Part:BBa_K3634004:Design

4-DG Pathway (E.coli Optimised)

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

Due to the two-phase nature of our project, the selected promoter and RBS used may be subject to change after wet lab application in 2021 to maximise shinorine production. Golden gate assembly methods will facilitate this optimisation if necessary. It is also worth noting that the third enzyme (ATPG) may be included on plasmid A with NRPS instead of on plasmid B such that mycosporine glycine, which also absorbs UV radiation to a certain degree, is not produced in a bacteria which by chance has taken up plasmid B and would thereafter exploit UV resistance to confer a survival advantage in the environment.

Source

The initial genomic sequence came from Anabaena variabilis ATCC 29413 which can be obtained from parts BBa_K814000, BBa_K814002 and K8140001 (Minnesota iGEM, 2012) before being optimised using the IDT optimisation tool. The promoter and RBS were selected from each respective Anderson catalogue based on their relative strength. The terminator BBa_B0015 sequence can also be taken from the iGEM registry.

References

Minnesota iGEM 2012 - https://parts.igem.org/Part:BBa_K814000

Minnesota iGEM 2012 - https://parts.igem.org/Part:BBa_K814002

Balskus, E.P, Walsh, C.T. 2010. The Genetic and Molecular Basis for Sunscreen Biosynthesis in Cyanobacteria. Science. 329 (5999): p1653-1656. DOI: 10.1126/science.1193637