pYT-GCAA home page

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

pYT-GCAA 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 In this mutant the amino acid acceptor stem of the yeast-phe tRNA has been replaced by a small "thermal stable" tetraloop with the sequence 5'CGAA3'.

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

The sequence of the yeast-phe GCAA/AA tRNA mutant is:
5' G CGG ATT TAG CTC AGT TGG GAG AGC GCG CAA GAG GTC CTG TGT TCG ATC CAC AGA ATT CGC ACC A 3'

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