Simulated annealing not working

GROMACS version: 2020.3
GROMACS modification: No

Edit: All fixed now! I accidentally set the 2nd annealing time to the last time step instead of the last time.

I’m trying to run simulated annealing but my temperatures don’t seem to be following a linear decrease and instead are fluctuating a lot. How can I fix this? Also, what should I set ref_t to be during simulated annealing?

Here’s the start of my .log file:

>                       :-) GROMACS - gmx mdrun, 2020.3 (-:
> 
>                             GROMACS is written by:
>      Emile Apol      Rossen Apostolov      Paul Bauer     Herman J.C. Berendsen
>     Par Bjelkmar      Christian Blau   Viacheslav Bolnykh     Kevin Boyd    
>  Aldert van Buuren   Rudi van Drunen     Anton Feenstra       Alan Gray     
>   Gerrit Groenhof     Anca Hamuraru    Vincent Hindriksen  M. Eric Irrgang  
>   Aleksei Iupinov   Christoph Junghans     Joe Jordan     Dimitrios Karkoulis
>     Peter Kasson        Jiri Kraus      Carsten Kutzner      Per Larsson    
>   Justin A. Lemkul    Viveca Lindahl    Magnus Lundborg     Erik Marklund   
>     Pascal Merz     Pieter Meulenhoff    Teemu Murtola       Szilard Pall   
>     Sander Pronk      Roland Schulz      Michael Shirts    Alexey Shvetsov  
>    Alfons Sijbers     Peter Tieleman      Jon Vincent      Teemu Virolainen 
>  Christian Wennberg    Maarten Wolf      Artem Zhmurov   
>                            and the project leaders:
>         Mark Abraham, Berk Hess, Erik Lindahl, and David van der Spoel
> 
> Copyright (c) 1991-2000, University of Groningen, The Netherlands.
> Copyright (c) 2001-2019, The GROMACS development team at
> Uppsala University, Stockholm University and
> the Royal Institute of Technology, Sweden.
> check out http://www.gromacs.org for more information.
> 
> GROMACS is free software; you can redistribute it and/or modify it
> under the terms of the GNU Lesser General Public License
> as published by the Free Software Foundation; either version 2.1
> of the License, or (at your option) any later version.
> 
> GROMACS:      gmx mdrun, version 2020.3
> Executable:   /usr/local/gromacs/bin/gmx
> Data prefix:  /usr/local/gromacs
> Working dir:  /Users/profile1/Desktop/gromacs-related_work/NPs_BBB/PS/PSNP/PSNP2
> Process ID:   12331
> Command line:
>   gmx mdrun -deffnm SA -v -nt 1
> 
> GROMACS version:    2020.3
> Verified release checksum is c0599e547549c2d0ef4fc678dc5a26ad0000eab045e938fed756f9ff5b99a197
> Precision:          single
> Memory model:       64 bit
> MPI library:        thread_mpi
> OpenMP support:     enabled (GMX_OPENMP_MAX_THREADS = 64)
> GPU support:        OpenCL
> SIMD instructions:  AVX2_256
> FFT library:        fftw-3.3.8-sse2
> RDTSCP usage:       enabled
> TNG support:        enabled
> Hwloc support:      disabled
> Tracing support:    disabled
> C compiler:         /usr/local/bin/icc Intel 19.1.2.20200623
> C compiler flags:   -march=core-avx2 -std=gnu99 -ip -funroll-all-loops -alias-const -ansi-alias -no-prec-div -fimf-domain-exclusion=14 -qoverride-limits -O3 -DNDEBUG
> C++ compiler:       /usr/local/bin/icpc Intel 19.1.2.20200623
> C++ compiler flags: -march=core-avx2 -ip -funroll-all-loops -alias-const -ansi-alias -no-prec-div -fimf-domain-exclusion=14 -qoverride-limits -qopenmp -O3 -DNDEBUG
> OpenCL include dir: /Applications/Xcode.app/Contents/Developer/Platforms/MacOSX.platform/Developer/SDKs/MacOSX10.15.sdk/System/Library/Frameworks/OpenCL.framework
> OpenCL library:     /Applications/Xcode.app/Contents/Developer/Platforms/MacOSX.platform/Developer/SDKs/MacOSX10.15.sdk/System/Library/Frameworks/OpenCL.framework
> OpenCL version:     1.2
> 
> 
> Running on 1 node with total 4 cores, 4 logical cores, 0 compatible GPUs
> Hardware detected:
>   CPU info:
>     Vendor: Intel
>     Brand:  Intel(R) Core(TM) i5-4570R CPU @ 2.70GHz
>     Family: 6   Model: 70   Stepping: 1
>     Features: aes apic avx avx2 clfsh cmov cx8 cx16 f16c fma htt intel lahf mmx msr nonstop_tsc pcid pclmuldq pdcm pdpe1gb popcnt pse rdrnd rdtscp sse2 sse3 sse4.1 sse4.2 ssse3 tdt x2apic
>   Hardware topology: Only logical processor count
>   GPU info:
>     Number of GPUs detected: 1
>     #0: name: Iris Pro, vendor: Intel, device version: OpenCL 1.2 , stat: incompatible (please recompile with GMX_OPENCL_NB_CLUSTER_SIZE=4)
> 
> 
> ++++ PLEASE READ AND CITE THE FOLLOWING REFERENCE ++++
> M. J. Abraham, T. Murtola, R. Schulz, S. Páll, J. C. Smith, B. Hess, E.
> Lindahl
> GROMACS: High performance molecular simulations through multi-level
> parallelism from laptops to supercomputers
> SoftwareX 1 (2015) pp. 19-25
> -------- -------- --- Thank You --- -------- --------
> 
> 
> ++++ PLEASE READ AND CITE THE FOLLOWING REFERENCE ++++
> S. Páll, M. J. Abraham, C. Kutzner, B. Hess, E. Lindahl
> Tackling Exascale Software Challenges in Molecular Dynamics Simulations with
> GROMACS
> In S. Markidis & E. Laure (Eds.), Solving Software Challenges for Exascale 8759 (2015) pp. 3-27
> -------- -------- --- Thank You --- -------- --------
> 
> 
> ++++ PLEASE READ AND CITE THE FOLLOWING REFERENCE ++++
> S. Pronk, S. Páll, R. Schulz, P. Larsson, P. Bjelkmar, R. Apostolov, M. R.
> Shirts, J. C. Smith, P. M. Kasson, D. van der Spoel, B. Hess, and E. Lindahl
> GROMACS 4.5: a high-throughput and highly parallel open source molecular
> simulation toolkit
> Bioinformatics 29 (2013) pp. 845-54
> -------- -------- --- Thank You --- -------- --------
> 
> 
> ++++ PLEASE READ AND CITE THE FOLLOWING REFERENCE ++++
> B. Hess and C. Kutzner and D. van der Spoel and E. Lindahl
> GROMACS 4: Algorithms for highly efficient, load-balanced, and scalable
> molecular simulation
> J. Chem. Theory Comput. 4 (2008) pp. 435-447
> -------- -------- --- Thank You --- -------- --------
> 
> 
> ++++ PLEASE READ AND CITE THE FOLLOWING REFERENCE ++++
> D. van der Spoel, E. Lindahl, B. Hess, G. Groenhof, A. E. Mark and H. J. C.
> Berendsen
> GROMACS: Fast, Flexible and Free
> J. Comp. Chem. 26 (2005) pp. 1701-1719
> -------- -------- --- Thank You --- -------- --------
> 
> 
> ++++ PLEASE READ AND CITE THE FOLLOWING REFERENCE ++++
> E. Lindahl and B. Hess and D. van der Spoel
> GROMACS 3.0: A package for molecular simulation and trajectory analysis
> J. Mol. Mod. 7 (2001) pp. 306-317
> -------- -------- --- Thank You --- -------- --------
> 
> 
> ++++ PLEASE READ AND CITE THE FOLLOWING REFERENCE ++++
> H. J. C. Berendsen, D. van der Spoel and R. van Drunen
> GROMACS: A message-passing parallel molecular dynamics implementation
> Comp. Phys. Comm. 91 (1995) pp. 43-56
> -------- -------- --- Thank You --- -------- --------
> 
> 
> ++++ PLEASE CITE THE DOI FOR THIS VERSION OF GROMACS ++++
> https://doi.org/10.5281/zenodo.3923645
> -------- -------- --- Thank You --- -------- --------
> 
> Input Parameters:
>    integrator                     = md
>    tinit                          = 0
>    dt                             = 0.02
>    nsteps                         = 2500000
>    init-step                      = 0
>    simulation-part                = 1
>    comm-mode                      = Linear
>    nstcomm                        = 100
>    bd-fric                        = 0
>    ld-seed                        = -299389755
>    emtol                          = 10
>    emstep                         = 0.01
>    niter                          = 20
>    fcstep                         = 0
>    nstcgsteep                     = 1000
>    nbfgscorr                      = 10
>    rtpi                           = 0.05
>    nstxout                        = 0
>    nstvout                        = 0
>    nstfout                        = 0
>    nstlog                         = 1000
>    nstcalcenergy                  = 100
>    nstenergy                      = 100
>    nstxout-compressed             = 1000
>    compressed-x-precision         = 100
>    cutoff-scheme                  = Verlet
>    nstlist                        = 15
>    pbc                            = xyz
>    periodic-molecules             = false
>    verlet-buffer-tolerance        = 0.005
>    rlist                          = 1.1
>    coulombtype                    = Reaction-Field
>    coulomb-modifier               = Potential-shift
>    rcoulomb-switch                = 0
>    rcoulomb                       = 1.1
>    epsilon-r                      = 15
>    epsilon-rf                     = inf
>    vdw-type                       = Cut-off
>    vdw-modifier                   = Potential-shift
>    rvdw-switch                    = 0
>    rvdw                           = 1.1
>    DispCorr                       = No
>    table-extension                = 1
>    fourierspacing                 = 0.12
>    fourier-nx                     = 0
>    fourier-ny                     = 0
>    fourier-nz                     = 0
>    pme-order                      = 4
>    ewald-rtol                     = 1e-05
>    ewald-rtol-lj                  = 0.001
>    lj-pme-comb-rule               = Geometric
>    ewald-geometry                 = 0
>    epsilon-surface                = 0
>    tcoupl                         = Nose-Hoover
>    nsttcouple                     = 15
>    nh-chain-length                = 1
>    print-nose-hoover-chain-variables = false
>    pcoupl                         = No
>    pcoupltype                     = Isotropic
>    nstpcouple                     = -1
>    tau-p                          = 1
>    compressibility (3x3):
>       compressibility[    0]={ 0.00000e+00,  0.00000e+00,  0.00000e+00}
>       compressibility[    1]={ 0.00000e+00,  0.00000e+00,  0.00000e+00}
>       compressibility[    2]={ 0.00000e+00,  0.00000e+00,  0.00000e+00}
>    ref-p (3x3):
>       ref-p[    0]={ 0.00000e+00,  0.00000e+00,  0.00000e+00}
>       ref-p[    1]={ 0.00000e+00,  0.00000e+00,  0.00000e+00}
>       ref-p[    2]={ 0.00000e+00,  0.00000e+00,  0.00000e+00}
>    refcoord-scaling               = No
>    posres-com (3):
>       posres-com[0]= 0.00000e+00
>       posres-com[1]= 0.00000e+00
>       posres-com[2]= 0.00000e+00
>    posres-comB (3):
>       posres-comB[0]= 0.00000e+00
>       posres-comB[1]= 0.00000e+00
>       posres-comB[2]= 0.00000e+00
>    QMMM                           = false
>    QMconstraints                  = 0
>    QMMMscheme                     = 0
>    MMChargeScaleFactor            = 1
> qm-opts:
>    ngQM                           = 0
>    constraint-algorithm           = Lincs
>    continuation                   = true
>    Shake-SOR                      = false
>    shake-tol                      = 0.0001
>    lincs-order                    = 4
>    lincs-iter                     = 1
>    lincs-warnangle                = 30
>    nwall                          = 0
>    wall-type                      = 9-3
>    wall-r-linpot                  = -1
>    wall-atomtype[0]               = -1
>    wall-atomtype[1]               = -1
>    wall-density[0]                = 0
>    wall-density[1]                = 0
>    wall-ewald-zfac                = 3
>    pull                           = false
>    awh                            = false
>    rotation                       = false
>    interactiveMD                  = false
>    disre                          = No
>    disre-weighting                = Conservative
>    disre-mixed                    = false
>    dr-fc                          = 1000
>    dr-tau                         = 0
>    nstdisreout                    = 100
>    orire-fc                       = 0
>    orire-tau                      = 0
>    nstorireout                    = 100
>    free-energy                    = no
>    cos-acceleration               = 0
>    deform (3x3):
>       deform[    0]={ 0.00000e+00,  0.00000e+00,  0.00000e+00}
>       deform[    1]={ 0.00000e+00,  0.00000e+00,  0.00000e+00}
>       deform[    2]={ 0.00000e+00,  0.00000e+00,  0.00000e+00}
>    simulated-tempering            = false
>    swapcoords                     = no
>    userint1                       = 0
>    userint2                       = 0
>    userint3                       = 0
>    userint4                       = 0
>    userreal1                      = 0
>    userreal2                      = 0
>    userreal3                      = 0
>    userreal4                      = 0
>    applied-forces:
>      electric-field:
>        x:
>          E0                       = 0
>          omega                    = 0
>          t0                       = 0
>          sigma                    = 0
>        y:
>          E0                       = 0
>          omega                    = 0
>          t0                       = 0
>          sigma                    = 0
>        z:
>          E0                       = 0
>          omega                    = 0
>          t0                       = 0
>          sigma                    = 0
>      density-guided-simulation:
>        active                     = false
>        group                      = protein
>        similarity-measure         = inner-product
>        atom-spreading-weight      = unity
>        force-constant             = 1e+09
>        gaussian-transform-spreading-width = 0.2
>        gaussian-transform-spreading-range-in-multiples-of-width = 4
>        reference-density-filename = reference.mrc
>        nst                        = 1
>        normalize-densities        = true
>        adaptive-force-scaling     = false
>        adaptive-force-scaling-time-constant = 4
> grpopts:
>    nrdf:        3597
>    ref-t:         400
>    tau-t:           6
> annealing:      Single
> annealing-npoints:           2
> annealing-time [0]:	         0.0   1250000.0
> annealing-temp [0]:	       400.0       200.0
>    acc:	           0           0           0
>    nfreeze:           N           N           N
>    energygrp-flags[  0]: 0
> 
> Changing nstlist from 15 to 50, rlist from 1.1 to 1.205
> 
> Using 1 MPI thread
> Using 1 OpenMP thread 
> 
> System total charge: 0.000
> Reaction-Field:
> epsRF = 0, rc = 1.1, krf = 0.375657, crf = 1.36364, epsfac = 9.26236
> The electrostatics potential has its minimum at r = 1.1
> Potential shift: LJ r^-12: -3.186e-01 r^-6: -5.645e-01
> 
> Using SIMD 4x8 nonbonded short-range kernels
> 
> Using a dual 4x8 pair-list setup updated with dynamic pruning:
>   outer list: updated every 50 steps, buffer 0.105 nm, rlist 1.205 nm
>   inner list: updated every 33 steps, buffer 0.004 nm, rlist 1.104 nm
> At tolerance 0.005 kJ/mol/ps per atom, equivalent classical 1x1 list would be:
>   outer list: updated every 50 steps, buffer 0.211 nm, rlist 1.311 nm
>   inner list: updated every 33 steps, buffer 0.054 nm, rlist 1.154 nm
> 
> Using full Lennard-Jones parameter combination matrix
> 
> 
> Initializing LINear Constraint Solver
> 
> ++++ PLEASE READ AND CITE THE FOLLOWING REFERENCE ++++
> B. Hess
> P-LINCS: A Parallel Linear Constraint Solver for molecular simulation
> J. Chem. Theory Comput. 4 (2008) pp. 116-122
> -------- -------- --- Thank You --- -------- --------
> 
> The number of constraints is 1200
> 1200 constraints are involved in constraint triangles,
> will apply an additional matrix expansion of order 4 for couplings
> between constraints inside triangles
> There are: 1600 Atoms
> Center of mass motion removal mode is Linear
> We have the following groups for center of mass motion removal:
>   0:  rest
> 
> Started mdrun on rank 0 Sat Sep  5 18:06:21 2020
> 
>            Step           Time
>               0        0.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.44396e+03    4.75099e+03   -2.23206e+04    0.00000e+00   -1.61256e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     7.68328e+03   -8.44233e+03   -8.44233e+03    5.13809e+02   -3.57386e-02
>    Constr. rmsd
>     1.30156e-05
> 
>            Step           Time
>            1000       20.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.47773e+03    4.29683e+03   -2.24219e+04    0.00000e+00   -1.66473e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     6.74575e+03   -9.90155e+03   -7.73541e+03    4.51113e+02   -2.43294e-01
>    Constr. rmsd
>     9.53834e-06
> 
>            Step           Time
>            2000       40.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.31959e+03    3.81602e+03   -2.32187e+04    0.00000e+00   -1.80831e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     6.01271e+03   -1.20704e+04   -6.98541e+03    4.02092e+02   -7.65389e-02
>    Constr. rmsd
>     8.13347e-06
> 
>            Step           Time
>            3000       60.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.11215e+03    3.77865e+03   -2.37647e+04    0.00000e+00   -1.88739e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.61672e+03   -1.32572e+04   -6.32046e+03    3.75611e+02    1.72571e+00
>    Constr. rmsd
>     8.28727e-06
> 
>            Step           Time
>            4000       80.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.05270e+03    3.60942e+03   -2.36426e+04    0.00000e+00   -1.89805e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.43226e+03   -1.35482e+04   -5.76827e+03    3.63275e+02   -2.04870e+00
>    Constr. rmsd
>     7.11529e-06
> 
>            Step           Time
>            5000      100.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.16428e+03    3.58246e+03   -2.39778e+04    0.00000e+00   -1.92311e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.50359e+03   -1.37275e+04   -5.36259e+03    3.68046e+02    1.03050e+00
>    Constr. rmsd
>     7.23516e-06
> 
>            Step           Time
>            6000      120.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.19378e+03    3.61724e+03   -2.35766e+04    0.00000e+00   -1.87656e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.23937e+03   -1.35262e+04   -5.07204e+03    3.50376e+02   -5.53289e-01
>    Constr. rmsd
>     6.82184e-06
> 
>            Step           Time
>            7000      140.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.23539e+03    3.78676e+03   -2.35362e+04    0.00000e+00   -1.85140e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.64150e+03   -1.28725e+04   -4.65291e+03    3.77268e+02   -1.11789e+00
>    Constr. rmsd
>     7.08152e-06
> 
>            Step           Time
>            8000      160.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.28060e+03    4.02801e+03   -2.29844e+04    0.00000e+00   -1.76758e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     6.24316e+03   -1.14326e+04   -4.75908e+03    4.17503e+02    6.96880e-01
>    Constr. rmsd
>     9.27470e-06
> 
>            Step           Time
>            9000      180.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.29935e+03    4.02473e+03   -2.29541e+04    0.00000e+00   -1.76300e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     6.77080e+03   -1.08592e+04   -4.85127e+03    4.52789e+02   -9.72132e-01
>    Constr. rmsd
>     1.06542e-05
> 
>            Step           Time
>           10000      200.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.28415e+03    3.84730e+03   -2.32077e+04    0.00000e+00   -1.80762e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.69769e+03   -1.23785e+04   -4.95573e+03    3.81025e+02    1.49824e+00
>    Constr. rmsd
>     7.12534e-06
> 
>            Step           Time
>           11000      220.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.27041e+03    3.82537e+03   -2.34829e+04    0.00000e+00   -1.83871e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.86775e+03   -1.25194e+04   -5.03481e+03    3.92398e+02    2.52879e+00
>    Constr. rmsd
>     9.09290e-06
> 
>            Step           Time
>           12000      240.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.11491e+03    3.82147e+03   -2.35672e+04    0.00000e+00   -1.86309e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.48923e+03   -1.31416e+04   -5.08356e+03    3.67085e+02   -1.90958e+00
>    Constr. rmsd
>     7.73796e-06
> 
>            Step           Time
>           13000      260.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.23664e+03    3.82506e+03   -2.34781e+04    0.00000e+00   -1.84164e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.50849e+03   -1.29079e+04   -4.75688e+03    3.68373e+02    9.53551e-01
>    Constr. rmsd
>     7.10332e-06
> 
>            Step           Time
>           14000      280.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.41071e+03    3.86086e+03   -2.30838e+04    0.00000e+00   -1.78122e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     6.52360e+03   -1.12886e+04   -4.70929e+03    4.36258e+02   -8.29381e-01
>    Constr. rmsd
>     8.99320e-06
> 
>            Step           Time
>           15000      300.00000
> 
> Current ref_t for group System:    400.0
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.35228e+03    4.11305e+03   -2.30397e+04    0.00000e+00   -1.75744e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     6.63192e+03   -1.09424e+04   -4.75942e+03    4.43501e+02    1.04442e+00
>    Constr. rmsd
>     9.53887e-06
> 
>            Step           Time
>           16000      320.00000
> 
> Current ref_t for group System:    399.9
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.35592e+03    4.06219e+03   -2.29018e+04    0.00000e+00   -1.74837e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     6.49538e+03   -1.09883e+04   -4.87241e+03    4.34370e+02    6.34556e-01
>    Constr. rmsd
>     1.03111e-05
> 
>            Step           Time
>           17000      340.00000
> 
> Current ref_t for group System:    399.9
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.27899e+03    3.93051e+03   -2.34645e+04    0.00000e+00   -1.82550e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     6.17516e+03   -1.20799e+04   -4.85316e+03    4.12956e+02   -1.00637e+00
>    Constr. rmsd
>     1.03241e-05
> 
>            Step           Time
>           18000      360.00000
> 
> Current ref_t for group System:    399.9
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.31331e+03    3.77261e+03   -2.35246e+04    0.00000e+00   -1.84387e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.97483e+03   -1.24638e+04   -4.91147e+03    3.99559e+02   -8.68265e-01
>    Constr. rmsd
>     9.64900e-06
> 
>            Step           Time
>           19000      380.00000
> 
> Current ref_t for group System:    399.9
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.21508e+03    3.76006e+03   -2.35352e+04    0.00000e+00   -1.85601e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.71313e+03   -1.28469e+04   -4.92194e+03    3.82058e+02    7.41787e-01
>    Constr. rmsd
>     8.53789e-06
> 
>            Step           Time
>           20000      400.00000
> 
> Current ref_t for group System:    399.9
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.20786e+03    3.68793e+03   -2.37471e+04    0.00000e+00   -1.88513e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.41134e+03   -1.34400e+04   -4.87929e+03    3.61876e+02    3.37654e+00
>    Constr. rmsd
>     7.71699e-06
> 
>            Step           Time
>           21000      420.00000
> 
> Current ref_t for group System:    399.9
> 
>    Energies (kJ/mol)
>            Bond          Angle        LJ (SR)   Coulomb (SR)      Potential
>     1.16944e+03    3.60634e+03   -2.35745e+04    0.00000e+00   -1.87987e+04
>     Kinetic En.   Total Energy  Conserved En.    Temperature Pressure (bar)
>     5.64993e+03   -1.31487e+04   -4.82150e+03    3.77832e+02    9.86983e-01
>    Constr. rmsd
>     7.39476e-06

Here’s my temperature graph:
SA-temp.xvg (390.4 KB)

Thank you in advance!

Thanks for updating us!