Difference between revisions of "Part:BBa K3846300"
 
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<partinfo>BBa_K3846300 short</partinfo>
 
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In this inducible <b><u>expression cassette</u></b> for  <i>Synechocystis PCC 6803 </i> a CYP P450-reductase fusion protein is  expressed under the rhamnose-inducible rhaA promoter. The transcription activator rhaS is also present. Recent studies have shown that the rhamnose-inducible can achieve an up to 55x fold induction and is very suitable to be used for the expression of metabolic high burden proteins like alpha bisabolene synthase.  
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In this inducible <b><u>expression cassette</u></b> for  <i>Synechocystis PCC 6803 </i> a protein is  expressed under the rhamnose-inducible rhaA promoter. The transcription activator rhaS is also present. Recent studies have shown that the rhamnose-inducible can achieve an up to 55x fold induction and is very suitable to be used for the expression of metabolic high burden proteins like bisabolene synthase.  
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Latest revision as of 02:58, 20 October 2021


Alpha-bisabolene synthase expression cassette (rhamnose inducible)

In this inducible expression cassette for Synechocystis PCC 6803 a protein is expressed under the rhamnose-inducible rhaA promoter. The transcription activator rhaS is also present. Recent studies have shown that the rhamnose-inducible can achieve an up to 55x fold induction and is very suitable to be used for the expression of metabolic high burden proteins like bisabolene synthase.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 15
    Illegal NheI site found at 38
    Illegal NheI site found at 1070
    Illegal NheI site found at 1093
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



iGEM Hamburg 2021 part collection

Terpenoids are an important group of natural products used as biofuels, drugs or fragrances. Naturally occuring in plants it has been shown that microbial terpene production in microorganisms like yeast, E. coli or cyanobacteria is possible. Nevertheless iGEM projects seem to rarely focus on this interesting class of natural products which is correlated with a lack of useful parts inside the iGEM registry.

Fortunately we were able to change that and designed a novel golden gate based toolbox which allows.

  1. production of terpenoid precursors and simple terpenoids
  2. creation of CYP P450-reductase fusion enzymes to optimise processing of terpenoid precursors and production of bioactive target products
  3. modularity of the system to enable exchange of linker sequences/promoters/etc. (MoClo-compatible toolbox)

MoClo-based Part Design 2.0

To improve the usefulness of our parts, we then aimed to make our parts compatible with the MoClo standard of goten gate based IIS restriction enzyme assembly. Thereby we expanded the Common Genetic Syntax for fusion sites to allow the creation of a) fusion proteins connected by linker sequences and b) multiple CDS expressed in an operon. More useful information and an overview of all our parts can be found on our wiki.

Figure 1: MoClo syntax of the part collection.