Part:BBa_K3002012
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This basic part contains the Tub2-terminator (B6-C1) for Chlamydomonas reinhardtii and was built as a part of the Kaiser Collection. Combined with a CDS and a promoter, this level 0 construct leads to the expression of a target protein.
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Secondary Structure
Measurement
- [http://openwetware.org/wiki/Cconboy:Terminator_Characterization/Results How these parts were measured]
The Tubulin2 terminator was used as a terminator for the hygromycin resistance in the L2AB (BBa_K3002227) construct. The different terminator, compared to the one for the coding sequence, prevent repetition in the DNA which can decrease the expression of the constructs. The tubulin2 terminator is supposed to work well in combination with the tubulin2 promotor. The ligation of the resistance cassette was successful as well as the secretion of the substrate.
The Kaiser Collection
We are proud to present our very own MoClo part collection for C. reinhardtii - the Kaiser collection.
These 20 Parts are specifically designed and codon optimized for Chlamydomonas. Among them are regulatory elements, antibiotic resistances, resistance cassettes, secretion signals and tags. These parts were tested and optimized thoroughly and we can guarantee that they work 100%. With these, expression and secretion in Chlamy will be a success. Because this is a MoClo collection, the parts are highly standardized for worldwide application. The combination with other part collections works fast and easy. While in MoClo, nomenclature is a bit different from the iGEM BioBricks, it is quickly explained:
Level 0 parts are equivalent to basic parts, e.g. Promoters, coding sequences, etc.
Level 1 parts are combinations of basic parts and usually form functional transcription units.
Level 2 parts are combinations of Level 1 parts, in case you want to transfer multiple transcription units at once. For example, you can pair your gene of interest with a selection marker.
The great thing about the Kaiser Collection and MoClo is that the ligation works in a one pot, one step reaction, as the Type IIs restriction enzymes cut out their own recognition sites. This way, multiple constructs can be combined linearly in a fixed order to create complex structures. This is ensured by the standardized overlaps that assign the parts one of 10 positions in the final constructs. After trying MoClo once, you won’t go back to traditional ligation. It is incredibly easy and reliable. For this reason, we believe that our Kaiser Collection will strike a significant chord, as the future lies in standardized, easy to use methods such as MoClo. Visit our part collection site to get an overview over all parts of the Kaiser Collection
Usage and Biology
Team UVU-Utah-2, 2024
We obtained this part in the Chlamydomonas MoClo kit as described in Crozet et al. (2018). We utilized and characterized this part in composite part(s) K5078008. Please check out the documentation for that composite part for more details!
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 87
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 5
- 1000COMPATIBLE WITH RFC[1000]
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