Coding

Part:BBa_K642006

Designed by: Wilson Lam, Matthew Orton   Group: iGEM11_uOttawa   (2011-09-23)

cI repressor tagged with yBFP and a VP16 activation domain

This is the cI repressor from phage lambda tagged with yeast codon optimized BFP and a VP16 activation domain. cI will bind to two operators already present in the registry: BBa_R0051 & BBa_K105021. BFP is a monomeric fluorescent protein that has an excitation peak of 399 nm and an emission peak of 465 nm (1). It was yeast codon optimized through DNA synthesis for the purpose of expressing in S. cerevisiae tagged to various repressors and activators. VP16 is a transcription activation domain from the herpes simplex virus that will initiate transcription when fused to a repressor protein. (2)

References

(1) Subach, O. M., I. S. Gundorov, et al. (2008). "Conversion of red fluorescent protein into a bright blue probe." Chem Biol 15(10): 1116-24.

(2) Urlinger et al. (2000). "Exploring the sequence space for tetracycline-dependent transcriptional activators: novel mutations yield expanded range and sensitivity." Proc Natl Acad Sci U S A 97(14):7963-8.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 711
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
n/acI repressor tagged with yBFP and a VP16 activation domain