Reporter

Part:BBa_K1388001:Design

Designed by: Tom Geddes   Group: iGEM14_USyd-Australia   (2014-10-07)


aeBlue-aacC1 gene cassette for integron testing


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 28
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1075
    Illegal BamHI site found at 810
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

- attC placed at beginning of part to allow for the option of cointegration of plasmids using integron recombination (retaining aeBlue/aacC1 near attI site and any nearby promoters). - 5' spacer region included to allow for primer annealing; attC has significant secondary structure which can make primer design difficult. - NheI was placed to allow ELAN circularisation without SpeI without including unnecessary spacer region. - BamHI was placed to allow excision of aacC1 gene with BamHI/Spe digest.


Source

Synthesised as a complete gBlock gene fragment from Integrated DNA Technologies: attC sequence was sourced from pUS23 and aacC1 was sourced from pUCP24. aeBlue sequence was taken from part BBa_K864401.

References