Difference between revisions of "Part:BBa K3505034"

Revision as of 17:33, 27 October 2020

pAndersonJ23115:tetO:RBS-lacI-terminator

Usage and Biology

This parts consists of AndersonJ23115 promoter BBa_K3505012 , LacI BBa_K3505003 and Double Terminator BBa_K3505017. By this way ,as AndersonJ23115 is a strong constitutive promoter, LacI is constitutively expressed. LacI binds to lac operator and inhibits the expression of a desirable transcription unit.

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 alpha vector BBa_K3505009 and has overhangs compatible for Golden Braid cloning.

Figure 2. The level A module of Tet-Off system : AndersonJ23115:TetO:RBS-LacI-Double terminator

Verification of Cloning

Fig.1:(U=Uncut C=Cut) Restriction digestion of TL: AndersonJ23115:Tet0-LacI-double terminator(C5-C8 ) with: HindIII + NheI (C5-C18) , Expected bands : 2573+891+427,Positive result : C5,C6

Sequence and Features