Regulatory

Part:BBa_K640008:Design

Designed by: Emily Hicks   Group: iGEM11_Calgary   (2011-10-28)

Putative Naphthenic Acid - Sensitive Promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 136


Design Notes

According to the chromosomal map (Figure 10), fad-2 (gene coding for Enoyl coA hydratase) appears to be in a gene cluster or operon, where transcription of the genes in the operon would be theoretically controlled by a single promoter element upstream of the operon. Three intergenic regions in the vicinity of the genes were explored for possible promoter sequences that are responsive to naphthenic acid exposure. First, in the chromosomal map, the fad-2 gene is read from right to left, and thus the intergenic region (Tax ID: 220664) (Source: http://www.pseudomonas.com) on the right of fad-2 in the chromosomal map is directly upstream of fad-2. The sequence (178bp) of this region (in the right direction) was synthsized and biobricked.


Source

This promoter region was put together using Gibson Assembly.

References