Difference between revisions of "Part:BBa K2294007"

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===Usage and Biology===
 
===Usage and Biology===
  
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This promoter is build up fully modular. The promoter starts with an GAL4 binding site derived from <i>GAL1</i>. After this trnaskription factor binding site a neutral spacer is placed to avoid steric hindrances between GAL4 and the TATA-box-binding- proteins of the preinitiation complex. Following the TATA-box there is the core1 consiting of 30 random nucleotides. Whose porpose is as spacer between the TATA-box and the TSS sequence. In yeast the minimal gap between TATA-box and TSS should be 30 nucleotides. TSS, core1 and TATA-box are the minimal elements for an active promoter but without an additional activation element ( in this case the GAL4 binding site) the activity of the minmal promoter would be to low for efficient protein expression.  <br>  
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This promoter is build up fully modular. It starts with a GAL4 binding site derived from <i>GAL1</i>. After this transcription factor binding site a neutral spacer is placed to avoid steric hindrances between GAL4 and the TATA-box-binding-proteins of the preinitiation complex. Following the TATA-box there is the core1 consisting of 30 random nucleotides. It functions as a spacer between the TATA-box and the TSS sequence. In yeast the minimal gap between TATA-box and TSS should be 30 nucleotides. TSS, core1 and TATA-box are the minimal elements for an active promoter but without an additional activation element (in this case the GAL4 binding site) the activity of the minimal promoter would be to low for efficient protein expression.  <br><br>  
 
All information taken from Redden 2015.  
 
All information taken from Redden 2015.  
 
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Latest revision as of 12:01, 1 November 2017


Synthtic minimal Galactose induced promoter + Kozak sequence

A short synthtetic promoter which can be induced by the addition of Galactose. This promoter contains already a Kozak sequence for the expression of proteins. The promoter is only 160 bp long.


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
    COMPATIBLE WITH RFC[1000]

Usage and Biology

This promoter is build up fully modular. It starts with a GAL4 binding site derived from GAL1. After this transcription factor binding site a neutral spacer is placed to avoid steric hindrances between GAL4 and the TATA-box-binding-proteins of the preinitiation complex. Following the TATA-box there is the core1 consisting of 30 random nucleotides. It functions as a spacer between the TATA-box and the TSS sequence. In yeast the minimal gap between TATA-box and TSS should be 30 nucleotides. TSS, core1 and TATA-box are the minimal elements for an active promoter but without an additional activation element (in this case the GAL4 binding site) the activity of the minimal promoter would be to low for efficient protein expression.

All information taken from Redden 2015.

Promoter characterization

The BOKU-Vienna Team 2017 submitted and characterized this part by measuring the GFP expression using flow cytometry. For detais go to the Experience page.


References

Redden, H., & Alper, H. S. (2015). The development and characterization of synthetic minimal yeast promoters. Nature Communications, 6, 7810. https://doi.org/10.1038/ncomms8810