Generating and studying transgenes is a powerful technique for studying gene function in vivo. An expression vector capable of producing multiple proteins from a single transcript in equimolar amounts would be a useful molecular tool. Such a vector would allow for the co-expression of multiple genes, resulting in distinct protein products that retain their native function. This tool could be used as a means to identify successfully transfected embryos, provide a way to target proteins to various subcellular locations, drive expression of multiple genes in equimolar amounts, and facilitate production of stably transfected ascidian lines. Here, I developed a bicistronic vector using CHYSEL (cis-acting hydrolase element) technology and demonstrate successful expression of discrete protein products in equimolar amounts. Further, the capabilities of CHYSEL were compared to those of an operon system and to the single-gene, two-plasmid approach. CHYSEL was shown to out perform both the operon and two-plasmid approach in regards to the production of discrete protein products in equivalent amounts. Adoption of CHYSEL technology will supply our lab with a useful molecular biology tool, providing the ability to perform certain experiments previously unavailable to us.