Part:BBa_K2656101

sfGFP transcriptional unit 2

Constitutive expressed transcriptional unit assembled with a one-pot [http://2018.igem.org/Team:Valencia_UPV/Design Level 1] Golden Gate reaction using BsaI type IIS endonuclease.

This composite part of our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection#com BioArt DNA toolkit palette: Printone ], as it can be used to express a green bright color with medium intensity.

This transcriptional unit is composed of the following standardized parts from our [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Part Collection]:

• BBa_K2656004: the J23106 constitutive promoter in its Golden Braid compatible version
• BBa_K2656009: the B0030 strong ribosome biding site in its Golden Braid compatible version
• BBa_K2656013: the BBa_I746916 sfGFP sequence in its Golden Braid standardized version
• Terminator BBa_K2656026: the B0015 transcriptional terminator in its Golden Braid compatible version

This transcriptional unit has been used to characterize the RBS basic part BBa_K2656009. Following this [http://2018.igem.org/Team:Valencia_UPV/Modeling#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 the results of the next graphs have been obtained after an [http://2018.igem.org/Team:Valencia_UPV/Modeling#optimization optimization] and decision-making process, in which the optimal parameters have been selected and the constituent model for each objective has been simulated and compared with the experimental data.

Figure 1. RBS expression experiment with K2656008, K2656009 and K2656012 basic pieces

Figure 2. RBS expression experiment with K2656008 and K2656012 basic pieces

Table 1. Optimized parameters for the BBa_ K2656101.
Parameter	Value
Translation rate p	p = 0.2059 min-1
Dilution rate μ	μ = 0.01492 min-1
PoI degradation rate	RBS Experiment 2: dp = 0.00818 min-1

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 11
  Illegal NheI site found at 34
  
• 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 78