Composite

Part:BBa_K3257116

Designed by: Yi Qiao   Group: iGEM19_Fudan-TSI   (2019-10-13)

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 ChlR and non-functional ChlR_L158X. Plasmids containing target gene (ChlR 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 ChlR are able to grow on a solid plate with chloromphinicol (the right one) while cells containing nonsense mutated ChlR 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


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