Coding

Part:BBa_K4294215

Designed by: Aristotelis Anastopoulos   Group: iGEM22_Athens   (2022-09-30)


TetR

This is the coding sequence for the TetR repressor that regulates the pTet promoter. It is codon optimized for E.coli.

Design

TetR forms a dimer and binds to tet operators in pTet promoter and blocks the assembly of the transcription machinery. pTet is a medium-strength promoter that is constitutive “ON” in the absence of TetR. It contains two tet operators, one inside the core promoter sequence and the other after the -10 hexamer, leading to efficient repression. The system is efficiently induced via anhydrotetracycline hydrochloride (aTc), a tetracycline derivative which binds Tet R with an ~35-fold higher binding constant, being an effective inducer at very low concentrations. Additionally, its antibiotic activity is ~100-fold lower and has a minimal toxic effect on E.coli cells in the concentration used for induction.

As a team, we used it for our Senders' constructs.

Athens2022-Senders-Circuit.png

References

[1] Lutz, R., & Bujard, H. (1997). Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic acids research, 25(6), 1203–1210. https://doi.org/10.1093/nar/25.6.1203


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None