pAAV-Con/Fon-N2cG

AAV transfer plasmid was generated by the Columbia University Zuckerman Institute Molecular Tools Core Facility (RRID:SCR_026201) through modifications of existing plasmids using HIFI DNA assembly cloning (NEB). pAAV-Ef1a-Con/Fon N2cG, based on pAAV-Ef1a-Con/Fon oG (Addgene #131778), was made by replacing coding sequences from the rabies oG protein with orthologous sequences from N2cG (PCR amplified from Addgene #73481). High scoring predicted splice donor and splice acceptor sites on the N2cG sense sequence were mitigated by introduction of synonymous codon substitutions to enhance protein expression. This plasmid was put into an AAV8 and injected to label presynaptic inputs to SNr neurons in Foxp2-Cre;Vgat-Flp mice.