Table 3 Primer sequences used in this study

HA1-kanphleo

ATGCTATCATTTCCTTTGATATTGGATCATGGTA GAC AACCCTTAATATAACTTCGTA)

Cloning of antibiotic resistance gene cassettes. Sequences in italics are priming sites in pUG6/pUG66. For HA1-kanphleo, the 5′ sequence is homologous with sequence just downstream from the 2 μm ori pSPG1/pSPG2; the SalI site in pUG6 is destroyed, but the AccI site (underlined) remains for facile removal of the cassette. For HA2-kanphleo, the 5′ sequence is homologous with sequence at the end of the ADH1 terminator in pSPG1/pSPG2.

HA2-kanphleo

TGCTTTCTCAGGTATAGCATGAGGTCGCTCCTAGTGGATCTGATATCACC

2muDown

CCATTCCATGCGGGGTATCG

Screening colonies and sequencing for pCEV-G1-Ph, pCEV-G1-Km, pCEV-G2-Ph and pCEV-G2-Km

ADH1-T F1

TCGTTGGTAGATACGTTGTTGAC

GAL10LacF

TGATATCGAATTCCTGCAGCCCGGG GGATCC GTTTTTTCTCCTTGACGTTAAAGTA

Amplification of GAL10-GAL1 promoter region from S. cerevisiae S288C. The sequence in italics is homologous to the target vector pSF011 [22]. Restriction enzyme sites are underlined. The remaining sequence is the priming site for S. cerevisiae S288C genomic DNA.

Gal10LacR

GTAATCATGGTCATGGTGCGGCCG CTCTAGA GAATTTTCAAAAATTCTTACTTTTTTTTTG

GAL10P2B

GTGTGCGGCCGGCC GTTTTTTCTCCTTGACGTTAAAGTA

Amplification of the GAL promoter region from pGAL10lac. Restriction sites are underlined and priming sequence is italicised

GAL10P1A

GATCCCACTAGT GAATTTTCAAAAATTCTTACTTTTTTTTTG

HXT7P1B

GTGTGCGGCCGGCC CCGTGGAAATGAGGGGTATG

Amplification of the HXT7 promoter from pSF015 [22]. Restriction sites are underlined and priming sequence is italicised

HXT7P2A

GATCCCACTAGT TTTTTGATTAAAATTAAAAAAAC

CYC1-TR1

GGGACCTAGACTTCAGGTTGTC

Screening for promoter:lacZ yeast strains

Lac9434r

GAAGCCTGCGATGTCGGTTTC

ADH1-T R1

GGAGCGACCTCATGCTATACC

Screening for pCEV-G3 and pCEVG4 constructs

SFB018

GGATATGTATATGGTGGTAATGCC

Screening for pCEV-G3 constructs

SFB017

GAGACGATATATGCCAATACTTC

Screening for pCEV-G4 constructs