Part:BBa_K4235012

T7 promoter

Usage and biology

This T7 promoter is part of the Protein S expression circuit BBa_K4235030 which was generated by cloning our insert BBa_K4235011 into the vector BBa_K4235016 using LIC protocol. T7 promoter regulatory gene circuit employs transcriptional control by a repressor protein. The lac operon circuit consists of an operator site in the promoter region, to which the lacI repressor protein can bind and establish transcriptional repression. For initiating transcription, an inducer must bind to the lac repressor protein (lacI) which prevents the repressor from binding to the operator. The T7 promoter is induced using IPTG, which mimics allolactose and binds the lac repressor protein, allowing for the T7 RNA polymerase to bind to the promoter and induce transcription of the downstream gene sequence.

Mathematical modeling

We used MATLAB simulations to model our genetic circuits. We used a system of ODEs derived for both constitutive and regulatory genetic circuits(IPTG inducible lac operon) for our E coli expression system. We were able to input our system of ODE’s into MATLAB to predict our steady state concentrations prior to starting wet lab in order to determine the process that would yield more favorable results. Those plots can be found below:

E coli constitutive gene circuit model:

(1.) Protein vs time:

Figure 1:Plot showing Protein S production over time

(2.) mRNA vs time:

Figure 2: Plot showing mRNA production over time

(3.) Effects of altering DNA concentration on protein production:

Figure 3: Plot showing the effects of altering initial DNA concentrations on protein production

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]