Transformed E. coli were grown overnight at 37 °C on Luria Bertani Broth (Lennox) (LB) agar plates containing the appropriate antibiotics (50 μg/mL kanamycin and/or 100 μg/mL ampicillin). A single colony was used to inoculate 5 mL of liquid LB with the appropriate antibiotics and grown at 37 °C and 200 rpm for 20 h. Cells were harvested by centrifugation (4000 rcf 5 min rt) resuspended in 0.5 mL of ultrapure water and DNA was isolated using Zymo Research Zyppy Plasmid Miniprep Kit. Purified DNA was sent to plasmidsaurus for whole-plasmid Oxford Nanopore sequencing.
.dna files of plasmid maps can be opened with SnapGene Viewer a freely accessible software. https://www.snapgene.com/snapgene-viewer (accessed 2025-01-30)
.dna files of plasmid maps can be opened with SnapGene Viewer a freely accessible software. https://www.snapgene.com/snapgene-viewer (accessed 2025-01-30)