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pCSG-IBAwt2 is a large expression vector with universal features for transient expression and for the generation of stable mammalian cell lines. The vector carries an ampicillin resistance cassette for the selection of transformed E. coli cells and ColE1 origin for a high plasmid copy number. Extrachromosomal replication in mammalian cells could occur either by origin of replication from Epstein-Barr Virus (oriP) or SV40 ori. For origin of replication from oriP, the vector provides the EBNA-1 gene, and for the latter, the cell line has to be latently infected with SV40 or express the SV40 large T antigen, such as HEK293T, COS-1, COS-7.
The expressed protein is transferred into the medium due to the BM40 secretory signal peptide. During translocation from the cytosol, the signal peptide is removed from the protein by endogenous proteases.
The neomycin resistance gene (NeoR) can select stable cell lines. In addition, the human cytomegalovirus (CMV) immediate-early promoter enables a high-level expression in a wide range of mammalian cells. Please note that cloning into pCSG-IBA acceptor vectors compulsorily requires the restriction enzyme Esp3I since no other MCS for the integration of a gene of interest is available. Besides the direct cloning of the gene of interest into pCSG-IBA vectors with Esp3I, another option via an Entry Vector is possible.
