Table 2 Descriptions and sources of plasmids used in this study

pTrc99a

AmpR, pBR322-origin, trc promoter, 4.2 kb

[35, 38]

pCas

Kan^R, Cas9 nuclease expression plasmid, temperature-sensitive origin

[39, 40]

pTrc-ilvA*

AmpR, ilvA* cloned in the sac I and BamH I site of pTrc99A

This study

Gap-ilvA*

Amp^R, replacement of Trc promoter in pTrc-ilvA* with the Gap promoter

This study

Pbs-ilvA*

Amp^R, replacement of Trc promoter in pTrc-ilvA* with the Pbs promoter

This study

pTrc-ilvA* -BleuDH

Amp^R, BleuDH cloned in the Xba I and Hind III site of pTrc-ilvA*

This study

pTrc-ilvA*-leuDH

AmpR, leuDH cloned in the Xba I and Hind III site of pTrc-ilvA*

This study

pTrc-ilvA*-Trc-leuDH

Amp^R, leuDH carrying a Trc promoter and RBS cloned in the Xba I and Hind III site of pTrc-ilvA*

This study

pTrc-ilvA*-Gap-leuDH

Amp^R, leuDH carrying a Gap promoter and RBS cloned in the Xba I and Hind III site of pTrc-ilvA*

This study

pTrc-ilvA*-Pbs-leuDH

Amp^R, leuDH carrying a Pbs promoter and RBS cloned in the Xba I and Hind III site of pTrc-ilvA*

This study

Gap-ilvA*-leuDH

Amp^R, replacement of Trc promoter in pTrc-ilvA*-leuDH with the Gap promoter

This study

Gap-ilvA*-Trc-leuDH

Amp^R, leuDH carrying a Trc promoter and RBS cloned in the Xba I and Hind III site of Gap-ilvA*

This study

Gap-ilvA*-Gap-leuDH

Amp^R, leuDH carrying a Gap promoter and RBS cloned in the Xba I and Hind III site of Gap-ilvA*

This study

Gap-ilvA*-Pbs-leuDH

Amp^R, leuDH carrying a Pbs promoter and RBS cloned in the Xba I and Hind III site of Gap-ilvA*

This study

Pbs-ilvA*-leuDH

Amp^R, replacement of Trc promoter in pTrc-ilvA*-leuDH with the Pbs promoter

This study

Pbs-ilvA*-Trc-leuDH

Amp^R, leuDH carrying a Trc promoter and RBS cloned in the Xba I and Hind III site of Pbs-ilvA*

This study

Pbs-ilvA*-Gap-leuDH

Amp^R, leuDH carrying a Gap promoter and RBS cloned in the Xba I and Hind III site of Pbs-ilvA*

This study

Pbs-ilvA*-Pbs-leuDH

Amp^R, leuDH carrying a Pbs promoter and RBS cloned in the Xba I and Hind III site of pbs-ilvA*

This study

pTarget-X

A plasmid used to transcript sgRNA targeting the particular gene X in genome. X refers to the amino acid biosynthetic pathways

This study