Cloning of human TSHRv1.3

Because the entire coding sequence of the TSHRv1.3 was published (14), we synthesized the entire ORF through exon 8 along with the 63 divergent bases of intron 8 sequence tagged to either green fluorescent protein (GFP) or histidine (His) with a stop codon and polyadenylation signal of the vector. Because one of the vectors we used had a built-in N-terminal secretion signal of IgG κ, we made expression constructs with and without the endogenous TSHR signal peptide. The cDNA was flanked with restriction enzyme sites XhoI and BamH I and cloned into either pSecTag2His-Myc or pEGFP-N1 mammalian expression vectors and sequence verified.