The worldwide recognition of agroforestry as a climate change mitigation strategy has led to significant interest in assessing agroforestry systems for their carbon sequestration potential. However, there is limited knowledge regarding the carbon stocks present in Eucalyptus-based agroforestry land use systems within semi-arid regions of Pakistan. Therefore, the present study was undertaken in the semi-arid region in central Punjab (Tandlianwala), Pakistan, which is characterized by a significant extent of Eucalyptus-based agroforestry land use systems. Biomass and carbon storage were assessed and compared in three different Eucalyptus intercropping configurations (A: 130 trees ha-1, B: 256 trees ha-1, and C: 192 trees ha-1) and two cropping systems (maize and wheat cropping system) under semi-arid conditions. A significant difference in the carbon stock of Eucalyptus trees was observed among the three configurations, with the highest value of 60.55 Mg ha-1 in configuration B. Configuration A demonstrated the maximum biomass (39.84 Mg ha-1) and carbon stock (15.11 Mg ha-1) of crops, which was significantly greater than that of other configurations. Soil carbon stock was higher (63.25 Mg ha-1) at 0-60 cm soil depth in configuration B, and it was 8.88% and 18.01% greater than configuration C and configuration A, respectively. In conclusion, the results of this study suggest that configuration B exhibits the highest levels of biomass production and carbon stocks, making it the most favorable option in terms of both economic benefits and coping with climate change mitigation through carbon sequestration.