How do I clone my insert into a transfer vector with no viable restriction sites but compatible with the Gateway ® cloning system?
Gateway® compatible vectors use recombination in order to generate clones containing the insert of interest. In brief, the insert is first cloned into an entry vector at a region flanked by sequences (called attP1 and attP2) that allow the insert to recombine with the destination vector (in this case the destination vector would be the lentiviral transfer vector). The destination vector contains attB sequences with which the attP sequences recombine. More information on the Gateway® cloning system can be found at invitrogen’s website.
