Part:BBa_K1954004:Design

Nuclease from Staphylococcus aureus

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.

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]

Source

Our device was based on the sequence of AF154675.1

Design Notes

All restriction sites were removed by silent mutagenesis.

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