Table 1. Bacterial strains and plasmids used in this study.
| Strain or plasmid | JPUB Part IDa | Relevant Characteristics | Source or reference |
|---|---|---|---|
| E. coli strains | |||
| BL21(DE3) | F- ompT gal dcm lon hsdSB(rB- mB-) λ(DE3) | [17] | |
| DH10B | F- mcrA endA1 recA1 galE15 galK16 nupG rpsL ΔlacX74 Φ80lacZΔM15 araD139 Δ(ara leu)7697 Δ(mrr-hsdRMS-mcrBC) λ- | [18] | |
| DH5αZ1 | attB::[lacIq PN25-tetR SpR] F- Δ(argF-lac)169 Φ80dlacZ58(M15) ΔphoA8 glnX44(AS) λ- deoR481 gyrA96(NalR) recA1 endA1 thiE1 hsdR17 | [19] | |
| MG1655 | F- λ- rph-1 | [20] | |
| ladd-initial | JPUB_006782 | DH5αZ1with pPJ176 | This study |
| op3 final | JPUB_006764 | DH5αZ1with with pPJ167 | This study |
| op4 final | JPUB_006766 | DH5αZ1with with pPJ168 | This study |
| op5 final | JPUB_006768 | DH5αZ1with with pPJ169 | This study |
| op6 final | JPUB_006770 | DH5αZ1with with pPJ170 | This study |
| op7 final | JPUB_006772 | DH5αZ1with with pPJ171 | This study |
| op8 final | JPUB_006774 | DH5αZ1with with pPJ172 | This study |
| op10 final | JPUB_006776 | DH5αZ1with with pPJ173 | This study |
| Lyc-no-CrtI | JPUB_006743 | MG1655 with pPJ179 | This study |
| Lyc36 | JPUB_006744 | MG1655 with pPJ177 | This study |
| Lyc07 | JPUB_006745 | MG1655 with pPJ178 | This study |
| Lyc | JPUB_006810 | MG1655 with pLyc | [21] |
| MBP-3336 | JPUB_006784 | BL21(DE3) with pPJ180 | This study |
| MBP-3607 | JPUB_006786 | BL21(DE3) with pPJ181 | This study |
| MBP-CrtI | JPUB_006788 | BL21(DE3) with pPJ182 | This study |
| N-His-3336 | JPUB_006790 | BL21(DE3) with pPJ183 | This study |
| N-His-3607 | JPUB_006792 | BL21(DE3) with pPJ184 | This study |
| N-His-CrtI | JPUB_006794 | BL21(DE3) with pPJ185 | This study |
| C-His-3336 | JPUB_006796 | BL21(DE3) with pPJ186 | This study |
| C-His-3607 | JPUB_006798 | BL21(DE3) with pPJ187 | This study |
| C-His-CrtI | JPUB_006800 | BL21(DE3) with pPJ188 | This study |
| N-StrepII-3607 | JPUB_006806 | BL21(DE3) with pPJ219 | This study |
| C-StrepII-3607 | JPUB_006808 | BL21(DE3) with pPJ220 | This study |
| N-8xHis-StrepII-MBP-3607 | JPUB_006802 | BL21(DE3) with pPJ217 | This study |
| Plasmids | |||
| pFAB217 | KmR; p15a vector | [22] | |
| pBbA0k | KmR; p15a vector | [23] | |
| pBbE0k | KmR; ColE1 vector | [23] | |
| pBbE0a_mut | AmpR; ColE1 vector; BsaI site removed by mutagenesis | [23] | |
| pET24 | KmR; T7 promoter | Novagen | |
| pET28a | KmR; T7 promoter | Novagen | |
| pET28a-MBP | KmR; T7 promoter; maltose-binding protein (MBP) tag sequence | Footnote b | |
| pSKB3-EL3 | KmR; T7 promoter; 8xHis, StrepII, and MBP tag sequences | Footnote c | |
| pPJ174 | JPUB_006779 | KmR; operon 1 genes (kuste3603, kuste3605, kuste3606) with unique Ptet promoter, BCD, and terminator in pFAB217 | This study |
| pPJ175 | JPUB_006781 | KmR; operon 2 genes (kuste3604, kuste3607, kuste3608) with unique Ptet promoter, BCD, and terminator in pBbA0k | This study |
| pPJ176 | JPUB_006783 | KmR; EcoRI- and BamHI-digestion fragment of pPJ174 ligated into EcoRI and BglII sites of pPJ175 | This study |
| pPJ158 | JPUB_006747 | KmR; operon 3 genes (kuste2803, kuste2804) in pBbE0k | This study |
| pPJ159 | JPUB_006749 | KmR; operon 4 genes (kuste2802, kuste2805) in pBbE0k | This study |
| pPJ160 | JPUB_006751 | AmpR; operon 5 genes (kuste3340, kuste3350) in pBbE0a_mut | This study |
| pPJ161 | JPUB_006753 | KmR; operon 6 genes (kuste3346, kuste3348, kuste3349) in pBbE0k | This study |
| pPJ162 | JPUB_006755 | AmpR; operon 7 genes (kuste3338, kuste3347) in pBbE0a_mut | This study |
| pPJ163 | JPUB_006757 | AmpR; operon 8 genes (kuste3342, kuste3343) in pBbE0a_mut | This study |
| pPJ164 | JPUB_006759 | AmpR; operon 9 genes (kuste3345, kuste3351, kuste3352) in pBbE0a_mut | This study |
| pPJ165 | JPUB_006761 | KmR; operon 10 genes (kuste3336, kuste3339, kuste3341) in pBbE0k | This study |
| pPJ166 | JPUB_006763 | AmpR; operon 11 genes (kuste3335, kuste3344) in pBbE0a_mut | This study |
| pPJ149 | JPUB_006726 | KmR; new unique Ptet promoter and terminator for operon 3 in pBbA0k | This study |
| pPJ150 | JPUB_006728 | KmR; new unique Ptet promoter and terminator for operon 4 in pBbA0k | This study |
| pPJ151 | JPUB_006730 | KmR; new unique Ptet promoter and terminator for operon 5 in pBbA0k | This study |
| pPJ152 | JPUB_006732 | KmR; new unique Ptet promoter and terminator for operon 6 in pBbA0k | This study |
| pPJ153 | JPUB_006722 | KmR; new unique Ptet promoter and terminator for operon 7 in pBbA0k | This study |
| pPJ154 | JPUB_006734 | KmR; new unique Ptet promoter and terminator for operon 8 in pBbA0k | This study |
| pPJ146 | JPUB_006724 | KmR; old Ptet promoter, BCD, and terminator for operon 9 in pBbA0k | This study |
| pPJ156 | JPUB_006736 | KmR; new unique Ptet promoter and terminator for operon 10 in pBbA0k | This study |
| pPJ167 | JPUB_006765 | KmR; operon 3 genes in pPJ149 | This study |
| pPJ168 | JPUB_006767 | KmR; operon 4 genes in pPJ150 | This study |
| pPJ169 | JPUB_006769 | KmR; operon 5 genes in pPJ151 | This study |
| pPJ170 | JPUB_006771 | KmR; operon 6 genes in pPJ152 | This study |
| pPJ171 | JPUB_006773 | KmR; operon 7 genes in pPJ153 | This study |
| pPJ172 | JPUB_006775 | KmR; operon 8 genes in pPJ154 | This study |
| pPJ173 | JPUB_006777 | KmR; operon 10 genes in pPJ156 | This study |
| pLyc | CmR; BR322 vector with araC repressor; crtE, crtI, and crtB under PBAD promoter | [21] | |
| pPJ179 | JPUB_006738 | CmR; pLyc with crtI gene removed | This study |
| pPJ177 | JPUB_006740 | CmR; pLyc with kuste3336 replacing crtI | This study |
| pPJ178 | JPUB_006742 | CmR; pLyc with kuste3607 replacing crtI | This study |
| pPJ180 | JPUB_006785 | KmR; kuste3336 with N-terminal MBP tag sequence in pET28a-MBP | This study |
| pPJ181 | JPUB_006787 | KmR; kuste3607 with N-terminal MBP tag sequence in pET28a-MBP | This study |
| pPJ182 | JPUB_006789 | KmR; Pantoea agglomerans crtI with N-terminal MBP tag sequence in pET28a-MBP | This study |
| pPJ183 | JPUB_006791 | KmR; kuste3336 with N-terminal 6xHis tag sequence in pET28a | This study |
| pPJ184 | JPUB_006793 | KmR; kuste3607 with N-terminal 6xHis tag sequence in pET28a | This study |
| pPJ185 | JPUB_006795 | KmR; Pantoea agglomerans crtI with N-terminal 6xHis tag sequence in pET28a | This study |
| pPJ186 | JPUB_006797 | KmR; kuste3336 with C-terminal 6xHis tag sequence in pET24 | This study |
| pPJ187 | JPUB_006799 | KmR; kuste3607 with C-terminal 6xHis tag sequence in pET24 | This study |
| pPJ188 | JPUB_006801 | KmR; Pantoea agglomerans crtI with C-terminal 6xHis tag sequence in pET24 | This study |
| pPJ218 | JPUB_006805 | KmR; E. coli acpP with N-terminal StrepII tag sequence in pET24 | This study |
| pPJ219 | JPUB_006807 | KmR; kuste3607 with N-terminal StrepII tag sequence in pET24 | This study |
| pPJ220 | JPUB_006809 | KmR; kuste3607 with C-terminal StrepII tag sequence in pET24 | This study |
| pPJ217 | JPUB_006803 | KmR; kuste3607 with N-terminal 8xHis, StrepII, and MBP tag sequences in pSKB3-EL3 | This study |
a Accessible via public instance of the JBEI Registry (https://public-registry.jbei.org/folders/222)
b Provided by Andrew Hagen (JBEI).
c Provided by Aindrila Mukhopadhyay (JBEI).