<p>The popularity of pomegranates has increased recently due to the growing scientific evidence of their health benefits. The harvest location and cultivar significantly influence genetic diversity and postharvest fruit quality. The present study was undertaken to evaluate the influence of harvest location and cultivar on the physical properties, biochemical traits and antioxidant activity of pomegranate fruits. Commercially matured fruits of two pomegranate cultivars (‘Sandhora’ and ‘Kalehar’) were harvested from three districts of South Punjab, Pakistan—Rahim Yar Khan (RYK), Muzaffargarh, and Lodhran—and kept at shelf temperature (25 ± 2 °C and 55–60% relative humidity) for further analysis until marketable. The results revealed that cultivar and harvest location significantly influenced fruit quality. Among the physical parameters, higher fruit weight (213 g/fruit), diameter (77.86 mm), and 100-aril weight (33.11 g) were recorded in the ‘Sandhora’ cultivar harvested from RYK, whereas maximum fruit firmness (69.55 N) was noted in the ‘Kalehar’ cultivar harvested from RYK. Among the biochemical fruit quality parameters, higher total soluble solids (15.92°Brix), vitamin&#xa0;C (54.35 mg/100 mL juice), and ripening index (45.45) were detected in fruits harvested from RYK as compared to other locations. However, the maximum titratable acidity (0.52%) was recorded for ‘Kalehar.’ The contents of phytochemicals, including carotenoids (0.27 μg/g FW), anthocyanins (0.030 ∆Ag⁻¹FW), total antioxidants (76.17% inhibition), and total phenolics (214.48 mg GAE 100 g⁻¹), were greater in ‘Sandhora’ than in ‘Kalehar.’ In enzymatic studies, superoxide dismutase (67.61 U&#xa0;mg⁻¹ protein), peroxidase (3.62 U&#xa0;mg⁻¹ protein), and catalase activities (46.25 U&#xa0;mg⁻¹ protein) were found to be greater in the arils of ‘Kalehar’ cultivar as compared to ‘Sandhora’. Therefore, it was concluded that the ‘Sandhora’ cultivar harvested from RYK has the best postharvest fruit quality attributes.</p>