Coding

Part:BBa_K1499002:Design

Designed by: Raman Nelakanti   Group: iGEM14_StanfordBrownSpelman   (2014-08-27)

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

This gene was isolated from purified P. fluorescens genomic DNA using the following primers: 

wssH Forward: 5'-CTGACGCACTCGCTAACTGA-3'
wssH Reverse: 5'-CTTCACCATCAGCCAAACGC-3'

However, because this gene contained multiple restriction sites incompatible with RFC10, we decided to redesign and synthesize the whole construct from DNA2.0 for Biobricking.

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]