Table 1.
Strains, plasmids and primers used in this study
| Name | Description | Source/purpose |
|---|---|---|
| Strain | ||
| E. coli DH5α | supE44 Δ(lacZYA-argF) U169 (Φ80lacZ ΔM15) hsdR17 recA endA1 gyrA96 thi-1 relA1 | Invitrogen |
| E. coli BW 25113 | lacI q rrnB T14 ΔlacZ WJ16 hsdR514 ΔaraBAD AH33 ΔrhaBAD LD78 | [17] |
| E. coli DOPA-30N | L-DOPA overproducer | [16] |
| Plasmid | ||
| pBbB2k-GFP | Expressing plasmid, BglBrick vectors, tet promoter, pBBR1 ori, kanr, Addgene plasmid #35345 | [18] |
| pBbB2k-ipdC-L-feaB | pBbB2k derivatives harboring the fusion gene cluster of ipdC from Azospirillum brasilense and feaB from E. coli with a (GSG)2 linker | This study |
| pBbB2k-ARO10-L-feaB | pBbB2k derivatives harboring the fusion gene cluster of ARO10 from Saccharomyces cerevisiae and feaB from E. coli with a (GSG)2 linker | This study |
| pBbB2k-aas-L-feaB | pBbB2k derivatives harboring the fusion gene cluster of aas from Petroselinum crispum and feaB from E. coli with a (GSG)2 linker | This study |
| pBbB2k-tydc-L-tynA-L-feaB | pBbB2k derivatives harboring the fusion gene cluster of tydc from Pseudomonas putida KT2440, tynA and feaB from E. coli with a (GSG)2 linker | This study |
| pBbB2k-ARO10*- L-feaB* | pBbB2k derivatives harboring the fusion gene cluster of the evolved ARO106D5 from S. cerevisiae and feaB2E1 from E. coli with a (GSG)2 linker | This study |
| pBbB2k-ARO10*-feaB* | pBbB2k derivatives harboring the gene cluster of the evolved ARO106D5 from S. cerevisiae and feaB2E1 from E. coli, one operon | This study |
| pBbB2k-ARO10*-TIGR-feaB* | pBbB2k derivatives harboring the TIGR-mediated gene cluster of the evolved ARO106D5 from S. cerevisiae and feaB2E1 from E. coli | This study |
| pZBK | BglBrick/ePathBrick expression vector, pBBR1 ori, P37 promoter, kanr | [19] |
| pZBK-PesaSIC-PesaRAS | Quorum-sensing plasmid, pBBR1 ori, P37 promoter, kanr | This study |
| pZBK-PesaRAS-4HPAA | pZBK-PesaSIC-PesaRAS harboring the TIGR-mediated gene cluster of the evolved ARO106D5 from S. cerevisiae and feaB2E1 from E. coli | This study |
| Primer | ||
| FeaBF | CCGGAATTCAAAAGATCTGGTATGACAGAGCCGCATGTAGCAG (EcoRI, BglII) | PCR for feaB |
| FeaBR | CGCGGATCCTTAATACCGTACACACACCGACTTAGTTTCACACCAAC (BamHI) | |
| TynAF | CCGGAATTCAAAAGATCTGGTATGGGAAGCCCCTCTCTGTATTCTG (EcoRI, BglII) | PCR for tynA |
| TynAR | CGCGGATCCACCTGAACCCTTATCTTTCTTCAGCGCCC (BamHI) | |
| ARO10F | CCGGAATTCAAAAGATCTTTTCGGAATTAAGGAGGTAATAAATATGGCACCTGTTACAATTG (EcoRI, BglII) | PCR for ARO10 |
| ARO10R | CGCGGATCCACCACTACCTTTTTTATTTCTTTTAAGTG (BamHI) | |
| AROF | GATAGAGAAAAGAATTCAAAAGATCTTTTCGGAATTAAGGAGG (EcoRI, BglII) | error-prone PCR forARO10-feaB |
| FeaR | TACTCGAGTTTGGATCCTTAATACCGTACACACACCGACTT (BamHI) | |
| epAROF | ACCTTCGATTCCGACCTCATTAAGC | error-prone PCR for ARO10 |
| epAROR | CGCGGATCCTTAATACCGAACACACACCGACTTAGTTTCACACCAAC | |
| TIGRs-F(X) | CTAGCTAGCGCCTAGCAAGATCTCCTGATCCCGGTGC (NheI) | PCR for TIGRs |
| TIGRs-R(A) | TTTTCCTTTTGCGGCCGCGGATACAGTATCTGCGGTACCCTAG (NotI) | |
| Qaro10F | CGACACCTTGTTTCCAACCG | qRT-PCR for ARO10 |
| Qaro10R | TGAAGTCACCAGGAACACCG | |
| QfeaBF | GCCAACGCTGGTGGTAAATC | qRT-PCR for feaB |
| QfeaBR | GCCTCTTCTCCATCCGCTAC | |