Difference between revisions of "Part:BBa K786001:Design"
Rickyleung (Talk | contribs) (→Source) |
Rickyleung (Talk | contribs) (→Design Notes) |
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Linker was refereed and modified from previous research study of SRII fusion with HtrII. | Linker was refereed and modified from previous research study of SRII fusion with HtrII. | ||
+ | Restriction sites of HindIII and BamHI were added before and after the SRII gene respectively for two reasons. | ||
+ | 1. Enable further integration of other peptides (e.g.: His-tag), for the construction of a larger fusion protein. (HindIII for N-terminal while, BamHI for C-terminal) | ||
+ | 2. Enable for switching the sensory rhodopsin portion of the fusion protein. According to previous study. [2] A series of mutant sensory rhodopsins have been identified which covers a large variation of absorbing spectrum. These two restriction sites allow further switching of the sensing unit, so the light sensing system can be tuned for sensing different kinds of light source. | ||
+ | |||
+ | The speI RE site after the promoter was kept so that the constitutive promoter can be switched to strictly controlled promoters such as PTET, tetracycline-inducible promoter; PBAD, arabinose-inducible promoter. | ||
===Source=== | ===Source=== |
Revision as of 09:27, 26 September 2012
Sensory rhodopsin II (SRII) with HtrII & Tsr, sensitive to blue-green light
- 10INCOMPATIBLE WITH RFC[10]Unknown
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 37
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 37
Illegal NgoMIV site found at 140
Illegal NgoMIV site found at 398 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1139
Illegal SapI site found at 913
Illegal SapI.rc site found at 1919
Design Notes
Pfam (version 26.0) was used to predict the domain of fusion protein. Linker was refereed and modified from previous research study of SRII fusion with HtrII.
Restriction sites of HindIII and BamHI were added before and after the SRII gene respectively for two reasons. 1. Enable further integration of other peptides (e.g.: His-tag), for the construction of a larger fusion protein. (HindIII for N-terminal while, BamHI for C-terminal) 2. Enable for switching the sensory rhodopsin portion of the fusion protein. According to previous study. [2] A series of mutant sensory rhodopsins have been identified which covers a large variation of absorbing spectrum. These two restriction sites allow further switching of the sensing unit, so the light sensing system can be tuned for sensing different kinds of light source.
The speI RE site after the promoter was kept so that the constitutive promoter can be switched to strictly controlled promoters such as PTET, tetracycline-inducible promoter; PBAD, arabinose-inducible promoter.
Source
From genomic sequence of bacterial N. pharaonis (DSM 2160) and E.coli K12.