Project

Part:BBa_K1662000:Design

Designed by: Janet Standeven Jack Kwon   Group: iGEM15_Lambert_GA   (2015-08-26)


Construct 1 Bicistronic:PelB-CDA under tetR and GFP under P(Lac)IQ promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1081
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 163
    Illegal AgeI site found at 453
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 564
    Illegal BsaI.rc site found at 1724


Design Notes

We had to eliminate illegal restriction sites within the mature CDA gene and the eGFP gene. So we introduced wobbles (a change in the third base pair of the codon which still produced the same amino acid) so that the restriction enzymes would not cut our DNA sequence. DNA sequence was also created with help from one of our advisers who went to a camp and learned about creating DNA sequences. We added scars and spacers to make sure our DNA sequence would work.

Source

Mature CDA2 gene sequence came from saccharomyces cerevisiae which is a type of yeast. The PelB leader sequence was found on the iGEM registry and the eGFP sequence came from an adviser at Georgia Tech. The tetR promoter, ribosomal binding sites, P(Lac) IQ promoter, and terminator were all also found in the iGEM registry.

References