Part:BBa_K863104:Design
Cellulose binding Domain from C. Fimi with const. Promotor and GFP
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 37
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 37
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 37
Illegal NgoMIV site found at 65
Illegal AgeI site found at 1136 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1047
Design Notes
This part is was made by a mixture of Freiburg-assembly of BBa_K863101 and BBa_K863121 and suffix insertion into BBa_J61101. Its purpose was to analyze of the expression problem of a similar construct with a T7-promoter.
The Biobrick was designed to express the cellulose binding domain of Cellulomonas fimi exoglucanase (CBDcex) with a fused GFP as a reporter protein with a constitutive promoterBBa_J23100. 12 additional conserved bases (4 AS) were isolated upstream of the cellulose binding domain and 9 bases downstream of the exoglucanase gene. A short C-terminal (Glycine, Serine) linker and the Freiburg-scar connect the CBD to GFP. For easy capturing a His-tag was added to the C-terminal end of the GFP.
Source
The origin of this part is a cloning-vector from the fermentation group of Bielefeld University (the cellulose domain) and BBa_K863121 as template for the GFP-fusion protein.