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AK circular permutation using laboratory evolution. (A) Permuteposon insertion into the TnAK gene by MuA transposase yields vectors that express (B) circularly permuted TnAK. (C) P1 initiates translation before the R2R1 transposase recognition sequence and amends an eighteen-residue peptide to the N-termini of permuted AK. (D) P2 and P3 amend only two residues because they contain an RBS within the transposase recognition sequence.
