Background and Objectives: Some of the human papillomaviruses (HPVs) can infect genital tracts and are sometimes associated with anogenital tract cancers. HPVs induced cervical cancers through the expression of E6 and E7 genes by inactivating the tumor suppressor proteins. In this study, E6 and E7 genes were chosen in order to construct an expression vector which is able to express target proteins. Patients and Methods: This experimental investigation was performed in Virology Department of Tarbiat Modares University. An expression vectorcontaining human papillomavirus type 16 E6 and E7 genes was constructed. The accuracy of the plasmid was confirmed by polymerase chain reaction (PCR) and restriction enzyme analysis. The construct was transfected into the eukaryotic cells and its ability for protein production was confirmed by Western blotting. Results:The colonies containing desired plasmid have the fragment about 995 bp. For confirming the ability of the construct for protein production in eukaryotic cells, Western blotting was done using the lyses-cells as antigen and they showed the desired bands using monoclonal antibodies. Conclusion: The designed vector can consider as a based vaccine for construction a therapeutic vaccine in suitable vectors for gene therapy in order to administration in Iranian patients with cervical cancer.