the tag is:EEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSS.
About the Cleavage Tag:This plasmid also encodes a protease cleavage site that is designed to be positioned between your gene of interest and the tag to allow the removal of the tag following protein purification or isolation. This plasmid contains a TEV cleavage tag. The protein sequence of the cleavage tag is: ENLYFQG. Cleavage occurs between the Glu and Gly residues. TEV is often reported to have better specificity for its recognition site compared to EKT Thrombin or Faxtor Xa.
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.
Sequence
Analysis Note

form	buffered aqueous solution
mol wt	size 7274 bp
conjugate	maltose-binding protein (MBP) tagged
Origin of replication	pUC (500 copies)
Peptide cleavage	TEV
Peptide tag location	N-terminal
Promoter	Promoter name: CMV Promoter activity: constitutive Promoter type: mammalian
bacteria selection	kanamycin
mammalian cells selection	puromycin
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