Composite

Part:BBa_K424017:Design

Designed by: Ernesto Gmez   Group: iGEM10_Panama   (2010-10-11)
Revision as of 00:04, 28 October 2010 by Ernestopro (Talk | contribs) (→‎Source)

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Test plataform for rhamnosyltransferase BioBrick (Rh1AB_BB) expression in E. coli


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 220
    Illegal BamHI site found at 780
    Illegal XhoI site found at 956
    Illegal XhoI site found at 2242
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1135
    Illegal NgoMIV site found at 1856
    Illegal NgoMIV site found at 1969
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 445
    Illegal BsaI site found at 1485
    Illegal BsaI.rc site found at 629
    Illegal BsaI.rc site found at 3135


Design Notes

We were able to ligate successfully the promotor with the RBS into a plasmid backbone and the reporter with the terminator; and we get the rhamnosiltransferase 1 gene enzyme mutated to be compatible with the Assembly Standard 10. We need to ligate all the parts to test our plataform for rhamnosyltransferase expression.

Source

The promoter, RBS, GFP reporter and the terminator were given to us as a genetic tool kit by iGEM. The Standard Rh1AB part was designed by Team Panama 2010. The rhamnosyltransferase 1 complex (Rh1AB) was isolated from a Pseudomonas aeruginosa and mutated by a mutagenesis protocol from Stratagene to be compatible with the Assembly Standard Protocol 10.

References

Assembly Standard Protocol 10, www.parts.igem.org

QuikChange Lightning Multi Site-Directed Mutagenesis Kit Instruction manual, catalog #210515 from Stratagene.