Part:BBa_K2282012
mRFP under pL promoter
Usage and Biology
The pL promoter is used for driving our heat shock response. Without cI857, pL acts as a constitutive promoter. The promoter and the ribosome binding site allow the transcription of the mRFP, which is a Red Fluorescent Protein. The terminator permits the end of transcription. This part was used to test that the pL worked well both at low and high temperature without its repressor cI857 protein by allowing transcription of mRFP.
Source of this part
Go to https://www.addgene.org/browse/sequence_vdb/3001/ select the pL promoter from the plasmid pGW10. The size of the pL promoter is 244pb. The pL promoter was found using (Norma A Valdez-Cruz et al, 2010). The genomic sequence (RBS - mRFP - Double terminator) were found on the IGEM registry of standard biological parts and the part has been assembled. We added the nucleotide sequence “TACTAG” at the end of the RBS to create a spacer between the RBS and the mRFP so as to have a better transcription of the chromoprotein. The RBS was 12bp and after adding the sequence “TACTAG” the total size was 18pb.
Design consideration
We chose the promoter pL instead of an Anderson promoter, because we will use the construction for the sequence BBa_K22820013 and the pL phage lambda promoter is regulated by the thermolabile cI857 repressor and express above 37ºC (Norma A Valdez-Cruz et al, 2010).
References
Norma A Valdez-Cruz, Luis Caspeta, Néstor O Pérez, Octavio T Ramírez, Mauricio A Trujillo-Roldán - Production of recombinant proteins in E.Coli by the heat inducible expression system based on the phage lambda pL and/or pR promoters - Microbial Cell Factories 2010, 9:18.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 825
Illegal AgeI site found at 937 - 1000COMPATIBLE WITH RFC[1000]
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