Difference between revisions of "Part:BBa K3765015"

 
 
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<partinfo>BBa_K3765015 short</partinfo>
 
<partinfo>BBa_K3765015 short</partinfo>
  
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Desc.
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CDS of fusion protein consisting of SpyTag002 linked to OpdA (phosphotriesterase), which can covalently bind to SpyCatcher/SpyCatcher002. In our project, the protein encoded by this part is designed to get anchored to a [https://parts.igem.org/Part:BBa_K3765012 SpyCatcher002-dCBD] fusion protein.
  
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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This CDS part is the same as [https://parts.igem.org/Part:BBa_K3765014 OpdA-SpyTag002] excluding the OpdA signal peptide. It was generated by amplification of the rest of the CDS of the previous OpdA-SPyTag002 CDS by PCR.
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[[File:T--IISc-Bangalore--images--Map_C2(3.0).png|thumb|center|900px|Fig 1. Feature Map]]
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A tertiary structure for this fusion protein was predicted using [https://colab.research.google.com/github/deepmind/alphafold/blob/main/notebooks/AlphaFold.ipynb AlphaFold]. The OpdA module folds as in its native state, while the other stretches of amino acids exist as intrinsically disordered regions (IDRs). The SpyTag002/SpyCatcher002 binding is unaffected. Due to the absence of the considerably hydrophobic OpdA signal peptide which was predicted to exist as an intrinsically disordered region (IDR) by AlphaFold previously, the fusion protein encoded by this CDS is expected to be more stable and soluble in physiological conditions.
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[[File:T--IISc-Bangalore--images--C2(3.0)_CartoonModel.png|thumb|center|400px|Fig 2. Predicted cartoon model [N- to C- direction is blue to red]]]
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3765015 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3765015 SequenceAndFeatures</partinfo>

Latest revision as of 18:27, 20 October 2021


OpdA+22aaL+SpyTag002+10xHis

CDS of fusion protein consisting of SpyTag002 linked to OpdA (phosphotriesterase), which can covalently bind to SpyCatcher/SpyCatcher002. In our project, the protein encoded by this part is designed to get anchored to a SpyCatcher002-dCBD fusion protein.

Usage and Biology

This CDS part is the same as OpdA-SpyTag002 excluding the OpdA signal peptide. It was generated by amplification of the rest of the CDS of the previous OpdA-SPyTag002 CDS by PCR.

Fig 1. Feature Map

A tertiary structure for this fusion protein was predicted using AlphaFold. The OpdA module folds as in its native state, while the other stretches of amino acids exist as intrinsically disordered regions (IDRs). The SpyTag002/SpyCatcher002 binding is unaffected. Due to the absence of the considerably hydrophobic OpdA signal peptide which was predicted to exist as an intrinsically disordered region (IDR) by AlphaFold previously, the fusion protein encoded by this CDS is expected to be more stable and soluble in physiological conditions.

Fig 2. Predicted cartoon model [N- to C- direction is blue to red]

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1138
    Illegal AgeI site found at 1201
  • 1000
    COMPATIBLE WITH RFC[1000]