IN VIVO ASSEMBLY OF DNA VIA HOMOLOGOUS RECOMBINATION

摘要

According to the present invention, a DNA construct of interest is assembled from overlapping subfragments via an acceptor module which comprises the distal end of the construct at a position downstream from a promoter. The construct is assembled distal to proximal via homologous recombination events occurring in the span between that distal end of the construct and the upstream end of the promoter. These recombination events occur iteratively between the acceptor module and alternative donor modules. Successful recombination places one of at least two marker genes under the transcriptional control of an active form of the promoter. As a result of alternating use of two varieties of donor modules, as few as two selection markers may be used to produce a complex DNA construct.