You have inserted a 5-kb piece of eukaryotic DNA into a 3.5-kb plasmid at the BamHI site. When you digest the recombinant DNA with various restriction enzymes, you obtain the fragments listed below. Draw a circular map of the recombinant plasmid showing all the restriction sites. Is it possible to determine the orientation of the insert in the plasmid? If so, what information reveals this? If not, what restriction digest would you perform to determine the orientation of the insert?
Restriction enzyme Size of DNA fragments (kb)
EcoRI and BamHI..............................1, 1.25, 2.75, 3.5
PstI and BamHI.................................1.75, 3.25, 3.5
EcoRI, PstI, and BamHI........................0.75, 1, 1.25, 2, 3.5
SalI.................................................8.5
SalI and BamHI..................................0.5, 3, 5
BamHI.............................................3.5, 5