Difference between revisions of "Part:BBa K786001:Design"

(→‎Source)
(→‎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


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 37
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal SpeI site found at 37
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 785
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 37
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 37
    Illegal NgoMIV site found at 140
    Illegal NgoMIV site found at 398
  • 1000
    INCOMPATIBLE 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.

References