Coding
Part:BBa_K777000:Design
Designed by: Bianca Genenncher Group: iGEM12_Goettingen (2012-09-11)
Tar receptor
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1282
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 111
Design Notes
- Genomic sequence was amplified using the following primers.
- Primers PCR: Base pairs in caps represent the respective restriction sites followed by either prefix or suffix. Primers were provided by SIGMA.
- Fwd: Tar for + prefix: 5´ actGAATTCgcggccgctTCTAGAtgattaaccgtatccgcgtagtc 3´
- Rev: Tar rev + suffix: 5´ tcCTGCAGcggccgctACTAGTcaaaatgtttcccagtttggatc 3´
- Primers PCR: Base pairs in caps represent the respective restriction sites followed by either prefix or suffix. Primers were provided by SIGMA.
- One XbaI site was found at position 420. The restriction site was changed from TCTAGt to CCTAGt via QuikChange PCR.
- Primers QuikChange (QC): Primers were provided by SIGMA.
- Fwd: TarQC for: 5´ ACTGATTGATTAcCTAGATTATGGCAATACTGGAG 3´
- Rev: TarQC rev: 5´ TGCCATAATCTAGgTAATCAATCAGTTCAGTTAAC 3´
(the uncapitalized letter induces the mutation for removal of the XbaI site)
- Primers QuikChange (QC): Primers were provided by SIGMA.
Source
The part was amplified from genomic DNA of E. coli str. K-12 substr. DH10B, complete genome (CP000948.1).
References
- Derr P., Boder E. and Goulian M. Changing the specificity of a bacterial chemoreceptor. J. Mol. Biol. 2006. 355:923–932.