Difference between revisions of "Part:BBa K3893028"
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By using the same [http://2018.igem.org/Team:Valencia_UPV/Experiments#exp_protocol experimental protocol as Valencia_UPV team ], we have obtained the parameters for the following model:
 
By using the same [http://2018.igem.org/Team:Valencia_UPV/Experiments#exp_protocol experimental protocol as Valencia_UPV team ], we have obtained the parameters for the following model:
  
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[[File:T--Ecuador_UPV--Model_pLux_Charact_eq.png|500px|none|center|alt=REquation|]]
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[[File:T--Ecuador_UPV--Model_pLux_Charact_eq.png|300px|thumb|center|alt=equation|]]
  
 
{|class='wikitable'
 
{|class='wikitable'
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|colspan=4|center|Table 1. Parameters obtained from part K3893028 corresponding to K2656003.  
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|colspan=4|Table 1. Parameters obtained from part K3893028 corresponding to K2656003.  
 
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|'''Parameter'''
 
|'''Parameter'''
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|Effective translation rate 𝛼
 
|Effective translation rate 𝛼
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|p = 0.082 min-1
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|𝛼 = 0.082 min-1
 
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|Basal expression
 
|Basal expression
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|Dilution rate μ
 
|Dilution rate μ
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| μ = 0.01631 min-1
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| μ = 0.0115 min-1
 
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|}
  

Revision as of 18:44, 18 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 to express GFP together with a TU for the constitutive expression of LuxR. It was assembled with a one-pot Level 2 Golden Gate reaction using BsmBI type IIS endonuclease.

This device 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

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 the control of the pLux promoter and a TU that expresses LuxR constitutively. It is the first half of our population oscillator release device.

Characterization

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

We used this part to characterize the Golden Braid version (K2656003) of pLux promoter (R0062) made by [http://2018.igem.org/Team:Valencia_UPV Valencia_UPV 2018 Team]. Check our experimental page for more details.

By using the same [http://2018.igem.org/Team:Valencia_UPV/Experiments#exp_protocol experimental protocol as Valencia_UPV team ], we have obtained the parameters for the following model:

equation
Table 1. Parameters obtained from part K3893028 corresponding to K2656003.
Parameter Value
Effective translation rate 𝛼 𝛼 = 0.082 min-1
Basal expression 𝛽 = 0.1049
Dilution rate μ μ = 0.0115 min-1


RBS experiment 1.
Experimental data and simulation to characterize the pLux promoter





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]