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Overview of methods for random domain insertion with optional circular permutation of insert. (A) Methods for making a linear plasmid DNA containing the acceptor gene in which the site of linearization is created randomly throughout the plasmid by DNase I or S1 nuclease or targeted to occur in the acceptor gene by multiplex inverse PCR. (B) Methods for preparing the insert DNA either without circular permutation or with circular permutation via a PCR on a gene duplication fusion.
