Part:BBa_K2656106

GFP transcriptional unit 2

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 (Figure 1).

Figure 1. Assembled transcriptional unit. Golden Gate scars: standardized overhangs for promoter, RBS, CDS and terminator

It is a 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 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_K2656005: the J23102 constitutive promoter in its Golden Braid compatible version
• BBa_K2656009: the J61100 strong ribosome biding site in its Golden Braid compatible version
• BBa_K2656022: the BBa_K2656022 GFPmut3b sequence in its Golden Braid standardized version
• Terminator BBa_K2656026: the B0015 transcriptional terminator in its Golden Braid compatible version

It has been used to characterize the constitutive BBa_K2656005 promoter. 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 so the results of the next graph 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. Promoter expression experiment with composite parts BBa_K2656105, BBa_K2656106 and BBa_K2656107

Table 1. Optimized parameters for the constitutive model with TU BBa_K2656106.
Parameter	Value
Constitutive transcription rate CR	CR = 539.5 min-1
Dilution rate μ	μ = 0.0058 min-1
PoI degradation rate	Promoter Experiment: dp =

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
  COMPATIBLE WITH RFC[1000]