Schematic of the expression plasmid CD19R/HyTK-pMG.

This DNA vector was used to genetically modify primary human cytolytic T cells to coexpress the chimeric immunoreceptor specific for CD19, termed CD19R, and the bifunctional gene HyTK. Regulatory elements include the promoters hEFl-1 and hCMV-1A driving the expression of CD19R and HyTK genes, respectively. The polyadenylation signals from SV40 and bovine growth hormone are included after the termination codons of CD19Rand HyTK, respectively. The synthetic promoter EM7 drives the prokaryotic expression of hygromycin to select for bacterial drug resistance. The plasmid was linearized at the uniquePacI site (at base pair position 3643) prior to use in electroporation.