Difference between revisions of "Part:BBa K1725002"

 
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1725002 short</partinfo>
 
<partinfo>BBa_K1725002 short</partinfo>
−
 
−
The promoter is on normally, and off when the PhlF repressor (K1725041) 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 K1725020.
 
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 11: Line 8:
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1725002 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1725002 SequenceAndFeatures</partinfo>
 +
 +
This reporter was used to characterise the promoter K1725000.
 +
 +
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.
 +
 +
https://static.igem.org/mediawiki/2015/d/df/Glasgow_2015_Repressors_Promoter_Graph_2.png
 +
 +
<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>
  
  

Revision as of 20:02, 16 September 2015

PhlF repressible promoter + weak RBS + GFP

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 719

This reporter was used to characterise the promoter K1725000.

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.