This plasmid is designed to express tagged proteins in E. coli. The plasmid contains a constitutive promoter (OXB20) derived from the region upstream of the E. coli RecA gene. It does not require induction or any additional components for activity. It is the strongest of the bacterial promoters that we provide and this high level of expression can cause expression problems with some proteins with poor solubility. For this reason we sell a range of bacterial promoters with different expression levels (OXB1(low)>OXB20(high)) that can be provided with the peptide tags in this plasmid on request.
About the Peptide Tag:This plasmid contains an Outer Membrane Protein A (OmpA) secretory signal peptide (SP) to allow proteins to be exported from the cytosol. During translocation from the cytosol the signal peptide is removed from the protein by endogenous proteases. This plasmid also contains a secondary Strep protein tag. The sequence of this tag is: WSHPQFEKWe provide a range of dual peptide tag plasmids. This is because some peptide tags provide specific biological properties (e. g., small molecule affinity new epitopes solubility or protein secretion) that are not provided by others.
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 FXa cleavage tag. The protein sequence of the cleavage tag is: IEGR. It cleaves after the Arg residue but can also cleave less frequently at secondary basic sites. Its most common secondary cleave site is between the Gly and Arg residues although the frequency of these events is protein specific.
Promoter Expression Level: This plasmid contains a constitutive bacterial promoter that does not require induction. It is the strongest bacterial promoter we sell and this can cause solubility and expression problems with some proteins. We also offer a range of other bacterial promoters that are compatible with this plasmid and are available on request.
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.
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.
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