Part:BBa_K2995000

Paph promoter

Constitutive promoter for expression in E. coli and B. japonicum.

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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Characterization

(Waterloo iGEM 2019)

To characterize our promoter we cloned it upstream of the RBS in BBa_E0240 (RBS-GFP-Term) and compared it's GFP expression with the already well characterized BBa_J23101 promoter in BBa_I20260. To limit the impact of other variables we insured that BBa_J23101 and BBa_E0240 shared the same backbone, RBS and terminator. In addition to characterizing this promoter with respect to BBa_I20260 we also confirmed proper function of this promoter in the pRJPaph-bjGFP plasmid which was used for expression in Bradyrhizobium for our 2019 project.

Note: empty DH5alpha was used as a negative control for GFP expression

1. Cell were grown overnight in LB media at 37 degrees 2. The following morning GFP expression was quantified using the Synergy 4 BioTek plate reader.

The graph below indicates the results from this characterization experiment:

(Excitation: 485, Emmission: 525)