Nucleotide sequence of an essential region for autonomous replication of cloned yeast mitochondrial DNA

A 341 bp sequence from yeast mtDNA was cloned, which consisted of an upstream 98 bp AT stretch and a downstream 206 bp AT stretch separated by a single 37 bp GC cluster. Cleavage of this GC cluster did not cause loss of the autonomously replicating function of this sequence. The recloned first 98 bp AT stretch was incapable of replication, while the recloned 206 bp AT stretch could replicate. We were able to confine an essential sequence for autonomous replication within a 186 bp AT stretch. Sequencing data revealed a sequence of ATATAAAT and stem and loop structures within the AT stretch.