Composite

Part:BBa_K4194011

Designed by: Jiong Huang   Group: iGEM22_BUCT-China   (2022-09-24)


pF-RBS-mRFP1-LVA-RBS-TetR-LVA-terminator

The expression of mRFP1 and TetR is regulated by promoter pF, which can be specifically activated by δ Activator, an activator from phage phiR73. The mRFP1 was used as a reporter gene. The Tet-regulated gene expression system was established based on the Tet-resistant operon on the Tn10 transposon of Escherichia coli. TetR has specific affinity with DNA sequences in TetO. TetR binds to two tetracycline manipulation sites (tetO) in p (TetR) with high affinity and inhibits the transcription of downstream genes, which can be relieved by tetracycline. In addition, ssrA-LVA tag was added to the downstream of mRFP1 and TetR in order to allow the protein to be degraded in time.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 91
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 809
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 91
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 91
    Illegal AgeI site found at 18
    Illegal AgeI site found at 691
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None