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Table 4 Primers used in this study

From: Metabolic engineering design to enhance (R,R)-2,3-butanediol production from glycerol in Bacillus subtilis based on flux balance analysis

Name

Sequence (5′ → 3′)

Usage

P_ackA01

CGGTATCGGTGAAAACAGTGGTTTTAGAGCTAGAAATAGCAAGTT

Construction of pJOE_ackA sgRNA

P_ackA02

GAGAATTGAGTAAAATGTACCTACGCGGTATCGGTGAAAACAGTG

 

P_ackA03

GGTCGACGGCCAACGAGGCCCCCCTCTCAACCGTGTTTCTATTTT

PCR of homology arms for ackA deletion

P_ackA04

TCAAGAGAATGTGCTTTCATGCGATGATTGACGCTCCTTTATACTCTGTA

 

P_ackA05

TCAAGAGAATGTGCTTTCATGCGATGATTGACGCTCCTTTATACTCTGTA

 

P_ackA06

ATGAGCGAAGGAATGGCCTTCGTGGCCAATAAGGCCTTTCTAG

 

P_ackA07

CGACGGAAGTATCAAGACCTCCTGA

PCR of check ackA deletion

P_ackA08

ACTGGCAGATCACAAATGACCGTG

 

P_pta01

GAATTGAACCTTACGCTGGGCGGGTTTTAGAGCTAGAAATAGCAAGTT

Construction of pJOE_pta spacer

P_pta02

GAGAATTGAGTAAAATGTACCTACGGAATTGAACCTTACGCTGGGCGG

 

P_pta03

GGTCGACGGCCAACGAGGCCTGGATACAGCTTTTGCAGCT

PCR of homology arms for pta deletion

P_pta04

GAGCTGCCATTGTCTTCAATTTTAAATAAAACCTCCTCA

 

P_pta05

TTGTCTTCAATTTTAAATAAAACCTCCTCAAAAAGTTACAA

 

P_pta06

GGCCAAAAAGCTGATTTTATGGGCCAATAAGGCCTTTCTAG

 

P_pta07

GTCCCGGAGCCTTTCTCTCT

PCR of check pta deletion

P_pta08

GAAGATCTGGCAACGACAAC

 

P_lctE01

GATGTGATGGATTTAAACCAGTTTTAGAGCTAGAAATAGCAAGTT

Construction of pJOE_lctE spacer

P_lctE02

GAGAATTGAGTAAAATGTACCTACGGATGTGATGGATTTAAACCA

 

P_lctE03

GGTCGACGGCCAACGAGGCCCTTTCAATGATCAACTGACGATTAC

PCR of homology arms for lactE deletion

P_lctE04

GAAACATACCCTGGAAGGATGATCCGCAACTTTAGAGTAAAGGG

 

P_lctE05

GAAACATACCCTGGAAGGATGATCCGCAACTTTAGAGTAAAGGG

 

P_lctE06

GAATCAAACAGACATGGCCCGGCCAATAAGGCCTTTCTAG

 

P_lctE07

TGTAGCTGTGCTTTATCCTGATGATATTGC

PCR of check lactE deletion

P_lctE08

TGCTCGGGCCGGAATTAGCAAACATTTT

 

P_mmgA01

CTGAATGAGGTGAAACCATGGTTTTAGAGCTAGAAATAGCAAGTT

Construction of pJOE_mmgA spacer

P_mmgA02

GAGAATTGAGTAAAATGTACCTACGCTGAATGAGGTGAAACCATG

 

P_mmgA03

GGTCGACGGCCAACGAGGCCTCGGATGATTCATGCCCAGC

PCR of homology arms for mmgA deletion

P_mmgA04

TTCAAATCCCCTTTTTCATCATGTTGGTTTCACCTCATTCAGAAGATAGA

 

P_mmgA05

TTCAAATCCCCTTTTTCATCATGTTGGTTTCACCTCATTCAGAAGATAGA

 

P_mmgA06

TCCTTTAAGTGATGTCGTGCGATTAGGCCAATAAGGCCTTTCTAG

 

P_mmgA07

GGCCTGAGTGAAGGACTTCCATAAT

PCR of check mmgA deletion

P_mmgA08

ATCAAGCTCTATCCCATCCGCTCCG