Team:EPFL/Part Collection

Part
Collection

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