Part:BBa_K607001

PBad_cI-LVA

cI with a fast degradation tag (LVA) under the control of the pBAD/araC promoter. This part is a cI-LVA generator when it is induced by arabinose, but degrades faster compared to cI without LVA tag. The pBAD promotor is a promotor which belongs to the arabinose operon and its regulatory gene, araC.(1) This regulatory gene is both a positive and negative regulator (2), so in the presence of arabinose, the transcription from the pBAD will be turned on and in its absence, the transcription of the gene cloned downstream of the pBAD promotor occurs at very low levels. (1)
The cI gene codes for the bacteriophage λ repressor(3). This bacteriophage λ-repressor from phage lambda also contains a ssrA tag (small stable RNA tag). The ssrA tag is located at the C-terminal end of the incomplete protein product of a stalled ribosome. This tag is a target for proteolytic enzymes and cause degradation of the protein.(4)

References: (1) Luz-Maria Guzman, Dominique Belin, Michael J. Carson and Jon Beckwith. Tight Regulation, Modulation, and High-Level Expression by Vectors Containing the Arabinose PBAD Promoter. Journal of bacteriology, July 1995.
(2) Wanzhi Huang, Matthew McKevitt and Timothy Palzkill. Use of the arabinose pbad promoter for tightly regulated display of proteins on bacteriophage. Department of Molecular Virology & Microbiology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA
(3)Robert T. Sauer* and Robert Anderegg. Primary Structure of the λ Repressor. Biochemistry, 1978.
(4) Jeff Hasty, David McMillen, Farren Isaacs and James J. Collins. Computational studies of gene regulatory networks: in numero molecular biology. Nature 268

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1205
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1144
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 979
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 961