Part:BBa_K2656004

Constitutive promoter J23106

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 GoldenBraid assembly methods.

Following the Golden Gate assembly protocol , it can be combined with other GB compatible parts from our 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 Part Collection
• CDS BBa_K2656022: the E0040 GFPmut3b sequence in its Golden Braid standardized version from our Part Collection
• Terminator BBa_K2656026: the B0015 transcriptional terminator in its Golden Braid compatible version from our Part Collection

By using this experimental protocol, we have obtained the parameters to valide our constitutive model and so rationale choose its optimized values based for each promoter tested.

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]