Difference between revisions of "Part:BBa K2273108:Design"
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===DesignNotes===
 
===DesignNotes===
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This part has been codon optimized for expression in <i>Bacillus subtilis</i> using the online tool provided by IDT DNA. A Ribosome Binding Site (AGGAGG) specific for translation in <i>Bacillus subtilis</i> has been added upstream of the gene. Further the part features the RFC10 prefix and suffix:<br>
+
This part has been codon optimized for expression in <i>Bacillus subtilis</i> using the online tool provided by IDT DNA. A Ribosome Binding Site (AGGAGG) specific for translation in <i>Bacillus subtilis</i> has been added upstream of the gene followed by a seven nucleotide spacer. Further the part features the RFC10 prefix and suffix:<br>
 
<table>
 
<table>
 
<tr>
 
<tr>
 
<td width="70">Prefix with</td>
 
<td width="70">Prefix with</td>
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<td width="200"><span style="color:blue">EcoRI</span>, <span style="color:green">NotI</span>, <span style="color:red">XbaI</span> and <span style="color:silver">SD</span></td>
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<td width="200"><span style="color:blue">EcoRI</span>, <span style="color:green">NotI</span>, <span style="color:red">XbaI</span>, <span style="color:silver">RBS</span> and <span style="color:purple">spacer sequence</span></td>
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<td><span style="color:blue">GAATTC</span><span style="color:green">GCGGCCGC</span>T<span style="color:red">TCTAGA</span>T<span style="color:silver">AAGGAGG</span>TCAAAA</td>
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<td><span style="color:blue">GAATTC</span><span style="color:green">GCGGCCGC</span>T<span style="color:red">TCTAGA</span>T<span style="color:silver">AGGAGG</span><span style="color:purple">TGTCAAA</span></td>
 
</tr>
 
</tr>
 
<tr>
 
<tr>

Revision as of 13:03, 25 September 2017


BlaR1 Beta-Lactam Receptor derived from Staphylococcus aureus N315

The blaR1 gene is part of the Beta-Lactam Biosensor project of [http://2017.igem.org/Team:TU_Dresden iGEM Team TU Dresden 2017 (EncaBcillus - It's a trap!)].

This gene is part of the bla operon found in Staphylococcus aureus and encodes a receptor that localizes in the inner cell membrane and can bind beta-lactam antibiotics, Uniprot [http://www.uniprot.org/uniprot/P18357]).

DesignNotes

This part has been codon optimized for expression in Bacillus subtilis using the online tool provided by IDT DNA. A Ribosome Binding Site (AGGAGG) specific for translation in Bacillus subtilis has been added upstream of the gene followed by a seven nucleotide spacer. Further the part features the RFC10 prefix and suffix:

Prefix with EcoRI, NotI, XbaI, RBS and spacer sequence GAATTCGCGGCCGCTTCTAGATAGGAGGTGTCAAA
Suffix with SpeI, NotI and PstI TAATACTAGTAGCGGCCGCTGCAGA

Sites of restriction enzymes generating compatible overhangs are indicated by sharing one color. (EcoRI and PstI are marked in blue, NotI in green, XbaI and SpeI in red


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

This part was chemically synthesized as we needed a codon optimized version of the blaR1 gene from Staphylococcus aureus. The original sequence was taken from the S. aureus N315 genome sequence found in the NCBI database.

References

Llarrull, L., Prorok, M., Mobashery, S. (2011)