Part:BBa_K3781012

mCerulean, MocloMania B4

This part codes for the cyan fluorescent protein mCerulean. The protein was originally derived from the green fluorescent protein GFP which was first isolated from the bioluminescent jellyfish Aequorea victoria in 1962. It can be fused to a protein of interest, making it accessible to easy and non-invasive screening opportunities that involve fluorescence monitoring, such as fluorescence spectroscopy or microscopy.

size 26.8 kDa

function fluorescent tag

excitation wavelength 435 nm

emission wavelength 477 nm

cloning position B4

plasmid backbone pAGM1299 (BBa_K3781103)

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-translational 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
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

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

Reference Literature

R Heim, D C Prasher, R Y Tsien, "Wavelength mutations and posttranslational autoxidation of green fluorescent protein", Proceedings of the National Academy of Sciences Dec 1994, 91 (26) 12501-12504; DOI: 10.1073/pnas.91.26.12501

Mark A. Rizzo, David W. Piston, High-Contrast Imaging of Fluorescent Protein FRET by Fluorescence Polarization Microscopy, Biophysical Journal, Volume 88, Issue 2, 2005, Pages L14-L16, ISSN 0006-3495, https://doi.org/10.1529/biophysj.104.055442.