Translational_Unit

Part:BBa_K523002:Design

Designed by: Sylvia Ispasanie, Mun Ching Lee   Group: iGEM11_Edinburgh   (2011-07-19)

RBS + bglX (E. coli perisplasmic β-glucosidase)


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 AgeI site found at 1658
    Illegal AgeI site found at 1880
    Illegal AgeI site found at 2069
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The part was made using the strategy outlined in BBa_K523000, and therefore contains 4 extra bases at the 5' end which generate a BglII restriction site.

The part was created from a lab E. coli strain using primers:

  • Forward: ACT AGATCT gcc acg tcg ggc aac
    • adds BglII site; starts upstream of gene to catch RBS; RFC10 compliant after replacement of BBa_K523000 in a normal BioBrick vector.
  • Reverse: ACT ACTAGT A TTA tta cag caa ctc aaa ctc gcc
    • adds SpeI site; RFC10 compliant after insertion into vector; has double stop codon.


A PstI site was later removed by site-specific mutagenesis ([http://openwetware.org/wiki/Cfrench:MABEL MABEL protocol]).

Source

The source E. coli genome sequence can be seen using GenBank U00096.2

References