You want to insert an ORF into pT5 plasmid. The DNA fragment is an open reading frame with ATG on the 5’end. pT5 plasmid contains a bacterial promoter that spans from 0.5 to 0.8 kb and goes clockwise, an EcoRI site at 0/6 kb, a HindIII site at 1 kb, a BamHI site at 3 kb. Your ORF is a HindIII fragment (2 kb long) with an EcoRI site at 1 kb and a BamHI site at 1.5 kb. Required:
a. Draw the recombinant plasmid to show the right orientation of the ORF in the plasmid.
b. Which enzyme can you use to find out the orientation of the ORF after ligation?