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Generation of clones with mutations close to each other by overlapping mutagenesis primers. Sequences of overlapping primers, parent clone MCon3 and resulting mutants are aligned. The codon in which the mutation was introduced is underlined. The mutation positions are indicated with black dots. (A) Mutagenesis primers overlap each other but do not overlap the actual mutation bases. (B) Mutagenesis primers overlap each other and the mutations.
