Coding

Part:BBa_K2656022

Designed by: Adrian Requena Gutierrez, Carolina Ropero   Group: iGEM18_Valencia_UPV   (2018-09-22)


GFPmut3b Coding Sequence

domestication.
Figure 1. DNA basic parts domestication. Third construction corresponds with CDS Basic Part adaptation into the GoldenBraid grammar.

Part BBa_K2656022 is Green Fluorescent Protein coding sequence BBa_E0040 standarized into the Golden Gate assembly method. Thus, it is a coding sequence compatible with both Biobrick and GoldenBraid 3.0 assembly methods.

It can be combined with other standarized parts from our Valencia UPV IGEM 2018 Printeria Collection to assemble transcriptional units in a one-step BsaI endonuclease reaction with the Golden Gate assembly protocol .

The characterization of this protein (and by extension of all the other part that codify for the GFP) was performed with our transcriptional unit BBa_K2656105. This transcriptional unit was assembled in a Golden Braid alpha1 plasmid including the following parts:

In order to carry out a correct characterization of the protein and to be able to use it to make measurements of the different transcriptional units that we assembled with it, we have obtained the emission and excitation spectra in the conditions of our equipment. By using this [http://2018.igem.org/Team:Valencia_UPV/Experiments#spectra with the parameters of Table 1, Figure 1 was obtained.


Parameter Value
Number of samples 6
Excitation Wavelength measurement range (nm) [430-520]
Emission wavelenght (nm) 545
Emission Wavelength measurement range (nm) [490-580]
Excitation wavelenght (nm) 475
Gain (G) 70
Table 1. Parameters used to obtain the spectra


GFP spectra.
Figure 1. GFP emission and excitation spectra


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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None