Difference between revisions of "Part:BBa K2539500"
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This construct uses the inducible promoter PalcA (BBa_K2092002). PalcA is activated in the presence of both ethanol and a transcription factor, alcR (BBa_K2092001), and was originally isolated from the fungus Aspergillus nidulans (Panozzo et al., 1997). GFP is used as a visible reporter so we can characterize the function of the promoter PalcA. | This construct uses the inducible promoter PalcA (BBa_K2092002). PalcA is activated in the presence of both ethanol and a transcription factor, alcR (BBa_K2092001), and was originally isolated from the fungus Aspergillus nidulans (Panozzo et al., 1997). GFP is used as a visible reporter so we can characterize the function of the promoter PalcA. | ||
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The PalcA promoter is placed in front of a strong RBS (BBa_B0034), GFP (BBa_E0040), and double terminator (BBa_B0015). This places the expression of GFP under the control of the PalcA promoter. | The PalcA promoter is placed in front of a strong RBS (BBa_B0034), GFP (BBa_E0040), and double terminator (BBa_B0015). This places the expression of GFP under the control of the PalcA promoter. | ||
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https://static.igem.org/mediawiki/parts/a/a2/T--TAS_Taipei--300pcr.jpg | https://static.igem.org/mediawiki/parts/a/a2/T--TAS_Taipei--300pcr.jpg | ||
− | <b>PCR check for BBa_K2539500 using VF2 and VR primers. | + | <b>PCR check for BBa_K2539500 using VF2 and VR primers. Using these primers, PCR produced a band at the expected size of 2 kb.</b> |
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Revision as of 05:40, 11 October 2018
PalcA-Regulated GFP Expression Construct
This construct uses the inducible promoter PalcA (BBa_K2092002). PalcA is activated in the presence of both ethanol and a transcription factor, alcR (BBa_K2092001), and was originally isolated from the fungus Aspergillus nidulans (Panozzo et al., 1997). GFP is used as a visible reporter so we can characterize the function of the promoter PalcA.
Construct Design
The PalcA promoter is placed in front of a strong RBS (BBa_B0034), GFP (BBa_E0040), and double terminator (BBa_B0015). This places the expression of GFP under the control of the PalcA promoter.
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_K2539500 using VF2 and VR primers. Using these primers, PCR produced a band at the expected size of 2 kb.
References
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]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 274
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1513