Types | DnaRegion
|
Roles | Reporter
engineered_region
|
Sequences | BBa_K216015_sequence (Version 1)
|
Description
This is the nitrate- and nitrite-responsive promoter PyeaR of Escherichia coli with the luciferase of firefly Photinus pyralis (deposited by Slovenia iGEM 2007) with the addition of a strong bacterial ribosome binding site. It should generate light in the presence of nitrate/nitrite and luciferin.
Notes
No special considerations. Note that sequencing shows an unexpected insertion of 6 bases, ACCACC, after the scar between the ribosome binding site and the ATG of the luciferase coding sequence; that is, the sequence reads ACTAGACCACCATG rather than ACTAGATG. If this is correct, these 6 bases must be present in luciferase BioBrick BBa_I712019, or else were introduced by a random event during construction. This makes the RBS 12 bases from the ATG, somewhat more than the optimum, but with a strong RBS (like J15001) should still give about 25% of the expression level expected for optimum spacing in E. coli (Vellanoweth & Rabinowitz, 1992, Molecular Microbiology 6, 1105-1114).
Source
Made by joining PyeaR BioBrick BBa_K216005 to a PCR fusion product of strong synthetic ribosome binding site J15001 and firefly luciferase BioBrick BBa_I712019.