Difference between revisions of "Part:BBa K3038001:Design"

Alcohol dehydrogenase - Geobacillus stearothermophilus_Cmyc and 6His tags_Cterminal

Design Notes

C Myc and 6his Tags have been added to the Cterminal part of the ADR sequence.

The sequence is flanked with the sequence of EcoRI and XbaI on the 5'end and the SpeI and PstI on the 3' end.

Design of the ADR-C-term gene with the Geneious Software.
The design show restriction enzymes like EcoRI, XbaI, SpeI and PstI. The gene includes pBAD promoter and its terminater. Finally 6-His and C-myc tag are present in C-term.

The sequence is cloned in pSB1A3 . This design was necessary because the construction will be inserted later on in a chloramphenicol and kanamycin resistant strain.

The RFC10 standard was followed.

THe GC content has been optimized for the synthesis by IDT

Source

It comes from a synthetic gene, with the RFC10 standard, synthesized by IDT.

References

Gene structure and amino acid sequences of alcohol dehydrogenases of Bacillus stearothermophilus. Robinson,G.A., Bailey,C.J. and Dowds,B.C. JOURNAL Biochim. Biophys. Acta 1218 (3), 432-434 (1994)

Overexpression, Purification, and Biochemical Characterization of the Thermostable NAD-dependent Alcohol Dehydrogenase from Bacillus stearothermophilus. SHIM, EUN-JUNG, SANG-HOON JEON, AND KWANG-HOON KONG. J. Microbiol. Biotechnol. (2003), 13(5), 738–744