Motivating example

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BNGL Version

# Signal Transduction with receptor internalization and transcriptional reg.  #
# vanilla BNGL code: justin.s.hogg@gmail.com                                  #
# conception: Leonard A. Harris, Justin S. Hogg, James R. Faeder              #
# 6 June 2009                                                                 #
#                                                                             #
# Vanilla version requires +15 rules (and context) compared to cBNGL.         #
# A motivating example for Winter Simulation Conference 2009 invited paper.   #

begin model
begin parameters
  nEndo           5      # mean number of endosomes

  vol_EC        	20.0   # volumes
  vol_CP          4.0
  vol_NU          1.0
  vol_EN          0.1*nEndo

  sa_PM           0.4    # membrane surface areas
  sa_NM           0.1
  sa_EM           0.01*nEndo
 
  eff_width       1.0    # effective surface width

  L0              1000   # initial species counts (extensive units: quantity, not concentration)
  R0              200     
  TF0             200
  DNA0            2
  Im0             40
  NP0             4

  kp_LR            0.1    # kinetic parameters (2nd order reaction params in vol/time units)
  km_LR           1.0    
  kp_LL           0.1
  km_LL           1.0
  k_R_endo        1.0
  k_recycle       0.1
  k_R_transphos   1.0
  k_R_dephos      0.1
  kp_R_TF         0.1
  km_R_TF         0.1
  kp_R_TFp        0.1
  km_R_TFp     	 10.0
  k_TF_transphos  1.0
  k_TF_dephos     1.0
  kp_TF_TF        0.1
  km_TF_TF        1.0
  kp_TF_p1        0.1
  km_TF_p1        1.0
  k_transcribe    1.0
  k_translate     1.0
  k_mRNA_to_CP    1.0	 # volume to volume species transport.
  k_mRNA_deg      1.0
  k_P_deg         0.1
  k_Im_bind_CP    0.1
  k_Im_unbind_CP  0.1
  k_Im_bind_NU    0.1
  k_Im_unbind_NU  10.0
  k_Im_enters_NP  0.1
  k_Im_exits_NP   1.0
  k_Im_cross_NP   1.0
  kp_P1_p2        0.1
  km_P1_p2        1.0
  
  eVol_PM         sa_PM * eff_width		# effective surface volumes
  eVol_NM         sa_NM * eff_width
  eVol_EM         sa_EM * eff_width
end parameters

begin molecule types
  L(r,d,loc~EC~EN)               # Ligand w/ receptor binding and dimer sites.
  R(l,tf~Y~pY,loc~PM~EM)         # Receptor with ligand and TF binding sites.
  TF(r,d~Y~pY,dna,im,loc~CP~NU)  # Transcription factor (monomer) w/ receptor, DNA, and importin binding sites; and dimerization domain.
  DNA(p1,p2)                     # DNA molecule with two promoter sites. 
  mRNA1(loc~CP~NU)               # mRNA transcript for Protein 1.
  mRNA2(loc~CP~NU)               # mRNA transcript for Protein 2.
  P1(im,dna,loc~CP~NU)           # Protein 1 w/ importin and DNA binding site.
  P2()                           # Protein 2.
  Im(fg,cargo,loc~CP~NU)         # nuclear importin mol. w/ hydrophobic domain (fg) that interacts with nuclear pore. 
  NP(fg)                         # nuc. pore complex w/ hydrophobic FG repeat.
  Sink()                         # a place for deleted molecules.  
end molecule types

begin species
  L(r,d,loc~EC)            L0
  R(l,tf~Y,loc~PM)         R0
  TF(r,d~Y,dna,im,loc~CP)  TF0
  DNA(p1,p2)               DNA0
  Im(fg,cargo,loc~CP)      Im0
  NP(fg)                   NP0
  Sink()                   0
end species

begin reaction rules
  # ligand dimerization (1 -> 6 rules)
  #
  # @EC: neither ligand bound to PM through Rec
  Rule1_1:  L(d,r,loc~EC) + L(d,r,loc~EC)  \
    		<->  L(d!1,r,loc~EC).L(d!1,r,loc~EC)	   kp_LL/vol_EC,  km_LL

  # @EN: neither ligand bound to EM through Rec
  Rule1_2:  L(d,r,loc~EN) + L(d,r,loc~EN)  \
      	<->  L(d!1,r,loc~EN).L(d!1,r,loc~EN)      kp_LL/vol_EN,  km_LL  

  # @EC: one ligand bound to PM through Rec
  Rule1_3:  L(d,r!+,loc~EC) + L(d,r,loc~EC)  \
    		<->  L(d!1,r!+,loc~EC).L(d!1,r,loc~EC)    kp_LL/vol_EC,  km_LL

  # @EN: one ligand bound to EM through Rec
  Rule1_4:  L(d,r!+,loc~EN) + L(d,r,loc~EN)  \
    		<->  L(d!1,r!+,loc~EN).L(d!1,r,loc~EN)    kp_LL/vol_EN,  km_LL

  # @PM: both ligands bound to PM through Rec  
  Rule1_5:  L(d,r!+,loc~EC) + L(d,r!+,loc~EC)  \
    		<->  L(d!1,r!+,loc~EC).L(d!1,r!+,loc~EC)  kp_LL/eVol_PM, km_LL

  # @EM: both ligands bound to EM through Rec
  Rule1_6:  L(d,r!+,loc~EN) + L(d,r!+,loc~EN)  \
    		<->  L(d!1,r!+,loc~EN).L(d!1,r!+,loc~EN)  kp_LL/eVol_EM, km_LL

  # receptor-ligand binding
  Rule2_1:  L(r,loc~EC) + R(l,loc~PM)  <->  L(r!1,loc~EC).R(l!1,loc~PM)	kp_LR/vol_EC,  km_LR  exclude_reactants(1,R)
  Rule2_2:  L(r,loc~EN) + R(l,loc~EM)  <->  L(r!1,loc~EN).R(l!1,loc~EM) kp_LR/vol_EN,  km_LR  exclude_reactants(1,R)
  Rule2_3:  L(r,loc~EC) + R(l,loc~PM)  <->  L(r!1,loc~EC).R(l!1,loc~PM)	kp_LR/eVol_PM, km_LR  include_reactants(1,R)
  Rule2_4:  L(r,loc~EN) + R(l,loc~EM)  <->  L(r!1,loc~EN).R(l!1,loc~EM) kp_LR/eVol_EM, km_LR  include_reactants(1,R)

  # receptor-dimer internalization
  Rule3:  R(loc~PM).L(loc~EC).L(loc~EC).R(loc~PM) -> R(loc~EM).L(loc~EN).L(loc~EN).R(loc~EM)  k_R_endo 

  # receptor,ligand recycling
  Rule4_1:  R(l,loc~EM)  ->  R(l,loc~PM)  						  k_recycle
  Rule4_2:  R(l!1,loc~EM).L(r!1,d,loc~EN) ->  R(l!1,loc~PM).L(r!1,d,loc~EC)  	  k_recycle
  Rule4_3:  R(l!1,loc~EM).L(r!1,d!2,loc~EN).L(r,d!2,loc~EN) -> R(l!1,loc~PM).L(r!1,d!2,loc~EC).L(r,d!2,loc~EC)  k_recycle
  Rule4_4:  R(l!1,loc~EM).L(r!1,d!2,loc~EN).L(r!3,d!2,loc~EN).R(l!3,loc~EM)  \
        	-> R(l!1,loc~PM).L(r!1,d!2,loc~EC).L(r!3,d!2,loc~EC).R(l!3,loc~PM)  k_recycle   

  Rule5_1:  L(r,d,loc~EN)  ->  L(r,d,loc~EC)                   			  k_recycle
  Rule5_2:  L(r,d!1,loc~EN).L(r,d!1,loc~EN) -> L(r,d!1,loc~EC).L(r,d!1,loc~EC)  k_recycle

  # receptor transphosphorylation
  # Rule6 (1 -> 1 rule)
  R.R(tf~Y) -> R.R(tf~pY)          k_R_transphos

  # receptor dephosphorylation
  Rule7:  R(tf~pY) -> R(tf~Y)      k_R_dephos
 
  # receptor--transcriptionFactor binding
  Rule8:  R(tf~pY) + TF(d~Y,r,loc~CP) <-> R(tf~pY!1).TF(d~Y,r!1,loc~CP)    kp_R_TF/vol_CP, km_R_TF

  # Rule9 (1 -> 1 rule)
  Rule9:  R(tf~pY) + TF(d~pY,r,loc~CP) <-> R(tf~pY!1).TF(d~pY,r!1,loc~CP)  kp_R_TFp/vol_CP, km_R_TFp

  # transcription factor trans-phosphorylation
  Rule10:  TF.R.R.TF(d~Y) -> TF.R.R.TF(d~pY)   k_TF_transphos

  # transcription factor dephosphorylation
  Rule11:  TF(d~pY,loc~CP) -> TF(d~Y,loc~CP)   k_TF_dephos

  # transcription factor dimerization
  Rule12_1:  TF(r,d~pY,dna,loc~CP) + TF(r,d~pY,dna,loc~CP)  \
        	 <->  TF(r,d~pY!1,dna,loc~CP).TF(r,d~pY!1,dna,loc~CP)    kp_TF_TF/vol_CP, km_TF_TF

  Rule12_2:  TF(r,d~pY,dna,loc~NU) + TF(r,d~pY,dna,loc~NU)  \
        	 <->  TF(r,d~pY!1,dna,loc~NU).TF(r,d~pY!1,dna,loc~NU)    kp_TF_TF/vol_NU, km_TF_TF

  # TF dimer binds promoters
  Rule13:  TF(dna,im,loc~NU).TF(dna,im,loc~NU) + DNA(p1)  \
           <->  TF(dna!1,im,loc~NU).TF(dna!2,im,loc~NU).DNA(p1!1!2)  kp_TF_p1/vol_NU, km_TF_p1

  # transcription
  Rule14:  DNA(p1!+)  ->  DNA(p1!+) + mRNA1(loc~NU)            k_transcribe
  Rule15:  DNA(p2!+)  ->  DNA(p2!+) + mRNA2(loc~NU)            k_transcribe

  # mRNA transport to cytoplams
  Rule16:  mRNA1(loc~NU)  ->  mRNA1(loc~CP)                    k_mRNA_to_CP
  Rule17:  mRNA2(loc~NU)  ->  mRNA2(loc~CP)                    k_mRNA_to_CP

  # mRNA translation to protein
  Rule18:  mRNA1(loc~CP)  ->  mRNA1(loc~CP) + P1(im,dna,loc~CP)    k_translate
  Rule19:  mRNA2(loc~CP)  ->  mRNA2(loc~CP) + P2()                 k_translate

  # mRNA degradation (2 -> 2 rules)
  Rule20:  mRNA1  ->  Sink()                        k_mRNA_deg  DeleteMolecules
  Rule21:  mRNA2  ->  Sink()                        k_mRNA_deg  DeleteMolecules

  # Protein degradation (2 -> 2 rules)
  Rule22:  P1     ->  Sink()                        k_P_deg  DeleteMolecules
  Rule23:  P2     ->  Sink()                        k_P_deg  DeleteMolecules

  # importin binds TF dimer (tends to pick up in CP, drop off in NU).
  Rule24:  TF(im,dna,r,loc~CP).TF(im,dna,r,loc~CP) + Im(cargo,loc~CP)  \
      	<-> TF(im!1,dna,r,loc~CP).TF(im!2,dna,r,loc~CP).Im(cargo!1!2,loc~CP)  k_Im_bind_CP/vol_CP, k_Im_unbind_CP

  Rule25:  TF(im,dna,r,loc~NU).TF(im,dna,r,loc~NU) + Im(cargo,loc~NU)  \
      	<-> TF(im!1,dna,r,loc~NU).TF(im!2,dna,r,loc~NU).Im(cargo!1!2,loc~NU)  k_Im_bind_NU/vol_NU, k_Im_unbind_NU

  # importin binds P1 (tends to pick up in CP, drop off in NU).
  Rule26:  P1(im,dna,loc~CP) + Im(cargo,loc~CP) <-> P1(im!1,dna,loc~CP).Im(cargo!1,loc~CP)  k_Im_bind_CP/vol_CP, k_Im_unbind_CP
  Rule27:  P1(im,dna,loc~NU) + Im(cargo,loc~NU) <-> P1(im!1,dna,loc~NU).Im(cargo!1,loc~NU)  k_Im_bind_NU/vol_NU, k_Im_unbind_NU

  # importin enters nuclear pore
  Rule28_1:  Im(fg,loc~CP) + NP(fg) <-> Im(fg!1,loc~CP).NP(fg!1)  k_Im_enters_NP/vol_CP, k_Im_exits_NP
  Rule28_2:  Im(fg,loc~NU) + NP(fg) <-> Im(fg!1,loc~NU).NP(fg!1)  k_Im_enters_NP/vol_NU, k_Im_exits_NP

  # importin traverses nuclear pore (with any cargo)
  Rule29_1:  Im(cargo,loc~CP).NP <-> Im(cargo,loc~NU).NP  k_Im_cross_NP, k_Im_cross_NP
  Rule29_2:  TF(loc~CP).TF(loc~CP).Im(loc~CP).NP <-> TF(loc~NU).TF(loc~NU).Im(loc~NU).NP  k_Im_cross_NP, k_Im_cross_NP
  Rule29_3:  P1(loc~CP).Im(loc~CP).NP <-> P1(loc~NU).Im(loc~NU).NP  k_Im_cross_NP, k_Im_cross_NP

  # P1 binds promoter 2
  Rule30:  P1(im,dna,loc~NU) + DNA(p2) <-> P1(im,dna!1,loc~NU).DNA(p2!1)  kp_P1_p2/vol_NU, km_P1_p2  
end reaction rules  

begin observables
  Molecules  Tot_L         L
  Molecules  Tot_R         R
  Molecules  Tot_TF        TF
  Molecules  Tot_DNA       DNA
  Molecules  Tot_mRNA1     mRNA1
  Molecules  Tot_mRNA2     mRNA2
  Molecules  Tot_P1        P1
  Molecules  Tot_P2        P2
  Molecules  Tot_NP        NP
  Molecules  Tot_Im        Im

  Species    L_Dimers_EC   L(r,loc~EC).L(r,loc~EC)
  Species    L_Dimers_PM   L.L.R(loc~PM)
  Species    L_Dimers_EN   L(r,loc~EN).L(r,loc~EN)
  Species    L_Dimers_EM   L.L.R(loc~EM)
  
  Species    L_Bound_PM    L(d).R(loc~PM), L.L.R(loc~PM), L.L.R(loc~PM)
  Species    L_Bound_EM    L(d).R(loc~EM), L.L.R(loc~EM), L.L.R(loc~EM)

  Species    R_Dimers_PM   R(loc~PM).R(loc~PM)
  Species    R_Dimers_EM   R(loc~EM).R(loc~EM)

  Molecules  Catalytic_R   R(tf~pY!?)
  Molecules  Catalytic_TF  R(tf~pY!1).TF(r!1)
  Molecules  Phos_TF       TF(d~pY!?)

  Species    TF_Dimer_CP   TF(d~pY!1,loc~CP).TF(d~pY!1,loc~CP)
  Species    TF_Dimer_NU   TF(d~pY!1,loc~NU).TF(d~pY!1,loc~NU)

  Species    Bound_prom1   DNA(p1!+)
  Species    Bound_prom2   DNA(p2!+)

  Species    P1_NU         P1(loc~NU)
  Species    P1_CP         P1(loc~CP)

  Species    Im_NU         Im(loc~NU)
  Species    Im_CP         Im(loc~CP)
  
  Species    Im_Cargo_NP   Im(fg!+,cargo!+)

  Species    P1_NU_free    P1(im,dna,loc~NU)
  Species    P1_NU_dna     P1(im,dna!+,loc~NU)
end observables
end model

# actions #
generate_network({overwrite=>1,TextReaction=>1});
simulate_ode({suffix=>'ode',t_start=>0,t_end=>200,n_steps=>100});

cBNGL Version

# Signal transduction with receptor internalization and transcriptional reg.  #
#  cBNGL code: justin.s.hogg@gmil.com                                         #
# conception: Leonard A. Harris, Justin S. Hogg, James R. Faeder              #
# 6 June 2009                                                                 #
#                                                                             #
# Demonstrates features of cBNGL in a biologically relevant scenario.         #
# A motivating example for Winter Simulation Conference 2009 invited paper.   #

begin model
begin parameters
  nEndo           5      # mean number of endosomes

  vol_EC   		   20.0   # volumes
  vol_CP          4.0
  vol_NU          1.0
  vol_EN          0.1*nEndo

  sa_PM           0.4    # membrane surface areas
  sa_NM           0.1
  sa_EM           0.01*nEndo

  eff_width       1.0    # effective surface width

  L0              1000   # initial species counts (extensive units: quantity, not concentration)
  R0              200     
  TF0             200
  DNA0            2
  Im0             40
  NP0             4

  kp_LR           0.1    # kinetic parameters (2nd order reaction params in vol/time units)
  km_LR           1.0    
  kp_LL           0.1
  km_LL           1.0
  k_R_endo        1.0
  k_recycle       0.1
  k_R_transphos   1.0
  k_R_dephos      0.1
  kp_R_TF         0.1
  km_R_TF         0.1
  kp_R_TFp        0.1
  km_R_TFp       10.0
  k_TF_transphos  1.0
  k_TF_dephos     1.0
  kp_TF_TF        0.1
  km_TF_TF        1.0
  kp_TF_p1        0.1
  km_TF_p1        1.0
  k_transcribe    1.0
  k_translate     1.0
  k_mRNA_to_CP    1.0
  k_mRNA_deg      1.0
  k_P_deg         0.1
  k_Im_bind_CP    0.1
  k_Im_unbind_CP  0.1
  k_Im_bind_NU    0.1
  k_Im_unbind_NU  10.0
  k_Im_enters_NP  0.1
  k_Im_exits_NP   1.0
  k_Im_cross_NP   1.0
  kp_P1_p2        0.1
  km_P1_p2        1.0
end parameters

begin compartments
  EC  3  vol_EC 
  PM  2  sa_PM * eff_width          EC
  CP  3  vol_CP                     PM
  NM  2  sa_NM * eff_width          CP
  NU  3  vol_NU                     NM
  EM  2  sa_EM * eff_width          CP
  EN  3  vol_EN                     EM
end compartments

begin molecule types
  L(r,d)               # Ligand w/ receptor binding and dimerization sites.
  R(l,tf~Y~pY)         # Receptor with ligand and TF binding sites.
  TF(r,d~Y~pY,dna,im)  # Transcription factor (monomer) with receptor, DNA, and importin binding sites; and dimerization domain.
  DNA(p1,p2)           # DNA molecule with two promoter sites. 
  mRNA1()              # mRNA transcript for Protein 1.
  mRNA2()              # mRNA transcript for Protein 2.
  P1(im,dna)           # Protein 1 with importin and DNA binding domains.
  P2()                 # Protein 2.
  Im(fg,cargo)         # nuclear importin molecule with hydrophobic domain (fg) that interacts with nuclear pore. 
  NP(fg)               # nuclear pore complex w/ hydrophobic FG repeat domain.
  Sink()               # a place for deleted molecules.
end molecule types

begin species
  L(r,d)@EC            L0
  R(l,tf~Y)@PM         R0
  TF(r,d~Y,dna,im)@CP  TF0
  DNA(p1,p2)@NU        DNA0
  Im(fg,cargo)@CP      Im0
  NP(fg)@NM            NP0
  
  # Arbitrarily assign compartment CP to the abstract molecule "Sink".  
  Sink()@CP            0
end species

begin reaction rules
  # receptor-ligand binding.
  Rule1:  L(r) + R(l)  <->  L(r!1).R(l!1)            kp_LR, km_LR

  # ligand dimerization.
  Rule2:  L(d) + L(d)  <->  L(d!1).L(d!1)            kp_LL, km_LL  

  # Rule3: receptor-dimer internalization.
  Rule3:  @PM:R.R       ->  @EM:R.R                  k_R_endo

  # receptor, ligand recycling.
  Rule4:  @EM:R  ->  @PM:R                           k_recycle
  Rule5:  @EN:L  ->  @EC:L                           k_recycle

  # receptor transphosphorylation.
  Rule6:  R.R(tf~Y)  ->  R.R(tf~pY)                  k_R_transphos

  # receptor dephosphorylation.
  Rule7:  R(tf~pY)   ->  R(tf~Y)                     k_R_dephos

  # receptor-TF binding.  favor binding if TF(dim~Y), unbinding if TF(dim~pY).
  Rule8:  R(tf~pY) + TF(d~Y,r)   <->  R(tf~pY!1).TF(d~Y,r!1)	   kp_R_TF,  km_R_TF
  Rule9:  R(tf~pY) + TF(d~pY,r)  <->  R(tf~pY!1).TF(d~pY,r!1)     kp_R_TFp, km_R_TFp

  # transcription factor trans-phosphorylation.
  Rule10:  TF.R.R.TF(d~Y)   ->  TF.R.R.TF(d~pY)      k_TF_transphos

  # transcription factor dephosphorylation (CP only).
  Rule11:  TF(d~pY)@CP      ->  TF(d~Y)@CP           k_TF_dephos

  # transcription factor dimerization.
  Rule12:  TF(r,d~pY,dna) + TF(r,d~pY,dna) <-> TF(r,d~pY!1,dna).TF(r,d~pY!1,dna)  	  kp_TF_TF, km_TF_TF

  # TF dimer binds promoter 1.
  Rule13:  TF(dna,im).TF(dna,im) + DNA(p1) <-> TF(dna!1,im).TF(dna!2,im).DNA(p1!1!2)  kp_TF_p1, km_TF_p1

  # transcription.
  Rule14:  DNA(p1!+) ->  DNA(p1!+) + mRNA1()@NU      k_transcribe
  Rule15:  DNA(p2!+) ->  DNA(p2!+) + mRNA2()@NU      k_transcribe

  # mRNA transport to cytoplams.
  Rule16:  mRNA1@NU  ->  mRNA1@CP                    k_mRNA_to_CP
  Rule17:  mRNA2@NU  ->  mRNA2@CP                    k_mRNA_to_CP

  # mRNA translation to protein.
  Rule18:  mRNA1@CP  ->  mRNA1@CP  +  P1(im,dna)@CP  k_translate
  Rule19:  mRNA2@CP  ->  mRNA2@CP  +  P2()@CP        k_translate

  # mRNA degradation.
  Rule20:  mRNA1     ->  Sink()@CP                  k_mRNA_deg  DeleteMolecules
  Rule21:  mRNA2     ->  Sink()@CP                  k_mRNA_deg  DeleteMolecules

  # protein degradation.
  Rule22:  P1        ->  Sink()@CP                  k_P_deg  DeleteMolecules
  Rule23:  P2        ->  Sink()@CP                  k_P_deg  DeleteMolecules

  # importin binds TF dimer (tends to pick up in CP, drop off in NU).
  Rule24:  TF(im,dna,r).TF(im,dna,r) + Im(cargo)@CP <-> TF(im!1,dna,r).TF(im!2,dna,r).Im(cargo!1!2)@CP	k_Im_bind_CP, k_Im_unbind_CP
  Rule25:  TF(im,dna,r).TF(im,dna,r) + Im(cargo)@NU <-> TF(im!1,dna,r).TF(im!2,dna,r).Im(cargo!1!2)@NU	k_Im_bind_NU, k_Im_unbind_NU

  # importin binds P1 (tends to pick up in CP, drop off in NU).
  Rule26:  P1(im,dna) + Im(cargo)@CP    <->  P1(im!1,dna).Im(cargo!1)@CP	   k_Im_bind_CP, k_Im_unbind_CP
  Rule27:  P1(im,dna) + Im(cargo)@NU    <->  P1(im!1,dna).Im(cargo!1)@NU      k_Im_bind_NU, k_Im_unbind_NU

  # importin enters nuclear pore.
  Rule28:  Im(fg) + NP(fg)  <->  Im(fg!1).NP(fg!1)            k_Im_enters_NP, k_Im_exits_NP

  # importin traverses nuclear pore (with any cargo).
  Rule29:  Im(fg!1)@CP.NP(fg!1)  <->  Im(fg!1)@NU.NP(fg!1)    k_Im_cross_NP, k_Im_cross_NP  MoveConnected

  # P1 binds promoter 2.
  Rule30:  P1(im,dna) + DNA(p2)  <->  P1(im,dna!1).DNA(p2!1)  kp_P1_p2, km_P1_p2  
end reaction rules  

begin observables
  Molecules  Tot_L         L
  Molecules  Tot_R         R
  Molecules  Tot_TF        TF
  Molecules  Tot_DNA       DNA
  Molecules  Tot_mRNA1     mRNA1
  Molecules  Tot_mRNA2     mRNA2
  Molecules  Tot_P1        P1
  Molecules  Tot_P2        P2
  Molecules  Tot_NP        NP
  Molecules  Tot_Im        Im

  Species    L_Dimers_EC   @EC:L.L
  Species    L_Dimers_PM   @PM:L.L
  Species    L_Dimers_EN   @EN:L.L
  Species    L_Dimers_EM   @EM:L.L

  Molecules  L_Bound_PM    @PM:L
  Molecules  L_Bound_EM    @EM:L

  Species    R_Dimers_PM   @PM:R.R
  Species    R_Dimers_EM   @EM:R.R

  Molecules  Catalytic_R   R(tf~pY!?)
  Molecules  Catalytic_TF  R(tf~pY!1).TF(r!1)
  Molecules  Phos_TF       TF(d~pY!?)

  Species    TF_Dimer_CP   TF(d~pY!1)@CP.TF(d~pY!1)@CP
  Species    TF_Dimer_NU   TF(d~pY!1)@NU.TF(d~pY!1)@NU

  Species    Bound_prom1   DNA(p1!+)
  Species    Bound_prom2   DNA(p2!+)

  Species    P1_NU         P1@NU
  Species    P1_CP         P1@CP

  Species    Im_NU         Im@NU
  Species    Im_CP         Im@CP

  Species    Im_Cargo_NP   Im(fg!+,cargo!+) 
  Species    P1_NU_free    P1(im,dna)@NU
  Species    P1_NU_dna     P1(im,dna!+)@NU
end observables
end model

# actions #
generate_network({overwrite=>1,TextReaction=>1});
simulate_ode({suffix=>'ode',t_start=>0,t_end=>200,n_steps=>100});

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