Gene G of bacteriophage φX174 DNA has been characterized by three techniques. (1) Transcription by RNA polymerase of appropriate DNA fragments from restriction enzyme digestion. (2) Use of such DNA fragments as primers for DNA polymerase. (3) Amino acid sequencing of the gene product. The combined results give a DNA sequence of 195 nucleotides which codes for the N-terminal 65 amino acids of the protein sequence.