The insect system can use the pFastBacDual vector. After inserting one sequence, reverse complementarity is required before inserting another sequence.
Mammalian systems can construct two plasmids and transfect them together, with no requirements for vectors or cells.
The yeast system can construct two plasmids. The pPIC9K plasmid is first transformed into the GS115 strain, and if the protein is expressed, GS115 is prepared into a competent state and then transferred into the pPICZαA plasmid.