Difference between revisions of "Part:BBa K1659300:Design"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1659300 short</partinfo>
 
<partinfo>BBa_K1659300 short</partinfo>
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===Design Notes===
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===Design Notes and Sources===
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to be updated
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<html>
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    <figure>
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        <img src="https://static.igem.org/mediawiki/parts/6/61/Oxford15_blastdnase.png" alt="DNase BLAST" width="856" height="123">
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        <figcaption>
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        <p style="font-size:11px">
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BLAST search using translated sequence of <a href="https://parts.igem.org/Part:BBa_K729004>BBa_K729004</a>, from which only the nucleotide sequence that coded for the DNase was identified
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        </p>
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        </figcaption>
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    </figure>
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</html>
  
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The gene sequence for micrococcal DNase was obtained from [https://parts.igem.org/Part:BBa_K729004 BBa_K729004], a part comprising DsbA 2-19 fused to the N-terminal of micrococcal nuclease made by [http://2012.igem.org/Team:University_College_London Team UCL 2012]. The part sequence was not annotated, and as such we ran a [http://blast.ncbi.nlm.nih.gov/blast/Blast.cgi?PROGRAM=blastp&PAGE_TYPE=BlastSearch&LINK_LOC=blasthome Protein BLAST search] using the part's translated sequence to identify the bases that coded for our desired peptide [1]. The sequence for micrococcal DNase was adopted as-is from said part, without further codon optimization.
  
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===Source===
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A Hisx6 tag is added at the C-terminus for ease of protein purification using metal-affinity chromatography.
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to be updated
 
  
 
===References===
 
===References===
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[1] Stephen F. Altschul, Thomas L. Madden, Alejandro A. Schäffer, Jinghui Zhang, Zheng Zhang, Webb Miller, and David J. Lipman, 1997. "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs", Nucleic Acids Res. 25:3389-3402.

Revision as of 01:24, 15 September 2015

Micrococcal Nuclease (DNase)


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 and Sources

DNase BLAST

BLAST search using translated sequence of BBa_K729004, a part comprising DsbA 2-19 fused to the N-terminal of micrococcal nuclease made by [http://2012.igem.org/Team:University_College_London Team UCL 2012]. The part sequence was not annotated, and as such we ran a [http://blast.ncbi.nlm.nih.gov/blast/Blast.cgi?PROGRAM=blastp&PAGE_TYPE=BlastSearch&LINK_LOC=blasthome Protein BLAST search] using the part's translated sequence to identify the bases that coded for our desired peptide [1]. The sequence for micrococcal DNase was adopted as-is from said part, without further codon optimization.

A Hisx6 tag is added at the C-terminus for ease of protein purification using metal-affinity chromatography.


References

[1] Stephen F. Altschul, Thomas L. Madden, Alejandro A. Schäffer, Jinghui Zhang, Zheng Zhang, Webb Miller, and David J. Lipman, 1997. "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs", Nucleic Acids Res. 25:3389-3402.