Generation of SORL1, ERBB2, and ERBB3 Expression Vectors

SORL1 ORF was ordered from Origene in the pLenti-C-Myc-DDK plasmid (Origene, PS100064). The SORL1 ORF was digested using EcoRI and XhoI and ligated into the pLenti-C-GFP plasmid (Origene, PS100065) between EcoRI and XhoI restriction sites. pDEST-SorLA-v1 (Addgene, #154892) was generated by first PCR amplifying the SorLA coding sequence from the pLenti-SorLA-C-GFP vector using the primers 5’-GGTACTCGAGGCCACCatggcgacacggagcagcaggaggga-3’ and 5’-GGTCGAATTCggctatcaccatggggacgtcatctgaaaatccag-3’. PCR fragments were then subcloned into the pDEST-ORF-v1 (a kind gift from Darren Saunders, Addgene, #73637) [32 (link)] vector using the XhoI/EcoRI restriction enzymes. For the pDEST-ERBB2-v2 (Addgene, #154895), pDEST-ERBB3-v1 (Addgene, #154893) and pDEST-ERBB3-v2 (Addgene, #154894) vectors, LR reactions (LR clonase II, ThermoFisher Scientific) were performed using the pDEST-ORF-v1 and pDEST-ORF-v2 (Addgene, #73638) [32 (link)] destination vectors, and pDONR223-ERBB2 (a kind gift from William Hahn & David Root, Addgene, #23888) [62 (link)] and pDONR223-ERBB3 (Addgene, #23874) [62 (link)] shuttle vectors. pENTR2b-mVenus was LR subcloned into pEF.DEST51 (ThermoFisher Scientific, #12285011) to generate the expression plasmid, pEF.DEST51-mVenus. All vectors were verified by analytical digests and sequencing.