Transformed colonies were verified by PCR with the abovementioned primers. The PsaD–ble cassette was extracted with KpnI and XhoI restriction enzymes from extracted plasmids using the Bioneer AccuPrep Plasmid Mini extraction kit. Restricted PsaD–ble was ligated into KpnI–XhoI restricted pBS–pds S or Land transformed into E. coli DH5α. Transformed colonies were verified by restriction assay using EcoRI and BglII and DNA
sequencing.