Cotton plant faces several stresses during its life cycle that limit seed cotton yield and fiber production. Lack of availability of potential genotypes is one of the stress main factors that the farmer community faces. A complete diallel mating design was used to study combining ability of a set of upland cotton genotypes developed in Pakistan. Five parents namely, IR-NIBGE-3, FH-166, KZ-189, MS-40 and B-557 were hybridized in a glasshouse and F1 seeds with five parents were planted in field conditions. The combining ability analysis revealed that IR-NIBGE-3 is a good general combiner for gin turnout and fiber strength, while KZ-189 and FH-166 are good general combiners for number of bolls per plant, yield of seed cotton and fiber length. The combination of FH-166 × B-557 and MS-40 × B- 557 exhibited the best specific combining ability for number of bolls per plant, yield of seed cotton and gin turnout; whereas combination of KZ-189 × B-557 and MS-40 × KZ-189 were the best for fiber length and fiber strength, respectively. Positive and significant reciprocal effects signify the role of female parents for their use in hybridization. SCA variance was greater than GCA variance for all traits indicating the importance of non-additive effects in genetic control of these traits. This study indicates that this population can be used for selection of transgressive segregants for fiber and yield related traits and superior parents can be used in hybridization programs.