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Part:BBa_K1804001:Design
GFP under the control of constitutive promoter lacI
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 734
Design Notes
plac-gfp was constructed by prepending the KpnI sequence and appending the XhoI sequence to the start and stop codons of gfp (BBa_E0040 [1]) respectively. This was done via extension polymerase chain reaction (PCR). Restriction enzyme digest with KpnI and XhoI was then carried out to replace esaR and esaI with the PCR product in the pAC-esaR-esaI plasmid (Figure 1). This placed gfp under the control of the plac promoter. The resultant pAC-plac-gfp plasmid encoded gfp under the control of plac.
Source
The lacI promoter (plac) was obtained from the pAC-EsaR-EsaI plasmid (Shong & Collins, 2013), which was a gift from Cynthia Collins (Addgene plasmid # 47660).
The Green Fluorescence Protein (GFP) gene was obtained from BioBricks Part BBa_E0040 (https://parts.igem.org/Part:BBa_E0040).
References
Shong, J., & Collins, C. H. (2013). Engineering the esaR promoter for tunable quorum sensing-dependent gene expression. ACS synthetic biology, 2(10), 568-575.