Difference between revisions of "Part:BBa K2926003"

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==Characterisation==
 
==Characterisation==
 
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We followed the standard <a href="https://www.protocols.io/view/calibration-protocol-plate-reader-fluorescence-cal-6zrhf56">iGEM protocoll for GFP fluorescence calibration</a>,
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We followed the standard <a href="https://www.protocols.io/view/calibration-protocol-plate-reader-fluorescence-cal-6zrhf56">iGEM protocoll for GFP fluorescence calibration</a>, measuring our cultures against Flourescin.
measuring our cultures against Flourescin.
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  We performed the analysis with <i>E. coli DH5α</i> as well as <i>S. cerevisiae</i> cultures.
 
  We performed the analysis with <i>E. coli DH5α</i> as well as <i>S. cerevisiae</i> cultures.
 
  <br>
 
  <br>
  See <a href="XXX">BBa_K2926073</a> for the characterization results.
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  <b>Please see <a href="XXX">BBa_K2926073</a> for the characterization results of this part.</b>
 
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Revision as of 21:05, 22 September 2019


GALL yeast promoter


Usage and Biology

GALL is a shortend version of the natural GAl1 promotor from Saccharomyces cerevisiae, which is regulating the gene encoding the galactokinase. The GAL1 promotor is tightly repressed by glucose and strongly induced by galactose. While the GAL1 Promotor is composed of the 461bp upstream of the gal1 gene, GALL is shortend by 31bt to 430bp upstream of the gene. Thats because the GALL Promotor is lacking one of the three upstream activating sequences, which are required for full induction by galactose.


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 47
  • 1000
    COMPATIBLE WITH RFC[1000]


Plasmid Design

For the analysis and characterization of the GALL Promotor XXXX in combiantion with the TPs1 Terminaror XXXX, mCherry XXXX was cloned beteen them using Gibson Assembly. The GALL Promotor and TPs1 Terminator where optained as a gBlock from IDT.


Sequencing Results

The biobrick was analysed with Sanger Sequenceing to confirm its correct base sequence.

Characterisation

We followed the standard iGEM protocoll for GFP fluorescence calibration, measuring our cultures against Flourescin. We performed the analysis with E. coli DH5α as well as S. cerevisiae cultures.
Please see BBa_K2926073 for the characterization results of this part.


References