Part:BBa_K2229400

Proteorhodopsin-Expressing Construct

A Proteorhodopsin-based construct that employs the strong promoter/strong RBS (K880005) combination to up-regulate expression of Proteorhodopsin, a membrane protein with two lysine residue sites (57 and 59) used to bind citrate through electrostatic interactions.

Characterization

Bba_K2229400 expresses Proteorhodopsin (BBa_K2229450), a membrane protein capable of binding to citrate. We obtained the DNA sequence of pR (Syed 2011) and modified it to remove three internal cutting sites (EcoRI, PstI, and SpeI). The sequence of pR was then flanked by an upstream strong promoter and strong ribosome combination (BBa_K880005), and a downstream double terminator (BBa_B0015) to maximize expression of PR protein. PCR checks and sequencing results from Tri-I Biotech confirmed that our final pR construct is correct.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]