The present study was conducted to uncover effects of partitioning treatment on Cu–Ni–Mo alloyed ductile iron (DI) austempered at different temperatures. For this purpose, the DI samples, produced via sand casting, were austenitized at 900 °C for 60 min, followed by austempering at the temperatures of 275–325–375 °C for 120 min and afterwards a partitioning treatment was applied at 200 °C for 15 min. In the characterization studies, dilatometer, image analysis, JMat-Pro, mechanical tests, XRD, optical microscope, and scanning electron microscope (SEM) equipped with EBSD detector were utilized. Characterization studies showed that the effects of partitioning treatment were directly correlated with austempering temperature and high carbon austenite volume fraction changed in the range of 19.48–35.45%. That redistribution of carbon (C) between bainitic ferrite and high carbon austenite occurred, in turn, the carbon content of high carbon austenite increased with the partitioning treatment irrespective of austempering temperature were uncovered. Furthermore, the partitioning treatment considerably changed the grain morphologies of both high carbon austenite and banitic ferrite. As a consequence of these microstructural differences, the highest tensile strength of 1489.2 MPa was established in the sample austempered at 275 °C and partitioned at 200 °C, whereas the highest ductility of 5.61% acquired at the austempering temperature of 375 °C.