Bacterial Antibiotic Resistance Screening

The following working concentrations of antibiotics were used: carbenicillin (Solarbio, 50 μg/ml), kanamycin (Solarbio, 50 μg/ml), spectinomycin (Macklin, 50 μg/ml), chloramphenicol (Macklin, 25 μg/ml). PHANTA 2x mix (Vazyme) was used for cloning PCR, and Flash 2x mix (Vazyme) was used for verification PCR and Sanger sequencing (Tsingke Bioscience). All cloning fragments were assembled by Golden Gate assembly (New England Biolabs) or ClonExpress assembly (Vazyme) methods. Plasmids were cloned in DH5α competent cells (HT Health). Synthetic genes were ordered from Tsingke Bioscience. Cloned plasmids were extracted by Tiangen DNA extraction kit. E. coli strain S2060^{55 (link)} was used in all aspects of the EvoScan process, including system construction, evolution, and plaque assays. The DH5α strain was used for flow cytometry experiments. Detailed information on the plasmids and selection phage (SP) used in this work is given in Supplementary Table 6.