Difference between revisions of "Part:BBa K3505013"

Revision as of 16:15, 27 October 2020

AndersonJ23115:LacO:RBS GB compatible A1-B2

Constitutive Anderson BBa_J23115with Lac Operator and RBS BBa_B0034

Usage and Biology

This part is a constitutive promoter from Andesron family , J23115 , having downstream with a Lac Operator and RBS. The activity of this part is regulated by LacI BBa_K3505003. LacI binds to Lac Operator and inhibits the expression of the desirable transcription unit.

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 Golden Braid cloning. This has position A1-B3.

Fig.1:The overhangs of this part in the Golden Braid Grammar

Fig.2:The level 0 module :pupd2- AndersonJ23115:LacO:RBS I (illustration from SnapGene)

Verification of cloning

Fig.3:((U=Uncut , C= Cut)Restriction digestion of LacO (C3 & C4) with : EcoRV + HindIII (C3 & C4) , Expected bands : 2173bp ,Positive result : C3 + C4

Experimental Use and Experience

This part is used in BBa_K3505032

Sequence and Features