Abstract A plasmid, pBP614 (5.647 kb), has been isolated from the New Zealand Bacillus popilliaestrain 17 and characterized by physical mapping, cloning, and sequencing. An open reading frame was found which could encode a protein with homology to the replication (Rep) proteins of rolling-circle plasmids. The predicted B. popilliaeRep protein shows closest homology to Rep proteins from Bacillusplasmids of the pC194 family. Sequences homologous to plus- and minus-strand replication origins were found. The minus-strand origin shows similarities to the pal-T type family. Taken together, these observations suggest that pBP614 is a rolling-circle plasmid, the first such plasmid characterized from B. popilliae.