Translational_Unit

Part:BBa_K1321334:Design

Designed by: Laura de Arroyo Garcia   Group: iGEM14_Imperial   (2014-10-08)

pSB-AcsAB


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1831
    Illegal BglII site found at 2545
    Illegal BglII site found at 4468
    Illegal BamHI site found at 841
    Illegal BamHI site found at 3835
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1817
    Illegal AgeI site found at 1894
    Illegal AgeI site found at 2450
    Illegal AgeI site found at 3251
    Illegal AgeI site found at 3343
    Illegal AgeI site found at 3449
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The AcsAB coding sequence was extracted from NCBI (refer to Source). Some native regulation was removed by codon-optimisation for expression in E.coli. The native ribosomal binding sites (RBS) were predicted to be weak by translation rate calculators (Sallis Lab RBS calculator and Postech UTR designer), therefore they were replaced by the B0034 strong RBS. Furthermore, the six base pairs downstream and upstream from this RBS were edited with the Sallis Lab RBS calculator so as to tune RBS strength and try to preserve the native stoichiometry.

Source

  • http://www.ncbi.nlm.nih.gov/nuccore/X54676.1
  • https://salis.psu.edu/software/
  • http://sbi.postech.ac.kr/utr_designer

    References

    Lee, K.Y; Guldum, G.; Mantalaris, A.; Bismarck, A.; (2014) “More than meets the eye in Bacterial Cellulose: Biosynthesis, Bioprocessing and Applications in Advanced Fiber Composites” Available on: http://onlinelibrary.wiley.com/store/10.1002/mabi.201300298/asset/mabi201300298.pdf?v=1&t=i1i6d96u&s=ca0d79f542dfb621dcecf8e90feefb7dd06d031c