Difference between revisions of "Part:BBa K1954004:Design"

Revision as of 16:30, 19 October 2016

Mutacin III biosynthetic device

The biosynthetic locus of mutacin III was designed by our team in a form allowing for high-yield and fine-tuned expression of the peptide (Fig. 1). We placed a strong T7 promoter upstream of mutA to obtain high levels of the propeptide, a repressible pTet promoter upstream of the mutBCDP co-transcription unit and an inducible araBAD promoter for the mutT gene, coding for the ATP-binding-cassette-like transporter of mutacin III. All illegal restriction sites were removed from the endogenous sequences by silent mutagenesis.

Fig. 1. Simplified diagram of the mutacin III biosynthetic locus designed by UCL iGEM 2016.

</html>

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1294
  Illegal BglII site found at 2915
  Illegal BglII site found at 3097
  Illegal BglII site found at 3763
  Illegal BglII site found at 3835
  Illegal BglII site found at 4976
  Illegal BglII site found at 5507
  Illegal BamHI site found at 1742
  Illegal BamHI site found at 8141
  Illegal BamHI site found at 8625
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 7976
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 9690
  Illegal SapI site found at 1161
  Illegal SapI site found at 7958
  Illegal SapI.rc site found at 3194
  

Design Notes

All restriction sites were removed by silent mutagenesis.

Source

Our device was based on the sequence of AF154675.1

References