Composite

Part:BBa_K3505036

Designed by: Venetios Michelioudakis, Magdalini Koroxenidou, Foteini Papadaki   Group: iGEM20_Thessaly   (2020-10-20)
Revision as of 15:02, 26 October 2020 by Venetios (Talk | contribs) (Experimental Use and Experience)


pFliC:RBS-LacI-Terminator- pAndersonJ23115:LacO:RBS-ECFP-terminator

Fig.1: Activation of eCFP in absence of SCFAs,Based on NOT GATE
Fig.2: The NOT GATE


Usage and Biology

2 Trancription Units

This way we take signal in the Absence of SCFAs


Design Notes

The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is present in BBa_K3505010 and has overhangs compatible for Golden Braid cloning.



Verification of cloning

Fig.3:U2 C2 pFliC:RBS-LacI-Terminator-pAndersonJ23115:LacO:RBS-ECFP-terminator EcoRV, HindIII. Expected bands 3734, 1699.

Experimental Use and Experience

Fig.3:J23115-eCFP-Terminator


Fig.4:J23115-eCFP-Terminator


Fig.5:J23115-eCFP-Terminator


Fig.6:J23115-eCFP-Terminator

Sequnce and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1321
    Illegal NheI site found at 1344
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]
[edit]
Categories
Parameters
None