. 2015 Sep 2;201(3):989–1016. doi: 10.1534/genetics.115.177725

Table 2. Plasmids, primers, and oligonucleotides used in this work.

Plasmid^a	Residue alterations	Primer sets
pRR536	Wild-type Gln3_1–730-Myc¹³	5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
pRR536	Wild-type Gln3_1–730-Myc¹³	5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR658^b	Gln3_{NES mam.}	5′-CAATCGGCCGCTGGAAAAGACCGCAGGCATTGCAAAGAGT-3
		5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
		5′-CAATCGGCCGTCGGACGTTATCAAAAAGAGGATTTCAAAG-3
		5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT- 3′
		5′-GGCCGAATGAATTAGCCTTGAAATTAGCAGGTCTTGATATCAACAAGACAC-3′
		5′-GGCCGTGTCTTGTTGATATCAAGACCTGCTAATTTCAAGGCTAATTCATTC-3′
pRR702^b	Gln3_10–20Δ	5′-CAATCGGCCGCTTCGAATTTTCGGGGTCGTCTTGCATTTG-3′
		5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
		5′-CAATCGGCCGCATGGTCGAAGTAATGAAGAGCCGAG-3′
		5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR714^b	Gln3_64–73Δ	5′-CAATCGGCCGACGGACTTCGTGTCTCCTTTTACAGCAGC-3′
		5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
		5′-CAATCGGCCGCATTGAGTCGAATGTGCCGCCGTTTAATCC-3′
		5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR752^c	Gln3_{L64D,L67R,L71D,F73D}-Myc¹³	5′-CAATCGGCCGGATGATgacTATgacACGGACTTCGTG-3′
		5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
		5′-CAATCGGccgTGCCTCgtcCATTGAGTCGAATGTGCCGCC-3′
		5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR787	Gln3_{L332D,L336D,M340D,L343D}-Myc¹³	5′-CAATCGGCCGatcGGTACCATGgtcTTTCTGGAAatcACCGCAGGCATTGCAAAGAG-3′
		5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
		5′-CAATCGGCCGgacTCCTTAAAATCGGACGTTATCAAAAAG-3′
		5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR1090^c	Gln3_{L64A,L67R,L71A,F73A}-Myc¹³	5′-CAATCGGCCGGATGATgcaTATgctACGGACTTCGTGTCTCC-3′
		5′-CGCGGATCCTATACCAAATTTTAACCAATCCAATTCGTCAGCAATTGCT-3′
		5′-CAATCGGccgTGCCTCtgcCATTGAGTCGAATGTGCCGCCG-3′
		5′-ATCCCCGCGGGACGTCAACTCCATAGAAGTGACTTTTCCG-3′
pRR1215	Gln3_{M340D,L343D,L345D}-Myc¹³	5′-TTACATGGTACCgacAGGCCAgacTCCgacAAATCGGACGTTATCAAAAAG-3′
pRR1215	Gln3_{M340D,L343D,L345D}-Myc¹³	5′-CGGAACAACAGATCTGGATGAAGATTTACTGGAACTTG-3′
pRR1217	Gln3_{M340A,L343A,L345A}-Myc¹³	5′-TTACATGGTACCgcgAGGCCAgcaTCCgcaAAATCGGACGTTATCAAAAAG-3′
pRR1217	Gln3_{M340A,L343A,L345A}-Myc¹³	5′-CGGAACAACAGATCTGGATGAAGATTTACTGGAACTTG-3′
DAL3-5	DAL3 promoter oligonucleotide	5′-tcgacTGGATTGGCAAATAAATGGGGAAAGATAAGCGAGATAAGACTGATAAGAAG
		CATATGCGGTCTATTCATGg-3′
		5′-cCATGAATAGACCGCATATGCTTCTTATCAGTCTTATCTCGCTTATCTTTCCCCATT
		TATTTGCCAATCCAgtcga-3′
pAA15	NLS-LexA-URE2	Structure in Kulkarni et al. (2001)
pRS316	Vector	Structure in http://genome-www.stanford.edu/vectordb/vector_descrip/COMPLETE/PRS316.SEQ.html

All plasmids contain full-length gln3 genes driven by the native GLN3 promoter.

An EagI site, encoding arginine and proline, replaced the deleted amino acids.

The L67R substitution was required for the cloning strategy we employed.