The related phenomena of ColE1 DNA replication control and plasmid incompatibility are analysed in detail. Replication is negatively regulated in different ways by two gene products, RNA I and gene rop-coded polypeptide, while plasmid incompatibility is specified only by RNA I. Thus, copy-number mutants in the RNA I region usually have altered incompatibility and vice versa. RNA I is transcribed from the opposite strand to RNA II; the processed RNA II transcript serves as the primer for DNA replication. The base-pairing interaction between RNA I and RNA II controls DNA synthesis and incompatibility by preventing primer formation. A model is proposed to explain the phenotype of RNA I mutants in terms of the changes they cause in the secondary structure of RNA I or RNA II.