Coding

Part:BBa_K2510109

Designed by: Aya Gomaa   Group: iGEM17_Paris_Bettencourt   (2017-10-27)


pdDronpa mut TetR

This part encodes for TetR in mutant dronpa where the mutant version of dronpa showed a better optical control of TetR than wtdronpa

In our project we developed a logic gate at the promoter level by creating dually repressed promoters using different combinations of the operators for TetR, P22 c2 and HK CI, the logic gate was tested with Dronpa-repressors fusions

By illuminating our system with violet light, caging increase the repression strength while uncaging with cyan light decreases the repression strength. Our results also show that the mutDronpa can better control the activity of TetR than the wtDronpa. However, HKC1 and C2P22 are better controlled by the wtDronpa. 


                      <img src="96_conditions.png">

                            Figure 1 :  the experiment conducted with the repressors caged with Dronpa
                                        
                                   </span>

                                     <img src="Repression_strength_of_closed_state.png ">

                                      Figure 2 :  An over view of the results of the cell-free experiment. Each 
                                       promoter was tested with its cognate repressors. Our results show that in 90% of our 
                                       constructs caging the repressors with Dronpa has increased the repression strength. 
                                       We have also obtained a wide linear range of repression strength-indicated by the red 
                                       slope-.
                                   </span>
                                   <img src="TetR_dronpa.png">
                                   <img src="P22c2_dronpa.png">
                                    <img src="HKCI_caged_dronpa.png"> 
                                   
                                           Figure 3: detailed Results of the cell-free experiment. Each promoter was 
                                           tested with its cognate repressors. Top: Testing with TetR caged with 
                                           either wt-Dronpa (BBa_K2510108) or a mutated version(BBa_K2510109)Middle: 
                                           Testing with P22c2 caged with either wt-Dronpa (BBa_K2510112) or a mutated 
                                            version(BBa_K2510113).Bottom: Testing with HKcI caged with either wt-
                                           Dronpa (BBa_K2510110) or a mutated version(BBa_K2510111)           

                                  <img src="Wtvs_mut_repressors.png" >
                                   Figure 4: the wtDronpa has a better control of HKC1 and C2P22 repressors while the mutDronpa has a better control of TetR repressors </span>

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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