Aiming at developing a highly efficient gene therapy for hemophilia B, we constructed a promoterless donor vector containing the human coagulation factor IX cDNA, carrying the V86A, E277A, and R338L-Padua hyperactive mutations (Simioni et al., 2009; Lin et al., 2010; Kao et al., 2013). This evidence concerns the gene F9 and hemophilia B.