Difference between revisions of "Part:BBa K398029:Design"
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===Design Notes=== | ===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 | ||
| + | |||
| + | According to the authors [1], the purified protein has an activity of dodecanal dehydrogenase equal to 22.995 U/mg protein (4e-7 kat/mg protein). This part was expressed using the promoter-rbs combination J23100-J61117 and also J13002. Thus generating the parts: [https://parts.igem.org/Part:BBa_K398029 BBa_K398029] and [https://parts.igem.org/Part:BBa_K398030 BBa_K398030], respectively. | ||
===Source=== | ===Source=== | ||
Revision as of 15:13, 27 October 2010
ALDH generator (low expression)
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 37
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 37
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 37
- 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
According to the authors [1], the purified protein has an activity of dodecanal dehydrogenase equal to 22.995 U/mg protein (4e-7 kat/mg protein). This part was expressed using the promoter-rbs combination J23100-J61117 and also J13002. Thus generating the parts: BBa_K398029 and BBa_K398030, respectively.
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/]