Difference between revisions of "Part:BBa K2151666"

 
 
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<partinfo>BBa_K2151666 short</partinfo>
 
<partinfo>BBa_K2151666 short</partinfo>
  
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<p>This is a plasmid backbone consisting of a modified version of the <i>S. thermophilus</i> expression vector pMG36ET. The multiple cloning site was replaced with the biobrick prefix and suffix. The backbone contains the pWV01 broad-host range origin of replication, making it a suitable shuttle vector for use within <i>E. coli</i> and lactic acid bacteria such as <i>S. thermophilus</i>. The backbone also contains an erythromycin resistance gene as selectable marker.</p>
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<p<We were able to transform both <i>E. coli</i> and <i>S. thermophilus</i> with this plasmid, resulting in erythromycin-resistant colonies. By using this plasmid as a shuttle vector, we were able to clone BioBrick constructs in E. coli, and then purify the DNA for transformation of <i>S. thermophilus</i>. We successfully transformed <i>S. thermophilus</i> with an amilCP BioBrick construct in this shuttle vector,confirming that the plasmid is suitable for use in both <i>E. coli</i> and <i>S. thermophilus</i></p>
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<p>The part contains BBa_J04450 as is required by iGEM for submission of plasmid backbones.</p>
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[[File:T--Glasgow--shuttlevectors.jpeg]]
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<p><b>Figure 1:</b> Left: empty pMG36ET with BioBrick restriction sites highlighted Right: pMG36ET with J04450 BioBrick construct inserted between existing EcoRI and PstI restriction sites.</p>
  
 
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<!-- Add more about the biology of this part here

Latest revision as of 05:16, 29 October 2016


pMG36ET

This is a plasmid backbone consisting of a modified version of the S. thermophilus expression vector pMG36ET. The multiple cloning site was replaced with the biobrick prefix and suffix. The backbone contains the pWV01 broad-host range origin of replication, making it a suitable shuttle vector for use within E. coli and lactic acid bacteria such as S. thermophilus. The backbone also contains an erythromycin resistance gene as selectable marker.

<p<We were able to transform both E. coli and S. thermophilus with this plasmid, resulting in erythromycin-resistant colonies. By using this plasmid as a shuttle vector, we were able to clone BioBrick constructs in E. coli, and then purify the DNA for transformation of S. thermophilus. We successfully transformed S. thermophilus with an amilCP BioBrick construct in this shuttle vector,confirming that the plasmid is suitable for use in both E. coli and S. thermophilus</p>

The part contains BBa_J04450 as is required by iGEM for submission of plasmid backbones.

T--Glasgow--shuttlevectors.jpeg

Figure 1: Left: empty pMG36ET with BioBrick restriction sites highlighted Right: pMG36ET with J04450 BioBrick construct inserted between existing EcoRI and PstI restriction sites.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3639
    Illegal NheI site found at 2426
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 3645
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3639
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3639
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3639
    Plasmid lacks a suffix.
    Illegal XbaI site found at 3654
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal AgeI site found at 447
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.