Coding

Part:BBa_K3634000

Designed by: Laurence Seeley   Group: iGEM20_St_Andrews   (2020-07-20)
Revision as of 15:13, 20 July 2020 by Registry (Talk | contribs)

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3-Dehydroquinate Synthase (DHQS) (Codon Optimised for E.coli)

Codon optimised 3-dehydroquinate synthase (DHQS) CDS for use in E.coli as part of the shinorine synthesis gene cluster. The original gene for DHQS (Ava_3858) can be taken from the cyanobacteria species Anabaena variabilis ATCC 29413. The enzyme catalyses the conversion of sedoheptulose 7-phosphate, an intermediate of the pentose phosphate pathway (PPP), to (R)-demethyl-4-deoxygadusol.

This part has been previously categorised by the Minnesota iGEM team of 2012 who also sought to express the enzyme in E.coli (BBa_K814000). Here, by using the IDT codon optimisation tool, we have optimised the GC content of the sequence taken from A.variabilis to improve production efficiency of the final product shinorine. We have further made the part biobrick assembly standard RFC[10] & RFC[1000] compatible by removing EcoR1 and SapI restriction sites introduced by this optimisation step.

The part should be used alongside the additional optimised parts (BBa_K3634001, BBa_K3634002 and BBa_K3634003) responsible for ultimate conversion of the substrate sedoheptulose 7-phosphate to the final product shinorine.

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]


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Categories
Parameters
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