Difference between revisions of "Part:BBa K3038000:Design"

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===Design Notes===
 
===Design Notes===
  
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C MYC and  6His Tags have been added on the Nterminal part of the sequence.  
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C MYC and  6His Tags have been added on the Nterminal part of the 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.  
 
The sequence is flanked with the sequence of EcoRI and XbaI on the 5'end and the SpeI and PstI on the 3' end.  
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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
 
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
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Latest revision as of 15:32, 21 October 2019


Alcohol dehydrogenase - Geobacillus stearothermophilus cMYC and 6His N-terminal tags


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

C MYC and 6His Tags have been added on the Nterminal part of the 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.

T--Poitiers--TDesignADRNter.png
Design of the ADR-N-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 N-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

Sequence synthesis 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