Our study herein focuses on the creation of multivalent vaccine vectors presenting HIV antigens in the context of the Ad capsid protein hexon, as well as expressing HIV antigens as a transgene. Specifically, these novel vectors utilize the hexon HVR2 as an incorporation site for a twenty-four amino acid region (EKNE-KELLELDKWASLWNWFDITN) of the HIV membrane proximal ectodomain region (MPER), derived from HIV glycoprotein gp41 (gp41). In addition, to presenting MPER within HVR2 these vectors express HIV-specific protein Gag from the Ad5 DE1 region. Our study herein illustrates that our multivalent anti-HIV vectors elicit a humoral and cellular anti-HIV response. Furthermore, vaccinations with these vectors, which present model antigens at HVR2, elicit a HIV epitope-specific humoral immune response.