Difference between revisions of "Part:BBa K1351036:Design"
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agrA-fwd: gatcgaattccgcggccgcttctagataaggaggagccggcATGAAAATTTTCATTTGCGAAGACG | agrA-fwd: gatcgaattccgcggccgcttctagataaggaggagccggcATGAAAATTTTCATTTGCGAAGACG | ||
agrA-rev: gatcctgcagcggccgctactagtattaaccggtTATTTTTTTAACGTTTCTCACCG | agrA-rev: gatcctgcagcggccgctactagtattaaccggtTATTTTTTTAACGTTTCTCACCG | ||
| − | agrC-fwd: gatcgaattccgcggccgcttctagataaggaggagccggcTTGATTCCAACTTTTTCATCTATC | + | agrC-II-fwd: gatcgaattccgcggccgcttctagataaggaggagccggcTTGATTCCAACTTTTTCATCTATC |
| − | agrC-rev: gatcctgcagcggccgctactagtattaaccggtGTTGTTAATAATTTCAACTTTTTGAATAAAG | + | agrC-II-rev: gatcctgcagcggccgctactagtattaaccggtGTTGTTAATAATTTCAACTTTTTGAATAAAG |
| − | The two intermediated parts agrA and agrC were then fused by using the restriction sites EcoRI and SpeI for agrA, as well as XbaI and PstI for agrC, and ligation into EcoRI and PstI cut pSB1C3, thus creating the composite part | + | The two intermediated parts agrA and agrC-II were then fused by using the restriction sites EcoRI and SpeI for agrA, as well as XbaI and PstI for agrC, and ligation into EcoRI and PstI cut pSB1C3, thus creating the composite part agrC-IIA. |
===References=== | ===References=== | ||
Revision as of 19:29, 16 October 2014
Quorum sensing two component system (AIP-II sensing histidine kinase agrC, response regulator agrA)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1139
Illegal AgeI site found at 1114 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 475
Design Notes
This part contains the native sequences of agrC-II and agrA from Staphylococcus aureus N315. Since these two parts are forming a two component system, this is actually a composite part. The basic parts agrCII and agrA can be yielded by using the NgoMIV or AgeI restriction sites with appropiate restriction sites in the pre- or suffix.
Source
This part is derived from Staphylococcus aureus N315 gDNA by PCR using primers with modified RFC25 prefix and suffix:
agrA-fwd: gatcgaattccgcggccgcttctagataaggaggagccggcATGAAAATTTTCATTTGCGAAGACG agrA-rev: gatcctgcagcggccgctactagtattaaccggtTATTTTTTTAACGTTTCTCACCG agrC-II-fwd: gatcgaattccgcggccgcttctagataaggaggagccggcTTGATTCCAACTTTTTCATCTATC agrC-II-rev: gatcctgcagcggccgctactagtattaaccggtGTTGTTAATAATTTCAACTTTTTGAATAAAG
The two intermediated parts agrA and agrC-II were then fused by using the restriction sites EcoRI and SpeI for agrA, as well as XbaI and PstI for agrC, and ligation into EcoRI and PstI cut pSB1C3, thus creating the composite part agrC-IIA.