Abstract: This work is focused on preparation of the plant transformation vector pZP6 containing “self-excision” Cre/loxP system. The T-DNA of binary vector consists of the cre recombinase gene driven by the Arabidopsis DLL promoter and the nptII expression unit flanked by two loxP sites in direct orientation. The gus reporter gene controlled by the double CaMV 35S promoter was placed out of the loxP embedded DNA. To confirm functionality of the Cre/loxP system, the pZP6 was analyzed for correct removal of the loxP embedded sequence in E. coli. The pZP6 was transformed into two bacterial strains A. tumefaciens AGLO and LBA 4404. Its stability in agrobacteria was evaluated by restriction analyses.