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Probing DNA binding by random mutagenesis: Random mutagenesis PCR was carried out on the Stl coding sequence generating various single mutations. The PCR product was cloned into pKW08 vector and electroporated into the Stl switch system. The depicted mutants showed blue phenotype in the designed Stl switch system, suggesting defective DNA-binding ability. (A) The location of the hits in the Stl full-length protein; (B) Mutants mapped on the Phyre2 structural model of Stl [8].
