Difference between revisions of "Part:BBa K3505032"

Revision as of 13:34, 27 October 2020

pAndersonJ23115:lacO:RBS-eCFP -terminator

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

Fig.1:J23115-eCFP-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 , 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) (U=Uncut , C= Cut) Restriction digestion of LE: AndersonJ23115:Lac0-EGFP-double terminator(C1-C 4) with : HindIII + BtgZI (C1-C4) , Expected bands : 3200+ 597bp ,Positive result: C1,C2

Experimental Use and Experinece

This part is used in BBa_K3505036

Extra Engineering Use

Fig.3:Validation that J23115-TetO BBa_K3505044 and J23115-LacO fusion promoters are able to drive expression of ECFP.

Fig.4:NOT-GATE-regulated ECFP fluorescence after in the presence or absence of 2mM acetate.

Fig.5:NOT-GATE-regulated ECFP fluorescence after in the presence or absence of 2mM propionate.

Sequence and Features