| Competition: | Cypriot First Division |
| Season: | 2000–01 |
| Winners: | Omonia (18th title) |
| Relegated: | Nea Salamina Digenis Aris |
| Continentalcup1: | Champions League |
| Continentalcup1 Qualifiers: | Omonia (2nd qualifying round) |
| Continentalcup2: | UEFA Cup |
| Continentalcup2 Qualifiers: | Olympiakos (qualifying round) Apollon (qualifying round; via Cypriot Cup) |
| Continentalcup3: | Intertoto Cup |
| Continentalcup3 Qualifiers: | Anorthosis (1st round) |
| League Topscorer: | Rainer Rauffmann (30 goals)[1] |
| Matches: | 182 |
| Total Goals: | 646 |
| Prevseason: | 1999–2000 |
| Nextseason: | 2001–02 |
The 2000–01 Cypriot First Division was the 62nd season of the Cypriot top-level football league. Omonia won their 18th title.
Fourteen teams participated in the 2000–01 Cypriot First Division. All teams played against each other twice, once at their home and once away. The team with the most points at the end of the season crowned champions. The last three teams were relegated to the 2001–02 Cypriot Second Division.
The champions ensured their participation in the 2001–02 UEFA Champions League and the runners-up in the 2001–02 UEFA Cup.
The teams had to declare their interest to participate in the 2001 UEFA Intertoto Cup before the end of the championship. At the end of the championship, the higher placed team among the interested ones participated in the Intertoto Cup (if they had not secured their participation in any other UEFA competition).
Teams received three points for a win, one point for a draw and zero points for a loss.
Ethnikos Assia, Anagennisi Deryneia and Alki Larnaca were relegated from previous season and played in the 2000–01 Cypriot Second Division. They were replaced by the first three teams of the 1999–2000 Cypriot Second Division, Digenis Morphou, Aris Limassol and Doxa Katokopias.
Before the start of the season, APOP Paphos and Evagoras Paphos were merged forming AEP Paphos, which took the place of APOP Paphos in the Cypriot First Division.