Difference between revisions of "Part:BBa K2656005"
Line 23: Line 23:
 
By using this [http://2018.igem.org/Team:Valencia_UPV/Experiments#exp_protocol experimental protocol], we have obtained the parameters to valide our  [http://2018.igem.org/Team:Valencia_UPV/Modeling#models constitutive model] and so rationale choose its optimized values based on each promoter tested.
 
By using this [http://2018.igem.org/Team:Valencia_UPV/Experiments#exp_protocol experimental protocol], we have obtained the parameters to valide our  [http://2018.igem.org/Team:Valencia_UPV/Modeling#models constitutive model] and so rationale choose its optimized values based on each promoter tested.
  
−
[[File:T--Valencia_UPV--optimization_GFP_expUPV2018.png|900px|thumb|none|alt=RBS experiment.|Figure 2. Promoter expression experiment with  BBa_K2656004, K2656005 and BBa_K2656007 basic pieces]]
+
[[File:T--Valencia_UPV_promoter_UPV2018.png|900px|thumb|none|alt=RBS experiment.|Figure 2. Promoter expression experiment with  BBa_K2656004, K2656005 and BBa_K2656007 basic pieces]]
  
  

Revision as of 23:04, 17 October 2018

Constitutive promoter J23102

domestication.
Figure 1. DNA basic parts domestication. First construction refers to promoter GB domestication.

Part BBa_K2656004 is the constitutive promoter BBa_J23102 adapted into the Golden Gate grammar. Thus, it is compatible with both Biobrick and [http://2018.igem.org/Team:Valencia_UPV/Design GoldenBraid] assembly methods.

Following the [http://2018.igem.org/Team:Valencia_UPV/Protocols Golden Gate assembly protocol], it can be combined with other GB compatible parts from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Valencia UPV IGEM 2018 Printeria Collection] to assemble transcriptional units in a BsaI Type IIS one-pot reaction.

Characterization of the this part was performed with the transcriptional unit BBa_K2656105, which was used in a comparative promoters expression experiment with composite parts BBa_K2656105 and BBa_K2656107. They all were assembled in a Golden Braid alpha1 plasmid using the same RBS, CDS and terminator:

  • RBS BBa_K2656009: the B0030 RBS in its Golden Braid compatible version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]
  • CDS BBa_K2656022: the E0040 GFPmut3b sequence in its Golden Braid standardized version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]
  • Terminator BBa_K2656026: the B0015 transcriptional terminator in its Golden Braid compatible version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]



By using this [http://2018.igem.org/Team:Valencia_UPV/Experiments#exp_protocol experimental protocol], we have obtained the parameters to valide our [http://2018.igem.org/Team:Valencia_UPV/Modeling#models constitutive model] and so rationale choose its optimized values based on each promoter tested.

RBS experiment.
Figure 2. Promoter expression experiment with BBa_K2656004, K2656005 and BBa_K2656007 basic pieces


Table 1. Optimized parameters for the BBa_K2656005 promoter.
Parameter Value
Constitutive transcription rate CR CR = 539.5 min-1
Dilution rate μ μ = 0.0058 min-1

We have also calculated the relative force between the different promoters, taking BBa_K2656005 BBa_K2656005 promoter as a reference. Likewise, a ratio between p parameters of the different promoters and p parameter of the reference one has been calculated.

Table 2. BBa_K2656005 relative strength and p ratio.
Parameter Value
Relative strength 1
p parameter ratio 1

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 11
    Illegal NheI site found at 34
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]