Construction of the pBD003 binary vector. Construction procedure of the mini binary vector - pBD003 contains an agrobacterium replication origin (pSa) and an E. coli. replication origin (Ori) for the plasmid replication in agrobacterium and E. coli, respectively. The Cauliflower mosaic virus (CaMV) 35S promoter (35S), the ribozyme (RZ) sequence, and the nopaline synthase terminator (nos) were constructed and inserted between the left border (LB) and right border (RB) on pBD003. The restriction sites frequently used in cloning on the vector backbone were mutated by PCR-mediated mutagenesis. The Kpn I restriction site was created between the CaMV 35S promoter and ribozyme sequence for further cloning or construction.