Difference between revisions of "Part:BBa K792012"

Line 10: Line 10:
 
* '''''BamHI''''' restriction site ''(introduced to facilitate cloning into yeast expression vectors (pEG202, pCM180 series) - '''this is not essential''' for the device function)''
 
* '''''BamHI''''' restriction site ''(introduced to facilitate cloning into yeast expression vectors (pEG202, pCM180 series) - '''this is not essential''' for the device function)''
 
* '''''Kozak''''' consensus sequence for initiation of translation
 
* '''''Kozak''''' consensus sequence for initiation of translation
* '''''Signal peptide''''' that targets the product of the gene for secretion
+
* '''''Signal''''' peptide that targets the product of the gene for secretion
 
* '''''Trojan''''' peptide, to increase internalization in target cell ''(Polyarginine)''
 
* '''''Trojan''''' peptide, to increase internalization in target cell ''(Polyarginine)''
 
* '''''Payload''''': this is the exported '''Tryptophan''' rich domain of the protein ''(a custom designed peptide that we called PolyWb)''
 
* '''''Payload''''': this is the exported '''Tryptophan''' rich domain of the protein ''(a custom designed peptide that we called PolyWb)''

Revision as of 18:59, 25 September 2012

Yeast exportable Trp-rich peptide w/enhanced import (2)

This composite device produces and exports a Tryptophan rich peptide, that is import enhanced (thanks to a trojan peptide). This device is intended to be used with Yeast chassis, so it is composed by subparts designed also to be use in yeast.


The high level structure of the device is:

  • BamHI restriction site (introduced to facilitate cloning into yeast expression vectors (pEG202, pCM180 series) - this is not essential for the device function)
  • Kozak consensus sequence for initiation of translation
  • Signal peptide that targets the product of the gene for secretion
  • Trojan peptide, to increase internalization in target cell (Polyarginine)
  • Payload: this is the exported Tryptophan rich domain of the protein (a custom designed peptide that we called PolyWb)


The device's sequence has inherited HindIII restriction site from the parts it's made of.


Important note: this part has been submitted to the registry using a direct synthesis DNA sample, so it does not contain scars from BB assembly standard.
See also: Another Trp-rich export and enhanced import composite device Part:BBa_K792010.
Similar design His-rich export and enhanced import composite device Part:BBa_K792011.
The device was originally designed and built to be used in [http://2012.igem.org/Team:Buenos_Aires iGEM BsAs 2012 project] to implement a cross-feeding regulatory system between two different yeast strains with specific auxotrophy. [http://2012.igem.org/Team:Buenos_Aires Visit our wiki] to read details about the designing process and implementation details. Also feel free to contact us for any question regarding this part. Igem.bsas.png


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]