The most simple way of generating new constructs in MacVector using restriction enzyme sites is to:
MacVector is intelligent enough to understand compatible sticky ends during this procedure and will prompt if they do not match. For a simple cloning (e.g. inserting a copied EcoRI fragment into the EcoRI site of a vector), by default, MacVector inserts the fragment in the “direct” orientation. However, if you wish to insert the fragment in the “reverse” orientation, there is a simple trick: simply hold down the [option] key when you choose the Paste menu item and the fragment will insert in the opposite orientation. You can see this in action by clicking on the Edit menu and holding down the [option] key – the Paste menu item changes to Paste Flipped;