Part:BBa_K642005

LacI repressor tagged with yBFP and a VP16 activation domain

This is the LacI repressor from E.coli tagged with yeast codon optimized BFP and a VP16 activation domain. LacI will bind with high specificity to the the operator lacO and is inhibited in the presence of IPTG. (1) BFP is a monomeric fluorescent protein that has an excitation peak of 399 nm and an emission peak of 465 nm (2). 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. (3)

References

(1) Yansura DG, Henner DJ. (1984). "Use of the Escherichia coli lac repressor and operator to control gene expression in Bacillus subtilis." Proc Natl Acad Sci U S A 81(2): 439-43.

(2) 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.

(3) 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 1084
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]