Table 2.

Primers used in this study

PrimerSequencea
P15′-GGTCTAGAATTAAAGAGGAGAAATTAAGC-3′
   XbaI
P25′-GAAGGCCTAGCGGCGGATTTGTCCT-3′
   StuI
P35′-CAGGTACCATTTATCAGGGTTATTGTCT-3′
   KpnI
P45′-GGTCTAGATGTGTGAAATTGTTATCCG-3′
   XbaI
Pgfp(up) 5′-ATATAGCATGC GTAAAGGAGAAGAACTTTTCA-3′
   SphI
Pgfp(down) 5′-CTCTCAAGCTTATTTGTATAGTTCATCCATGC-3′
   HindIII
Pgfp(down,LAA) 5′-CTCTCAAGCTTATTAAGCTGCTAAAGCGTAGTTTTCGTCGTTTGCTGCAGGCCT TTTGTATAGTTCATCCATGCCA-3′
          HindIII  A  A  L  A  Y  N  E  D  N  A  A  P  R   ◂—
                                                  StuI
Pgfp(down,AAV) 5′-CTCTCAAGCTTATTAAACTGCTGCAGCGTAGTTTTCGTCGTTTGCTGCAGGCCT TTTGTATAGTTCATCCATGCCA-3′
          HindIII V  A  A  A  Y  N  E  D  N  A  A  P  R   ◂—
                                                  StuI
Pgfp(down,LVA) 5′-CTCTCAAGCTTATTAAGCTACTAAAGCGTAGTTTTCGTCGTTTGCTGCAGGCCT TTTGTATAGTTCATCCATGCCA-3′
          HindIII A  V  L  A  Y  N  E  D  N  A  A  P  R   ◂—
                                                  StuI
Pgfp(down,ASV) 5′-CTCTCAAGCTTATTAAACTGATGCAGCGTAGTTTTCGTCGTTTGCTGCAGGCCT TTTGTATAGTTCATCCATGCCA-3′
          HindIII V  S  A  A  Y  N  E  D  N  A  A  P  R   ◂—
                                                  StuI
  • a Relevant restriction sites are underlined. Regions hybridizing to gfpmut3 (8) are shown in boldface, while regions encoding the C-terminal peptide tags (protease targets) are shown in italics. The arrows indicate the orientation by which the amino acid sequences of the resulting C-terminal peptide tags should be read.