The development of high-entropy alloys (HEAs) has been historically constrained by trade-off between strength and ductility. In this study, the Al0.25FeCoNiV duplex HEA showed a yield strength of 625 ± 35.2 MPa and a fracture elongation of 48 ± 1.5 % after rolling and annealing. These values are 43 % and 60 % higher, respectively, than the as-cast specimens, demonstrating an excellent synergy between strength and ductility. Through in-situ electron backscatter diffraction (EBSD) tensile experiments and transmission electron microscopy characterization of the as-cast and annealed specimens, respectively, it is found that the superior mechanical properties of the annealed specimens are primarily attributable to the multilevel strengthening and strain-hardening coupling mechanism established by the synergistic action of phase boundaries (PBs), twin boundaries (TBs), and high-angle grain boundaries. This mechanism effectively suppresses strain localization and prevents premature fracture caused by local stress concentration at PBs. The strengthening mechanism of TBs on the mechanical properties of HEAs was investigated by examining the interaction mechanisms between annealed TBs and dislocations, as well as the coordinated plastic deformation mechanism of twins. This study provides novel insights into the development of innovative HEAs with enhanced properties and advances the understanding of the role of TBs in the strengthening mechanisms of materials.