Quikchange lightning 2 cloning kit

All plasmids were prepared using commercial MiniPrep kits (Qiagen, Hilden, Germany). All enzymes were obtained from New England Biolabs and used per the manufacturer’s recommendations. PCR reactions were carried out using Q5 High-Fidelity Polymerase (New England Biolabs, Ipswich, MA) per the manufacturer’s protocols. Primers were ordered from Genosys (Sigma-Aldirch, Burlington, MA) or Integrated DNA Technologies (Coralville, IA). Except where noted, genomic DNA was extracted using QuikExtract DNA reagent (Lucigen, Middleton, WI). Except where noted, non-viral plasmids were maintained in competent DH5-alpha E. coli, prepared by the Vanderbilt Molecular Cell Biology Resource (MCBR), lentiviral and retroviral plasmids were maintained in Stbl2 E. coli (ThermoFisher, Waltham, MA), and cloned plasmids were initially transformed into competent cells included with kits. Gibson assemblies were carried out using the NEB Gibson Assembly Cloning kit. Site Directed Mutagenesis was carried out using the QuikChange Lightning II cloning kit (Agilent, Santa Clara, CA). PCR cloning was carried out using the NEB PCR cloning kit. All Sanger sequencing was completed by Genewiz, Inc (South Plainfield, NJ).