Difference between revisions of "Team:UCLouvain/Basic Part"

pBAD/araC (I0500)

The pBAD promoter is a very common inducible one that is activated by L-arabinose and repressed by araC.

Terminators are necessary to stop the transcription at the end of a coding sequence.

Efficient ribosome binding site characterized by IIT Madras in iGEM 2016.

Fluorescent proteins are very common reporter genes. This particular one comes from Discosoma striata and was engineered to be monomeric. The excitation peak is at 584 nm, and the emission peak at 607 nm.

iGEM plasmid consisting of araC, a pBAD/AraC promoter (I0500), an RBS (B0034), an RFP (E1010), and two terminators (B0010 and B0012) contained inside one biobrick (B0015)

Protein originally found in Streptococcus thermophilius where it regulates natural competence. The protein activation and binding to specific promoters (ComR Box; BBa_K2341001) is only possible in presence of a short peptide, ComS (LPYFAGCL). Binding to the promoter will induce the downstream sequence expression.

Promote expression of the sequence downstream in presence of ComR (BBa_K2341000) and ComS (LPYFAGCL). Sequence containing the palindrome motif needed to bind ComR (= ComR box).

RNA guide to use in combination with CRISPR/CAS9. Cut inside the tyrA gene so that deletion by homologous recombination can occur if a homologous DNA strand is also added. RNA guide under constitutive expression by BBa_J23119.

Plasmid consisting of ComR (BBa_K2341000), the p1655 promoter (containing the ComR box, BBa_K2341001), GusA and an erythromycin resistance.

GusA encodes the beta-glucuronidase gene from E. coli. This is a popular reporter gene that uses glucuronides as substrates, the most common of which are X-gluc (5-bromo-4-chloro-3-indolyl glucuronide) and para-nitrophenyl glucuronide.

Here, it is under the control of the ComRS pathway and uses the ComS peptide from S. thermophilius (LPYFAGGL).