Table 2 Primer sets used to construct plasmids employed in this work
PlasmidsSequence changesPrimer sets
pRR536Gln31–730 (F.L. Wild Type)5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
Rai et al. (2013)
pRR609Gln31–4965′-CGCGGATCCGGATGAAGATTTACTGGAACTTGAGGTG-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR610Gln31–4765′-CGCGGATCCAAAATTAGGAGTAACAGTGTTCGAATTGTG-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR611Gln31–3845′-CGCGGATCCTGTAGTGGTTACTGAAGTTGAGGCAGTTGC-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR612Gln31–4005′-CGCGGATCCAGAGTTTTGTTGTAGTGATTTTTTCCTCGA-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR613Gln31–5425′-CGCGGATCCAATTCTTGGTGAGGATGCGACACTATTTCC-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
Rai et al. (2013)
pRR620Gln31–3505′-CGCGGATCCGATAACGTCCGATTTTAAGGATAATGGCC-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR622Gln31–5845′-CGCGGATCCCGATGAGGAGTACGATGCATTGCGCGAC-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR624Gln31–4865′-CGCGGATCCTGAAGTACTACTTCGTCTTGAAGACCTTCT-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR656Gln3244–296Δ5′-CAATCGGCCGGATAACGTCCGATTTTAAGGATAATGGCCTCATGG-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
5′-CAATCGGCCGCAAACGGACCCAAACATTGCACAAAAT-3′
5′-CGCGGATCCAGAGTTTTGTTGTAGTGATTTTTTCCTCGA-3′
pRR723Gln3388–395Δ5′-CAATCGGCCGTTTAGCATTTGTAGTGGTTACTGAAG-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
5′-CAATCGGCCGCTACAACAAAACTCTTTATCTAGAGTG-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATT GCT-3′
pRR725Gln3262–271Δ5′-CAATCGGCCGATTTGTTGTGTTGGATGAAGATAATCC-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
5′-CAATCGGCCGATGTCGTCCACGTCCCTGGCCAATTTC-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
pRR754Gln3R264D,K265D,S267D5′-CAATCGGCCGATGTCGTCCACGTCCCTGGCCAATTTC-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
5′-CAATCGGCCGGATAAGgtcGTTgtcgtcTACAGAATTTG-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR772Gln3K281D,K282D,S285D5′-CAATCGGCCGgatGTATCTTCCAGTATATCCAAT-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
5′-CAATCGGCCGatcatcGAAATTGGCCAGGGACGT-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR787Rai et al. (2015)
pRR789Gln3K352D,R353D,K356D,K357D5′-CAATCGGCCGTTTGTTTGGCgtcatcCTTTGAAATgtcatcTTTGATAACGTCCGATTTTAAGG-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
5′-CAATCGGCCGCAAACATTGCACAAAATACTCCAAGTGCAC-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
pRR792Gln3P388D,R392D,K393D5′-TCACTCTAGATAAAGAGTTTTGTTGTAGTGATTTgtcatcCGATCGTATgtcTTTAGCATTTG-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
5′-TCTTTATCTAGAGTGATACCTGAAGAAATCATTAGAG-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
pRR823Gln3S255D,S256D,S257D,S273D,S274D,S275D,S276D,S287D,S288D,S289D,S291D5′-CAATCGGCCGTTTATTTTGTCCTGATGGTTCCATATTggcTATagcggcagcTACTGAGGCAGCTCTTTTGAAATTGGCCAGggcagctgcagcCATTGGCTTGATAAGTGAGTTTTTTCTTACAGAATTTGTTGTGTTggctgcagcTAATCCTATCGAATCGC-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
5′-CAATCGGCCGCTGATACAATGTTTCAATTGTAAAAC-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATT GCT-3′
pRR826Gln3S255A,S256A,S257A,S273A,S274A,S275A,S276A,S287A,S288A,S289A,S291A5′-CAATCGGCCGTTTATTTTGTCCTGATGGTTCCATATTgtcTATatcgtcatcTACTGAGGCAGCTCTTTTGAAATTGGCCAGgtcatcgtcatcCATTGGCTTGATAAGTGAGTTTTTTCTTACAGAATTTGTTGTGTTgtcatcatcTAATCCTATCGAATCGC-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
5′-CAATCGGCCGCTGATACAATGTTTCAATTGTAAAAC-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
pRR834Gln3S249A,S251A,S262A,S267A5′-CAATCGGCCGTTTCTGAAGAAGgctGATgcgATAGGATTATCTTCATCCAACACAACAAAtgcTGTAAGAAAAAACgcaCTTATCAAG-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
5′-CAATCGGCCGCTGGATATTACTATTGTTGCTATT-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR835Gln3S249D,S251D,S262D,S267D5′-CAATCGGCCGTTTCTGAAGAAGgacGATgatATAGGATTATCTTCATCCAACACAACAAAtgaTGTAAGAAAAAACgatCTTATCAAG-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
5′-CAATCGGCCGCTGGATATTACTATTGTTGCTATT-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR1034Gln3S576D5′-GTACGATGCATTGCGgtcCATCTTTCTTCTTGGTATATTC-3′
5′-CATCCAGATCTGTTGTTCCGATATTACCAAAACC-3′
Rai et al. (2014)
pRR1037Gln3S576A5′-GTACGATGCATTGCGcgcCATCTTTCTTCTTGGTATATTC-3′
5′-CATCCAGATCTGTTGTTCCGATATTACCAAAACC-3′
Rai et al. (2014)
pRR1059Gln3S355D5′-CGGAACAACAGATCTGGATGAAGATTTACTGGAACTTG-3′
5′-CATGGTACCATGAGGCCATTATCCTTAAAATCGGACGTTATCAAAAAGAGGATTgacAAGAAGAGAGCC-3′
pRR1060Gln3S355A5′-CGGAACAACAGATCTGGATGAAGATTTACTGGAACTTG-3′
5′-CATGGTACCATGAGGCCATTATCCTTAAAATCGGACGTTATCAAAAAGAGGATTgcaAAGAAGAGAGCC-3′
pRR1293Gln3R264A,K265A,S267A5′-CAATCGGCCGGATAAGtgcGTTtgctgcTACAGAATTTGTTGTG-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR1295Gln3K281A,K282A,S285A5′-CAATCGGCCGATGTCGTCCACGTCCCTGGCCAATTTCgcagcaGCTGCCgcaGTATCTTCCAGTATATC-3′
5′-CGCGGATCC TATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
5′-CAATCGGCCGGATAAGTGAGTTTTTTCTTACAGAATTTGTTG-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR1345Gln3R572D,R573D,K574D5′-CATCCAGATCTGTTGTTCCGAGATATTACCAAAACC-3′
5′-GTACGATGCATTGCGCGACATgtcgtcatcTGGTATATTCATTCCTTGAC-3′
pRR1366Gln3S249D,S251D,S256D,S257D5′-CAGGACTAGTGGACGACATTGGCTTGATAAGTGAGTTTTTTCTTACAGAATTTGTTGTGTTgtcgtcAGATAATCCTATgtcATCgtcCTTCTTCAGAAAAG-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
5′-CAGGACTAGTCTGGCCAATTTCAAAAGAGCTGCCTCAG-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
pRR1368Gln3S267D,S273D,S274D,T275D,S276D5′-CAGGACTAGTgtcgtcCATTGGCTTGATAAGgtcGTTTTTTCTTACAG-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
5′-CAGGACTAGTCTGGCCAATTTCAAAAGAGCTGCCTCAG-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
pRR1370Gln3S285D,S287D,S288D,S289D5′-CAGGACTAGTCTGGCCAATTTCAAAAGAGCTGCCgacGTAgacgacgacATATCCAATATGGAACC-3′
5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
5′-CAGGACTAGTGGACGACATTGGCTTGATAAGTGAG-3′
5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRS316Vector
pRR482Kulkarni et al. (2001)
pKA10Kulkarni et al. (2001)
pKA14Kulkarni et al. (2001)
pKA36Kulkarni et al. (2001)
pKA38Kulkarni et al. (2001)
pKA53Kulkarni et al. (2001)