Coding

Part:BBa_K3505005

Designed by: Asteria Tsapadikou   Group: iGEM20_Thessaly   (2020-10-09)
Revision as of 20:29, 27 October 2020 by Asteriat (Talk | contribs) (→‎Design Notes)


TetR- Tet repressor GB compatible with B3-B5


TetR- tetR Coding region for Tet repressor without an RBS .


Usage and Biology

Design Notes

The sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 BBa_K3505007 and has overhangs compatible for GoldenBraid cloning. This has position B3-B5.

Figure 1.The overhangs of this part in the GoldenBraid Grammar.
Figure 2. The level 0 module : pupd2- TetR (illustration from SnapGene)

Verification of Cloning

Fig.3:(U=Uncut C=Cut) Restriction digestion of TetR (C2 -C4) with : SpeI + EcoRI (C2 - C4) , Expected bands : 2047 + 680 bp Positive result : C4

Source

Synthesized by IDT.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 4
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 4
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 4
  • 1000
    COMPATIBLE WITH RFC[1000]

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Categories
Parameters
None