The article introduces a novel method called PERMutation Using Transposase Engineering (PERMUTE) for creating libraries of circularly permuted proteins. PERMUTE utilizes the transposase MuA to randomly insert a minitranposon into a plasmid containing the open reading frame (ORF) being permuted. The process involves selecting bacteria that have acquired the integrated minitranposon, excising the ensemble of ORFs with integrated minitranposons, and circularizing these ORFs using intramolecular ligation. The method was tested on Thermotoga neapolitana adenylate kinase (TnAK), and the resulting library of circularly permuted proteins showed distinct sequence diversity compared to existing methods. Selections of this library for variants that complement Escherichia coli growth at a temperature-sensitive site identified functional proteins with novel architectures, suggesting that PERMUTE is useful for directed evolution of proteins with new functions. The article also discusses the construction of the target vector, minitranposon synthesis, library construction, and the characterization of the unselected and selected libraries.The article introduces a novel method called PERMutation Using Transposase Engineering (PERMUTE) for creating libraries of circularly permuted proteins. PERMUTE utilizes the transposase MuA to randomly insert a minitranposon into a plasmid containing the open reading frame (ORF) being permuted. The process involves selecting bacteria that have acquired the integrated minitranposon, excising the ensemble of ORFs with integrated minitranposons, and circularizing these ORFs using intramolecular ligation. The method was tested on Thermotoga neapolitana adenylate kinase (TnAK), and the resulting library of circularly permuted proteins showed distinct sequence diversity compared to existing methods. Selections of this library for variants that complement Escherichia coli growth at a temperature-sensitive site identified functional proteins with novel architectures, suggesting that PERMUTE is useful for directed evolution of proteins with new functions. The article also discusses the construction of the target vector, minitranposon synthesis, library construction, and the characterization of the unselected and selected libraries.