Sequence of Plasmid/ORF Junctions
5' C TAT CTA TTC GAT GAT GAA GAT ACC CCA CCA AAC CCA AAA AAA GAG ATC GAA TTC CAG CTG
ACC ACC ATG XXX XXX XXX XXX XXX XXX *** CAT GGC AAT TCC CGG GGA TCC GTC GAC CTG CAG
AGA TCT ATG AAT CGT AGA TAC TGA AAA ACC CCG CAA G 3'
Where
ATG represents the actual start codon of the yeast ORF.
XXX's represent the ORF-specific sequences.
***'s represent one of the 3 stop codons.
ACC ACC ATG XXX XXX XXX XXX XXX XXX *** CAT GGC AAT TCC CGG GGA TCC GTC GAC CTG CAG
AGA TCT ATG AAT CGT AGA TAC TGA AAA ACC CCG CAA G 3'
Where
ATG represents the actual start codon of the yeast ORF.
XXX's represent the ORF-specific sequences.
***'s represent one of the 3 stop codons.
Primers and Re-PCR Primers Used for Amplifying Yeast ORFs and Their "Common Tails"
Forward primer
5' AATTCCAGCTGACCACCATG 3'
Reverse primer
5' GATCCCCGGGAATTGCCATG 3'
Re-PCR Forward primer
5' CTATCTATTCGATGATGAAGATACCCCACCAAACCCAAAAAAAGAGATCGAATTCCAGCTGACCACCATG 3'
Re-PCR Reverse primer
5' CTTGCGGGGTTTTTCAGTATCTACGATTCATAGATCTCTGCAGGTCGACGGATCCCCGGGAATTGCCATG 3'
5' AATTCCAGCTGACCACCATG 3'
Reverse primer
5' GATCCCCGGGAATTGCCATG 3'
Re-PCR Forward primer
5' CTATCTATTCGATGATGAAGATACCCCACCAAACCCAAAAAAAGAGATCGAATTCCAGCTGACCACCATG 3'
Re-PCR Reverse primer
5' CTTGCGGGGTTTTTCAGTATCTACGATTCATAGATCTCTGCAGGTCGACGGATCCCCGGGAATTGCCATG 3'
