Difference between revisions of "Part:BBa K200018"
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K200018 short</partinfo>
 
<partinfo>BBa_K200018 short</partinfo>
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This BioBrick comprises a ligation of the registry parts for the promoter PcstA ([[Part:BBa_K118011 |BBa_K118011]]) and the RBS ([[Part:BBa_B0034 |BBa_B0034]]) to GFP ([[Part:BBa_E0040|BBa_E0040]]) and a double terminator ([[Part:BBa_B0015|BBa_B0015]]).
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The PcstA promoter is cAMP activated. Under low glucose concentrations, there is increased activity by adenylate cyclase. This results in cAMP binding to the cAMP receptor protein, and activating the promoter for downstream expression (more information on this part can be found on its registry page [[Part:BBa_K118011 |here]]). <br><br>
  
 
This part contains a GFP (green fluorescent protein) reporter in the functional transcriptional unit. As such, activity of the pCstA promoter can be assessed by the expression of GFP.
 
This part contains a GFP (green fluorescent protein) reporter in the functional transcriptional unit. As such, activity of the pCstA promoter can be assessed by the expression of GFP.
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===Usage and Biology===
 
===Usage and Biology===
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This construct was used by Imperial College's 2009 iGEM team for [http://2009.igem.org/Team:Imperial_College_London <i>The E.ncapsulator</i>] project. It was ligated to [https://parts.igem.org/Part:BBa_J5526 BBa_J5526] to produce the final construct [https://parts.igem.org/wiki/index.php/Part:BBa_K200019 BBa_K200019].
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This construct was used by Imperial College's 2009 iGEM team for [http://2009.igem.org/Team:Imperial_College_London <i>The E.ncapsulator</i>] project using the following subparts: [https://parts.igem.org/wiki/index.php/Part:BBa_K118011 BBa_K118011], [https://parts.igem.org/wiki/index.php/Part:BBa_B0034 BBa_B0034], [https://parts.igem.org/wiki/index.php/Part:BBa_E0040 BBa_E0040] and [https://parts.igem.org/wiki/index.php/Part:BBa_B0015 BBa_B0015]. It was ligated to [https://parts.igem.org/Part:BBa_J5526 BBa_J5526] to produce the final construct [https://parts.igem.org/wiki/index.php/Part:BBa_K200019 BBa_K200019].
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===Characterisation===
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<br>
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[[Image:II09_CRP-GFP fluor different media.jpg|500px|right]]
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Cells with BBa_K200018 were grown overnight in various different media, and the GFP fluorescence was measured. <br>
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After overnight culture, the corrected fluorescence of glucose is almost negligible, showing that glucose represses the PcstA promoter strongly.  <br>
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For all the other secondary carbon sources, 10% Casamino Acids in M9 shows the highest corrected fluorescence at 22000.<br>
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<br>
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For more information, go to the Imperial iGEM 2009 <i>E.ncapsulator</i> project page on
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[http://2009.igem.org/Team:Imperial_College_London/Wetlab/Results/CRP_and_Media BBa_K200018 testing results] <br>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K200018 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K200018 SequenceAndFeatures</partinfo>

Latest revision as of 00:11, 22 October 2009

pCstA+RBS+GFP+TT

This BioBrick comprises a ligation of the registry parts for the promoter PcstA (BBa_K118011) and the RBS (BBa_B0034) to GFP (BBa_E0040) and a double terminator (BBa_B0015).

The PcstA promoter is cAMP activated. Under low glucose concentrations, there is increased activity by adenylate cyclase. This results in cAMP binding to the cAMP receptor protein, and activating the promoter for downstream expression (more information on this part can be found on its registry page here).

This part contains a GFP (green fluorescent protein) reporter in the functional transcriptional unit. As such, activity of the pCstA promoter can be assessed by the expression of GFP.


Usage and Biology

This construct was used by Imperial College's 2009 iGEM team for [http://2009.igem.org/Team:Imperial_College_London The E.ncapsulator] project using the following subparts: BBa_K118011, BBa_B0034, BBa_E0040 and BBa_B0015. It was ligated to BBa_J5526 to produce the final construct BBa_K200019.


Characterisation


II09 CRP-GFP fluor different media.jpg

Cells with BBa_K200018 were grown overnight in various different media, and the GFP fluorescence was measured.

After overnight culture, the corrected fluorescence of glucose is almost negligible, showing that glucose represses the PcstA promoter strongly.

For all the other secondary carbon sources, 10% Casamino Acids in M9 shows the highest corrected fluorescence at 22000.

For more information, go to the Imperial iGEM 2009 E.ncapsulator project page on [http://2009.igem.org/Team:Imperial_College_London/Wetlab/Results/CRP_and_Media BBa_K200018 testing results]





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 801