Difference between revisions of "Part:BBa K2656010"
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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 on each RBS tested.
 
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 on each RBS tested.
  
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[[File:T--Valencia+UPV--optimization+exp1+RBS+graphUPV2018&ie=UTF-8&oe=UTF-8|900px|thumb|none|alt=RBS experiment 1.|Figure 1. RBS expression experiment with K2656009, K26560010 and K2656011 RBS basic parts]]
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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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Revision as of 21:18, 14 October 2018

Weak Ribosome Binding Site B0032

BBa_K2656010 is the BBa_B0032 ribosome binding site standardized into the Golden Braid assembly method. Thus, it is weak RBS 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 this part was performed with the transcriptional unit BBa_K2656102, which was used in a comparative RBS expression experiment with composite parts BBa_K2656103 and BBa_K2656101. They all were assembled in a Golden Braid alpha1 plasmid using the same promoter, coding sequence 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 on each RBS tested.

File:T--Valencia+UPV--optimization+exp1+RBS+graphUPV2018&ie=UTF-8&oe=UTF-8
Figure 1. RBS expression experiment with K2656009, K26560010 and K2656011 RBS basic parts


Table 1. Optimized parameters for the BBa_K2656010 RBS.
Parameter Value
Translation rate p p = 0.01 min-1
Dilution rate μ μ = 0.01641 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_K2656010 (GB B0030 RBS) relative strength and p ratio.
Parameter Value
Relative strength 0.045
p parameter ratio (pRBS/pref) 0.048


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]