Plasmid Assembly and DNA Preparation

All plasmids and oligonucleotide primers used in this study are listed in Supplementary Tables 1 and 2, respectively. Plasmids were assembled using either the USER cloning method (Bitinaite et al., 2007 (link)), NEBuilder HiFi DNA assembly method (New England Biolabs) or restriction enzyme-based cloning techniques (Sambrook et al., 1989 ). Plasmid DNA preparation was carried out using the QIAprep® Spin Miniprep Kit (Qiagen). Gel purified linearized DNA was extracted using the QIAquick® Gel Extraction Kit (Qiagen). Genomic DNA was isolated with the GenElute™ Bacterial Kit (Sigma-Aldrich). All restriction endonucleases, T4 DNA ligase and NEBuilder® HiFi DNA Assembly Master Mix were acquired from New England Biolabs. DNA sequences were verified by Sanger sequencing (Eurofins Genomics). A detailed assembly description for each plasmid is provided in the Supplementary information.