def hybridization(options, mySeq, myTrials=0, doFirstPassage=False):
# Using domain representation makes it easier to write secondary structures.
onedomain = Domain(name="itall", sequence=mySeq)
top = Strand(name="top", domains=[onedomain])
bot = top.C
# Note that the structure is specified to be single stranded, but this will be over-ridden when Boltzmann sampling is turned on.
startTop = Complex(strands=[top], structure=".")
startBot = Complex(strands=[bot], structure=".")
# Turns Boltzmann sampling on for this complex and also does sampling more efficiently by sampling 'trials' states.
if (myTrials > 0):
setBoltzmann(startTop, myTrials)
setBoltzmann(startBot, myTrials)
# Stop when the exact full duplex is achieved.
success_complex = Complex(strands=[top, bot], structure="(+)")
stopSuccess = StopCondition(
Options.STR_SUCCESS, [(success_complex, Options.exactMacrostate, 0)])
# Declare the simulation unproductive if the strands become single-stranded again.
failed_complex = Complex(strands=[top], structure=".")
stopFailed = StopCondition(Options.STR_FAILURE,
[(failed_complex, Options.dissocMacrostate, 0)])
options.start_state = [startTop, startBot]
# Point the options to the right objects
if not doFirstPassage:
options.stop_conditions = [stopSuccess, stopFailed]
else:
options.stop_conditions = [stopSuccess]
def changeComplex(options, expirement_type=NORMAL, trials=500):
# Easiest to use full dot paren here
# See SI Document for these sequences and lengths!
toehold_length = 7
h_length = 15
helper = Strand(name="Helper_AAq",
sequence="TTTCCTAATCCCAATCAACACCTTTCCTA")
produce_bot = Strand(
name="Produce_BOT_CApAq",
sequence="GTAAAGACCAGTGGTGTGAAGATAGGAAAGGTGTTGATTGGGATTAGGAAACC")
ap = Strand(name="ap",
sequence="CATCACTATCAATCATACATGGTTTCCTATCTTCACACCACTGG")
aq = Strand(name="aq",
sequence="CATCACTATCAATCATACATGGTTTCCTAATCCCAATCAACACC")
# Offset of two to account for clamp domains
produce_struct = "." * toehold_length + "(" * (
len(produce_bot.sequence) -
toehold_length) + "+" + '.' * (toehold_length + h_length - 2) + ')' * (
len(ap.sequence) - toehold_length - h_length + 2) + "+" + '.' * (
toehold_length + h_length) + ')' * (len(ap.sequence) -
toehold_length - h_length)
if expirement_type == WITHOUT_GG:
ap = Strand(name="ap",
sequence="CATCACTATCAATCATACATTTTCCTATCTTCACACCACTGG")
# bot, aq, ap
# Offsets due to a) clamp domains b) two b.p. removal
produce_struct = "." * toehold_length + "(" * (
len(produce_bot.sequence) - toehold_length) + "+" + '.' * (
toehold_length + h_length - 2) + ')' * (
len(ap.sequence) - toehold_length - h_length +
4) + "+" + '.' * (toehold_length + h_length - 2) + ')' * (
len(ap.sequence) - toehold_length - h_length + 2)
elif expirement_type == WITHOUT_G:
ap = Strand(name="ap",
sequence="CATCACTATCAATCATACATGTTTCCTATCTTCACACCACTGG")
# bot, aq, ap
# Offsets due to a) clamp domains b) one b.p. removal
produce_struct = "." * toehold_length + "(" * (
len(produce_bot.sequence) - toehold_length) + "+" + '.' * (
toehold_length + h_length - 2) + ')' * (
len(ap.sequence) - toehold_length - h_length +
3) + "+" + '.' * (toehold_length + h_length - 1) + ')' * (
len(ap.sequence) - toehold_length - h_length + 1)
elif expirement_type == HELPER_WITHOUT_CC:
# only modify helper sequence here - remove the two 3' most 'C'.
helper = Strand(name="Helper_AAq",
sequence="TTTCCTAATCCCAATCAACACCTTTTA")
# No change required elsewhere - we check for the release of strands rather than the complicated
# leak complex formed for simplicity. We should really check for ANY free strands here i.e. Ap OR Aq
# but it is hard to imagine a mechanism which results in the release of Ap in this simulation
produce_complex = Complex(name="produce",
strands=[produce_bot, aq, ap],
structure=produce_struct)
helper_complex = Complex(name="helper",
strands=[helper],
structure='.' * len(helper.sequence))
leak_complex = Complex(name="leak",
strands=[aq],
structure='.' * len(aq.sequence))
if trials > 0:
setBoltzmann(produce_complex, trials, 75)
setBoltzmann(helper_complex, trials, 75)
success_stop_cond = StopCondition(
Literals.success, [(leak_complex, Options.dissoc_macrostate, 0)])
# the leak has failed if we end up with our initial complexes again.
# check if we end up with a free helper complex
failure_stop_cond = StopCondition(
Literals.failure, [(helper_complex, Options.dissoc_macrostate, 0)])
options.start_state = [produce_complex, helper_complex]
options.stop_conditions = [success_stop_cond, failure_stop_cond]
def machinek2014(options, selector, trialsIn):
# we only allow first step mode at this point.
# these are the sequences we need to build the dot-parens
incumbent = ""
target = ""
invader = ""
toeholdSelect = selector / 12
mismatchSelect = selector % 12
positionSelector = [0, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14]
mismatchSelect = positionSelector[mismatchSelect]
# decide on toehold sequence
toeholdSeq = "ATGTGG" # 6 nt toehold option
if toeholdSelect == 1:
toeholdSeq = "ATGTGGA" # 7 nt toehold option
if toeholdSelect == 2:
toeholdSeq = "ATGTGGAGGG" # 10 nt toehold option
# determine the incumbent, target and invader sequences
# FD: copy-pasting supplementary Table 6 directly
if mismatchSelect == 0 or mismatchSelect == 2 or mismatchSelect == 12 or mismatchSelect == 14:
incumbent = "TGGTGTTTGTGGGTGTGGTGAGTTTGAGGTTGA"
target = "CCCTCCACATTCAACCTCAAACTCACC"
if mismatchSelect == 0: # perfect
invader = "GGTGAGTTTGAGGTTGA"
if mismatchSelect == 2:
invader = "GGTGAGTTTGAGGTTCA"
if mismatchSelect == 12:
invader = "GGTGACTTTGAGGTTGA"
if mismatchSelect == 14:
invader = "GGTCAGTTTGAGGTTGA"
if mismatchSelect == 3:
incumbent = "TGGTGTTTGTGGGTGTGGTGAGTTTGAGGTGAT"
target = "CCCTCCACATATCACCTCAAACTCACC"
invader = "GGTGAGTTTGAGGTCAT"
if mismatchSelect == 4:
incumbent = "TGGTGTTTGTGGGTGTGGTGAGTTTGAGTGAGT"
target = "CCCTCCACATACTCACTCAAACTCACC"
invader = "GGTGAGTTTGAGTCAGT"
if mismatchSelect == 5:
incumbent = "TGGTGTTTGTGGGTGTGGTGAGTTTGATGAGGT"
target = "CCCTCCACATACCTCATCAAACTCACC"
invader = "GGTGAGTTTGATCAGGT"
if mismatchSelect == 6:
incumbent = "TGGTGTTTGTGGGTGTGGTGAGTTTGTGAAGGT"
target = "CCCTCCACATACCTTCACAAACTCACC"
invader = "GGTGAGTTTGTCAAGGT"
if mismatchSelect == 7:
incumbent = "TGGTGTTTGTGGGTGTGGTGAGTTTTGAGAGGT"
target = "CCCTCCACATACCTCTCAAAACTCACC"
invader = "GGTGAGTTTTCAGAGGT"
if mismatchSelect == 8:
incumbent = "TGGTGTTTGTGGGTGTGGTGAGTTTGATGAGGT"
target = "CCCTCCACATACCTCATCAAACTCACC"
invader = "GGTGAGTTTCATGAGGT"
if mismatchSelect == 9:
incumbent = "TGGTGTTTGTGGGTGTGGTGAGTTGATTGAGGT"
target = "CCCTCCACATACCTCAATCAACTCACC"
invader = "GGTGAGTTCATTGAGGT"
if mismatchSelect == 10:
incumbent = "TGGTGTTTGTGGGTGTGGTGAGTGATTTGAGGT"
target = "CCCTCCACATACCTCAAATCACTCACC"
invader = "GGTGAGTCATTTGAGGT"
invader = invader + toeholdSeq
# set up the actual complexes
strandIncumbent = Strand(name="incumbent", sequence=incumbent)
strandTarget = Strand(name="target", sequence=target)
strandInvader = Strand(name="invader", sequence=invader)
intialDotParen = '.' * 16 + '(' * 17 + "+" + '.' * 10 + ')' * 17
intialInvaderDotParen = '.' * len(invader)
successDotParen = '.' * 33
initialComplex = Complex(strands=[strandIncumbent, strandTarget],
structure=intialDotParen)
initialInvader = Complex(strands=[strandInvader],
structure=intialInvaderDotParen)
successComplex = Complex(strands=[strandIncumbent],
structure=successDotParen)
# Turns Boltzmann sampling on for this complex and also does sampling more efficiently by sampling 'trials' states.
if (trialsIn > 0):
setBoltzmann(initialComplex, trialsIn)
setBoltzmann(initialInvader, trialsIn)
initialComplex.boltzmann_supersample = 25
initialInvader.boltzmann_supersample = 25
stopSuccess = StopCondition(
Options.STR_SUCCESS, [(successComplex, Options.dissocMacrostate, 0)])
stopFailed = StopCondition(Options.STR_FAILURE,
[(initialComplex, Options.dissocMacrostate, 0)])
# actually set the intial and stopping states
options.start_state = [initialComplex, initialInvader]
options.stop_conditions = [stopSuccess, stopFailed]
def show_interesting_trajectories(result_lists, seqs, type='fastest'):
mintimes = []
maxtimes = []
slowseeds = []
fastseeds = []
for n in range(len(result_lists)):
times = 1e6 * np.array([
i.time for i in result_lists[n]
]) # convert from seconds to microseconds units.
slowseeds.append(result_lists[n][np.argmax(
times)].seed) # find the seed used for the slowest simulation
fastseeds.append(result_lists[n][np.argmin(
times)].seed) # find the seed used for the fastest simulation
mintimes.append(np.min(times))
maxtimes.append(np.max(times))
# taken from hairpin_trajectories.py
def print_trajectory(o):
print o.full_trajectory[0][0][3] # the strand sequence
print o.start_state[0].structure
for i in range(len(o.full_trajectory)):
time = 1e6 * o.full_trajectory_times[i]
state = o.full_trajectory[i][0]
struct = state[4]
dG = state[5]
print struct + ' t=%11.9f microseconds, dG=%6.2f kcal/mol' % (time,
dG)
# take a look at the fastest folds. to take a look at the more interesting slowest folds,
# change "mintimes" to "maxtimes" and change "fastseeds" to "slowseeds"; do this based on 'type' argument
seeds = fastseeds if type == 'fastest' else slowseeds
times = mintimes if type == 'fastest' else maxtimes
for (seq, seed, time) in zip(seqs, seeds, times):
s1 = Strand(name="hairpin", sequence=seq)
c1 = Complex(strands=[s1], structure=16 * '.') # hard-coded length 16
c2 = Complex(strands=[s1], structure="((((((....))))))"
) # hard-coded stem length 6, loop length 4
sc = StopCondition("CLOSED", [(c2, Literals.exact_macrostate, 0)])
# For future reference, StopConditions and Macrostates (same thing) are provided as a list of match conditions,
# all of which must be matched. I.e. there is an implicit AND being evaluated. E.g.
# sc = StopCondition( "EXAMPLE", [(c2,Loose_Macrostate,8), (c1,Loose_Macrostate,4)]
# would specify the intersection of those two macrostates, i.e. any 2 base pairs of the helix (and maybe more).
o = Options(
temperature=310.15,
dangles='Some',
start_state=[c1],
simulation_time=
0.1, # 0.1 seconds (lots more time than hairpin_trajectories, to accommodate slow folds)
num_simulations=1, # don't play it again, Sam
output_interval=1, # record every single step
rate_method=
'Metropolis', # the default is 'Kawasaki' (numerically, these are 1 and 2 respectively)
rate_scaling=
'Calibrated', # this is the same as 'Default'. 'Unitary' gives values 1.0 to both.
simulation_mode='Trajectory'
) # numerically 128. See interface/_options/constants.py for more info about all this.
o.stop_conditions = [sc] # don't wait for the time-out
o.initial_seed = seed # start with the same random seed as before...
s = SimSystem(o)
s.start()
print_trajectory(o)
print "Original run's time: %g microseconds" % time
