Antibiotic and Nuclease-free Reagents for Molecular Biology

Antibiotics (Gold Biotechnology) were used at the following working concentrations: carbenicillin 50 μg/mL, spectinomycin 50 μg/mL, chloramphenicol 25 μg/mL, kanamycin 50 μg/mL, tetracycline 10 μg/mL, streptomycin 50 μg/mL. Nuclease-free water (Qiagen) was used for PCR reactions and cloning. For all other experiments, water was purified using a MilliQ purification system (Millipore). Unless otherwise noted, Phusion U Hot Start or Phusion Hot Start II DNA polymerase (Thermo Fisher Scientific) were used for all PCRs. Unless otherwise noted, plasmids and SPs were cloned by USER assembly^{26 (link)}. Genes were obtained as synthesized gene fragments from Twist Bioscience. Plasmids were cloned and amplified using either Mach1 (Thermo Fisher Scientific) or Turbo (New England BioLabs) cells. Plasmid or SP DNA was amplified using the Illustra Templiphi 100 Amplification Kit (GE Healthcare Life Sciences) prior to Sanger sequencing. Strain S2060^{25 (link)} was used in all luciferase, phage propagation, and plaque assays, and in all PACE experiments. A description of plasmids, SP, primers, and protospacer sequences used in this work is provided in Table S4.