Difference between revisions of "Part:BBa K2656009"

Revision as of 22:39, 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 on each RBS tested.

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]

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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.
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