Difference between revisions of "Part:BBa K2332051"

Revision as of 23:29, 22 October 2017

Intimin'-SpyCatcher (constitutive)

This biobrick is the fusion of a truncated version of Intimin (159kDa) and SpyCatcher (138 amino acids, 15 kDa) for the cell surface display of SpyCatcher. To trigger cell adhesion, we decided to make use of the SpyTag and SpyCatcher, two binding partners that form rapid covalent bonds, fused to outer membrane surface proteins. SpyTag (13 amino acids) and SpyCatcher (138 amino acids, 15 kDa) come from CnaB2 (immunoglobulin-llike collagen adhesin domain) of the FbaB protein, found in the invasive strains of S. pyogenes. We decided to fuse SpyTag and SpyCatcher to outer membrane surface proteins in distinct cell populations that would then aggregate once mixed together. Intimin'-SpyCatcher and Intimin'-SpyTag will be placed under the control of our blue light inducible promoter Pblind to guide cell structure formation using blue light along with our SpyTag variants.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 880
Illegal NgoMIV site found at 1621
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K2332051 short</partinfo>
-This biobrick is for the constitutive expression of Intimin'-SpyCatcher as part of or bacterial cell aggregation system.  This biobrick is the fusion of a truncated version of Intimin and SpyCatcher for the cell surface display of SpyCatcher under the control of of a constitutive promoter. When cells expressing this construct are mixed with cells expressing Intimin'-SpyTag aggregation is expected to occur.
+This biobrick is the fusion of a truncated version of Intimin (159kDa) and SpyCatcher (138 amino acids, 15 kDa) for the cell surface display of SpyCatcher. To trigger cell adhesion, we decided to make use of the SpyTag and SpyCatcher, two binding partners that form rapid covalent bonds, fused to outer membrane surface proteins. SpyTag (13 amino acids) and SpyCatcher (138 amino acids, 15 kDa) come from CnaB2 (immunoglobulin-llike collagen adhesin domain) of the FbaB protein, found in the invasive strains of S. pyogenes. We decided to fuse SpyTag and SpyCatcher to outer membrane surface proteins in distinct cell populations that would then aggregate once mixed together. Intimin'-SpyCatcher and [https://parts.igem.org/Part:BBa_K2332010 Intimin'-SpyTag] will be placed under the control of our blue light inducible promoter [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2332002 Pblind] to guide cell structure formation using blue light along with our SpyTag variants.
-SpyTag (13 amino acids) and SpyCatcher (138 amino acids, 15 kDa) are binding partners that originate from CnaB2 (immunoglobulin-llike collagen adhesin domain) of the FbaB protein, found in the invasive strains of S. pyogenes.
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