Part:BBa_K3781003

mVenus, MoCloMania B3_B4

This part codes for the yellow fluorescent protein with the poetic name mVenus. It is a deviation of the famous green fluorescent protein GFP originally isolated from the bioluminescent jellyfish Aequorea victoria in 1962. By coupling a protein of interest with a fluorescent tag such as mVenus, the target protein can be easily and non-invasively detected within living cells as well as cell preparations with the help of fluorescence spectroscopy and microscopy.

size 26.9 kDa

function fluorescent tag

excitation wavelength 515 nm

emission wavelength 537 nm

cloning position B3_B4

plasmid backbone pAGM1287 (BBa_K3781101)

The MocloMania collection

This basic part is part of the MocloMania collection, a collection of genetic parts specifically designed and optimized for Modular Cloning assembly and recombinant protein expression in the protozoan parasite Leishmania tarentolae.

Are you trying to express complexly glycosylated proteins? Large antibody side chains? Human proteins that require accurate post-translation modification? Then Leishmania might be just the right organism for you! Leishmania tarentolae’s glycosylation patterns resemble those of human cells more closely than any other microbial expression host, while still delivering all the benefits of microbial production systems like easy transfection and cultivation. So instead of relying on mammalian cell lines, try considering Leishmania as your new expression host of choice!

Our MocloMania collection will allow you to easily modify your protein of choice to make it suitable for downstream detection and purification procedures. We furthermore provide a specifically domesticated Leishmania expression vector, named weird_plex, which will package your desired fusion protein into a functional transcriptional unit that is optimized for high expression in Leishmania.

This part collection is the very first one to establish the Modular Cloning system in Leishmania. Finally! Employing Modular Cloning for your cloning operations will safe you a lot of time and frustration in the long run. Because MoClo utilizes type IIS restriction enzymes and because every basic part is equipped with a specified set of overhangs that assign it to its desired position within the transcriptional unit, restriction and ligation of your construct of choice can all happen simultaneously in a simple one-step, one-pot reaction.

Do we have your attention? Feel free to check out our wiki to find more information on Leishmania and Modular Cloning as well as to understand how this basic part integrates into our part collection. See you there!

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 202
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 202
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 202
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 202
  Illegal NgoMIV site found at 679
  
• 1000
  COMPATIBLE WITH RFC[1000]

Reference Literature

Cyan and Yellow Super Fluorescent Proteins with Improved Brightness, Protein Folding, and FRET Förster Radius, Gert-Jan Kremers, Joachim Goedhart, Erik B. van Munster, and Theodorus W. J. Gadella Biochemistry 2006 45 (21), 6570-6580 DOI: 10.1021/bi0516273

M.A. Rizzo, D.W. Piston High-contrast imaging of fluorescent protein FRET by fluorescence polarization microscopy Biophys. J., 88 (2005), pp. L14-L16