TABLE 2.

Primers and PCR products used in this study

Primer or PCR product	Nucleotide sequence (5′-3′)a or PCR product descriptionb	Fragment(s) amplified
Primers
Alfa1-HindIII	AAGCTTACGATGAGATTTCCTTCAATTTTTACTGC	α1
Alfa3-XbaI	TCTAGAAAGCTGGCGGCCGCCGCGGCTC	α1
Alfa4-SmaI	CCCGGGACGATGAGATTTCCTTCAATTTTTACTG	α-L50B
L50A1-XhoI	CTCGAGAAAAGAATGGGAGCAATCGCAAAATTAGTAGCAAAG	L50A
L50A2-XbaI	TCTAGAGTATAACCCCCGGGATTTTAAATATGTTTTTTAATCCACTCAATG	L50A
L50B1-XhoI	CTCGAGAAAAGAATGGGAGCAATCGCAAAACTAGTGAC	L50B
L50B2-XbaI	TCTAGAAACATTAATGTCTTTTTAGCCATTTTTCAATTTGATC	L50B, α-L50B
pYES2-F	AACCCCGGATCGGACTACTAG	α2
pYES2-R	GGCGTGAATGTAAGCGTGAC	α2
PCR products
Fragment α1	345-bp HindIII-XbaI fragment containing the Kozak sequence fused to MFα1s
Fragment α2	442-bp fragment containing the Kozak sequence fused to MFα1s
Fragment L50A	170-bp XhoI-XbaI fragment containing MFα1s fused to entL50A
Fragment L50B	154-bp XhoI-XbaI fragment containing MFα1s fused to entL50B
Fragment α-L50A	422-bp HindIII-XbaI fragment containing MFα1s fused to entL50A
Fragment α-L50B	406-bp SmaI-XbaI fragment containing MFα1s fused to entL50B

^aCleavage sites for restriction enzymes are underlined; Kozak sequence (ACGATGA), and the nucleotides encoding the Kex2 signal cleavage site (AAAAGA) of the mating pheromone α-factor 1 secretion signal (MFα1_s) are shown in bold.

^bMFα1_s refers to the yeast gene region encoding MFα1_s including the Kex2 cleavage site.