Generator

Part:BBa_K814000

Designed by: Nolan Kennedy, Michelle Goettge, Grayson Wawrzyn   Group: iGEM12_Minnesota   (2012-10-03)

dehydroquinate synthase (DHQS) generator

Our research has focused on two novel biosynthetic pathways found in two distinct algal species. A pathway ending in the production of two UV-protective compounds, shinorine and mycosporine-glycine, was cloned from Anabaena varibalis. DHQS catalyzes the first step in the pathway, converting sedoheptulose-7-phosphate into dehydroquinate.

This part includes a modified constitutive lac promoter (lacP'), and RBS and the open reading frame of DHQS. This part can be used to express DHQS in E. coli.


Dhqs.png

Figure 1. Colony PCR screen confirming cloning of DHQS into E. coli pUCBB plasmid.

Balskus and Walsh, 2010. [1]

BBa_K3634000

Codon optimised 3-dehydroquinate synthase (DHQS) CDS for use in E.coli as part of the shinorine synthesis gene cluster. By using the IDT codon optimisation tool, St Andrews iGEM 2020 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
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 250
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 250
    Illegal NotI site found at 1373
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 250
    Illegal BglII site found at 134
    Illegal XhoI site found at 1381
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 250
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 250
  • 1000
    COMPATIBLE WITH RFC[1000]


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