Difference between revisions of "Part:BBa K1954004:Design"
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<br><br>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.
 
<br><br>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.
  
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<partinfo>BBa_K1954004 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1954004 SequenceAndFeatures</partinfo>
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__NOTOC__
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<partinfo>BBa_K729004 short</partinfo>
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<partinfo>BBa_K729004 SequenceAndFeatures</partinfo>
  
  
 
===Design Notes===
 
===Design Notes===
 
All restriction sites were removed by silent mutagenesis.
 
All restriction sites were removed by silent mutagenesis.
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Revision as of 16:32, 19 October 2016



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.



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


Nuclease from Staphylococcus aureus


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]


Design Notes

All restriction sites were removed by silent mutagenesis.


Source

Our device was based on the sequence of AF154675.1

References