FIG 6.
Unaltered sequences and mutation regions of editing templates for ugtP, hasSE, and amyE KOs and schematic representation of the KI of the HA biosynthetic operon at the amyE locus. (A) The native (ugtP) and modified (editing template) sequences of the mutation region for ugtP KO at ugtP.P395T are in uppercase, and the adjacent 20 bp of flanking homology is in lowercase. In the native sequence, the PAM site is underlined and the two base pairs between which the DSB occurs are in bold. The BspHI restriction site is italicized in the modified sequence, and a summary of HLs analyzed during editing template (ET) HL optimization is shown. (B) The native (amyE) and modified (editing template) sequences of the mutation regions for amyE KO at amyE.P25NT, amyE.P330T, amyE.P636T, and amyE.P1344T are in uppercase, and the adjacent 18 to 21 bp of flanking homology is in lowercase. In the native sequences, PAM sites are underlined and the two base pairs between which the DSBs occur are in bold. The XhoI (amyE.P25NT and amyE.P330T), ScaI (amyE.P636T), and BamHI (amyE.P1344T) restriction sites are italicized in the modified sequences. (C) The unaltered (hasSE) and modified (editing template) sequences of the mutation region for hasSE KO at hasSE. P394T are in uppercase, and the adjacent 18 bp of flanking homology is in lowercase. In the unaltered sequence, the PAM site is underlined, and the two base pairs between which the chromosomal DSB occurs are in bold.