Part:BBa_K125100:Design
nir promoter from Synechocystis sp. PCC6803
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The nir promoter was synthesized as 4 different oligonucleotides that were annealed, then ligated together.
| Primer name | Primer | Length | G/C content | Tm |
|---|---|---|---|---|
| pnir1_forward | CTAGAGCTAAATGCGTAAACTGCATATGCCTTCGCTGAGTGTAATTTACGTTACA | 55 | 40% | 72.5C |
| pnir1_reverse | GTAAATTACACTCAGCGAAGGCATATGCAGTTTACGCATTTAGCT | 45 | 40% | 70.6C |
| pnir2_forward | AATTTTAACGAAACGGGAACCCTATATTGATCTCTACTACTAGTAGCGGCCGCTGCA | 57 | 43.9% | 74.3C |
| pnir2_reverse | GCGGCCGCTACTAGTAGTAGAGATCAATATAGGGTTCCCGTTTCGTTAAAATTTGTAAC | 59 | 42.4% | 73.0C |
Source
The nirA promoter originates from Synechocystis sp. PCC6803. The Ntc-relevant sequence used here was identified by Ivanikova.
References
Aichi, Makiko, Takatani, Nobuyuki and Omata, Tatsuo. "Role of NtcB in Activation of Nitrate Assimilation Genes in the Cyanobacterium Synechocystis sp. Strain PCC 6803." Journal of Bacteriology 2001 (183): 5840-5847.
Ivanikova, Natalia Valeryevna. "Lake Superior phototrophic picoplankton: nitrate assimiliation measured with a cyanobacterial nitrate-responsive bioreporter and genetic diversity of the natural community." Dissertation. Bowling Green State University, 2006.