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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− | + | Our device was based on the sequence of AF154675.1 | |
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===Design Notes=== | ===Design Notes=== | ||
All restriction sites were removed by silent mutagenesis. | All restriction sites were removed by silent mutagenesis. | ||
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− | === | + | <center> <img src ="https://static.igem.org/mediawiki/2016/0/07/T--UCL--bacteriocindevice.png" width="50%" height="50%" </center></html> |
− | + | <p> <center> Fig. 1. Simplified diagram of the mutacin III biosynthetic locus designed by UCL iGEM 2016. </center> </p> | |
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Latest revision as of 16:36, 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.
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE 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 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 7976
- 1000INCOMPATIBLE 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
Source
Our device was based on the sequence of AF154675.1
Design Notes
All restriction sites were removed by silent mutagenesis.