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Table 2 Oligonucleotides used in this study

From: Modular systems metabolic engineering enables balancing of relevant pathways for l-histidine production with Corynebacterium glutamicum

Oligonucleotide Sequence Purpose
hisG1 5′-CGCGGATCCCAGCTCGATTTGGGTATCACCG-3′ Outer primer #1 for introduction of hisGG233H−T235Q, BamHI site underlined
hisG2 5′-CCAGTTGTCGCGTGCCAGTGGGGATACCTGTGGATGGGATAAGCCTGGGGTTACTGC-3′ Inner primer #1 for introduction of hisGG233H−T235Q, affected triplets underlined
hisG3 5′-GCAGTAACCCCAGGCTTATCCCATCCACAGGTATCCCCACTGGCACGCGACAACTGG-3′ Inner primer #2 for introduction of hisGG233H−T235Q, affected triplets underlined
hisG4 5′-CGCGGATCCGTTGATGGTGGTTCGTGAGATTTGG-3′ Outer primer #2 for introduction of hisGG233H−T235Q, BamHI site underlined
hisGseq 5′-GGTATCCATCAAGCTTGG-3′ Sequencing primer for hisGG233H−T235Q
hisD1 5′-CCCAAGCTTCGGTGTCGCTGAAGTTAAGTTCTG-3′ Outer primer #1 for exchange of native promoter with Ptuf in operon hisDhisChisBcg2302cg2301, HindIII site underlined
hisD2 5′-CGCAGGTCAGTGACATTCAACATATGTATTATGCATCTCGACCACCCCAGATTTACCTG-3′ Inner primer #1 for exchange of native promoter with Ptuf in operon hisDhisChisBcg2302cg2301, NdeI and NsiI sites underlined
hisD3 5′-CAGGTAAATCTGGGGTGGTCGAGATGCATAATACATATGTTGAATGTCACTGACCTGCG-3′ Inner primer #2 for exchange of native promoter with Ptuf in operon hisDhisChisBcg2302cg2301, NdeI and NsiI sites underlined
hisD4 5′-CGCGGATCCCTCATCAACACCCAAGATGGAAC-3′ Outer primer #2 for exchange of native promoter with Ptuf in operon hisDhisChisBcg2302cg2301, BamHI site underlined
tuf1 5′-AACTGCAGAACCAATGCATCCACAGGGTAGCTGGTAGTTTG-3′ Primer #1 for amplification of Ptuf, NsiI site underlined
tuf2 5′-GGGAATTCCATATGATGTCCTCCTGGACTTCGTGGTGGC-3′ Primer #2 for amplification of Ptuf, NdeI site underlined
hisH1 5′-CCCAAGCTTCGGAGTGAAATGAGGTCCTTGGTC-3′ Outer primer #1 for exchange of native promoter with Ptuf in operon hisHhisAimpAhisFhisIcg2294, HindIII site underlined
hisH2 5′-GAGAAGGGCGACAGTTTTGGTCATATGTATTATGCATCTCCCTTCCTGAGAATGACGGCTAGTCG-3′ Inner primer #1 for exchange of native promoter with Ptuf in operon hisHhisAimpAhisFhisIcg2294, NdeI and NsiI sites underlined
hisH3 5′-CGACTAGCCGTCATTCTCAGGAAGGGAGATGCATAATACATATGACCAAAACTGTCGCCCTTCTC-3′ Inner primer #1 for exchange of native promoter with Ptuf in operon hisHhisAimpAhisFhisIcg2294, NdeI and NsiI sites underlined
hisH4 5′-CGCGGATCCGCACGAAGTAGAAACGCTCATCAGG-3′ Inner primer #2 for exchange of native promoter with Ptuf in operon hisHhisAimpAhisFhisIcg2294, BamHI site underlined
hisN1 5′-CCCAAGCTTCGCATGAGCATTTCAGCCCAGTCC-3′ Outer primer #1 for exchange of native promoter with Ptuf in operon cg0911hisN, HindIII site underlined
hisN2 5′-GCTCTGGATTAGTCATGCCTTCCATATGTATTATGCATCTCGGTGGCGTTCTAAGAATCACAATCG-3′ Inner primer #1 for exchange of native promoter with Ptuf in operon cg0911hisN, NdeI and NsiI sites underlined
hisN3 5′-CGATTGTGATTCTTAGAACGCCACCGAGATGCATAATACATATGGAAGGCATGACTAATCCAGAGC-3′ Inner primer #2 for exchange of native promoter with Ptuf in operon cg0911hisN, NdeI and NsiI sites underlined
hisN4 5′-CGCGGATCCCACCGAACCACGTATAACCCATGG-3′ Outer primer #2 for exchange of native promoter with Ptuf in operon cg0911hisN, BamHI site underlined
hisE1 5′-CCCAAGCTTCTGACAGAGTTGAAGGCCCTCGAG-3′ Outer primer #1 for exchange of native promoter with PdapA–A16 and change of the translational start codon from GTG to ATG in operon hisEhisG, HindIII site underlined
hisE2 5′-CGTACAGCGAGTCAAATGTCTTCATATGTATTATGCATCTCGGAGGATATTAGTCGAATAATTTC-3′ Inner primer #1 for exchange of native promoter with PdapA–A16 and change of the translational start codon from GTG to ATG in operon hisEhisG, NdeI and NsiI sites underlined, base exchange bold
hisE3 5′-GAAATTATTCGACTAATATCCTCCGAGATGCATAATACATATGAAGACATTTGACTCGCTGTACG-3′ Inner primer #2 for exchange of native promoter with PdapA–A16 and change of the translational start codon from GTG to ATG in operon hisEhisG, NdeI and NsiI sites underlined, base exchange bold
hisE4 5′-CGCGGATCCGCAACGTAGATGGCGATATCTTTAGG-3′ Outer primer #1 for exchange of native promoter with PdapA–A16 and change of the translational start codon from GTG to ATG in operon hisEhisG, BamHI site underlined
dapA1 5′-AACTGCAGAACCAATGCATTGGTTCTGCAGTTATCACACCC-3′ Primer #1 for amplification of PdapA–A16, NsiI site underlined
dapA2 5′-GGGAATTCCATATGAGGCTCCTTTTAAATCGAGCGGCTCCGGTCTTAGCTGTTAAACC-3′ Primer #2 for amplification of PdapA–A16, NdeI site underlined
hisF1 5′-CGCAAGCTTCTCCATGCCCATGCTGGGTAAACGC-3′ Primer #1 for integration of stop codon and PsodA in front of hisF, HindIII site underlined
hisF2 5′-GTCACAAGCCCGGAATAATTGGCAGTTATTTACTTGTACTCCTCATTTAACG-3′ Primer #2 for integration of stop codon and PsodA in front of hisF, stop codon bold
sodA1 5′-CGTTAAATGAGGAGTACAAGTAAATAACTGCCAATTATTCCGGGCTTGTGAC-3′ Primer #3 for integration of stop codon and PsodA in front of hisF, stop codon bold
sodA2 5′-GAATAACTCGAATTGCCACGCCCATGGGTAAAAAATCCTTTCGTAGGTTTC-3′ Primer #4 for integration of stop codon and PsodA in front of hisF
hisF3 5′-GAAACCTACGAAAGGATTTTTTACCCATGGGCGTGGCAATTCGAGTTATTC-3′ Primer #5 for integration of stop codon and PsodA in front of hisF
hisF4 5′-CGCGGATCCCGCCGCGCTTTGCCCACTCGATTGC-3′ Primer #6 for integration of stop codon and PsodA in front of hisF, BamHI site underlined
hisFseq 5′-CTGACACCGAAGGCCATC-3′  
tuf2_1 5′-GATCAGCGACGCCGCAGGGTCTAGACCACAGGGTAGCTGGTAGTTTGAAAATC-3′ Primer #1 for amplification of Ptuf, XbaI site underlined
tuf2_2 5′-CATTGAGCGCCGACAATAACGATTGCAGCCATTGTATGTCCTCCTGGACTTCG-3′ Primer #2 for amplification of Ptuf
purA1 5′-CGAAGTCCAGGAGGACATACAATGGCTGCAATCGTTATTGTCGGCGCTCAATG-3′ Primer #1 for amplification of purA (cg3063)
purA2 5′-GATCTTCTTTTTATCAGCCACTGTATGTCCTCCTGGACTTCCTAGTTGTCAGCTAGTACG-3′ Primer #2 for amplification of purA (cg3063)
purB1 5′-CGTACTAGCTGACAACTAGGAAGTCCAGGAGGACATACAGTGGCTGATAAAAAGAAGATC-3′ Primer #1 for amplification of purB (cg2876)
purB2 5′-CTTCTCTCATCCGCCAAAACAGTTAAAGAATCTCACCTGGTCGGTAGTC-3′ Primer #2 for amplification of purB (cg2876)
rrnB1 5′-GACTACCGACCAGGTGAGATTCTTTAACTGTTTTGGCGGATGAGAGAAG-3′ Primer #1 for amplification of terminator region of rrnB (E. coli)
rrnB2 5′-CTGCAGATATCCATCACACTGGCGGCCGCAGGAGAGCGTTCACCGACAAACAAC-3′ Primer #2 for amplification of terminator region of rrnB (E. coli), NotI site underlined
ABseq1 5′-GCGATTGAAGACCGTC-3′ Sequencing primer for pJC4 purA purB
ABseq2 5′-GCAATCGGCACCACCGGC-3′ Sequencing primer for pJC4 purA purB
ABseq3 5′-GCCTGCATGGGACGAAG-3′ Sequencing primer for pJC4 purA purB
ABseq4 5′-GTCACCGAACTGCTCAAC-3′ Sequencing primer for pJC4 purA purB
ABseq5 5′-GATGGAAGCAGGATCGCG-3′ Sequencing primer for pJC4 purA purB
pgi1 5′-CCCAAGCTTCAGCGTTGCGTGACGCACTCATTG-3′ Outer primer #1 for introduction of pgiGTG, HindIII site underlined
pgi2 5′-CCAAACCTGGGTGGTCGAAATGTCCGCCACGAAAACTCCTTTATTGTCG-3′ Inner primer #1 for introduction of pgiGTG, nucleotide exchange bold
pgi3 5′-CGACAATAAAGGAGTTTTCGTGGCGGACATTTCGACCACCCAGGTTTGG-3′ Inner primer #2 for introduction of pgiGTG, nucleotide exchange bold
pgi4 5′-CGCGGATCCCCAATGTTGACGATCTTCTTGATCG-3′ Outer primer #2 for introduction of pgiGTG, BamHI site underlined
pgiseq 5′-CAAGCGTTGGGTTAAGGAGGA-3′ Sequencing primer for pgiGTG
gcv_Cjk_start_EcoRV 5′-ATCGATATCCGAGAGGAGACACAACATGTCTTCTGCAGCTACTCGC-3′ Primer #1 for amplification of gene cluster gcvPTH from C. jeikeium genome
gcv_Cjk_MluI_XbaI 5′-CAGTCTAGAACGCGTGGAACCGACCATAGGGTCTTG-3′ Primer #2 for amplification of gene cluster gcvPTH from C. jeikeium genome
lipB-Cjk_start-EcoRV 5′-GCGGATATCATGGGATTCCAGCAAGGC-3′ Primer #1 for amplification of gene cluster lipAB from C. jeikeium genome
lipA-Cjk_stop-XbaI 5′-GCGTCTAGATCCTTCGCCATGGATTCAAC-3′ Primer #2 for amplification of gene cluster lipAB from C. jeikeium genome
pK19-fw 5′-TAATGCAGCTGGCACGAC-3′ Primer #1 for sequencing the pK19mobsacB inserts
pK19-rev 5′-GTAGCTGACATTCATCCG-3′ Primer #2 for sequencing the pK19mobsacB inserts