OnePot PURE
Mainly our collection consists of parts needed to produce the PURE cell-free translation-transcription system. PURE systems requires 36 different purified proteins, so in our collection we included the previously non documented gene sequences that encode for said proteins equipped with affinity tags, as well as the composite parts which allow their expression in E.coli under the regulation of lac operon. We believe that our collection is going to be very useful for the synthetic biology community, as it enables researchers to produce their one cell-free expression system at a negligible cost, compared to the commercially available solutions.
The table below contains all the Protein Sequence parts.The information provided on the table are: part name that links to the part page in the registry, part type, short description, and length.
| Name | Type | Description | Length |
| BBa_K2916000 | Coding | ArgRS protein equipped with a 10x HIS affinity tag | 1800 |
| BBa_K2916001 | Coding | GlnRS protein equipped with a 6x HIS affinity tag | 1706 |
| BBa_K2916002 | Coding | GluRS protein equipped with a 6x HIS affinity tag | 1457 |
| BBa_K2916003 | Coding | GlyRS protein equipped with a 6x HIS affinity tag | 3032 |
| BBa_K2916004 | Coding | IleRS protein equipped with a 6x HIS affinity tag | 2894 |
| BBa_K2916005 | Coding | LeuRS protein equipped with a 6x HIS affinity tag | 2624 |
| BBa_K2916006 | Coding | LysRS protein equipped with a 6x HIS affinity tag | 1559 |
| BBa_K2916007 | Coding | PheRS protein equipped with a 6x HIS affinity tag | 3483 |
| BBa_K2916008 | Coding | EF-G protein equipped with a 6x HIS affinity tag | 2148 |
| BBa_K2916009 | Coding | EF-Tu protein equipped with a 6x HIS affinity tag | 1218 |
| BBa_K2916010 | Coding | CK protein equipped with a 6x HIS affinity tag | 1185 |
| BBa_K2916011 | Coding | MK protein equipped with a 6x HIS affinity tag | 627 |
| BBa_K2916012 | Coding | NDK protein eqquiped with a 6x HIS affinity tag | 501 |
| BBa_K2916013 | Coding | PPiase protein equipped with a 6x HIS affinity tag | 905 |
The table below contains all the Protein Expression parts. The information provided on the table are: part name that links to the part page in the registry, part type, short description, and length.
| Name | Type | Description | Length |
| BBa_K2916014 | Composite | Expression of AlaRS in E.coli | 2888 |
| BBa_K2916015 | Composite | Expression of ArgRS in E.coli | 1950 |
| BBa_K2916016 | Composite | Expression of AsnRS in E.coli | 1658 |
| BBa_K2916017 | Composite | Expression of AspRS in E.coli | 1968 |
| BBa_K2916018 | Composite | Expression of CysRS in E.coli | 1581 |
| BBa_K2916019 | Composite | Expression of GlnRS in E.coli | 1856 |
| BBa_K2916020 | Composite | Expression of GluRS in E.coli | 1607 |
| BBa_K2916021 | Composite | Expression of GlyRS in E.coli | 3182 |
| BBa_K2916022 | Composite | Expression of HisRS in E.coli | 1470 |
| BBa_K2916023 | Composite | Expression of IleRS in E.coli | 3044 |
| BBa_K2916024 | Composite | Expression of LeuRS in E.coli | 2774 |
| BBa_K2916025 | Composite | Expression of LysRS in E.coli | 1709 |
| BBa_K2916026 | Composite | Expression of MetRS in E.coli | 2229 |
| BBa_K2916027 | Composite | Expression of PheRS in E.coli | 3695 |
| BBa_K2916028 | Composite | Expression of ProRS in E.coli | 1914 |
| BBa_K2916029 | Composite | Expression of SerRS in E.coli | 1488 |
| BBa_K2916030 | Composite | Expression of ThrRS in E.coli | 2186 |
| BBa_K2916031 | Composite | Expression of TrpRS in E.coli | 1200 |
| BBa_K2916032 | Composite | Expression of TyrRS in E.coli | 1470 |
| BBa_K2916033 | Composite | Expression of ValRS in E.coli | 3051 |
| BBa_K2916034 | Composite | Expression of IF1 in E.colI | 482 |
| BBa_K2916035 | Composite | Expression of IF2 in E.colI | 2930 |
| BBa_K2916036 | Composite | Expression of IF3 in E.colI | 800 |
| BBa_K2916037 | Composite | Expression of EF-G in E.coli | 2360 |
| BBa_K2916038 | Composite | Expression of EF-Tu in E.coli | 2360 |
| BBa_K2916039 | Composite | Expression of EF-Ts in E.coli | 1109 |
| BBa_K2916040 | Composite | Expression of RF1 in E.coli | 1340 |
| BBa_K2916041 | Composite | Expression of RF2 in E.coli | 1293 |
| BBa_K2916042 | Composite | Expression of RF3 in E.coli | 1847 |
| BBa_K2916043 | Composite | Expression of RFF in E.coli | 815 |
| BBa_K2916044 | Composite | Expression of MTF in E.coli | 1143 |
| BBa_K2916045 | Composite | Expression of CK in E.coli | 1403 |
| BBa_K2916046 | Composite | Expression of MK in E.coli | 777 |
| BBa_K2916047 | Composite | Expression of NDK in E.coli | 713 |
| BBa_K2916048 | Composite | Expression of PPiase in E.coli | 1055 |
| BBa_K2916049 | Composite | Expression of T7 RNA in E.coli | 2912 |
Functionality
In addition to the components used for PURE system, we also designed a series of toeholds needed for the realization of our project. Those toeholds regulate transcription for three different sequences (for Endogenous Control, Flavesence Doree and Bois Noir), and were designed using different parameters In the following tables you will find the sequences of the toeholds and their implementation to reporter devices with super folding Green Fluorescent Protein and CDO:
The table below contains all the Toehold Sequence parts. The information provided on the table are: part name that links to the part page in the registry, part type, short description, and length.
| Name | Type | Description | Length |
| BBa_K2916050 | Regulatory | Toehold for Grapevine Endogenous Control 1.1 | 125 |
| BBa_K2916051 | Regulatory | Toehold for Grapevine Endogenous Control 2.1 | 122 |
| BBa_K2916052 | Regulatory | Toehold for Grapevine Endogenous Control 2.2 | 122 |
| BBa_K2916053 | Regulatory | Toehold for Grapevine Endogenous Control 2.3 | 122 |
| BBa_K2916054 | Regulatory | Toehold for Grapevine Endogenous Control 2.4 | 122 |
| BBa_K2916055 | Regulatory | Toehold for Bois Noir 1.1 | 125 |
| BBa_K2916056 | Regulatory | Toehold for Bois Noir 1.2 | 125 |
| BBa_K2916057 | Regulatory | Toehold for Bois Noir 1.3 | 125 |
| BBa_K2916058 | Regulatory | Toehold for Bois Noir 1.4 | 125 |
| BBa_K2916059 | Regulatory | Toehold for Bois Noir 2.1 | 122 |
| BBa_K2916060 | Regulatory | Toehold for Bois Noir 2.2 | 122 |
| BBa_K2916061 | Regulatory | Toehold for Bois Noir 2.3 | 122 |
| BBa_K2916062 | Regulatory | Toehold for Bois Noir 2.4 | 122 |
| BBa_K2916063 | Regulatory | Toehold for Flavescence Doree 1.1 | 125 |
| BBa_K2916064 | Regulatory | Toehold for Flavescence Doree 2.1 | 122 |
| BBa_K2916065 | Regulatory | Toehold for Flavescence Doree 2.2 | 122 |
| BBa_K2916066 | Regulatory | Toehold for Flavescence Doree 2.3 | 122 |
| BBa_K2916067 | Regulatory | Toehold for Flavescence Doree 2.4 | 122 |
The table below contains all the Toehold switches with sf GFP parts. The information provided on the table are: part name that links to the part page in the registry, part type, short description, and length.
| Name | Type | Description | Length |
| BBa_K2916068 | Composite | Toehold switch for Grapevine Endogenous control 1.1 with superfolding GFP | 988 |
| BBa_K2916069 | Composite | Toehold switch for Bois Noir 1.1 with superfolding GFP | 988 |
| BBa_K2916070 | Composite | Toehold switch for Bois Noir 1.2 with superfolding GFP | 988 |
| BBa_K2916071 | Composite | Toehold switch for Bois Noir 1.3 with superfolding GFP | 988 |
| BBa_K2916072 | Composite | Toehold switch for Bois Noir 14 with superfolding GFP | 988 |
| BBa_K2916073 | Composite | Toehold switch for Bois Noir 2.1 with superfolding GFP | 985 |
| BBa_K2916074 | Composite | Toehold switch for Bois Noir 2.2 with superfolding GFP | 985 |
| BBa_K2916075 | Composite | Toehold switch for Bois Noir 2.3 with superfolding GFP | 985 |
| BBa_K2916076 | Composite | Toehold switch for Bois Noir 2.4 with superfolding GFP | 985 |
| BBa_K2916077 | Composite | Toehold switch for Flavescence Doree 1.1 with superfolding GFP | 988 |
The table below contains all the Toehold Switch with CDO parts. The information provided on the table are: part name that links to the part page in the registry, part type, short description, and length.
| Name | Type | Description | Length |
| BBa_K2916078 | Composite | Toehold switch for Grapevine Endogenous control 2.1 with CDO | 1052 |
| BBa_K2916079 | Composite | Toehold switch for Grapevine Endogenous control 2.2 with CDO | 1052 |
| BBa_K2916080 | Composite | Toehold switch for Grapevine Endogenous control 2.3 with CDO | 1052 |
| BBa_K2916081 | Composite | Toehold switch for Grapevine Endogenous control 2.4 with CDO | 1052 |
| BBa_K2916082 | Composite | Toehold switch for Bois Noir 2.1 with CDO | 1052 |
| BBa_K2916083 | Composite | Toehold switch for Flavescence Doree 2.1 with CDO | 1055 |
| BBa_K2916084 | Composite | Toehold switch for Flavescence Doree 2.2 with CDO | 1052 |
| BBa_K2916085 | Composite | Toehold switch for Flavescence Doree 2.3 with CDO | 1052 |
| BBa_K2916086 | Composite | Toehold switch for Flavescence Doree 2.4 with CDO | 1052 |