Difference between revisions of "Part:BBa K2332315"
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|'''RFC standard''' | |'''RFC standard''' | ||
− | |[https://parts.igem.org/Help:Assembly_standard_10 RFC10], [https://parts.igem.org/Help: | + | |[https://parts.igem.org/Help:Assembly_standard_10 RFC10], [https://parts.igem.org/Help:Standards/Assembly/RFC12 RFC12], [https://parts.igem.org/Help:Assembly_standard_23 RFC23] & [https://parts.igem.org/Help:Assembly_standard_25 RFC25] compatible |
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|'''Backbone''' | |'''Backbone''' | ||
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− | This coding region translates a transmembrane domain which is destined to go to the plasma membrane via the secretory pathway. While doing so it delivers any 'cytosolic domain insert' that has been cloned into this BioBrick in a plug-and-play via the KpnI-BamHI restriction sites. | + | This coding region translates a transmembrane domain which is destined to go to the plasma membrane via the secretory pathway. While doing so it delivers any 'cytosolic domain insert' that has been cloned into this BioBrick in a plug-and-play fashion (via the KpnI-BamHI restriction sites) to the plasma membrane. The delivered protein is localised to the plasma membrane but activation of the photocleavable linker PhoCl with 400 nm, violet light allows irreversible breakage of the connection with the PM and releases the 'cytosolic domain insert' into the cytoplasm. |
+ | For characterisation purposes mCherry has been chosen 'cytosolic domain insert' in this BioBrick. | ||
Latest revision as of 23:34, 22 October 2017
Plasma membrane delivery system (PMDS, mammalian)
Plasma Membrane Delivery System (PMDS) | |
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Function | Delivery of protein (e.g. mCherry) to the PM |
Use in | Mammalian cells |
Abstraction Hierarchy | Part |
RFC standard | RFC10, RFC12, RFC23 & RFC25 compatible |
Backbone | pSB1C3 |
Submitted by | [http://2017.igem.org/Team:UCL UCL iGEM 2017] |
This coding region translates a transmembrane domain which is destined to go to the plasma membrane via the secretory pathway. While doing so it delivers any 'cytosolic domain insert' that has been cloned into this BioBrick in a plug-and-play fashion (via the KpnI-BamHI restriction sites) to the plasma membrane. The delivered protein is localised to the plasma membrane but activation of the photocleavable linker PhoCl with 400 nm, violet light allows irreversible breakage of the connection with the PM and releases the 'cytosolic domain insert' into the cytoplasm.
For characterisation purposes mCherry has been chosen 'cytosolic domain insert' in this BioBrick.
Usage and Biology
As stated in the original paper:
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 2485
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 704
Illegal BsaI.rc site found at 1471
Functional Parameters
Protein data table for BioBrick BBa_ automatically created by the BioBrick-AutoAnnotator version 1.0 | ||||||||||||||||||||||||||||||||||||||||||||||
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Nucleotide sequence in RFC 10: (underlined part encodes the protein) ACCATGCGGGCC ... GTCGGATCCTAG ORF from nucleotide position 4 to 2490 (excluding stop-codon) | ||||||||||||||||||||||||||||||||||||||||||||||
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)
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Sequence features: (with their position in the amino acid sequence, see the list of supported features)
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Amino acid composition:
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Amino acid counting
| Biochemical parameters
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Plot for hydrophobicity, charge, predicted secondary structure, solvent accessability, transmembrane helices and disulfid bridges | ||||||||||||||||||||||||||||||||||||||||||||||
Codon usage
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Alignments (obtained from PredictProtein.org) There were no alignments for this protein in the data base. The BLAST search was initialized and should be ready in a few hours. | ||||||||||||||||||||||||||||||||||||||||||||||
Predictions (obtained from PredictProtein.org) | ||||||||||||||||||||||||||||||||||||||||||||||
There were no predictions for this protein in the data base. The prediction was initialized and should be ready in a few hours. | ||||||||||||||||||||||||||||||||||||||||||||||
The BioBrick-AutoAnnotator was created by TU-Munich 2013 iGEM team. For more information please see the documentation. If you have any questions, comments or suggestions, please leave us a comment. |
Reference
Zhang W, Lohman AW, Zhuravlova Y, Lu X, Wiens MD, Hoi H, Yaganoglu S, Mohr MA, Kitova EN, Klassen JS, Pantazis P, Thompson RJ, Campbell RE. Optogenetic control with a photocleavable protein, PhoCl. Nat Methods. 14(4):391-394 (2017) NCBI