The replication behaviour of a number of ARS1-based plasmids was investigated on propagation in Saccharomyces cerevisiae grown with either glucose or galactose as carbon source. Growth on galactose results in reduced plasmid stability, as well as in reduced replication efficiency, when the entire 1.5-kb TRP1-ARS1 fragment is present on a plasmid. The galactose sensitivity is mediated by a 0.13-kb fragment harbouring part of the GAL3 promoter. This fragment exerts its effect when situated either 5' or 3' to the ARS core consensus at distances up to 0.9 kb. The endogenous 2 microns plasmid remained unaffected by the choice of carbon source.