Ez10 spin column plasmid miniprep kit

A detailed list of oligonucleotide sequences is found in Supplementary Table 3. Cloning vectors were prepared using EZ10-Spin Column Plasmid Miniprep kit (Bio Basic) whereas genomic DNA (gDNA) and conjugative plasmids were extracted using the Quick-gDNA miniprep (Zymo Research) according to the manufacturer’s instructions. PCR amplifications were performed using Veraseq DNA polymerase (Enzymatics) or TaqB (Enzymatics) for DNA parts amplification and screening, respectively. Digestion with restriction enzymes (NEB) were incubated for 1 h at 37 °C following manufacturer’s recommendations. Plasmids were constructed by Gibson assembly^{38 (link)} using the NEBuilder Gibson Assembly mix (NEB) following manufacturer’s protocol. These procedures were applied to the construction of pPilS that are further detailed in the Supplementary methods section.

The EZ10‐Spin Column Plasmid Miniprep kit (Bio Basic) was used to extract small DNA vectors, whereas conjugative plasmids and genomic DNA were extracted using the Quick‐gDNA miniprep (Zymo Research) according to the manufacturer’s instructions. PCR amplifications were performed using Veraseq DNA polymerase (Enzymatics). Preparation of DNAseq libraries was performed using the NEBNext Ultra II FS DNA Library Prep Kit for Illumina (NEB). Purification of DNA was performed between each step of DNAseq and plasmid assembly using Agencourt AMPure DNA XP DNA binding beads (Beckman Coulter) following the manufacturer’s guidelines. After purification, DNA concentration and purity were routinely assessed using a NanoDrop spectrophotometer (Thermo Fisher Scientific).