Difference between revisions of "Part:BBa K3484004"
Line 7: Line 7:
 
This system allows a fast report on the AHL concentrations in the medium, something important to know the amount of protein. The composite part is Biobrick compatible.
 
This system allows a fast report on the AHL concentrations in the medium, something important to know the amount of protein. The composite part is Biobrick compatible.
  
−
This composite part includes all the following parts:
+
This composite part is composed by all the following parts:
  
−
(<html><a href="https://parts.igem.org/Part:BBa_B0015">BBa_B0015</a></html>): A reliable double terminator that consists on BBa_B0010 and BBa_B0012.
+
<html><a href="https://parts.igem.org/Part:BBa_B0015">BBa_B0015</a></html>: A reliable double terminator that consists on BBa_B0010 and BBa_B0012.
 
   
 
   
−
(<html><a href="https://parts.igem.org/Part:BBa_B0033">BBa_B0033</a></html>): A weak RBS. The RBS.4 (derivative of BBa_0030).
+
<html><a href="https://parts.igem.org/Part:BBa_B0033">BBa_B0033</a></html>: A weak RBS. The RBS.4 (derivative of BBa_0030).
  
−
(<html><a href="https://parts.igem.org/Part:BBa_B0032">BBa_B0032</a></html>): A medium RBS. The RBS.3 (medium) (derivative of BBa_0030)
+
<html><a href="https://parts.igem.org/Part:BBa_B0032">BBa_B0032</a></html>): A medium RBS. The RBS.3 (medium) (derivative of BBa_0030
  
−
(<html><a href="https://parts.igem.org/Part:BBa_R0062">BBa_R0062</a></html>): LuxR & HSL regulated promoter.
+
<html><a href="https://parts.igem.org/Part:BBa_R0062">BBa_R0062</a></html>: LuxR & HSL regulated promoter.
  
−
(<html><a href="https://parts.igem.org/Part:BBa_J72005">BBa_J72005</a></html>): Ptet Promoter. Is activated by the AHL-LuxR complex
+
<html><a href="https://parts.igem.org/Part:BBa_J72005">BBa_J72005</a></html>: Ptet Promoter. Is activated by the AHL-LuxR complex
  
−
+
<html><a href="https://parts.igem.org/Part:BBa_K3484006">BBa_K3484006</a></html>: sfGFP+ASV coding sequence
−
(<html><a href="https://parts.igem.org/Part:BBa_K3484006">BBa_K3484006</a></html>): sfGFP+ASV coding sequence
+
−
 
+
−
 
+
−
(<html><a href="https://parts.igem.org/Part:BBa_B0015">BBa_B0015</a></html>) followed by the sfGFP with the ASV degradation tag (This reporter sequence <html><a href="https://parts.igem.org/Part:BBa_K3484006">BBa_K3484006</a></html>) . A control with the original sequence (<html><a href="https://parts.igem.org/Part:BBa_I746916">BBa_I746916</a></html>), with the same promoter, double terminator and RBS, was done to see how the tag affected the final fluorescence.
+
  
 
[[File:UPF_Barcelona--Reporter1.png|600px|thumb|center|Figure 1. Scheme of the interactions on the AHL sfGFP+ASV tag reporter sensor cell.]]
 
[[File:UPF_Barcelona--Reporter1.png|600px|thumb|center|Figure 1. Scheme of the interactions on the AHL sfGFP+ASV tag reporter sensor cell.]]

Revision as of 11:33, 26 October 2020


AHL sfGFP+ASV reporter

This composite part produces sfGFP when a large enough concentration of lactone (AHL) is present in the medium. In this case, it constitutively produces LuxR, which binds to the lactone present in the medium and together, they form a complex that activates the pLux promoter, generating the sfGFP whose fluorescence can be measured. The schematics of this composite part can be seen in Figure 1.

This system allows a fast report on the AHL concentrations in the medium, something important to know the amount of protein. The composite part is Biobrick compatible.

This composite part is composed by all the following parts:

BBa_B0015: A reliable double terminator that consists on BBa_B0010 and BBa_B0012.

BBa_B0033: A weak RBS. The RBS.4 (derivative of BBa_0030).

BBa_B0032): A medium RBS. The RBS.3 (medium) (derivative of BBa_0030

BBa_R0062: LuxR & HSL regulated promoter.

BBa_J72005: Ptet Promoter. Is activated by the AHL-LuxR complex

BBa_K3484006: sfGFP+ASV coding sequence

Figure 1. Scheme of the interactions on the AHL sfGFP+ASV tag reporter sensor cell.


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 1249
    Illegal AgeI site found at 1372
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1142