Regulatory

Part:BBa_K2656004

Designed by: Adrian Requena Gutierrez, Carolina Ropero   Group: iGEM18_Valencia_UPV   (2018-09-16)

Constitutive promoter J23106

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

Part BBa_K2656004 is the constitutive low promoter BBa_J23106 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 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_K2656106 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 for each promoter tested.

RBS experiment.
Figure 1. Promoter expression experiment with BBa_K2656004, K2656005 and BBa_K2656007 basic pieces


Table 1. Optimized parameters for the BBa_K2656004 promoter.
Parameter Value
Constitutive transcription rate CR CR = 171.15 min-1
Dilution rate μ μ = 0.01001 min-1

We have also calculated the relative force between the different promoters, taking BBa_K2656005 promoter as a reference. Likewise, the promoter ratio (p parameter ratio) of the different promoters referring to the stronger one has been calculated.

Table 2. BBa_K2656004 relative strength and p ratio.
Parameter Value
Relative strength 0.2603
p parameter ratio 0.3172


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]


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Categories
Parameters
None