Composite

Part:BBa_K4194013

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


p(TetR)-RBS-EGFP-LVA-RBS-pleD-LVA-terminator

The expression of EGFP and pleD is regulated by promoter p (TetR), which can be repressed by protein TetR. EGFP 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 is added to the downstream of EGFP and pleD to ensure timely degradation of the protein.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 1359
    Illegal NotI site found at 1508
    Illegal NotI site found at 1611
    Illegal NotI site found at 2258
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1597
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1811
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None