Cryptococcus neoformans is an opportunistic fungal pathogen responsible for serious meningitis. Although many useful molecular tools have been developed for its study, there are currently few inducible promoters available for general use. To address this need, we have constructed expression plasmids incorporating upstream elements of the C. neoformans copper transporter gene CTR4, and tested them in C. neoformans serotypes A and D. In response to copper deprivation, these plasmids mediate high-level expression of a reporter protein. This expression can be completely repressed using physiologically low concentrations of copper. Notably, this new family of copper-sensing promoters demonstrates excellent expression in serotype A, contrasting with other available promoters. These plasmids therefore offer efficient and regulated expression for both serotypes A and D, and should be valuable tools for the C. neoformans research community. Copyright © 2004 John Wiley & Sons, Ltd.