Difference between revisions of "Part:BBa K243013:Design"

(→‎Design Notes)
(→‎Design Notes)
 
Line 6: Line 6:
  
 
===Design Notes===
 
===Design Notes===
−
The choice of the linker, lipocalin tag and purification tag allows us serval different combinations.
+
We applied the His tag to enable a simultaneous purification of constructs with a Strep tag. The used FluoresceinA tag allows the measurement by quenching and the coupling to a fluorescein linked oligo. Emanating from our 3D modeling this combination of FluA tagged oligo and the construct containing the protein domain Fok_i is more efficient than the use of a combination of DigA tagged oligo with a construct containing Fok_i. To get FluA and Fok_i together and therefore bring them in proximity to the DNA, we used the short linker. The linker itself has no influence on the connected parts.  
 
[https://static.igem.org/mediawiki/parts/2/2f/Freiburg09_His-FluA-SL-Fok_i.txt Commented GenBank file]
 
[https://static.igem.org/mediawiki/parts/2/2f/Freiburg09_His-FluA-SL-Fok_i.txt Commented GenBank file]
  

Latest revision as of 02:22, 22 October 2009

His-FluA-Short 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 272
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We applied the His tag to enable a simultaneous purification of constructs with a Strep tag. The used FluoresceinA tag allows the measurement by quenching and the coupling to a fluorescein linked oligo. Emanating from our 3D modeling this combination of FluA tagged oligo and the construct containing the protein domain Fok_i is more efficient than the use of a combination of DigA tagged oligo with a construct containing Fok_i. To get FluA and Fok_i together and therefore bring them in proximity to the DNA, we used the short linker. The linker itself has no influence on the connected parts. Commented GenBank file

Source

Combined the parts by serial cloning steps.

References