Table 1.

Oligonucleotide primers used in this study. Lowercase bases in ALID2018-ALID2019 indicate the chimerical regions for recombination in pPZP-201BK. Lowercase bases in GI114 indicate the region for recombination in pGI3, with the right border in bold and underlined the three bases that usually integrate in the genome; lowercase bases in GI115 are the reverse complement of ai290 used to amplify NAT; lowercase bases in ALID2147 indicate the region for recombination into pGI3. Lowercase bases in MAI0170 and MAI0171 are for recombination into pPZP-NEO11, and in MAI0172 and MAI0175 into pPZP-201BK.

Name	Sequence (5´-3´)	Purpose
ALID2018	GCTGCACTGAACGTCAGAAGgaaaagtgccacctgaacga	pGI3 generation
ALID2019	CCATTTGTATGTGCATGCCAggtaataactgatataatta	pGI3 generation
GI114	tcttaggtttacccgccaatatatcctgtcaTCTCAGGAGAGAATTGAG TGC	pGAL7 amplification for pGI7 construction
GI115	gcttctgtgagtcctcccGATCAGGCGCTGGCTGTGAGTTG	pGAL7 amplification for pGI7 construction
ai290	GGGAGGACTCACATAAGC	NAT amplification for pGI7 construction
ALID2147	tgattacgaattcttaattaagatatcgagGAAGAGATGTAGAAACGA	NAT amplification for pGI7 construction
GI120	CGTCATGTTGTCAATTGAGC	Inverse PCR to identify T-DNA insertion sites.
ai077	AGAGGCGGTTTGCGTATTGG
MAI0170	taaaacgacggccagtgccaGCTGTCCATACATCACAGAC	Amplify wild type copy of PRE3 for complementation
MAI0171	ggcgaattcttaattaagatGGAGTTTAGGTGTTACAAGG
MAI0172	aacagctatgacatgattacgTCGGTAGGCAGTATGAGG	Amplify 5´ side of PRE3 for gene replacement
MAI0173	GCTTATGTGAGTCCTCCCCGCACCAATGACAACACC
MAI0174	CTCGTTTCTACATCTCTTGGTTTTGGGAAGGCAAGG	Amplify 3´ side of PRE3 for gene replacement
MAI0175	gtaaaacgacggccagtgccaTGATTTCTGCTGCCTCAACG
ai006	GAAGAGATGTAGAAACGAG	Amplify NAT cassette with ai290
ai037	ATGGCGGCCGCCACTCTTGAC	Confirm PRE3 deletion on the 5´ side
MAI0184	GGTGATGACGGAGATACGAG
ai270	ACTTCTCGCAAAGTGAATCC	Confirm PRE3 deletion on the 3´ side
MAI0185	CTTCTCCTTTCTTGCTTCAC
ai144	GAAAAGGTTACGACAGAGG	Amplify MATα
ai145	TGGGAAGAACTATCAGAGG
ai150	ACGCAGTGTCAACTGGTC	Amplify MATa
ai151	CGCACCTTTTCTGTAGGG