Hepatitis C virus (HCV) causes chronic and acute hepatitis infections . As there is extreme variability in the HCV genome, no approved HCV vaccine has been available so far . An effective polypeptide vaccine based on the functionally conserved epitopes will be greatly helpful in curing disease . For this purpose, an immuno-informatics study is performed based on the published HCV subtype-3a from Pakistan . First, the virus genome was translated to a polyprotein followed by a subsequent prediction of T-cell epitopes . Non-allergenic, IFN-γ producer, and antigenic epitopes were shortlisted, including 5 HTL epitopes and 4 CTL, which were linked to the final vaccine by GPGPG and AAY linkers, respectively . Beta defensin was included as an adjuvant through the EAAAK linker to improve the immunogenicity of the polypeptide . To ensure its safety and immunogenicity profile, antigenicity, allergenicity, and various physiochemical attributes of the polypeptide were evaluated . Molecular docking was conducted between TLR4 and vaccine to evaluate the binding affinity and molecular interactions . For stability assessment and binding of the vaccine-TLR4 docked complex, molecular dynamics (MD) simulation and MMGBSA binding free-energy analyses were conducted . Finally, the candidate vaccine was cloned in silico to ensure its effectiveness . The current vaccine requires future experimental confirmation to validate its effectiveness . The vaccine construct produced might be useful in providing immune protection against HCV-related infections.