Part:BBa_K3257116

T7 Promoter-lox5171-R-PPT-RBS-ChlR_L158X-PBS-U5-R-loxP-T7 Terminator

This part includes the target gene, which is ChlR_L158X (https://parts.igem.org/Part:BBa_K3257039) in this part, and a set of necessary regulatory factor for the reverse transcription and recombination, such as the primer binding site (PBS https://parts.igem.org/Part:BBa_K3257063), the R region (https://parts.igem.org/Part:BBa_K3257061), the U5 region (https://parts.igem.org/Part:BBa_K3257062), the polypurine tract (PPT https://parts.igem.org/Part:BBa_K3257060) and lox sites (https://parts.igem.org/Part:BBa_K3257064 https://parts.igem.org/Part:BBa_K3257075). Together they function as a target of the reverse transcription and recombination process, making them vital and fundamental for our R-Evolution system.

Fig. 1 The verification of well-functional Chl^R and non-functional Chl^R_L158X. Plasmids containing target gene (Chl^R and its L158X mutant) and other necessary factors for the reverse transcription and recombination process are transformed into E.coli BL21(DE3). We can see that cells containing normal Chl^R are able to grow on a solid plate with chloromphinicol (the right one) while cells containing nonsense mutated Chl^R cannot grow on a solid plate with chloromphinicol (the left one).

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1107
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 945
  Illegal BsaI.rc site found at 966