Part:BBa_K2656013

sfGFP Coding Sequence

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 [http://2018.igem.org/Team:Valencia_UPV/Design Golden Braid 3.0] assembly methods. It can be combined with other compatible parts from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection 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 [http://2018.igem.org/Team:Valencia_UPV/GB3 Golden Braid alpha1 plasmid] including the following parts:

• BBa_K2656004: the J23106 promoter in its Golden Braid compatible version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]
• BBa_K2656009: the B0030 ribosome biding site in its Golden Braid compatible version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]
• BBa_K2656013: This coding sequence
• BBa_K2656026: the B0015 transcriptional terminator in its Golden Braid compatible version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]

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 [http://2018.igem.org/Team:Valencia_UPV/Experiments#spectra 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

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 14