Coding

Part:BBa_K1499002:Design

Designed by: Raman Nelakanti   Group: iGEM14_StanfordBrownSpelman   (2014-08-27)
Revision as of 05:41, 24 October 2014 by Raman2004 (Talk | contribs) (→‎Source)

wssH gene in wss operon


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 366
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

wssH was initially amplified out of pure genomic DNA using the following primers:

wssH F: 5'-CTGACGCACTCGCTAACTGA-3' wssH R: 5'-CTTCACCATCAGCCAAACGC-3'

However, we realized that this contained multiple BioBrick restriction sites that would be too hard to remove using site-directed mutagenesis. We ultimately decided to synthesize the whole construct from DNA2.0

Source

The wss operon was extracted from P. fluorescens, an organism that produces cellulose acetate as part of a biofilm that allows it to colonize the air-liquid interface [1]. For BioBricking, the gene was synthesized using DNA2.0.

References

[1] Spiers AJ et al. (2013) Cellulose Expression in Pseudomonas fluorescens SBW25 and Other Environmental Pseudomonads in Cellulose - Medical, Pharmaceutical, and Electronic Applications. DOI: [http://cdn.intechopen.com/pdfs-wm/45637.pdf 10.5772/53736]