Coding

Part:BBa_K2656013

Designed by: Adrián Requena Gutiérrez, Carolina Ropero   Group: iGEM18_Valencia_UPV   (2018-09-20)


sfGFP Coding Sequence

domestication.
Figure 1. DNA basic parts domestication. Third construction refers to CDS GB domestication.

Part BBa_K2656013 is the superfolder Green Fluorescent Protein coding sequence BBa_I746916 compatible with both Biobrick and Golden Braid 3.0 assembly methods. It can be combined with other compatible parts from our Valencia UPV IGEM 2018 Printeria Collection to assemble transcriptional units in a one-step BsaI reaction with the Golden Gate assembly protocol .

The characterization of this protein (and by extension of all the other part that codify for the sfGFP) was performed with our transcriptional unit BBa_K2656101. 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 have assembled with it, we have obtained the emission and excitation spectra in the conditions of our equipment. By using this protocol with the parameters of Table 1, Figure 1 has been obtained.


Table 1. Parameters used to obtain the spectra
Parameter Value
Number of samples 6
Excitation Wavelength measurement range (nm) [420-525]
Emission wavelenght (nm) 545
Emission Wavelength measurement range (nm) [470-580]
Excitation wavelenght (nm) 450
Gain (G) 50


sfGFP spectra.
Figure 2. sfGFP 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]


[edit]
Categories
//chassis/prokaryote/ecoli
//function/reporter
//function/reporter/fluorescence
//test
Parameters
None