Part:BBa_K398406:Design
Solvent tolerance cluster
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 37
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 37
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 37
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The original sequences can be found in [http://www.ncbi.nlm.nih.gov/nuccore THIS LINK], these sequences were enhanced for their expression on E. coli K12. Additionally, the restriction sites for the enzymes XbaI, PstI, EcoRI and SpeI were removed from the CDS and the stop codon was replaced by the codon TAA in order to avoid the formation of possible ORF's. For this process we used the [http://www.jcat.de/ jcat] software tool.
An extra XbaI site can be found between RBS and Promoter so you can increase the expression level of the protein by changing the promoter.
Prefoldin alpha aa sequence 16.97 kDa:
MIRMAQNNKELEKLAYEYQVLQAQAQILAQNLELLNLAKAEVQTVRETLENLKKIEEEKPEILVPIGAGSFLKGVIVDKNNAIVSVGSGYAVERSIDEAISFLEKRLKEYDEAIKKTQGALAELEKRIGEVARKAQEVQQKQSMTSFKVKK*
Prefoldin beta aa sequence 13.28 kDa:
MQNIPPQVQAMLGQLDTYQQQLQLVIQQKQKVQADLNEAKKALEEIETLPDDAQIYKTVGTLIVKTTKEKAVQELKEKIETLEVRLNALNRQEQKINEKVKELTQKIQAALRPPTAG*
Source
Genomic sequence obtained from NCBI.
Prefoldin alpha accession number: [http://www.ncbi.nlm.nih.gov/gene/1444414 PH0527]
Prefoldin beta accession number: [http://www.ncbi.nlm.nih.gov/gene/1444421 PH0532]