Difference between revisions of "Part:BBa K2671000"

Revision as of 15:02, 16 October 2018

AraC-pBAD-mNG

Liu iGEM2017s plasmid (BBa_K2474000) with an improvement. A stop codon was mutated behind mNeonGreen (mNG). The original part was mNG fused to Aβ1-42, where the fusion protein had its stop codon after Aβ1-42. Another mutation was made in the GS-linker sequence where a SpeI-site was added and the sequence for Aβ1-42 was cleaved off. With these improvements mNG can now be used as a reporter in a fully functional operon with the pBAD-AraC inducible system.

Usage and Biology

Figure 1. From left to right: Reference, mutated version (this biobrick) and the unmutated version (BBa_K2474000).

Figure 3. Comparing the sequencing results of the mutated version (this biobrick) and the template (BBa_K2474000). Showing a successful addition of an stop codon. Comparing the length of the sequence, both done with the VR primer, a loss of ~170 bases. This suggests a successful removal of the Aβ1-42-tail.

Verification As seen in fig 1. the improved version has a much stronger color, which indicates that the Aß1-42 fusion was successfully terminated. And as seen in fig 2. the sequencing results confirms this.

Figure 2. Sequencing results analyzed in benchling, annotations added for clarity. The sequencing data attached suggests a successful removal of Aβ1-42 and addition of a stop codon. Sequencing was done with the VR primer.

File:T--Linkoping Sweden--BBa K2671000seq.zip Sequencing data files. The analyzed data in figure 2. is from these files. These contain full information on the sequencing including a chromatogram.

Sequence and Features