Difference between revisions of "Part:BBa K863112:Design"

(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
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The Biobrick was designed to express the cellulose binding domain of ''Clostridium Cellulovorans'' binding protein (CBDclos) under the control of a T7-promoter <partinfo>BBa_K525998</partinfo>. 6 additional conserved bases (2 AS) were isolated upstream of the cellulose binding domain and 6 bases downstream of the binding protein gene and also a short C-terminal (Glycine, Serine) linker was added. The sequence ends with the ''Age''I-restriction-site of the Freiburg-Suffix, to make in-frame ([https://parts.igem.org/Assembly_standard_25 Standard 25]) assembly possible.
 
Added the regulatory unit BBa_K525998 (T7+RBS) in front of the CBD and a short GS-Linker behind the domain and used Freiburg-Standard to be able to fuse proteins to the domain.
 
Added the regulatory unit BBa_K525998 (T7+RBS) in front of the CBD and a short GS-Linker behind the domain and used Freiburg-Standard to be able to fuse proteins to the domain.
  

Latest revision as of 20:44, 26 September 2012

Cellulose binding domain of C. cellulovorans with T7, RBS, GS-Linker (Freiburg-Standard)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 329
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The Biobrick was designed to express the cellulose binding domain of Clostridium Cellulovorans binding protein (CBDclos) under the control of a T7-promoter BBa_K525998. 6 additional conserved bases (2 AS) were isolated upstream of the cellulose binding domain and 6 bases downstream of the binding protein gene and also a short C-terminal (Glycine, Serine) linker was added. The sequence ends with the AgeI-restriction-site of the Freiburg-Suffix, to make in-frame (Standard 25) assembly possible. Added the regulatory unit BBa_K525998 (T7+RBS) in front of the CBD and a short GS-Linker behind the domain and used Freiburg-Standard to be able to fuse proteins to the domain.

Primers that were used to make this BioBrick:

CBDclos_T7RBS CCGCTTCTAGAGTAATACGACTCACTATAGGGAAAGAGGAGAAATAATGTCAGTTGAATTTTACAACTCTAAC
CBDclos_2ASlink_compl CTGCAGCGGCCGCTACTAGTATTAACCGGTGCTGCCTGCAAATCCAAATTCAACATATGTATC

Source

Cloning-vector

References