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Table 2 Primers used in the present study

From: NADH-dependent biosensor in Saccharomyces cerevisiae: principle and validation at the single cell level

Name

Description

Sequence

TRP1_gpd1Over_f

Forward primer for amplification of TRP1 (p424 as template) with GPD1 overhangs

TCCACAAACACAAATATTGATAATATAAAGAACGACATTACTATATATATAATATAG

TRP1_gpd1Over_r

Reverse primer for amplification of TRP1 (p424 as template) with GPD1 overhangs

AGTATGATATGTTATCTTTCTCCAATAAATAGGCAAGTGCACAAACAATAC

TRP1_gpd1_veri_f

Amplification of the 5' region of GPD1 (CEN.PK113-7D as template)

TTCCATTCACATATCGTCTTTGG

+ Verification of GPD1 deletions

5-OEGpd1_r

Amplification of the 5' region of GPD1 (CEN.PK113-7D as template)

TATATTATCAATATTTGTGTTTGTGG

3-OEGpd1_f

Amplification of the 3' region of GPD1 (CEN.PK113-7D as template)

TTTATTGGAGAAAGATAACATATCATAC

3-OEGpd1_r

Amplification of the 3' region of GPD1 (CEN.PK113-7D as template)

ATTTTCTTAGGACGCCGCAAAATATC

TRP1_gpd1_veri_r

Verification of GPD1 deletions

GAGGAACTCTTGGTATTCTTGCC

LEU2_gpd2Over_f

Forward primer for amplification of LEU2 (p425 as template) with GPD2 overhangs

TTCCTTTTCCTTCGCTCCCCTTCCTTATCATCGACTACGTCGTAAGGCCG

LEU2_gpd2Over_r

Forward primer for amplification of LEU2 (p425 as template) with GPD2 overhangs

GATCAGAGGGGGAGGGGGGGGGAGAGTGTCGAGGAGAACTTCTAGTATATC

LEU2_gpd2OO_f

Forward primer for nested PCR for adding homology regions to the GPD2 deletion casette

GTATTTTGGTAGATTCAATTCTCTTTCCCTTTCCTTTTCCTTCGCTCCC

LEU2_gpd2OO_r

Reverse primer for nested PCR for adding homology regions to the GPD2 deletion casette

AAATTGGTTGGGGGAAAAAGAGGCAACAGGAAAGATCAGAGGGGGAGGG

LEU2_gpd2_veri_f

Verification of GPD2 deletions

CGTGTATCTTCTAAGATTCAGTC

LEU2_gpd2_veri_r

Verification of GPD2 deletions

CTAATGGCTCAACGTGATAAGG

GPD2p_XbaI_r

Cloning of GPD2 from CEN.PK113-7D

CTTCTAGATTGATAAGGAAGGGGAG

GPD2p_SacI_f

Cloning of GPD2 from CEN.PK113-7D

TCGAGCTCCGCAATGTTTCGTTGG

yEGFP_XbaI_f

Cloning of yEGFP3 from pyEGFP3

AATCTAGAAGCATTAAAAAATGTCTAAAGGTGAAG

yEGFP_PstI_r

Cloning of yEGFP3 from pyEGFP3

TTCCTGCAGAGTTATTTGTACAATTCATCCATAC

HIS3_flank_f

Cloning of HIS 3 - Forward primer

CCAGGTATCGTTTGAACACGG

HIS3_flank_r

Cloning of HIS 3 - Reverse primer

GCTCAGTTCAGCCATAATATG

LEU2_flank_f

Cloning of LEU2 - Forward primer

GGATAATTATACTCTATTTCTCAAC

LEU2_r

Cloning of LEU2 - Reverse primer

TTAAGCAAGGATTTTCTTAAC

TRP1_f

Cloning of TRP1 - Forward primer

ATGTCTGTTATTAATTTCAC

TRP1_r

Cloning of TRP1- Reverse primer

CTATTTCTTAGCATTTTTG

YIplac211_f

Cloning of GPD2 p-yEGFP3-PGK 1t

TTTATCTTCGTTTCCTGC

YIplac211_r

Cloning of GPD2 p-yEGFP3-PGK1 t

AAAACTGTATTATAAGTAAATG

HIS3_O_f

Cloning of HIS3- expression cassette, for change of auxotrophic marker.

TTATAATACAGTTTTCCAGGTATCGTTTGAACACGG

HIS3_O_r

Cloning of HIS3-expression cassette, for change of auxotrophic marker.

GGAAACGAAGATAAATCGCTCAGTTCAGCCATAATATG