Various short autonomously replicating sequences from the yeast seemingly without canonical consensus.

Eukaryotic autonomously replicating sequences (ARSs) are composed of three domains, A, B, and C. Domain A is comprised of an ARS consensus sequence (ACS), while the B domain has the DNA unwinding element and the C domain is important for DNA-protein interactions. In and ARS101, the ACS is commonly composed of 11 bp, 5'-(A/T)AAA(C/T)ATAAA(A/T)-3'. This core sequence is essential for and ARS activity. In this study, we identified ARS-containing sequences from genomic libraries of the yeast DMKU3-1042 and validated their replication activities. The identified DMKU3-1042 ARSs (ARSs) have very effective replication ability but their sequences are divergent and share no common consensus. We have carried out point mutations, deletions, and base pairs substitutions within the sequences of some of the ARSs to identify the sequence(s) that influence the replication activity. Consensus sequences same as the 11 bp ACS of and were not found in all minimum functional ARSs reported here except ARS7. Moreover, partial sequences from different ARSs are interchangeable among each other to retain the ARS activity. We have also specifically identified the essential nucleotides, which are indispensable for replication, within some of the ARSs. Our deletions analysis revealed that only 21 bp in ARS18 could retain the ARS activity. The identified ARSs in this study are unique compared to other yeasts' ARSs, do not share common ACS, and are interchangeable.