Composite

Part:BBa_K3002122

Designed by: Marlene Schlosser   Group: iGEM19_TU_Kaiserslautern   (2019-10-21)
Revision as of 16:40, 13 December 2019 by MSchlosser (Talk | contribs)


Level 1 Mutant PETase + GLE + sp20-His

This composite part contains the PAR-promoter (BBa_K3002027) in combination with the RPL23-Terminator (BBa_K3002006), the SP20-His-tag (BBa_K3002018), the secretion signal GLE (BBa_K3002008) and the coding sequence of the Mutant PETase (BBa_K3002014) plus the MoClo connectors for positions B1-B2 (BBa_K3002302), B2-B3 (BBa_K3002303), B4-B5 (BBa_K3002304) and B5-B6 (BBa_K3002305).

Since the Mut-PETase in combination with the secretion signal GLE, the SP20-module and HA-tag showed secretion by C.reinhardtii we designed this construct harboring a His-tag instead of a HA-tag. His-tag is designed to allow purification of MUT-PETase via Ni-NTA column and to differentiate between HA-tagged MHETase and His-tagged MUT-PETase.

The Chlamy Yummy Project Collection

We are proud to present our MoClo part collection for C. reinhardtii - the Chlamy Yummy project collection.

These 67 parts are all parts used during our project and were specifically designed and codon optimized for Chlamydomonas. Among them are basic parts (L0) of a novel mutant of the PETase (BBa_K3002014), the wildtype PETase and MHETase as well as a variety of functional composite parts (L1+2). Containing different tags as well as selection markers, this collection serves as a perfect base for plastic degradation projects with Chlamydomonas. These parts were tested and optimized thoroughly and we can guarantee that they work 100%. 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.

Sequence and Features BBa_K3002122 SequenceAndFeatures

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