Regulatory

Part:BBa_K2384009

Designed by: Size Cai   Group: iGEM17_FAFU-CHINA   (2017-10-23)


TetR

This part is combinded with trc promoter+lac operator+RBS+CDS+rrnB T1 terminator.


Usage and Biology

Tetracycline (tc) is a broad family of antibiotics to which bacteria have evolved resistance. Tc normally kills bacteria by binding to the bacterial ribosome and halting protein synthesis. The expression of tc resistance genes is regulated by Tet Repressor Protein, called TetR. More specifically, TetR represses the expression of TetA, a membrane protein that pumps out substances toxic to the bacteria, by binding the TetA operator. In tc resistant bacteria, TetA will pump out tc before it can bind to the ribosome. Therefore, TetR may have an important role in helping scientists to better understand mechanisms of antibiotic resistance and how to treat antibiotic-resistant bacteria. TetR is one of many proteins in the TetR protein family, which is so named because TetR is the most well-characterized member.

Figure 1:Toggle switch.

This part is the combination of other two parts(BBa_K2384011) and (BBa_K2384010) , which could work together as a kill switch that uses a transcription-based monostable toggle design to provide rapid and robust target cell killing, dubbed ‘Toggle switch’.

This part has made a condon optimization.

Sequence optimization information:

Expression System: Bacillus megaterium

Gene Length:624(bp)

Show the relative codon used distribution


Figure 2 :Codon Used Distribution after optimization.



Figure 3:GC Content after optimization.
Figure 4: Digested with EcoRI//.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 726
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//biosafety
Parameters
None