Difference between revisions of "Part:BBa K3407004:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | + | Fox-1 amino acid sequence was retrieved from NCBI, and residues 109–208 corresponding to the RNA Binding Domain (RBD) were used following the Literature’s materials and methods <html><a href="#1">[1]</a></html>. As an N-HisTag from pET-28(a) was used in materials and methods, its amino acid sequence was added in the N-Terminal of Fox-1 RBD. The Fox-1 RBS sequence was codon-optimized for expression in <i>Escherichia coli</i> using the GenSmartTM Codon Optimization Tool. The forbidden restriction enzymes sites were the following: BioBrick forbidden restriction sites (EcoRI, BglII, XhoI, BamHI). They were designed to be cloned with Gibson Assembly into pBbB7a backbone. | |
− | + | ||
===Source=== | ===Source=== | ||
− | + | Plasmids used were from BglBrick repository. pBbB7a-GFP was a gift from Jay Keasling (Addgene plasmid # 35358; http://n2t.net/addgene:35358 ; RRID:Addgene_35358) <html><a href="#2">[2]</a></html>. | |
+ | Please refer to the Addgene page for more information about licences associated with the use of the plasmid. | ||
===References=== | ===References=== | ||
+ | <html> | ||
+ | |||
+ | <head> | ||
+ | |||
+ | <title>Ordered List</title> | ||
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+ | </head> | ||
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+ | <style> | ||
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+ | ol li { | ||
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+ | counter-increment: OrderedList; | ||
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+ | list-style: none; | ||
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+ | } | ||
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+ | ol li :before { | ||
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+ | content: "[" counter(OrderedList) "]"; | ||
+ | |||
+ | } | ||
+ | |||
+ | </style> | ||
+ | <html> | ||
+ | <ol> | ||
+ | |||
+ | <li> | ||
+ | <a id="1" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1356361/" target="_blank"> | ||
+ | Auweter, S., Fasan, R., Reymond, L., Underwood, J., Black, D., Pitsch, S. and Allain, F., 2020. Molecular Basis Of RNA Recognition By The Human Alternative Splicing Factor Fox-1.</a> | ||
+ | </li> | ||
+ | |||
+ | <li> | ||
+ | <a id="2" href="https://jbioleng.biomedcentral.com/articles/10.1186/1754-1611-5-12" target="_blank"> | ||
+ | BglBrick vectors and datasheets: A synthetic biology platform for gene expression. Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK, Keasling JD. J Biol Eng. 2011 Sep 20;5:12. 10.1186/1754-1611-5-12 PubMed 21933410</a> | ||
+ | </li> | ||
+ | </ol> | ||
+ | </html> |
Latest revision as of 03:07, 28 October 2020
Fox-1 RBD: a protein domain binding strongly and specifically to RNA sequence UGCAUGU.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Fox-1 amino acid sequence was retrieved from NCBI, and residues 109–208 corresponding to the RNA Binding Domain (RBD) were used following the Literature’s materials and methods [1]. As an N-HisTag from pET-28(a) was used in materials and methods, its amino acid sequence was added in the N-Terminal of Fox-1 RBD. The Fox-1 RBS sequence was codon-optimized for expression in Escherichia coli using the GenSmartTM Codon Optimization Tool. The forbidden restriction enzymes sites were the following: BioBrick forbidden restriction sites (EcoRI, BglII, XhoI, BamHI). They were designed to be cloned with Gibson Assembly into pBbB7a backbone.
Source
Plasmids used were from BglBrick repository. pBbB7a-GFP was a gift from Jay Keasling (Addgene plasmid # 35358; http://n2t.net/addgene:35358 ; RRID:Addgene_35358) [2]. Please refer to the Addgene page for more information about licences associated with the use of the plasmid.
References
- Auweter, S., Fasan, R., Reymond, L., Underwood, J., Black, D., Pitsch, S. and Allain, F., 2020. Molecular Basis Of RNA Recognition By The Human Alternative Splicing Factor Fox-1.
- BglBrick vectors and datasheets: A synthetic biology platform for gene expression. Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK, Keasling JD. J Biol Eng. 2011 Sep 20;5:12. 10.1186/1754-1611-5-12 PubMed 21933410