Whole genome sequencing, COG annotation, and secondary metabolites of B-4359

Bacterial genomic DNA was extracted using a Solg Genomic DNA Prep Kit (Solgent, Daejeon, South Korea), following the manufacturer’s instructions. The concentration of the extracted DNA was determined using a Qubit 2.0 fluorometer (Invitrogen, Carlsbad, CA, United States). DNA contamination of the cultured strain was tested by sequencing the 16S rRNA gene using an ABI 3730 DNA sequencing machine (Applied Biosystems, Foster City, CA, USA). The integrity of the gDNA was verified using agarose gel electrophoresis and quantified using a Qubit 2.0 fluorometer (Invitrogen). Sequencing libraries were prepared according to the manufacturer’s instructions for 20 kb template preparation using the BluePippin Size-Selection System and PacBio DNA Template Prep Kit 1.0. Briefly, 10 μg of the gDNA was sheared to 20 kb using g-tubes (Covaris, Woburn, MA, United States); they were then purified, end-repaired, and the blunt-end SMARTbell adapters were ligated. The libraries were quantified using a Qubit 2.0 fluorometer (Invitrogen) and qualified using a DNA 12,000 chip (Agilent Technologies, Waldbronn, Germany). Subsequently, the libraries were sequenced using PacBio P6C4 chemistry in an 8-well SMART Cell v3 at PacBio RSII.