The origin of replication (Ori) was successfully located in all chromosomes. To demonstrate how we visually located the Ori, we selected the DNA Walk (A,B) and mfold (C,D) analysis of chromosome Pac8 (A,C) and Pac4 (B,D). The DNA Walk shows abrupt changes in base composition bias that are characteristic of Oris, and the mfold analysis shows stem-loop configurations containing T-rich loops that are also characteristics of Oris. For Pac8, the Ori was located at position 4,950, while for Pac4, the Ori was located at position 7,850 (both Oris are denoted by a black box). DNA Walker window size: 50.