Difference between revisions of "Part:BBa K777002:Design"
(New page: __NOTOC__ <partinfo>BBa_K777002 short</partinfo> <partinfo>BBa_K777002 SequenceAndFeatures</partinfo> ===Design Notes=== * Genomic sequence was amplified using the following primers. *...) |
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===References=== | ===References=== | ||
*Derr P., Boder E. and Goulian M. Changing the specificity of a bacterial chemoreceptor. J. Mol. Biol. 2006. 355:923–932. | *Derr P., Boder E. and Goulian M. Changing the specificity of a bacterial chemoreceptor. J. Mol. Biol. 2006. 355:923–932. | ||
− | *[https://parts.igem.org/Part: | + | *[https://parts.igem.org/Part:BBa_J23104 Anderson promoters] from the 2006 Berkeley group |
Latest revision as of 10:38, 21 September 2012
Tar receptor under the control of constitutive promoter J23104
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1323
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 152
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).
- J23104 information was taken from the parts.igem and physical DNA from the 2012 distribution kit.
References
- Derr P., Boder E. and Goulian M. Changing the specificity of a bacterial chemoreceptor. J. Mol. Biol. 2006. 355:923–932.
- Anderson promoters from the 2006 Berkeley group