Nanopore sequencing of S. cerevisiae after Recombination: 5xUAS_C, 5xUAS_D
Genomic DNA of the recombined yeast cultures (strains Y2010, Y2012)  was extracted using the YeaStar Genomic DNA kit (Zymo Research, Freiburg, Germany). The recombined promoters derived from the 5xUAS_C-D promoters were amplified using the KAPA HiFI HotStart ReadyMix PCR Kit (Roche, Basel, Schweiz) with the appropriate primer pair. The PCR products were purified using Agencourt AMPure XP beads (Beckmann Coulter) and analyzed on a 2 % agarose gel. Further, the amplicons were prepared for Nanopore sequencing according to the Ligation Sequencing DNA V14 (SQK-LSK114) protocol provided by Oxford Nanopore Technologies. The library was loaded onto Flongle flowcells (FLO-FLG114). The sequencing run lasted 24 h and generated 461 k reads, with N50 of 637 b, respectively. Basecalling of the Nanopore data was performed by the High-accuracy model v4.3.0, 400 bp