[The naive] decompression added 909 nucleotides to the wild-type genome. We next addressed practical constraints arising from the length of DNA that can be physically packaged within a øX174 capsid without impacts to reproductive fitness. Previous work has shown that the length of a øX174 genome, when packaged in vitro, must be kept within a few percent of the 5386 nucleotide wild-type length in order to avoid any significant fitness decrease ( Aoyama and Hayashi, 1985). Similar results were shown in vivo ( Russell and Muller, 1984). To reduce the decompressed genome length we removed the first 916 nucleotides of gene F, encoding the coat protein ( Air et al., 1978). We chose gene F because a plasmid containing a restriction fragment encoding wild-type gene F was able to complement two conditional gene F mutations ( Avoort et al., 1983). Additionally, the gene F coding sequence is greater than the total of the combined increases needed to implement the øX174.1 genome design. The truncated gene F version of the decompressed genome was named øX174.1f. To complement øX174.1f when transformed into host cells we designed a medium copy vector expressing gene F under control of a rhamnose-inducible promoter ( Fig. S1).