The final constructs, in theory

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The final constructs, in theory Promoter: R0010 RBS: B0032 Signal: Lpp(1-29) Membrane: OmpA(46-66) OmpA(46-159) Streptavidin: Monomeric StrepW StrepH StrepD pTSA13 Dimeric SCD-NM C2 E2

BioBricking dimeric streptavidin 250 550 1 825 PstI 620 QuikChange mutagenesis to constructs in original pET22, PCR, Topo cloning Sequencing Missing His codon “CAC” from C-term His6 SpeI site mutated “ACTAGT” to “ATTAGT” Another colony? Another QuikChange?

Assembly progress Lpp(1-29)-OmpA(46-66) and Lpp(1-29)-OmpA(46-159) completed with correct sequence New pTSA13 and dimeric streptavidin BioBricks R0010-B0032 and Lpp-OmpA-Strep assemblies have been performed; to be checked for insert by colony PCR and then sequenced