Part:BBa_K5206003
Palsl-alsR
This part is composed of BBa_K5206000, BBa_K5206001 and BBa_K731721. BBa_K5206000 (alsR) is derived from Escherichia coli BL21(DE3), which is terminated by BBa_K731721 (T7 terminator). In the E. coli genome, AlsR is responsible for the control of six genes related to D-allose metabolism, aISRBACEK and its bidirectional promoter BBa_K5206001(PalsI). The gene downstream of the bidirectional promoter is also regulated by AlsR. When D-allose is present in the environment, the AslR transcriptional regulatory protein binds preferentially to D-allose and deregulates downstream genes.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1263
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Results
Plasmid construction
The plasmid vector pETDuet-1 is used to construct a recombinant plasmid pETDuet-1-PalsI-alsR to verify the effect of BBa_K5206003. We amplified alsR and the PalsI region using primers alsR-For and alsR-Rev, and amplified the vector using pETDuet1-gj(alsR)-For and pETDuet1-gj(alsR)-Rev. Subsequently, Gibson assembly was used to ligate the vector and the target gene fragment.
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