Composite

Part:BBa_K3182000

Designed by: Oliver Hild Walett   Group: iGEM19_Linkoping_Sweden   (2019-07-08)
Revision as of 05:26, 12 July 2019 by Olle111 (Talk | contribs)


pT7-CBDcipA-AsPink

A cellulose binding domain (CBD) from Clostridium thermocellum which can be used to purify or attach proteins to cellulose. The part also has a flexible GS-linker with a thrombin site added at the end, clevage with thrombin will add one glycine and one serine to the N-terminal of the fusion protein. The part has a AsPink (Codon opt. BBa_K1033933) fused to the CBD to ensure easy trackable expression and characterization ([http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5946454 Liljeruhm et al. 2018]) of the CBD. The part has a very strong expression with a T7 promotor (BBa_I719005) as well as a 5'UTR (BBa_K1758100) region which has been shown to further increase expression in E. coli (BBa_K1758106), ([http://www.ncbi.nlm.nih.gov/pubmed/2676996 Olins et al. 1989]), ([http://www.ncbi.nlm.nih.gov/pubmed/23927491 Takahashi et al. 2013]).

Figure 1. E. coli BL21 cells expressing this biobrick, incubated for 16 hours at 16 degrees Celsius.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 580
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None