A growth-coupling strategy for improving the stability of terpenoid bioproduction in Escherichia coli

Table 2 Plasmids used in the study

Plasmid	Description	Source
pSIMcpf1	pSC101^T^S ori; Hyg^R; P_R: cI^TS; P_L: Gam-Beta-Exo; P_BAD: AraC, crRNA_pMB1-T_{BBa_B1006}; P_JS23151: AsCpf1-T_{BBa_B1002}	[38]
pTF	pMB1 ori; Stm^R; P_JS23119	[38]
pTF-dxr	pMB1 ori; Stm^R; P_JS23119: crRNA_dxr-T_rrnBT1-T7Te; dDNA_FRT-dxr-FRT	This work
pCP20	pSC101^T^S ori; Amp^R, Cam^R; P_R: cI^TS, Flp	[39]
pTet-RFP	pSC101 ori; Cam^R; P_tet: TetR, mRFP1-T_rrnBT1-T7Te	pBbS2c-RFP [40]
pTet-Flp	pSC101 ori; Cam^R; P_tet: TetR, Flp-T_T7Te	This work
pTrc-RFP	p15A ori; Amp^R; P_lacIq: LacI; P_trc: mRFP1-T_rrnBT1-T7Te	pBbA1a-RFP [40]
pMVA-Lin	p15A ori; Amp^R; P_lacI: LacI; P_lacUV5: AACT-HMGS-HMGR; P_J23116: MK-PMK-PMD-IDI-T_{BBa_B1002}; P_trc: GPPS-LinS-T_rrnBT1-T7Te	pMVA-GLinS NR2 [28]
pLMVA-Lin	p15A ori; Amp^R; P_lacI: LacI; P_J23116: MK-PMK-PMD-IDI-T_{BBa_B1002}; P_trc: GPPS-LinS-T_ECK120015170	This work

P and T: promoters and terminators respectively
^TS: temperature-sensitive; FRT: Flp recombinase target sequences; Amp^R, Cam^R, Stm^R, Hyg^R: enzyme for resistance to ampicillin/carbenicillin, chloramphenicol, streptomycin and hygromycin respectively; crRNA: CRISPR RNA array recognised by the AsCpf1 Cas12a; dDNA: donor DNA used for repair
Refer to Figure S2 for plasmid maps and full gene names

ISSN: 1475-2859