pCSG-IBA145 vector

pCSG-IBA145 is a large expression vector with universal features for transient expression of target proteins with an N-terminal Twin-Strep-tag(R) and C-terminal His-tag as well as for generation of stable mammalian cell lines. The vector carries an ampicillin resistance cassette for 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 by SV40 ori. For the former 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 (e.g. HEK293T, COS-1, COS-7). Stable cell lines can be selected by the neomycin resistance gene (NeoR). 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 to the direct cloning of the gene of interest into pCSG-IBA vectors with Esp3I, another option via a so-called entry-vector is possible.