Patent ID: 8815600
Filing Date: 2014-08-26
Classification: C12N

Abstract:
1. A method of cloning a donor DNA molecule into an acceptor vector at a predetermined location, the method comprising: a) preparing an extended donor DNA molecule by adding to the 5′-end and the 3′-end of the donor DNA molecule a first sequence and a second sequence, respectively, wherein each of the first and second sequences, independently, is at least 12 nucleotides in length and is at least 90% identical to a first region and a second region of the acceptor vector, respectively; b) providing a reaction mixture comprising: c) incubating the reaction mixture to obtain an intermediate product; d) transforming a cell with the intermediate product to obtain a transformed cell; and e) culturing the transformed cell under conditions to produce a recombinant DNA molecule comprising the donor DNA located between the first region and the second region.