Difference between revisions of "Part:BBa K398006:Design"
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The original CDS can be found [http://www.ncbi.nlm.nih.gov/nuccore/AB047106.1 in this link], the file reports a cluster of proteins induced by alkanes. We took the ORF contained from 1213 to 2706. | The original CDS can be found [http://www.ncbi.nlm.nih.gov/nuccore/AB047106.1 in this link], the file reports a cluster of proteins induced by alkanes. We took the ORF contained from 1213 to 2706. | ||
Nucleotide sequence optimized for expression in ''E.coli'' K12. We used [http://www.jcat.de/ jcat] for this purpose. Moreover, we deleted the restriction sites for EcoRI, PstI, SpeI and XbaI and we change the stop codon for a TAA. | Nucleotide sequence optimized for expression in ''E.coli'' K12. We used [http://www.jcat.de/ jcat] for this purpose. Moreover, we deleted the restriction sites for EcoRI, PstI, SpeI and XbaI and we change the stop codon for a TAA. | ||
+ | |||
+ | The expected aminoacid sequence is: | ||
+ | |||
+ | MTMISAQTTEADLWIDGEWRPAASGERFDVIDPATGEVTARVAN | ||
+ | AGEDDVDAAVAIAEEAFSVRRWLAISPLERGRILRRIAELIRQHHCELAQLMTRENGM | ||
+ | PINLALFIEIPLADCFDFFASLVVKPQGEVLPFSVAGSAPDYMAWTMKEPIGVAGLIT | ||
+ | PWNFPLLMPTWKIAPALAAGCTMVVKPAPETPLTALKLAEICHEAGVPEGVINVLPGL | ||
+ | DEAGKALVRHPRVPKIAFTGETETGRHILQAAAPHIKRVTLELGGKSPNIIFADADLE | ||
+ | QAAKSALFGVFYNSGQVCQAGSRILVERTVYEPFVERLAERAKKLKVGPGTNPRSDLG | ||
+ | PVISREQYEKVLRYIEIGKQEGARLAAGGRALDGGGGGYFIEPTVFADVSPSMRIACE | ||
+ | EIFGPVAAVIPFADEEEAVRIANGTMYGLAAAVWTNDIKRALRLARRVKSGTVWLNTY | ||
+ | QVLSPTVPFGGYKQSGLGRELGMQALDAYLETKTVICDLNDRPMTLF | ||
===Source=== | ===Source=== |
Revision as of 13:39, 27 October 2010
Medium-Chain Aldehyde Dehydrogenase
Assembly Compatibility:
- 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 original CDS can be found [http://www.ncbi.nlm.nih.gov/nuccore/AB047106.1 in this link], the file reports a cluster of proteins induced by alkanes. We took the ORF contained from 1213 to 2706. Nucleotide sequence optimized for expression in E.coli K12. We used [http://www.jcat.de/ jcat] for this purpose. Moreover, we deleted the restriction sites for EcoRI, PstI, SpeI and XbaI and we change the stop codon for a TAA.
The expected aminoacid sequence is:
MTMISAQTTEADLWIDGEWRPAASGERFDVIDPATGEVTARVAN AGEDDVDAAVAIAEEAFSVRRWLAISPLERGRILRRIAELIRQHHCELAQLMTRENGM PINLALFIEIPLADCFDFFASLVVKPQGEVLPFSVAGSAPDYMAWTMKEPIGVAGLIT PWNFPLLMPTWKIAPALAAGCTMVVKPAPETPLTALKLAEICHEAGVPEGVINVLPGL DEAGKALVRHPRVPKIAFTGETETGRHILQAAAPHIKRVTLELGGKSPNIIFADADLE QAAKSALFGVFYNSGQVCQAGSRILVERTVYEPFVERLAERAKKLKVGPGTNPRSDLG PVISREQYEKVLRYIEIGKQEGARLAAGGRALDGGGGGYFIEPTVFADVSPSMRIACE EIFGPVAAVIPFADEEEAVRIANGTMYGLAAAVWTNDIKRALRLARRVKSGTVWLNTY QVLSPTVPFGGYKQSGLGRELGMQALDAYLETKTVICDLNDRPMTLF
Source
Genbank accession number: [http://www.ncbi.nlm.nih.gov/nuccore/AB047106 AB047106]
References
- Kato T. et al. "Gene cloning and characterization of an aldehyde dehydrogenase from long-chain alkane-degrading Geobacillus thermoleovorans B23.", Extremophiles. 2010 Jan;14(1):33-9. Epub 2009 Sep 29.
- National Center for Biotechnology Information [http://www.ncbi.nlm.nih.gov/]