Composite

Part:BBa_K3505032

Designed by: Asteria Tsapadikou , Magdalini Koroxenidou, Foteini Papadaki   Group: iGEM20_Thessaly   (2020-10-20)
Revision as of 10:28, 25 October 2020 by Venetios (Talk | contribs)


pAndersonJ23115:lacO:RBS-eCFP -terminator


eCFP BBa_K3505020uder control of a constitutive promoter (andersonJ23115 with a lac operator) BBa_K2924013.

File:T--Thessaly--lacoecfpmap.png
Fig.1:pFliC-LacI-Terminator

Usage and Biology

This Trancriscription Unit (TU) is continuesly activated exressing the eCFP protein as a reporter. The Lac operator that is downsteam the anderson exists for the lac regulated inhibition.


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 cloned in both a1R BBa_K3505008and a2 BBa_K3505009 and has overhangs compatible for Golden Braid cloning.

Verification of cloning

Fig.2:(U=Uncut C=Cut) Restriction Enzyme with HindIII. Expected bands in bp 2847 + 804 + 146

Experimental Use and Experinece

This part is used in BBa_K3505036


Extra Engineering Use

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 11
    Illegal NheI site found at 34
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]
[edit]
Categories
Parameters
None