Abstract: Described herein are cell lines for recombinant adeno-associated virus (AAV) production, cell lines for AAV-implemented protein production, and cell lines for use in titering AAV; methods of making each of those cell lines; and methods of using each of those cell lines. In some aspects, the cell lines disclosed herein may be used in a manufacturing process that is seed virus-free, helper virus-free, and transfection-free that uses synthetic elements to control viral genes in a stable cell line.