Difference between revisions of "Part:BBa K3893028"
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===Usage and Biology===
 
===Usage and Biology===
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==Characterization==
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=Ecuador 2021's characterization=
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We characterized the Golden Braid version of this promoter ([https://parts.igem.org/Part:BBa_K2656003 K2656003]) made by [http://2018.igem.org/Team:Valencia_UPV Valencia_UPV 2018 Team] using the composite part [https://parts.igem.org/Part:BBa_K3893028 K3893028] that we built by assembling together
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<html>
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<ul>
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<li></html>[https://parts.igem.org/Part:BBa_K2656122 BBa_K2656122]: a Level 1 transcriptional unit expresing GFP under the control of the pux promoter (Golden Braid compatible)<html></li>
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<li></html>[https://parts.igem.org/Part:BBa_K2656114 BBa_K2656114]: a Level 1 transcriptional unit constitutively expressing luxR gene (Golden Braid compatible) <html></li>
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</ul>
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</html>
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Device [https://parts.igem.org/Part:BBa_K3893028 K3893028] allows us to measure GFP expression levels for different AHL induction concentrations.
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[[File:T--Ecuador--K3893028_sbolv.png|500px|thumb|center|alt=domestication.|Figure 1. SBOL Visual representation of the device [https://parts.igem.org/Part:BBa_K3893028 K3893028] from [https://2021.igem.org/Team:Ecuador Ecuador 2021 Team] ]]
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Following a standard experimental procedure and the IGEM 2018 Interlab calibration for Particles and MEFL we obtained the temporal behavior of the device for different inducer concentrations.
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[[File:T--Ecuador--K3893028_time_response.png|600px|thumb|center|alt=domestication.|Experimental reuslts for characterization of pLux promoter.]]
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[[File:T--Ecuador--K3893028_static_response.png|600px|thumb|center|alt=domestication.|Final GFP expression levels for different AHL inducer concentrations.]]
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<!-- <html>
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<table style="width:50%">
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              <tr>
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                <th><p>Experiment parameters</p></th>
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                <th><p>Description</p></th>
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              </tr>
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              <tr>
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                <td><p>Time</p></td>
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                <td><p>06:00:00 (HH:MM:SS). Measurement interval: 05:00 (MM:SS)</p></td>
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              </tr>
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              <tr>
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                <td><p>Number of samples</p></td>
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                <td><p>8 samples for each TU colony</p></td>
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              </tr>
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              <tr>
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                <td><p>Number of medium samples</p></td>
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                <td><p>8 samples of M9 medium</p></td>
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              </tr>
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              <tr>
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                <td><p>Temperature</p></td>
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                <td><p>37 ºC</p></td>
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              </tr>
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              <tr>
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                <td><p>Shake</p></td>
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                <td><p>Double Orbital. Continuously</p></td>
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              </tr>
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                <tr>
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                <td><p>Absorbance. Optical Density (OD) measure</p></td>
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                <td><p>Wavelenght at 600 nm emission</p></td>
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              </tr>
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              <tr>
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                <td><p>Excitacion wavelength</p></td>
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                <td><p>Wavelenght at 485 nm</p></td>
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              </tr>
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              <tr>
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                <td><p>Emission wavelength</p></td>
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                <td><p>Wavelenght at  528 nm</p></td>
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              </tr>
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              <tr>
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                <td><p>Gain (G)</p></td>
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                <td><p>60</p></td>
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              </tr>
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          </table></html> -->
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Revision as of 17:32, 16 October 2021

AHL induced expression of GFP (including luxR constitutive expression)

domestication.
Figure 1. SBOL Visual representation of the device

This device was used to characterize the pLux promoter. It is an AHL induced transcriptional unit assembled with a one-pot Level 1 Golden Gate reaction using BsaI type IIS endonuclease.


This composite part, together with K3893029, is an intermediate composite part to assemble the release system device (K3893030) of our Agrobactory 593. It includes a TU that expresses GFP under rhe control of the pLux promoter and a TU that expresses LuxR constitutively. It is the first half of our population oscillator release device.

This transcriptional unit is composed of the following standardized composite parts from Valencia_UPV 2018 iGEM Team [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]:

  • BBa_K2656122: a Level 1 transcriptional unit expresing GFP under the control of the pux promoter (Golden Braid compatible)
  • BBa_K2656114: a Level 1 transcriptional unit constitutively expressing luxR gene (Golden Braid compatible)

Usage and Biology

Characterization

Ecuador 2021's characterization

We characterized the Golden Braid version of this promoter (K2656003) made by [http://2018.igem.org/Team:Valencia_UPV Valencia_UPV 2018 Team] using the composite part K3893028 that we built by assembling together

  • BBa_K2656122: a Level 1 transcriptional unit expresing GFP under the control of the pux promoter (Golden Braid compatible)
  • BBa_K2656114: a Level 1 transcriptional unit constitutively expressing luxR gene (Golden Braid compatible)

Device K3893028 allows us to measure GFP expression levels for different AHL induction concentrations.


domestication.
Figure 1. SBOL Visual representation of the device K3893028 from Ecuador 2021 Team

Following a standard experimental procedure and the IGEM 2018 Interlab calibration for Particles and MEFL we obtained the temporal behavior of the device for different inducer concentrations.

domestication.
Experimental reuslts for characterization of pLux promoter.
domestication.
Final GFP expression levels for different AHL inducer concentrations.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 956
    Illegal NheI site found at 979
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]