Difference between revisions of "Part:BBa K2273111:Design"
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===References=== | ===References=== | ||
| + | <a target="_blank" href ="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2942778/">[1]</a></td> Leticia I. Llarrull, Mary Prorok, and Shahriar Mobashery (2010) Binding of the Gene Repressor BlaI to the bla Operon in Methicillin-Resistant Staphylococcus aureus. Biochemistry 49(37), 7975–7977 | ||
Revision as of 17:16, 30 October 2017
PblaZ Promoter controlling gene expression of blaZ in S. aureus
- 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
This Promoter was designed to be cloned in front of the luxABCDE operon in the pBS3C vector backbone and therefore the RFC10 Standard Restriction sites have been added as prefix and suffix.
| Prefix with | EcoRI, NotI, XbaI | GAATTCGCGGCCGCTTCTAGA |
| Suffix with | SpeI, NotI and PstI | ACTAGTAGCGGCCGCTGCAGA |
Sites of restriction enzymes generating compatible overhangs are indicated by sharing one color. (EcoRI and PstI are marked in blue, NotI in green, XbaI and SpeI in red
As the overall promoter sequence was quite short, we created it by ordering to long Primers that have a 20 base pair overlap:
Forward Primer Sequence containing RFC10 prefix: 5`-GATCGAATTCGCGGCCGCTTCTAGATTCAAATATTTATAATAAACAATTGACATCAATATTACAATTGTAATATTATTG-3`
Revers Primer Sequence containing RFC10 suffix: 5`-GATCCTGCAGCGGCCGCTACTAGTATATTACAGTTGTAATTTTTATAAATCAATAATATTACAATTGTAATATTGATG-3`
Source
This part sequence was derived from the genome sequence of Staphylococcus aureus N315 found in the NCBI database.
References
<a target="_blank" href ="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2942778/">[1]</a></td> Leticia I. Llarrull, Mary Prorok, and Shahriar Mobashery (2010) Binding of the Gene Repressor BlaI to the bla Operon in Methicillin-Resistant Staphylococcus aureus. Biochemistry 49(37), 7975–7977