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Part:BBa_K1429002:Design

Designed by: Christal Gordon   Group: iGEM14_Genspace   (2014-09-29)


Cyan Fluorescent Protein (CFP) "Cindy Lou" coding region, intellectual property-free


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 74
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2


Design Notes

We started with the FPB-20-444 plasmid from DNA 2.0 which contains an "IP-Free" synthetic non-Aequorea CFP coding region downstream from a ribosome binding site (AGGAGG ). PCR primers were designed complementary to sequences flanking the RBS-CFP region, with a "tail" that added the biobrick prefix/suffix. The PCR product was digested with EcoR1 and Pst1 and ligated into the pSB1C3 plasmid backbone.

Source

DNA 2.0 "Protein Paintbox". Note that although the company claims this protein is "IP-Free", we discovered that there are limitations for its use. One of them appears to be that in order to use it in an application that will be commercialized, you have to purchase the gene directly from DNA 2.0. You can, however, use this part for pure research applications.

References