Difference between revisions of "Part:BBa K3182000"

Revision as of 14:33, 13 July 2019

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°C at 80 rpm in 1 litre of LB-miller.

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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 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. <partinfo>BBa_K1033933</partinfo>) 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 (<partinfo>BBa_I719005</partinfo>) as well as a 5'-UTR (<partinfo>BBa_K1758100</partinfo>) region which has been shown to further increase expression in E. coli (<partinfo>BBa_K1758106</partinfo>), ([http://www.ncbi.nlm.nih.gov/pubmed/2676996 Olins et al. 1989]), ([http://www.ncbi.nlm.nih.gov/pubmed/23927491 Takahashi et al. 2013]).
-[[File:T--Linkoping_Sweden--CBD-AsPink.jpeg|430px|thumb|center|<b>Figure 1.</b> E. coli BL21 cells expressing this biobrick, incubated for 16 hours at 16 degrees Celsius at 80 rpm in 1 litre of LB-miller.]]
+[[File:T--Linkoping_Sweden--CBD-AsPink.jpeg|430px|thumb|center|<b>Figure 1.</b> E. coli BL21 cells expressing this biobrick, incubated for 16 hours at 16°C at 80 rpm in 1 litre of LB-miller.]]
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