Coding

Part:BBa_K523004:Design

Designed by: Sylvia Ispasanie, Mun Ching Lee   Group: iGEM11_Edinburgh   (2011-07-29)

bglX for BioSandwich


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 1625
    Illegal AgeI site found at 1847
    Illegal AgeI site found at 2036
  • 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:

  • ACT AGATCT atg aaa tgg cta tgt tca gta gg
    • Forward; adds BglII site; starts at start codon.
  • ACT ACTAGT ACC cag caa ctc aaa ctc gcc ttt c
    • Reverse; adds SpeI site; BioSandwich compliant; no stop codons, adds a glycine.


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

Source

See BBa_K523002

References