First attempt to mutate failed. Suspect the problem is a low mutation rate due to results of positive control. May be a result of the mutagenesis kit being old. Second attempt to mutate failed. Abandoned,
Quadpart Amplifier (B0034.C0062.B0015.R0062)
Lambda cI QPI w/ strong RBS, hybrid promoter regulated by lambda cI and LuxR
Tet QPI with B0032 which according to preliminary experiments is a "medium" strength RBS.
Mutation has been sequenced. The rest of the part will be sequenced shortly.
LacI QPI with strong RBS, hybrid promoter
LacI QPI with B0032 which according to preliminary experiments is a "medium" strength RBS. Also uses hybrid promoter.
HindIII - Q04400 - NcoI Note that this part does NOT have the standard biobrick ends and is available only as a PCR product.
This biobrick contains the sequence homologies (50bp) to delete lpp gen (LPS moiety carrier) in E. coli with an ampicillin resistance and a Multiple Cloning Site (for the generation of genomic knock-ins of any desired gen in E. coli). The deletion of lpp gene and the replacement of that sequence with the Amp resistance. Gene deletions are performed with lambda-red recombination technology. FLP-FRT sequences are included to remove the antibiotic resistance with a specific recombinase.
This is the general structure 50bp homology Lpp // FRT // Amp R Promoter // RBS // AmpR // Terminator // FRT // MCS // 50 bp homology Lpp
The genetic design is based in the lpp deletion done by: Ni, Y., Reyes, J., & Chen, R. (2007). lpp Deletion as a Permeabilization Method. Biotechnology and Bioengineering, 1347-1356. It also contains some parts of the registry like BBa_K1073000, BBa_B0034, BBa_B0015
Tet QPI with B0030 which according to preliminary experiments is a "strong" strength RBS.
First attempt to mutate failed. Suspect the problem is a low mutation rate due to results of positive control. May be a result of the mutagenesis kit being old. Second attempt to mutate failed. Sent out to Blue Heron for synthesis.
Currently entered as a placeholder. This is a modified version of the tetR inverter Q04400 with a single base switch in the RBS: AAAGAGGAGAAA to AAAGAGGGGAAA
LacI QPI with B0030 which according to preliminary experiments is a "medium" strength RBS. Also uses hybrid promoter.