Abstract: The present invention provides a bacterial continuous evolution system, an orthogonal error-prone DNA polymerase, and a continuous evolution method. In the present invention, by combining an orthogonal DNA replication system and an orthogonal error-prone DNA polymerase, a continuous evolution method that includes all mutant types, enables long-DNA-fragment mutation, and is good in continuity and simple and convenient to operate is obtained. By inducing the opening and closing of DNA polymerase expression, switching between a linear plasmid error-prone mutation process and a high-fidelity replication process is realized, so as to achieve the efficient and continuous evolution of a target DNA sequence.