Composite

Part:BBa_K515102:Design

Designed by: Atipat Patharagulpong   Group: iGEM11_Imperial_College_London   (2011-09-07)
Revision as of 03:05, 22 September 2011 by Cs3109 (Talk | contribs) (→‎Source)

J23100 promoter - PA2652


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 710
    Illegal NgoMIV site found at 812
    Illegal AgeI site found at 118
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1019


Design Notes

We have codon optimized the PA2652 coding region for Escherichia coli. We have also added an insulator sequence between the RBS and the promoter. This insulator sequence has been designed to insulate the RBS from the promoter meaning that we can easily interchange the promoter without altering the RBS strength providing a new layer of modularity. Also, the insulator sequence has been designed to have no homology with the plasmid backbone meaning that a primer can be designed for it. This allows for an easy extraction of the coding sequence through PCR.

Source

We have codon optimized the PA2652 gene from Pseudomonas aeruginosa and had it synthesised by Eurofins as two fragments. We then assembled it through CPEC.

References