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Table 4 The sequences of the oligonucleotide primers used in this study

From: Impact of an energy-conserving strategy on succinate production under weak acidic and anaerobic conditions in Enterobacter aerogenes

Primer

Primer sequences (5′–3′)

Disruption of poxB, pflB, ptsG, and ptsI genes

 ΔpoxB-attL

5′-ACGTAACCTGTAGTTTCATCTAAGCTTGATAGCGTTATCACAAAAAGGAGATGGAAAACCTGAAGCCTGCTTTTTTATACTAAGTTGGC-3′

 ΔpoxB-attR

5′-TATCACTGCGAAGATCTATCACGGCATATCCTTGTTCTGATTCAGGGTGATAGCATTGTTTACTCAGCCTTATTTTTCAGATTTTATTCGG-3′

 ΔpflB-attL

5′-CATTAATGGTTGTCTCAGGCAGTAAATAAAAAATCCACTTAAGAAGGTAGGTGT TACATGTGAAGCCTGCTTTTTTATACTAAGTTGGC-3′

 ΔpflB-attR

5′-AAAAAGGCCCCACTGATGTGGGGCCTTTATTGTACGCTTTTTCAGTCAGACAG GGAATTACGCTCAAGTTAGTATAAAAAAGCTGAACGA-3′

 ΔptsG-attL

5′-ATGTTTAAGAATGCATTTGCTAACCTGCAGAAGGTCGGTAAATCGCTGATGCTGCCAGTATGAAGCCTGCTTTTTTATACTAAGTTGGC-3′

 ΔptsG-attR

5′-TTAGCTATTGCGGATGTACTCATCCATCTCGGTTTTCAGGTTATCGGACTTGGTGCCGAACGCTCAAGTTAGTATAAAAAAGCTGAACGAG-3′

 ΔptsI-attL

5′-TGAACTCGAGTAAGTTCACGGGTTCTTTTTAATATCAGTCACAAGTAAGGTAGGGTTATGTGAAGCCTGCTTTTTTATACTAAGTTGGC-3′

 ΔptsI-attR

5′-ATCGAACAAACCCATGATCTTCTCCTAAGCAGTAATTTGGGCCGCGCATCTCGTGGATTACGCTCAAGTTAGTATAAAAAAGCTGAACGA-3′

 ΔpoxB-CF

5′-GTCGATTAGCTTATGTAGACCG-3′

 ΔpoxB-CR

5′-TGGAGTTCATTGACGGTTGGG-3′

 ΔpflB-CF

5′-GGCTTTGAACACAGCATCGC-3′

 ΔpflB-CR

5′-CTGTTTTGACAGGTACCC-3′

 ΔptsG-CF

5′-TGAATGAAACGTGATAGCCG-3′

 ΔptsG-CR

5′-TCAACTATAGACTTAGCTGC-3′

 ΔptsI-CF

5′-CAAGAAGTTACCATTACCGC-3′

 ΔptsI-CR

5′-TTGCCAGTAGGTTTGATGGC-3′

 frdABCD-CF

5′-GCGGCGATATTACATCCTGA-3′

 frdABCD-CR

5′-TCTAAGTGTAGATAGCGGCA-3′

A. succinogenes PCK expression cassette

 pck-tacR

5′-CGAGTTTGTTTAAGTCAGTCATGGCAGTCTCCTTGTGTGAAATTGTTATCCG-3′

 tac-pckF

5′-CGGATAACAATTTCACACAAGGAGACTGCCATGACTGACTTAAACAAACTCG-3′

 ΔpoxB-pckR

5′-TATCACTGCGAAGATCTATCACGGCATATCCTTGTTCTGATTCAGGGTGATAGCATTGTTATCTCAGCCTTATTTTTCAGATTTTATTCGG-3′

Promoter replacement of frdABCD

 frdABCD-tacF

5′-AAATAAAATTGATAAATTAGCGCACGGATTGATAAAAAAATCGAACGCGTTGAAGCCTGCTTTTTTATACTAAGTTGGCA-3′

 frdABCD-tacR

5′-CCTGCTCCGCCTGCGCCAATGACGACAAGATCGGCTTGAAAAGTTTGCACGGCAGTCTCCTTGTGTGAAATTGTTATCCG-3′