Streptomyces linear plasmids start replication at centrally located loci, usually consisting of iterons and adjacent rep genes. Here, we identified four new replication loci from Streptomyces linear plasmids. A discontinuous locus, consisting of two genes and iterons separated by two nonessential genes, was required for replication of pRL2 in both linear and circular modes. A temperature-sensitive plasmid, pRL4, contained a replication locus, a noncoding sequence and a SAP1.35-like gene. A telomere-adjacent locus, another noncoding sequence and SAP1.1-like gene, was identified for replication of the large plasmid pFRL2. The replication locus of pSHK1 consisted of SCP1-rep-like genes and iterons. These results indicate an unexpected variety of components, positions and combinations of replication loci among Streptomyces linear plasmids.