1. 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

2. 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?

3. 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