TABLE 1.

Primers used in mutagenesis

For mutants, the first amino acid (single letter code) designates the wild-type residue at the numbered location (translation start methionine at position 1) that is replaced with the second amino acid. Capitalized nucleotide(s) denote(s) those are different from the wild-type nAChR α6 subunit to create the designated replacement. F, forward primer; R, reverse primer.

Mutant	Primer sequences (5′ → 3′)
hα5(V290S)-F	5′-gtctctgcacttcagtacttTCgtctttgactgtcttccttc-3′
hα5(V290S)-R	5′-gaaggaagacagtcaaagacGAaagtactgaagtgcagagac-3′
hβ3(V273S)-F	5′-cattatccacatcggTCttggtttctctgacagttttcc-3′
hβ3(V273S)-R	5′-ggaaaactgtcagagaaaccaaGAccgatgtggataatg-3′
hα6(N143D+M145V)-F	5′-ggcaaaacaaaagctcttcttaaatacGatggcGtgataacctggactccaccagctattt-3′
hα6(N143D+M145V)-R	5′-aaatagctggtggagtccaggttatcaCgccatCgtatttaagaagagcttttgttttgcc-3′