Part:BBa_K2835001

Constitutive reporter: strong promoter + RBS + mutant of mRFP1 (RFC25 compatible)

The protein coding sequence for iGEM11_Uppsala's highly engineered mutant RFP with strong RBS made compatible with the iGEM RFC 25 assembly standard. and thereby all iGEM assembly standards. This improvement was made by iGEM18_Stockholm .

The colonies will turn red after 18 hours.
This reporter consists of Translational unitBBa_K2835000 and strong constitutive promoter BBa_J23119. This part is compatible with all iGEM assembly standards.

Characterization

This part was composed in order to show that inserted mutations in BBa_K2835000 would not negatively affect the functionallity of the protein mRFP1. This complex was transformed into Top10 E.coli, resulting in red colonies. Liquid cultures of the inoculated red colonies showed a bright red color after one night incubation, as can bee seen in the figures below, where cultures with the original BioBrick (BBa_I13502, consisiting of BBa_E1010 and BBa_B0034) and our improved version (BBa_K2835000) are shown side by side. To prove that the introduced mutations did not change the fluorescence of the protein, absorbance measurements were performed. A wavelength of 600 nm was used to check the cell density, while 584 nm (excitation peak of mRFP1) was used to measure the red fluorescent protein. The results showed close to identical absorbance for both versions of mRFP1, with values of OD584/OD600 = 1.058 for the original BioBrick and OD584/OD600 = 1.068 for our mutated version. These results, together with sequencing results confirming the introduced mutations, prove that our mutated BioBrick is an improved version of the original part. It has equal fluorescent properties and is compatible with assembly standard RFC25.