The diamine, 9,9-bis[4-(4-amino-3-hydroxylphenoxy)phenyl]fluorene (BAHPPF) was synthesized by the modified two-step method. Then, a series of acetate-containing copoly(ether-imide)s were prepared by the copolymerization of BAHPPF, 9,9-bis(4-aminophenyl)fluorene (BAF) and 2,2′-bis(3,4-dicarboxyphenyl)hexafluoropropane dianhydride (6FDA) followed by chemical imidization. The structures and properties of the BAHPPF and copoly(ether-imide)s were characterized by nuclear magnetic resonance spectrometer (NMR), Fourier transform infrared spectrometer (FTIR), X-ray diffractometer (XRD), differential scanning calorimeter (DSC), thermogravimetric analyzer (TGA), ultraviolet-visible spectrophotometer (UV-VIS), and tensile testing. Single gas permeation performances of these copoly(ether-imide)s were also studied for five representative gases of interest including H2, O2, N2, CO2, and CH4. The experimental results showed that the copoly(ether-imide)s showed excellent optical properties with high light transmittance above 80.2% at 450 nm. The glass transition temperature of these copolymers were higher than 333°C. Their tensile strength and Young’s module also increased, and the elongation decreased with the decrease of BAHPPF. High gas permeabilities of copoly(ether-imide)s were obtained, and the ideal selectivity of CO2/CH4 was improved due to the introduction of acetate group and flexible ether linkage. These copoly(ether-imide)s could be applied to the field of optics and gas separation.