Highly effective and broad-spectrum influenza vaccines are urgently required to prevent influenza outbreaks. Hemagglutinin (HA), M2 ectodomain (M2e), and nucleoprotein (NP) are crucial target antigens for the development of universal influenza vaccines. To generate a novel multivalent influenza vaccine, the HA genes of influenza B Yamagata (BY) and Victoria (BV) strains, and the NP gene of H1N1 were cloned into the E1 region of the chimpanzee adenoviral vector, AdC68, and M2e epitopes of H1N1 and H3N2 were fused to the loop region of the AdC68 fiber, resulting in the recombinant adenoviral vector vaccine, AdC-Flu-Tet. The immunoprotective effects of AdC-Flu-Tet were evaluated in the mouse models. The results showed that AdC-Flu-Tet successfully induced robust humoral and cellular immune responses and conferred full protection against H1N1, H3N2, BY, and BV infections. In conclusion, AdC-Flu-Tet is a promising candidate as a novel influenza vaccine with high protective efficacy.