Difference between revisions of "Part:BBa K2273108"
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This gene is part of the <i>bla</i> operon found in <i>Staphylococcus aureus</i> and encodes a receptor that localizes in the inner cell membrane and can bind beta-lactam antibiotics, Uniprot [http://www.uniprot.org/uniprot/P18357]).<br><br>
 
This gene is part of the <i>bla</i> operon found in <i>Staphylococcus aureus</i> and encodes a receptor that localizes in the inner cell membrane and can bind beta-lactam antibiotics, Uniprot [http://www.uniprot.org/uniprot/P18357]).<br><br>
  
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===Beta Lactam Biosensor===
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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 followed by a seven nucleotide spacer. Further the part features the RFC10 prefix and suffix:<br>
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<table>
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Worldwide, multidrug-resistant germs are on the rise and provoke the intensive search for novel effective compounds. To simplify the search for new antibiotics and to track the antibiotic pollution in water samples, whole cell biosensors constitute a helpful investigative tool. In this subproject, we developed a functional and complete heterologous beta-lactam biosensor in Bacillus subtilis. By the time these specified cells sense a compound of the beta-lactam family, they will respond by producing a measurable luminescence signal. We further investigated the detection spectrum of the biosensor by testing different beta-lactam antibiotics from various subclasses. For increased control and easy handling of the biosensor strain during a potential field application, we demonstrate that the encapsulation of the cells into Peptidosomes is quite advantageous.  
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<tr>
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<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>, <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><span style="color:silver">AGGAGG</span><span style="color:purple">TGTCAAA</span></td>
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</tr>
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<tr>
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<td>Suffix with</td>
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<td><span style="color:red">SpeI</span>, <span style="color:green">NotI</span> and <span style="color:blue">PstI</span></td>
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<td><span style="color:red">ACTAGT</span>A<span style="color:green">GCGGCCG</span><span style="color:blue">CTGCAG</span>A</td>
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</tr>
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</table>
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Sites of restriction enzymes generating compatible overhangs are indicated by sharing one color. (<span style="color:blue">EcoRI</span> and <span style="color:blue">PstI</span> are marked in blue, <span style="color:green">NotI</span> in green, <span style="color:red">XbaI</span> and <span style="color:red">SpeI</span> in red
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<partinfo>BBa_K2273108 SequenceAndFeatures</partinfo>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 13:29, 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]).

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 GAATTCGCGGCCGCTTCTAGAAGGAGGTGTCAAA
Suffix with SpeI, NotI and PstI ACTAGTAGCGGCCGCTGCAGA

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]

Sequence and Features


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]