Difference between revisions of "Part:BBa K2656013"
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K2656013 short</partinfo>
 
<partinfo>BBa_K2656013 short</partinfo>
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[[File:T--Valencia_UPV--im11UPV2018.png|250px|thumb|centro|alt=domestication.|Figure 1. DNA basic parts domestication. Third construction refers to CDS GB domestication. ]]
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Part BBa_K2656013 is the superfolder Green Fluorescent Protein coding sequence [https://parts.igem.org/Part:BBa_I746916 BBa_I746916] compatible with both Biobrick and [http://2018.igem.org/Team:Valencia_UPV/Design Golden Braid 3.0] assembly methods.
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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 <html><a href="https://static.igem.org/mediawiki/parts/b/bc/T--Valencia_UPV--protocols.pdf#page=27"> Golden Gate assembly protocol </a></html>.
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The characterization of this protein (and by extension of all the other part that codify for the sfGFP) was performed with our transcriptional unit [https://parts.igem.org/Part:BBa_K2656101 BBa_K2656101].
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This transcriptional unit was assembled in a [http://2018.igem.org/Team:Valencia_UPV/GB3 Golden Braid alpha1 plasmid] including the following parts:
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<html>
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<ul>
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<li></html>[https://parts.igem.org/Part:BBa_K2656004 BBa_K2656004]: the [https://parts.igem.org/Part:BBa_J23106 J23106] promoter in its Golden Braid compatible version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]<html></li>
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<li></html>[https://parts.igem.org/Part:BBa_K2656009 BBa_K2656009]: the [https://parts.igem.org/Part:BBa_B0030 B0030] ribosome biding site in its Golden Braid compatible version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]<html></li>
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<li></html>[https://parts.igem.org/Part:BBa_K2656013 BBa_K2656013]: This coding sequence<html></li>
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<li></html>[https://parts.igem.org/Part:BBa_K2656026 BBa_K2656026]: the [https://parts.igem.org/Part:BBa_B0015 B0015] transcriptional terminator in its Golden Braid compatible version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]<html></li>
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</ul>
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</html>
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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.
  
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Part BBa_I746916 standarized as Phytobrick. Super Folder Green Fluorescent Protein. It can be used to assemble transcriptional units with the Golden Gate assembly method, and also, it can be used as a Biobrick.
 
  
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We have obtained the emission and excitation spectra of this protein following this protocol ('''REF''') and with the parameters of the table 1 we have obtained the results of the graphic 1:
 
 
{|class='wikitable'
 
{|class='wikitable'
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|colspan=4|Table 1. Parameters used to obtain the spectra
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|-
 
|'''Parameter'''
 
|'''Parameter'''
 
|'''Value'''
 
|'''Value'''
 
|-
 
|-
−
|Sample number
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|Number of samples
−
|0
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|6
 
|-
 
|-
−
|Minimum excitation wavelenght (nm)
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|Excitation Wavelength measurement range (nm)
−
|0
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|[420-525]
−
|-
+
−
|Maximum excitation wavelenght (nm)
+
−
|0
+
 
|-
 
|-
 
|Emission wavelenght (nm)
 
|Emission wavelenght (nm)
−
|0
+
|545
 
|-
 
|-
−
|Minimum emission wavelenght (nm)
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|Emission Wavelength measurement range (nm)
−
|0
+
|[470-580]
−
|-
+
−
|Maximum emission wavelenght (nm)
+
−
|0
+
 
|-
 
|-
 
|Excitation wavelenght (nm)
 
|Excitation wavelenght (nm)
−
|0
+
|450
 
|-
 
|-
 
|Gain (G)
 
|Gain (G)
−
|0
+
|50
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|-
 
|}
 
|}
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Table 1.
 
  
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[[File:T--Valencia_UPV--sfGFP_spectrum.png|thumb|none|alt=sfGFP spectra.|Graphic 1.]]
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[[File:T--Valencia_UPV--sfGFP_spectrum.png|900px|thumb|none|alt=sfGFP spectra.|Figure 2. sfGFP emission and excitation spectra]]
  
  

Latest revision as of 02:08, 18 October 2018


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 [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


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 14