Difference between revisions of "Part:BBa K2656009"
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Using the [http://2018.igem.org/Team:Valencia_UPV/Protocols Golden Gate assembly protocol], it can be combined with other GB adapted basic parts, such as the ones from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Valencia UPV IGEM 2018 Printeria Collection], to assemble transcriptional units in a single one-pot reaction.  
 
Using the [http://2018.igem.org/Team:Valencia_UPV/Protocols Golden Gate assembly protocol], it can be combined with other GB adapted basic parts, such as the ones from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Valencia UPV IGEM 2018 Printeria Collection], to assemble transcriptional units in a single one-pot reaction.  
  
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Characterization of the this part was performed with the transcriptional unit [https://parts.igem.org/Part:BBa_K2656101 BBa_K2656101], which was used in a comparative RBS expression experiment with composite parts [https://parts.igem.org/Part:BBa_K2656102 BBa_K2656102 ] and [https://parts.igem.org/Part:BBa_K2656103 BBa_K2656103].  
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Characterization of the this part was performed with the transcriptional unit [https://parts.igem.org/Part:BBa_K2656101 BBa_K2656101], which was used in two comparative RBS expression experiments with composite parts [https://parts.igem.org/Part:BBa_K2656102 BBa_K2656102], [https://parts.igem.org/Part:BBa_K2656103 BBa_K2656103] and  [https://parts.igem.org/Part:],[https://parts.igem.org/Part:BBa_K2656100 BBa_K2656100], [https://parts.igem.org/Part:],[https://parts.igem.org/Part:BBa_K2656104 BBa_K2656104]
 
They all were assembled in a Golden Braid alpha1 plasmid using the same promoter, CDS and terminator:
 
They all were assembled in a Golden Braid alpha1 plasmid using the same promoter, CDS and terminator:
 
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By using this [http://2018.igem.org/Team:Valencia_UPV/Experiments#exp_protocol experimental protocol], we have obtained the parameters to valide our  [http://2018.igem.org/Team:Valencia_UPV/Modeling#models constitutive model]and rationale choose its optimized values based for each RBS.
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<img src="https://static.igem.org/mediawiki/2018/b/b1/T--Valencia_UPV--optimization_exp1_RBS_graphUPV2018.png" width="600" height="400"/>
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<img src="https://static.igem.org/mediawiki/2018/7/75/T--Valencia_UPV--optimization_exp2sfGFP_all_graphUPV2018.png" width="600" height="400"/>
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{|class='wikitable'
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|colspan=4|Table 1. Optimized parameters for the BBa_K2656009 RBS.
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|-
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|'''Parameter'''
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|'''Value'''
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|-
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|Translation rate p
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|p = 0.2059 min-1
 +
|-
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|Dilution rate μ
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| μ = 0.01492 min-1
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|}
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 +
We have also calculated the relative force between the different RBS, taking [https://parts.igem.org/Part:BBa_K2656009 BBa_K2656009 strong RBS] as a reference. Likewise, a ratio between p parameters of the different RBS parts and p parameter of the reference RBS has been calculated.
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{|class='wikitable'
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|colspan=4|Table 2. BBa_K2656008 (GB B0030 RBS) relative strength and p ratio.
 +
|-
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|'''Parameter'''
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|'''Value'''
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|-
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|Relative strength
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|1
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|-
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|p parameter ratio (pRBS/pref)
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|1
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|}
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Revision as of 22:38, 13 October 2018

Strong Ribosome Binding Site B0030

BBa_K2656009 is the BBa_B0030 RBS standardized into the Golden Braid assembly method. Thus, it is strong ribosome binding site compatible both with the BioBrick and Golden Gate grammar.

Using the [http://2018.igem.org/Team:Valencia_UPV/Protocols Golden Gate assembly protocol], it can be combined with other GB adapted basic parts, such as the ones from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Valencia UPV IGEM 2018 Printeria Collection], to assemble transcriptional units in a single one-pot reaction.

Characterization of the this part was performed with the transcriptional unit BBa_K2656101, which was used in two comparative RBS expression experiments with composite parts BBa_K2656102, BBa_K2656103 and [1],BBa_K2656100, [2],BBa_K2656104 They all were assembled in a Golden Braid alpha1 plasmid using the same promoter, CDS and terminator:

  • Promoter BBa_K2656004: the J23106 promoter in its Golden Braid compatible version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]
  • CDS BBa_K2656013: the BBa_K2656009 sfGFP sequence in its Golden Braid standardized version from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]
  • Terminator 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]
By using this [http://2018.igem.org/Team:Valencia_UPV/Experiments#exp_protocol experimental protocol], we have obtained the parameters to valide our [http://2018.igem.org/Team:Valencia_UPV/Modeling#models constitutive model]and rationale choose its optimized values based for each RBS.

Table 1. Optimized parameters for the BBa_K2656009 RBS.
Parameter Value
Translation rate p p = 0.2059 min-1
Dilution rate μ μ = 0.01492 min-1

We have also calculated the relative force between the different RBS, taking BBa_K2656009 strong RBS as a reference. Likewise, a ratio between p parameters of the different RBS parts and p parameter of the reference RBS has been calculated.

Table 2. BBa_K2656008 (GB B0030 RBS) relative strength and p ratio.
Parameter Value
Relative strength 1
p parameter ratio (pRBS/pref) 1


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]