Composite
406

Part:BBa_K398406:Design

Designed by: Hugo Federico Cueto Rojas and Ramon van der Valk   Group: iGEM10_TU_Delft   (2010-10-12)
Revision as of 16:50, 27 October 2010 by Hugo 87 (Talk | contribs) (→‎Design Notes)

Solvent tolerance cluster


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 37
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 37
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 37
  • 1000
    COMPATIBLE 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: 

 1 mirmaqnnke leklayeyqv lqaqaqilaq nlellnlaka evqtvretle nlkkieeekp
      61 eilvpigags flkgvivdkn naivsvgsgy aversideai sflekrlkey deaikktqga
     121 laelekrige varkaqevqq kqsmtsfkvk k

Prefoldin beta aa sequence 13.28 kDa: 


       1 mqnippqvqa mlgqldtyqq qlqlviqqkq kvqadlneak kaleeietlp ddaqiyktvg
      61 tlivkttkek avqelkekie tlevrlnaln rqeqkinekv keltqkiqaa lrpptag

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]

References