Difference between revisions of "Part:BBa K3846116"
 
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<partinfo>BBa_K3846116 short</partinfo>
 
<partinfo>BBa_K3846116 short</partinfo>
  
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This phytobrick features the HTH-type transcriptional activator RhaS. The RhaS protein with an helix-turn-helix (HTH) structure activates expression of rhaBAD and rhaT operons in presence of L-rhamnose. It contains a N-terminal domain required for dimerisation and response to L-Rhamnose and a C-terminal domain involved in DNA binding and transcription activation.
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This phytobrick features the codon-optimised coding sequence of alpha-bisabolene synthase from <i>Abies grandis </i>.  
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The enzyme converts farnesyl diphosphate to alpha-bisabolene, is involved in defensive oleoresin formation in conifers in response to insect attack or other injury and in the biosynthesis of sesquiterpene (C15) olefins
  
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Transcription activation of genes behind rhaA promoter in presence of L-rhamnose have been reported.
 
  
 
<b>5' fusion site:</b> AATG
 
<b>5' fusion site:</b> AATG

Latest revision as of 20:11, 19 October 2021


Alpha-bisabolene synthase (phytobrick) - with N-terminal 6xHis tag

This phytobrick features the codon-optimised coding sequence of alpha-bisabolene synthase from Abies grandis . The enzyme converts farnesyl diphosphate to alpha-bisabolene, is involved in defensive oleoresin formation in conifers in response to insect attack or other injury and in the biosynthesis of sesquiterpene (C15) olefins


5' fusion site: AATG

3' fusion site: GCTT


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 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.