Mutagenesis of Lk ARSs to identify sequences necessary for Lk ARS function. (A-C) the shortest functional fragments of the three Lk ARSs in Figure 4 were mutated and tested for ARS function. The mutated residues are underlined. Mutations that disrupted ARS function are colored in red. The motif logos correspond to the best match of the predicted 9 bp Lk ACS and the relevant sequence is colored blue. (D) Representative examples of ARS function. Lk ARS-E139 mutant plasmids transformed into L. kluyveri and plated on selective media. The numbers correspond to mutant ARS fragments in (A). 'Empty' denotes the empty vector negative control, 'WT' denotes the full length Lk ARS-E139 positive control.