Translational_Unit

Part:BBa_K118005:Design

Designed by: Andrew Hall   Group: iGEM08_Edinburgh   (2008-08-08)


rbs+crtI (phytoene dehydrogenase)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 87
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The native crtI sequence contains two internal PstI sites. Primers crtImutf1 and crtImutf2 were ordered so that the MABEL protocol (invented for this purpose) could be used to remove these sites. The mutagenic primers were as follows: 

* crtImutf1: gtt tac agt aag gtt gcc

* crtImutf2: agt aac tct ctg ttt gtg c

In both cases, the PstI site CTGCAG was converted to CTGCAA, a silent mutation.


Source

rbs: Escherichia coli JM109 genomic DNA. crtI: Pantoea ananatis genomic DNA

References