Fig. S4.
Sites 3.2 kb away from an antibiotic cassette are not sufficient for transformation. (A) Chromosome locations of GAATTC and RAATTY sites at gyrA. WT chromosome with RAATTY sites (green) and EcoRI sites (red) shown for DNA surrounding ctsM. ORFs are indicated by yellow arrows; those smaller than 200 aa are not shown. gyrA is shown in blue. Numbers under chromosomes indicate bases. SpeI and BglI sites are indicated in purple. A point mutation in gyrA confers naladixic acid (NA) resistance. (B) gDNA was purified from either WT NAR cells or ctsM::kan NAR cells. The DNA was then digested with either SpeI or BglI and in vitro methylated with M.EcoRI. The treated DNA was used to transform WT (blue) or ctsM::kan (yellow) cells. Transformation efficiency was measured as described in Materials and Methods. Error bars indicate SD. Data are the results of three biological replicates.