The LIC Duet™ Adaptors are designed to facilitate efficient, simultaneous and directional cloning of two protein coding sequences (ORFs) into an Ek/LIC vector - without the need for restriction digestion or ligation!

With LIC Adaptors the relative order of the two inserts can be specified and inserts can be expressed with different tags. All vector encoded sequences can be removed from purified proteins by protease cleavage.

The adaptor contains asymmetric GC-rich 12-base overhangs on each end. Compatible ORFs are created using specifically designed PCR primers. A post-PCR processing step creates the 12-base overhangs that are compatible with the adaptor. In an annealing reaction, one end of the adaptor anneals to the region encoding the C-terminus of ORF-1 and the other end anneals to the region encoding the N-terminus of ORF-2.

This strategy allows two protein coding regions to be simultaneously cloned into a single vector without the need for restriction digests or ligations. The cloning is highly efficient with minimal non-recombinant background because only correctly annealed LIC vector-inserts are formed during the annealing process.