Difference between revisions of "Part:BBa K243015:Design"

(→‎Design Notes)
(→‎Design Notes)
 
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===Design Notes===
 
===Design Notes===
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The choice of the linker, lipocalin tag and purification tag allows us several different combinations.
+
We applied the Streptavidine tag to enable a simultaneous purification of constructs with a His tag. Strep tag also shows a higher affinity towards Strep-Tactin than His tag. For that the purification with Strep-tag is more specific. The used DigA tag allows the coupling to an Digoxigenin linked oligo. Emanating from our 3D modeling this combination of DigA tagged oligo and the construct containing the protein domain Fok_i is not as efficient as the use of a combination of FluA tagged oligo with a construct containing Fok_i. To avoid interactions between the DigA tag with the connected protein domain Fok_i we applied the Split Linker. This linker is an improved part of the team Freiburg08 and a suitable linker for fusion proteins.  
 
[https://static.igem.org/mediawiki/parts/2/22/Freiburg09_strep-Dig-Split-Fok_as.txt Commented GenBank file]
 
[https://static.igem.org/mediawiki/parts/2/22/Freiburg09_strep-Dig-Split-Fok_as.txt Commented GenBank file]
  

Latest revision as of 02:16, 22 October 2009

Strep-DigA-Split Linker-Fok_i


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 278
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We applied the Streptavidine tag to enable a simultaneous purification of constructs with a His tag. Strep tag also shows a higher affinity towards Strep-Tactin than His tag. For that the purification with Strep-tag is more specific. The used DigA tag allows the coupling to an Digoxigenin linked oligo. Emanating from our 3D modeling this combination of DigA tagged oligo and the construct containing the protein domain Fok_i is not as efficient as the use of a combination of FluA tagged oligo with a construct containing Fok_i. To avoid interactions between the DigA tag with the connected protein domain Fok_i we applied the Split Linker. This linker is an improved part of the team Freiburg08 and a suitable linker for fusion proteins. Commented GenBank file

Source

Combined the parts by serial cloning steps.

References