Nanopore long-read sequencing data of synKmI segment 1.
Assemblies were extracted from yeast colonies using the Zymoprep Yeast Miniprep I kit (Zymo Research D2001) and electroporated into TransforMax EPI300 Electrocompetent E. coli (Lucigen EC300150). Transformants were recovered in SOC outgrowth medium for 1 hr with shaking and selected on LB agar plates containing chloramphenicol (30 g/ml). To prepare KmSeg1 assemblies for sequencing, plasmids were linearized using a single-cutter restriction enzyme AvrII. For Oxford Nanopore Technologies library preparation, we used the manufacturers SQK-LSK109 and EXP-NBD104 protocols. Samples were multiplexed and sequencing was performed on a Flongle flow cell (R10.4.1) with V14 chemistry. The raw sequencing data in .pod5 format were basecalled using Oxford Nanopore Technologies Dorado basecaller (v4.3.0, dna_r10.4.1_e8.2_400bps_sup). Resulting fastq files were demultiplexed and barcode-trimmed in Dorado using standard parameters.