Difference between revisions of "Part:BBa K1725020"
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<partinfo>BBa_K1725020 short</partinfo>
 
<partinfo>BBa_K1725020 short</partinfo>
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The promoter is on normally, and off when the SrpR repressor (K1725061) is bound to it. We measured the strength of the promoter with GFP and two different ribosome binding sites (B0032 and B0034), and compared to R0040 and K1725000.
 
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https://static.igem.org/mediawiki/2015/thumb/a/ad/2015-08-05_10.50.30_cropped.jpg/800px-2015-08-05_10.50.30_cropped.jpg
 
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From left to right: I13500(B0034.E0040); R0040.I13500; K1725000.I13500; K1725020.I13500 all in pSB3K3 in DH5a cells.
 
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1725020 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1725020 SequenceAndFeatures</partinfo>
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This promoter was characterised by driving expression of GFP with two different Ribosome Binding Sites, as found in the BioBricks E5501 and I13500.
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GFP fluorescence of K1725001, K1725002, K1725021, K1725022, K1725082, and E5504 with plasmid backbone pSB3K3 was measured to compare the relative strengths of promoters K1725000 and K1725020 to a promoter already well documented in the registry, R0040. The figure below shows the fluorescence scan image and a graph of approximate molecules of GFP per cell. These results indicated that K1725000 is a significantly stronger promoter than R0040 or K1725020.
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https://static.igem.org/mediawiki/2015/d/df/Glasgow_2015_Repressors_Promoter_Graph_2.png
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<b>All constructs with pSB3K3 plasmid backbone, in DH5α cells. Replicates of constructs and controls from three colonies, under the same conditions. Mean and standard deviation of replicates were calculated to give value and error bars.</b>
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K1725020 driven expression is repressed by K1725060 (<i>srpR</i> encoding SrpR repressor)
  
  

Revision as of 20:17, 16 September 2015

SrpR repressible promoter

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 20
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

This promoter was characterised by driving expression of GFP with two different Ribosome Binding Sites, as found in the BioBricks E5501 and I13500.

GFP fluorescence of K1725001, K1725002, K1725021, K1725022, K1725082, and E5504 with plasmid backbone pSB3K3 was measured to compare the relative strengths of promoters K1725000 and K1725020 to a promoter already well documented in the registry, R0040. The figure below shows the fluorescence scan image and a graph of approximate molecules of GFP per cell. These results indicated that K1725000 is a significantly stronger promoter than R0040 or K1725020.

Glasgow_2015_Repressors_Promoter_Graph_2.png

All constructs with pSB3K3 plasmid backbone, in DH5α cells. Replicates of constructs and controls from three colonies, under the same conditions. Mean and standard deviation of replicates were calculated to give value and error bars.

K1725020 driven expression is repressed by K1725060 (srpR encoding SrpR repressor)