The 2000–01 Cypriot Fourth Division was the 16th season of the Cypriot fourth-level football league.
Sourouklis Troullon won their 1st title.
Fourteen teams participated in the 2000–01 Cypriot Fourth Division.
The team with the most points at the end of the season crowned champions.
The first three teams were promoted to the 2001–02 Cypriot Third Division and the last three teams were relegated to regional leagues.