library_construction_protocol: Cells were disrupted using silica beads, followed by sonication with a Bioruptor UCD-200 (Diagenode) to fragment chromatin. Sonication was performed at high power for 15 minutes, with 30-second on/off cycles. For immunoprecipitation, S. cerevisiae chromatin was mixed with S. pombe spike-in chromatin prior to antibody incubation, with 1 µg of S. cerevisiae chromatin combined with 2.65 ng of S. pombe chromatin in 300 µL of buffer. Input samples consisted of 20 µL of chromatin, while 280 µL was used for immunoprecipitation. Immunoprecipitation was performed using 20 µL of Dynabeads Protein G (Invitrogen) pre-incubated with 2 µL of anti-FLAG M2 antibody (Sigma). Immunoprecipitation reactions were incubated with the antibody-conjugated beads for 90 minutes at room temperature with rotation. Beads were then washed under increasingly stringent conditions, and chromatin was eluted at 75°C. Crosslinks were reversed overnight at 65–75°C, followed by sequential RNase A and Proteinase K treatments. DNA was purified using a MinElute PCR Purification Kit (Qiagen), and concentrations were measured using PicoGreen quantification. The entire immunoprecipitation procedure was performed in three separate batches, with equivalent chromatin and bead amounts used across replicates. ChIP-seq libraries were prepared using the Ovation Ultralow System V2 (Tecan). Input libraries were generated from 66.7 ng of starting material, while IP libraries were generated from 1 µg of DNA. Libraries underwent 11 cycles of PCR amplification on a thermocycler and were purified with AMPure XP (Agencourt) beads to retain fragments greater than 100 bp. Final library quality was assessed using a TapeStation system. Libraries were sequenced on a NextSeq 2000 (Illumina) using a 50-bp paired-end configuration with single 8-bp indexes.