williams|4582|plasmid prep|restriction|pUC19|pDEFEND2

pYP is a plasmid that that encodes for a mutant of yeast-phe tRNA. It was constructed by restricting pDEFEND2 with BSA I and BstBI, followed by ligation of the appropriate sequence to the unique sticky ends. In this mutant the modifications of  yeast-phe tRNA have been removed.

The sequence of T7 RNA polymerase promotor is:
5' AA TTA TAA TAC GAC TCA CTA TA 3'

The sequence of the yeast-phe unmodified tRNA mutant is:

5’  CGG ATT TAG CTC AGT TGG GAG AGC GCC AGA CTG AAG ATC TGG AGG TCC TGT GTT CGA TCC ACA GAA TTC GCA CCA 3’
 

The sequence of the remainder of the polylinker is:
5' CGA AGA GCT GAA GCT TGC GTC ATC CTT 3'