SUMMARYThe versatility of insertional inactivation of P-galactosidase activity for subcloning and sequencing has been enhanced by combining a chemically synthesized oligonucleotide which specifies nine 6-bp-cutter restriction sites including Bg/II, XhoI, NruI, &I, Sac1 and EcoRV in various configurations with existing polylinkers to create a set of highly versatile cloning sites. These improved polylinkers have been inserted into plasmids (the pICs) for routine cloning of double-stranded DNA, and into chimeric phage/plasmids (the pICEMs) for biological production of single stranded DNA. The most versatile polylinker specifies 17 restriction sites in the p-galactosidase a-complementing gene fragment. One of the new polylinkers was inserted into Ml3 DNA to produce a vector (M13mIC7) with nine cloning sites.

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