Translational_Unit

Part:BBa_K1321335:Design

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

acsCD


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1837
    Illegal BglII site found at 4382
    Illegal BamHI site found at 1906
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 778
    Illegal AgeI site found at 1334
    Illegal AgeI site found at 1913
    Illegal AgeI site found at 2123
    Illegal AgeI site found at 2975
    Illegal AgeI site found at 3293
    Illegal AgeI site found at 3638
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The coding sequences of AcsC and AcsD were 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 flanking this RBS were edited with the Sallis Lab RBS calculator so as to tune RBS strength to try preserving the native stoichiometry.

Source

Protein coding sequence from genome of Gluconacetobacter xylinus ATCC 53582, available here:

  • http://www.ncbi.nlm.nih.gov/nuccore/X54676.1

    These parts were codon optimised for expression in E. coli and sent for gene synthesis ===References===