Basic Parts
During this project, our team designed the following basic parts.
Plasmid backbones
Part | Description |
BBa_K5061001 | Helper phage (HP) M13KO7-ΔgVI backbone |
BBa_K5061101 | Helper phage (HP) M13KO7-ΔgIII-ΔgVI backbone |
BBa_K5061002 | Accessory Plasmid (AP) and Test Plasmid (TP) backbone |
BBa_K5061003 | Mutagenesis Plasmid (MP) & Expression Plasmid (EP) backbone |
Coding sequences
Ribosomal binding sites (RBS)
Golden Gate adapters
Part | Description |
BBa_K5061005 | Golden Gate adapter for Helper phages (HP) M13KO7-ΔgVI and M13KO7-ΔgIII-ΔgVI Golden Gate cloning platforms |
BBa_K5061006 | Golden Gate adapter for Accessory Plasmids (AP) Golden Gate cloning platforms for M13 gVI expression |
BBa_K5061007 | Golden Gate adapter for Test Plasmid (TP) Golden Gate cloning platform |
BBa_K5061008 | Golden Gate adapter for Expression Plasmid (EP) Golden Gate cloning platform |
and one Promoter
Composite Parts
During this project, our team designed the following composite parts.
PHAGEVO Golden Gate cloning platforms
Part | Description |
BBa_K5061031 | Helper phage (HP) M13KO7-ΔgVI Golden Gate cloning platform |
BBa_K5061131 | Helper phage (HP) M13KO7-ΔgIII-ΔgVI Golden Gate cloning platform |
BBa_K5061032 | Accessory Plasmid (AP) Golden Gate cloning platform for M13 gVI expression |
BBa_K5061132 | Accessory Plasmid (AP) Golden Gate cloning platform for M13 gIII and gVI expression |
BBa_K5061033 | Expression Plasmid (EP) Golden Gate cloning platform |
BBa_K5061034 | Test Plasmid (TP) Golden Gate cloning platform |
PHAGEVO AP, TP and MP plasmids
Part | Description |
BBa_K5061041 | Mutagenesis Plasmid (MP) expressing the evoCDA1-BE4max - T7RNAP fusion protein from the pBad promoter |
BBa_K5061042 | Mutagenesis Plasmid (MP) expressing the ABE8.20-m - T7RNAP fusion protein from the pBad promoter |
BBa_K5061043 | Mutagenesis Plasmid (MP) expressing the evoCDA1-BE4max - T7RNAP-CGG-R12-KIRV fusion protein from the pBad promoter |
BBa_K5061044 | Mutagenesis Plasmid (MP) expressing the ABE8.20-m - T7RNAP-CGG-R12-KIRV fusion protein from the pBad promoter |
BBa_K5061045 | Accessory Plasmid (AP) expressing the M13 gVI and LuxAB from the PspA promoter |
BBa_K5061145 | Accessory Plasmid (AP) expressing the M13 gIII, M13 gVI and LuxAB from the PspA promoter |
BBa_K5061046 | Test Plasmid (TP) expressing XylS AI TPA from the pTAC promoter |
PHAGEVO HP, AP, EP and TP plasmids for XylS evolution and testing
Part |
Description |
BBa_K5061051 |
Helper phage (HP) M13KO7-ΔgVI carrying XylS K38R L224Q |
BBa_K5061151 |
Helper phage (HP) M13KO7-ΔgIII-ΔgVI carrying XylS K38R L224Q |
BBa_K5061052 |
Helper phage (HP) M13KO7-ΔgVI carrying XylS K38R L224Q in the PHAGEVO mutational region |
BBa_K5061152 |
Helper phage (HP) M13KO7-ΔgIII-ΔgVI carrying XylS K38R L224Q in the PHAGEVO mutational region |
BBa_K5061053 |
Helper phage (HP) M13KO7-ΔgVI carrying XylS W88C L224Q |
BBa_K5061153 |
Helper phage (HP) M13KO7-ΔgIII-ΔgVI carrying XylS W88C L224Q |
BBa_K5061054 |
Helper phage (HP) M13KO7-ΔgVI carrying XylS W88C L224Q in the PHAGEVO mutational region |
BBa_K5061154 |
Helper phage (HP) M13KO7-ΔgIII-ΔgVI carrying XylS W88C L224Q in the PHAGEVO mutational region |
BBa_K5061055 |
Accessory Plasmid (AP) expressing the M13 gVI and LuxAB from the Pm promoter |
BBa_K5061155 |
Accessory Plasmid (AP) expressing the M13 gIII, M13 gVI and LuxAB from the Pm promoter |
BBa_K5061056 |
Test Plasmid (TP) expressing sfGFP-LVAtag from the Pm promoter |
BBa_K5061160 |
XylS wt expression cassette in EP backbone |
BBa_K5061161 |
XylS K38R L224Q expression cassette in EP backbone |
BBa_K5061162 |
XylS W88C L224Q expression cassette in EP backbone |
BBa_K5061163 |
XylS AI PA expression cassette in EP backbone |
BBa_K5061164 |
XylS AI TPA expression cassette in EP backbone |
BBa_K5061165 |
XylS PHAGEVO-AI v1 PA expression cassette in EP backbone |
BBa_K5061166 |
XylS PHAGEVO-AI v1 TPA expression cassette in EP backbone |
BBa_K5061167 |
XylS PHAGEVO-AI v2 PA expression cassette in EP backbone |
BBa_K5061168 |
XylS PHAGEVO-AI v2 TPA expression cassette in EP backbone |
BBa_K5061169 |
XylS PHAGEVO-AI v3 r3m8 PA expression cassette in EP backbone |
BBa_K5061170 |
XylS PHAGEVO-AI v3 r5m8 PA expression cassette in EP backbone |
BBa_K5061171 |
XylS PHAGEVO-AI v3 r5m10 PA expression cassette in EP backbone |
BBa_K5061172 |
XylS PHAGEVO-AI v3 r5m11 PA expression cassette in EP backbone |
BBa_K5061173 |
XylS PHAGEVO-AI v3 r6m1 TPA expression cassette in EP backbone |
BBa_K5061174 |
XylS PHAGEVO-AI v3 r6m5 TPA expression cassette in EP backbone |
BBa_K5061175 |
XylS PHAGEVO-AI v3 r7m1 PA expression cassette in EP backbone |
Basic & Composite Parts for Cas9 & λ Red mediated recombineering
& also
During this project, our team also sequenced the M13KO7 helper phage from New England Biolabs (Cat#N0315S) in order to explain the differences observed upon cloning the HP plasmids listed above. The sequence was deposited in Parts Registry under the Acc number BBa_K5061000.
We have also constructed the E. coli S2060 ∆ung ∆nfi strain and sequenced its genome.