Maybe I'm overcomplicating this, but I'm having some issues with a construct.
So, I have three plasmids containing a total of eight DNA sequences that I need to fuse into one linear sequence. I plan to fuse the sequences using Gibson assembly. However, before I can fuse them, I need to amplify (PCR) them out of their plasmids.
I would like to use as few primers as possible, so my question is this; can I use the Gibson assembly primers to amplify them out of the plasmids and add the overlap sequences separately, then fuse them all together in a separate reaction without primers? Would the overlap sequences already added to each sequence separately be enough to act as primers for assembly? Even better (but I doubt it would work), could I amplify them out of their plasmids and fuse them in the same reaction?
If both these scenarios are unrealistic, I can still just order another round of primers to extract the sequences from their plasmids, but I would like to just use the primers I have if I can.