Part:BBa_K5323893

P_tac-RBS-golS-P_gol-RBS-cI-TT-PR-RBS-mazF-TT

Uses a factor strong RBS(BBa_J34801), golS (BBa_K4468005), P_gol(BBa_K4468001), cI (BBa_R1051), P_tac(BBa_K180000), TT(BBa_B0015), mazF(BBa_K1096002) and PR(BBa_K2229450). This part is an easy BioBrick.

Usage and Biology

This component can determine whether the mazF gene is expressed by specific test gold.The cI expression product in this component represses its downstream promoter P</sub>R</sub> in this component. When cI is inhibited, inhibition of the promoter P_R is lifted. When gold was detected, the downstream gene was not expressed, and when gold was not present, the downstream gene was expressed. The kill stwich designed in our project ensures that engineered bacteria can only survive in an environment containing Au[Ⅲ]. When Au[Ⅲ] is not present, the PR promoter is not inhibited, initiating transcription, and mazF expresses the toxic protein MazF to kill the engineered bacteria. Once Au[Ⅲ] is present, the promoter P_R is inhibited, mazF does not express MazF toxic protein, and the engineered bacteria survive. We successfully verified this result by using E.coli BL21 as the chassis cell.

Fig 1 The growth curves of E.coli BL21 in the presence of inducer IPTG and Au[Ⅲ], only inducer IPTG or Au[Ⅲ], and no inducer IPTG and Au[Ⅲ] respectively.

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 1635
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 3721
  Illegal AgeI site found at 1969
  Illegal AgeI site found at 2318
  Illegal AgeI site found at 3256
  
• 1000
  COMPATIBLE WITH RFC[1000]