TABLE 2.

Primers used for PCR amplification

Primer	Sequence (5′-3′)a	Target sequence
TatAoKO up for Hind	GATCAAGCTTCCCACCCACGAACAAATC	797 bp upstream of tatAo start codon, extension towards gene
TatAoKO up rev EcoRI	CATGGAATTCCATTACGTTCGGTTACGT	1 bp upstream of tatAo start codon, extension away from gene
TatAoKO down for Bam	GTACGGATCCTAGTACGCGTCTCCCCCG	tatAo stop codon, extension away from gene
TatAoKO down rev Xba	GTACTCTAGACCTCGCCTTCGTCCTCTC	797 bp downstream of tatAo stop codon, extension towards gene
TatAtKO up for Xho	GTACCTCGAGGAATCAGGTGCGCTACCG	798 bp upstream tatAt start codon, extension towards gene
TatAtKO up rev EcoR	GTACGAATTCCATCTATTTCTGCGTTACC	tatAt start codon, extension away from gene
TatAtKO down for Bam	GTACGGATCCTGACCACAGCTCACTGTC	tatAt stop codon, extension away from gene
TatAtKO down rev Xba	GTACTCTAGAGTGTTCCCACAGCCAGAC	819 bp downstream of tatAt stop codon, extension towards gene
TatCoKO up for Hind	GTACAAGCTTCGAAGATGCCGACGATGG	797 bp upstream of tatCo start codon, extension towards gene
TatCoKO up rev EcoRI	GTACGAATTCCATTCAGGTGTCCTACG	tatCo start codon, extension away from gene
TatCoKO down for Bam	GTACGGATCCACCAAACTGAAGGTCGCC	512 bp upstream of tatCo stop codon, extension away from gene
TatCoKO down rev Xba	GTACTCTAGAAGACAGGGCGACGAGGTC	136 bp downstream of tatCo stop codon, extension towards gene
TatCtKO up for Hind	GTACAAGCTTGAGCGCGTTGAGGATGTC	802 bp upstream of tatCt start codon, extension towards gene
TatCtKO up rev EcoRI	GTACGAATTCCATACGTACGTGGCGAAAGC	tatCt start codon, extension away from gene
TatCtKO down for Bam	GTACGGATCCGAGTGAGCACGGCGCACC	3 bp upstream tatCt stop codon, extension away from gene
TatCtKO down rev Xba	GTACTCTAGACGATGGCCTGACTCGTCG	793 bp downstream of tatCt stop codon, extension towards gene
ForFdx	GTAACGCCAGGGTTTTCC	199 bp upstream of pyrE2 start codon, extension towards pyrE2 gene
RevFdx	CATGGCTGCAGAGTTCGGCT	pyrE2 start codon, extension away from pyrE2 start codon
ForTatAo FdxOL	AGCCGAACTCTGCAGCCATGCTGGATTCCATTCCG	tatAo start codon, extension towards stop codon of tatAo
RevTatAo 6-His Xba	GATCTCTAGATCAGTGATGGTGATGGTGATGCGGGCCGCCTCTACTCTCGGCTTCCGT	1 bp upstream of tatAo stop codon, extension towards start codon of tatAo
ForTatAt FdxOL	AGCCGAACTCTGCAGCCATGTTCGAGACCATCACC	tatAt start codon, extension towards stop codon of tatAt
RevTatAt 6-His Xba	GATCTCTAGATCAGTGATGGTGATGGTGATGCGGGCCGCCGTTCGACGATTCCGTCGA	1 bp upstream of tatAt stop codon, extension towards start codon of tatAt
ForTatCo FdxOL	AGCCGAACTCTGCAGCCATGGCGGACGAGGAGCGT	tatCo start codon, extension towards stop codon of tatCo
RevTatCo 6-His Xba	GATCTCTAGATCAGTGATGGTGATGGTGATGCGGGCCGCCGTTCCCGGTCCAGCGTAA	1 bp upstream of tatCo stop codon, extension towards start codon of tatCo
ForTatCt FdxOL	AGCCGAACTCTGCAGCCATGTCCAGCGCCCTCGAC	tatCt start codon, extension towards stop codon of tatCt
ForTatCt FdxOL_NtermMyc	AGCCGAACTCTGCAGCCATGGAGCAGAAGCTCATCAGCGAGGAGGATCTGTCCAGCGCCCTCGAC	tatCt second codon, extension towards stop codon of tatCt
RevTatCt 6-His Hind	GATCAAGCTTTCAGTGATGGTGATGGTGATGCGGGCCGCCCTCCGCCGCGCCGCGGGC	1 bp upstream of tatCt stop codon, extension towards start codon of tatCt
Halocyanin For FdxOL	AGCCGAACTCTGCAGCCATGACGCAACGGGTTTCC	halocyanin1 start codon, extension towards stop codon of halocyanin1
Halocyanin RevMyc Hind	GAAAGCTTTTACAGATCCTCCTCGCTGATGAGCTTCTGCTCCTCGACGACGACGACGCCG	1 bp upstream of halocyanin1 stop codon, extension towards start codon of halocyanin1
HaloKK For	CAACGGGTTTCCAAGAAGTCGTTTCTCCACGCAACG	bp 25 of the halocyanin1 gene, extension towards stop codon of halocyanin1
HaloKK Rev	CTTCTTGGAAACCCGTTGCGTCAT	bp 18 of the halocyanin1 gene, extension towards start codon of halocyanin1

^aRestriction endonuclease sites are underlined. Regions required for overlap PCR addition of P_fdx or alteration of twin arginine codons are in bold. Sequences encoding 6His or Myc tags are italic.