pPAO2, plasmid map, and polylinker sequence. (A) The plasmid map of pPAO2. (B) The polylinker sequence between theHindIII and EcoRI sites of pUC119. The amino acid sequence of the in-frame construct is given above the DNA sequence. FS indicates a frameshift, which can only be overcome if an in-frame DNA fragment is cloned via the LIC approach. (C) The LIC cloning procedure in detail.