Figure 3. pFlpBtM architecture.
The multi-purpose vector pFlpBtM contains elements necessary for the use as a donor vector for Flp-recombinase mediated cassette exchange (FRT = Flp recombinase target sites) and Tn7-transposition sequences for the generation of recombinant bacmids. FRT-sites flank the region which is integrated into the RMCE locus in the master cell line. It contains the MCS and a downstream PGK-promoter for a selection trap method to screen for recombined cell clones. A larger section of the plasmid is integrated into the bacmids by TN7-based transposition. It contains the promoter region and a gentamicin-resistance gene for the selection of recombinant bacmids. The backbone of pFlpBtM-II additionally contains an Epstein-Barr virus oriP for increased nuclear transport and episomal replication in EBNA positive cell lines.