Difference between revisions of "Part:BBa K3046009"
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===Usage and Biology===
 
===Usage and Biology===
 
The bacterial promoter, BBa_K3046017, can be cut out using the Type IIs restriction enzyme as shown in the figure below. Following this, a promoter following the level 0 promoter + 5' UTR MoClo standard can be inserted and the assembled construct can be used for characterization of the promoter.  
 
The bacterial promoter, BBa_K3046017, can be cut out using the Type IIs restriction enzyme as shown in the figure below. Following this, a promoter following the level 0 promoter + 5' UTR MoClo standard can be inserted and the assembled construct can be used for characterization of the promoter.  
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<img style="padding:28px;" src="https://2019.igem.org/wiki/images/9/96/T--DTU-Denmark--PromEx.svg" alt="This figure shows a bacterial promoter in front of the fluorescent protein mCherry being replaced by a synthetic promoter when cut with BsaI, thus allowing for expression in our organism." class="safetyfirstimg">
 
<img style="padding:28px;" src="https://2019.igem.org/wiki/images/9/96/T--DTU-Denmark--PromEx.svg" alt="This figure shows a bacterial promoter in front of the fluorescent protein mCherry being replaced by a synthetic promoter when cut with BsaI, thus allowing for expression in our organism." class="safetyfirstimg">
 
<figcaption>The figure shows the general strategy for replacing the prokaryotic promoter BBa_K3046017 upstream of mCherry with a synthetic promoter using Golden Gate assembly.</figcaption>
 
<figcaption>The figure shows the general strategy for replacing the prokaryotic promoter BBa_K3046017 upstream of mCherry with a synthetic promoter using Golden Gate assembly.</figcaption>
 
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Revision as of 01:06, 22 October 2019


Fungal MoClo promoter test device

This is a device for testing promoters by expressing mCherry.


Usage and Biology

The bacterial promoter, BBa_K3046017, can be cut out using the Type IIs restriction enzyme as shown in the figure below. Following this, a promoter following the level 0 promoter + 5' UTR MoClo standard can be inserted and the assembled construct can be used for characterization of the promoter.

This figure shows a bacterial promoter in front of the fluorescent protein mCherry being replaced by a synthetic promoter when cut with BsaI, thus allowing for expression in our organism.
The figure shows the general strategy for replacing the prokaryotic promoter BBa_K3046017 upstream of mCherry with a synthetic promoter using Golden Gate assembly.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1158
    Illegal BamHI site found at 1418
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 232
    Illegal BsaI.rc site found at 6