Composite
CBDclos(T7

Part:BBa_K863112:Design

Designed by: Moritz Müller   Group: iGEM12_Bielefeld-Germany   (2012-09-20)

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