The context effect does not require a fourth base pair.

The translational activity of a transfer RNA at a codon varies at different message sites, although the codon does not vary. The source of this effect, which may help to determine the level of gene expression, is generally agreed to be in nearby message sequences. By making every possible nucleotide combination between position 33 of the transfer RNA and the major context nucleotide of the message, it was shown that base-pairing between the two nucleotides is not the source of this context effect on translation in vivo.