DNA duplex which will give rise to this tRNA precursor, was synthesized in the form of 26 small oligo nucleotide segments. These were then arranged, into six DNA duplex fragments having single stranded ends.

These six fragments give rise to presumed gene for E. coli tyrosine suppressor tRNA precursor. This gene, however, still lacked promoter region and other sequences essential for processing. Later, in 1979, Khorana reported completion of the total synthesis of a biologically functional tyrosine suppressor transfer RNA gene carrying all regulatory sequences,

This gene was 207 base pairs long and included the following:

(i) a 51 base pairs long DNA promoter region;

(ii) a 126 base pairs long DNA corresponding to the precursor tRNA and

(iii) a 25' base pairs long duplex DNA corresponding to 16 base pairs adjoining CCA end of tRNA and the remainder, a modified sequence including EcoRI endonuclease specific sequence.

The complete synthetic gene was cloned is the vector bacteriophage (lambda virus) by gene cloning method. On transformation of E. coli, the phage, could multiply with the doned gene.