Part:BBa_K2984030

L1c-Psad-ARS-YFP-SP20-Rbcs2

This vector is a part of the Chlamy-HUB-Collection. L1 construct containg arylsulfatase secretion Signal ARS, mVenus fluorescent marker, SP20 glycomodule. These parts are flanked by the PsaD promoter and the Rbcs2 terminator. The construct can be used for secretion analysis in C.reinhardtii. The glycomodule is supposed to ramp up secretion and post translational glycosilations.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1110
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 1110
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 4
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1110
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1110
  Illegal NgoMIV site found at 1587
  
• 1000
  COMPATIBLE WITH RFC[1000]

Characterization

We were able to demonstrate the presence of YFP mVenus in C. reinhardtii transformed with this construct. C. reinhardtii shows vesicles near to the membrane area containing high concentrations of YFP mVenus, which indicates the secretion mechanism driven by the secretion signal ARS and the glycomodule SP₂₀.

Fig. 1 - Presence of YFP mVenus in C. reinhardtii. For construct BBa K2984030 it can be seen that a vesicle with a high concentration of YFP mVenus was formed near the membrane. Taken with a confocal microscope.