Part:BBa_K2539300
AlcR Expression Construct
AlcR is a gene found in Aspergillus nidulans which encodes for a regulatory protein (Panozzo et al., 1997). AlcR can bind to the alcA and aldA promoters (PalcA and PaldA, respectively). In the presence both AlcR and ethanol or threonine, genes downstream of these promoters are expressed (Felenbok et al., 1988).
Construct Design
This construct constitutively express alcR, a regulatory protein necessary for the activation of PalcA. We use a strong promoter and strong RBS combination (BBa_K880005) to maximize protein production, the alcR sequence (BBa_K2092001), and a double terminator (BBa_B0015) to end transcription.
PCR Check Results
The part was confirmed by PCR using the primers VF2 and VR, as well as sequencing by Tri-I Biotech.
PCR check for BBa_K2539300 (A) using VF2 and VR primers. Using these primers, PCR produced a band around the expected size of 3.1 kb.
References
Felenbok B, Sequeval D, Mathieu M, Sibley S, Gwynne DI, Davies RW. (1988). The ethanol regulon in Aspergillus nidulans: characterization and sequence of the positive regulatory gene alcR. Gene, 73(2), 385–396.
Panozzo C, Capuano V, Fillinger S, Felenbok B. (1997). The zinc binuclear cluster activator AlcR is able to bind to single sites but requires multiple repeated sites for synergistic activation of the alcA gene in Aspergillus nidulans. J Biol Chem. 5;272(36):22859-65.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2611
Illegal BamHI site found at 1615
Illegal BamHI site found at 2353 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1319
Illegal AgeI site found at 1447
Illegal AgeI site found at 1978 - 1000COMPATIBLE WITH RFC[1000]
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