Difference between revisions of "Part:BBa K3866038"

Revision as of 09:24, 16 October 2021

J23115:sfGFP:Terminator

Usage and Biology

This TU includes the sfGFP gene placed under the control of the constitutive Anderson promoter J23115 BBa_K3866000. Initially, the level 1 cloning was performed using the trc promoter BBa_K3866001, though we weren't able to isolate a colony with the desired construct, as there is a possibility that the bacteria's metabolism went out of balance so no colonies survived to be chosen for screening. Instead we used the arabinose-indused promoter, which seemed to be working as expected.

Design Notes

The coding sequence was domesticated. We removed BsmBI and BsaI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in a PDGB3_α1 vector and has overhangs compatible for GoldenBraid cloning.

Figure 1. The first level α module of the Propionate Production: α1:ParaBAD:RBS-smb-Double terminator

Verification of Cloning

Figure 2.: (C=Cut, U=Uncut) Restriction digestion of α1-sbm with: EcoRI, Expected bands: 6345bp, 2947bp, 597bp, Positive result: C2

Experimental Use and Experience

This part showed functionality at the following parts: BBa_K3866029, BBa_K3866031

Sequence and Features