comparison scripts/S01_find_orf_on_multiple_alignment.py @ 2:0d2f72caea10 draft

planemo upload for repository https://github.com/abims-sbr/adaptsearch commit 44a89d5eeb82789bfc643b33c11f391281b6374b

1:567d5b771a90

    return(bash_codeUniversel)
###########################################################


######################################################################################################################
##### DEF 3 : Test if the sequence is a multiple of 3, and if not correct the sequence to become a multiple of 3 ##### 
######################################################################################################################
### WEAKNESS OF THAT APPROACH = I remove extra base(s) at the end of the sequence ==> I can lost a codon, when I test ORF (as I will decay the ORF)
def multiple3(seq):
    leng = len(seq)
    modulo = leng%3
    if modulo == 0:   # the results of dividing leng per 3 is an integer
        new_seq = seq        
    elif modulo == 1:    # means 1 extra nc (nucleotid) needs to be removed (the remaining of modulo indicate the part which is non-dividable per 3)
        new_seq = seq[:-1]  # remove the last nc
    elif modulo == 2:  # means 2 extra nc (nucleotid) needs to be removed (the remaining of modulo indicate the part which is non-dividable per 3)
        new_seq = seq[:-2]  # remove the 2 last nc
    len1 = len(new_seq)   
    return(new_seq, modulo)
##########################################################


#############################

        base1 = string.capitalize(base1)
        base2 = seq_dna[i+1]
        base2 = string.capitalize(base2)
        base3 = seq_dna[i+2]
        base3 = string.capitalize(base3)
        
        codon = base1+base2+base3
        codon = string.replace(codon, "T", "U")

        if codon in bash_codeUniversel.keys():
            aa = bash_codeUniversel[codon]
            seq_aa = seq_aa + aa            
        else:
            seq_aa = seq_aa +"?"    ### Take account for gap "-" and "N"
        i = i + 3

    return(seq_aa)
##########################################################


###### DEF 4 - Part 2 - ######
##############################
def find_good_ORF_criteria_3(bash_aligned_nc_seq, bash_codeUniversel):  
    
    ## 1 ## Get the list of aligned aa seq for the 3 ORF:
    bash_of_aligned_aa_seq_3ORF = {}
    bash_of_aligned_nuc_seq_3ORF = {}
    BEST_LONGUEST_SUBSEQUENCE_LIST_POSITION = []
    for fasta_name in bash_aligned_nc_seq.keys():
        ## 1.1. ## Get the raw sequence
        sequence_nc = bash_aligned_nc_seq[fasta_name]

        ## 1.2. ## Check whether the sequence is multiple of 3, and correct it if not:
        new_sequence_nc, modulo = multiple3(sequence_nc)                                  ### DEF 3 ###
        
        ## 1.3. ## Get the 3 ORFs (nuc) for each sequence
        seq_nuc_ORF1 = new_sequence_nc
        seq_nuc_ORF2 = new_sequence_nc[1:-2]
        seq_nuc_ORF3 = new_sequence_nc[2:-1]
        seq_reversed=ReverseComplement2(seq_nuc_ORF1)
        seq_nuc_ORF4=seq_reversed
        seq_nuc_ORF5=seq_reversed[1:-2]
        seq_nuc_ORF6=seq_reversed[2:-1]
        
        LIST_6_ORF_nuc = [seq_nuc_ORF1, seq_nuc_ORF2, seq_nuc_ORF3,seq_nuc_ORF4,seq_nuc_ORF5,seq_nuc_ORF6]
        bash_of_aligned_nuc_seq_3ORF[fasta_name] = LIST_6_ORF_nuc     ### For each seq of the multialignment => give the 6 ORFs (in nuc)

        ## 1.4. ## Get the 3 ORFs (aa) for each sequence
        seq_prot_ORF1 = simply_get_ORF(seq_nuc_ORF1,bash_codeUniversel)                ### DEF 4 - Part 1 - ##
        seq_prot_ORF2 = simply_get_ORF(seq_nuc_ORF2,bash_codeUniversel)                ### DEF 4 - Part 1 - ##
        seq_prot_ORF3 = simply_get_ORF(seq_nuc_ORF3,bash_codeUniversel)                ### DEF 4 - Part 1 - ##
        seq_prot_ORF4 = simply_get_ORF(seq_nuc_ORF4,bash_codeUniversel)                ### DEF 4 - Part 1 - ## 
        seq_prot_ORF5 = simply_get_ORF(seq_nuc_ORF5,bash_codeUniversel)                ### DEF 4 - Part 1 - ##
        seq_prot_ORF6 = simply_get_ORF(seq_nuc_ORF6,bash_codeUniversel)                ### DEF 4 - Part 1 - ##

        LIST_6_ORF_aa = [seq_prot_ORF1, seq_prot_ORF2, seq_prot_ORF3,seq_prot_ORF4,seq_prot_ORF5,seq_prot_ORF6]
        bash_of_aligned_aa_seq_3ORF[fasta_name] = LIST_6_ORF_aa     ### For each seq of the multialignment => give the 6 ORFs (in aa) 

    ## 2 ## Test for the best ORF (Get the longuest segment in the alignment with no codon stop ... for each ORF ... the longuest should give the ORF)
    BEST_MAX = 0
    for i in [0,1,2,3,4,5]:   ### Test the 6 ORFs 
        ORF_Aligned_aa = []
        ORF_Aligned_nuc = []
                

        ## 2.1 ## Get the alignment of sequence for a given ORF
        ## Compare the 1rst ORF between all sequence => list them in ORF_Aligned_aa // them do the same for the second ORF, and them the 3rd
        for fasta_name in bash_of_aligned_aa_seq_3ORF.keys(): 
            ORFsequence = bash_of_aligned_aa_seq_3ORF[fasta_name][i]
            aa_length = len(ORFsequence)
            ORF_Aligned_aa.append(ORFsequence)   ### List of all sequences in the ORF nb "i" =

        n = i+1
      
        for fasta_name in bash_of_aligned_nuc_seq_3ORF.keys(): 
            ORFsequence = bash_of_aligned_nuc_seq_3ORF[fasta_name][i]
            nuc_length = len(ORFsequence)
            ORF_Aligned_nuc.append(ORFsequence)   ### List of all sequences in the ORF nb "i" =

        ## 2.2 ## Get the list of sublist of positions whithout codon stop in the alignment
        ## For each ORF, now we have the list of sequences available (i.e. THE ALIGNMENT IN A GIVEN ORF)
        ## Next step is to get the longuest subsequence whithout stop
        ## We will explore the presence of stop "*" in each column of the alignment, and get the positions of the segments between the positions with "*"
        MAX_LENGTH = 0
        LONGUEST_SEGMENT_UNSTOPPED = ""        
        j = 0 # Start from first position in alignment       
        List_of_List_subsequences = []
        List_positions_subsequence = []
        while j < aa_length:                
                column = []
                for seq in ORF_Aligned_aa:
                    column.append(seq[j])
                j = j+1
                if "*" in column:
                    List_of_List_subsequences.append(List_positions_subsequence) ## Add previous list of positions
                    List_positions_subsequence = []                              ## Re-initialyse list of positions
                else:
                    List_positions_subsequence.append(j)
        
        ## 2.3 ## Among all the sublists (separated by column with codon stop "*"), get the longuest one (BETTER SEGMENT for a given ORF)
        LONGUEST_SUBSEQUENCE_LIST_POSITION = []
        MAX=0
        for sublist in List_of_List_subsequences:
            if len(sublist) > MAX and len(sublist) > MINIMAL_CDS_LENGTH:
                MAX = len(sublist)
                LONGUEST_SUBSEQUENCE_LIST_POSITION = sublist
        
        ## 2.4. ## Test if the longuest subsequence start exactly at the beginning of the original sequence (i.e. means the ORF maybe truncated)
        if LONGUEST_SUBSEQUENCE_LIST_POSITION != []:
            if LONGUEST_SUBSEQUENCE_LIST_POSITION[0] == 0:
                CDS_maybe_truncated = 1
            else:
                CDS_maybe_truncated = 0
        else:
            CDS_maybe_truncated = 0
            

        ## 2.5 ## Test if this BETTER SEGMENT for a given ORF, is the better than the one for the other ORF (GET THE BEST ORF)
        ## Test whether it is the better ORF
        if MAX > BEST_MAX:
            BEST_MAX = MAX

        ## 4 ## Get the corresponding position (START/END of BEST CODING SEGMENT) for nucleotides alignment
        pos_MIN_nuc = pos_MIN_aa * 3
        pos_MAX_nuc = pos_MAX_aa * 3

        BESTORF_bash_aligned_nc_seq = {}
        BESTORF_bash_aligned_nc_seq_CODING = {}    
        for fasta_name in bash_aligned_nc_seq.keys():
            seq = bash_of_aligned_nuc_seq_3ORF[fasta_name][index_BEST_ORF]
            seq_coding = seq[pos_MIN_nuc:pos_MAX_nuc]
            BESTORF_bash_aligned_nc_seq[fasta_name] = seq
            BESTORF_bash_aligned_nc_seq_CODING[fasta_name] = seq_coding

    ### Check whether their is a "M" or not, and if at least 1 "M" is present, that it is not in the last 50 aa
    ###########################################################################################################

    BESTORF_bash_of_aligned_aa_seq_CDS_with_M = {}
    BESTORF_bash_of_aligned_nuc_seq_CDS_with_M = {}
    
    Ortho = 0
    for fasta_name in BESTORF_bash_of_aligned_aa_seq_CODING.keys():
        seq_aa = BESTORF_bash_of_aligned_aa_seq_CODING[fasta_name]
        Ortho = detect_Methionine(seq_aa, Ortho)                                ### DEF6 ###

    if Ortho == 1:
        BESTORF_bash_of_aligned_aa_seq_CDS_with_M = BESTORF_bash_of_aligned_aa_seq_CODING
        BESTORF_bash_of_aligned_nuc_seq_CDS_with_M = BESTORF_bash_aligned_nc_seq_CODING

    ## CASE 2: in case the CDS is truncated, so the "M" is maybe missing:
    if Ortho == 0 and CDS_maybe_truncated == 1: 
        BESTORF_bash_of_aligned_aa_seq_CDS_with_M = BESTORF_bash_of_aligned_aa_seq_CODING
        BESTORF_bash_of_aligned_nuc_seq_CDS_with_M = BESTORF_bash_aligned_nc_seq_CODING

    ## CASE 3: CDS not truncated AND no "M" found in good position (i.e. before the last 50 aa):
        ## => the 2 bash "CDS_with_M" are left empty ("{}")
    
    return(BESTORF_bash_aligned_nc_seq,  BESTORF_bash_aligned_nc_seq_CODING, BESTORF_bash_of_aligned_nuc_seq_CDS_with_M, BESTORF_bash_of_aligned_aa_seq, BESTORF_bash_of_aligned_aa_seq_CODING, BESTORF_bash_of_aligned_aa_seq_CDS_with_M)    
##########################################################


##################################################################################################
###### DEF 5 : Detect all indices corresponding to all occurance of a substring in a string ######

    return listindex
######################################################


############################################################
###### DEF 6 : Detect if methionin in the aa sequence ###### 
############################################################
def detect_Methionine(seq_aa, Ortho):

    ln = len(seq_aa)
    nbre = sys.argv[2]
    CUTOFF_Last_50aa = ln - MINIMAL_CDS_LENGTH
    #Ortho = 0  ## means orthologs not found
    
    ## Find all indices of occurances of "M" in a string of aa
    list_indices = allindices(seq_aa, "M")                  ### DEF5 ###
         
    ## If some "M" are present, find whether the first "M" found is not in the 50 last aa (indice < CUTOFF_Last_50aa) ==> in this case: maybenot a CDS
    if list_indices != []:
        first_M = list_indices[0]        
        if first_M < CUTOFF_Last_50aa:
            Ortho = 1  ## means orthologs found

    return(Ortho)
###################################

############################################################
###### DEF 7 : Reverse complement DNA sequence ###### 
###### Reference: http://crazyhottommy.blogspot.fr/2013/10/python-code-for-getting-reverse.html
############################################################


def ReverseComplement2(seq):

#######################
##### RUN RUN RUN #####
#######################
import string, os, time, re, zipfile, sys

MINIMAL_CDS_LENGTH = int(sys.argv[3])  ## in aa number

## INPUT / OUTPUT
list_file = []
zfile = zipfile.ZipFile(sys.argv[1])
for name in zfile.namelist() :
    list_file.append(name)
    zfile.extract(name, "./")
 
F2 = open(sys.argv[2], 'r')

os.mkdir("04_BEST_ORF_nuc")
Path_OUT1 = "04_BEST_ORF_nuc"
os.mkdir("04_BEST_ORF_aa")
Path_OUT2 = "04_BEST_ORF_aa"

Path_OUT5 = "06_CDS_with_M_nuc"
os.mkdir("06_CDS_with_M_aa")
Path_OUT6 = "06_CDS_with_M_aa"


### Get Universal Code
bash_codeUniversel = code_universel(F2)  ### DEF2 ###
F2.close()

### Get the Bash corresponding to an alignment file in fasta format
count_file_processed = 0
count_file_with_CDS = 0
count_file_without_CDS = 0
count_file_with_CDS_plus_M = 0

for file in list_file:
    count_file_processed = count_file_processed + 1   
    fasta_file_path = "./%s" %file
    bash_fasta = dico(fasta_file_path)   ### DEF 1 ###
    BESTORF_nuc, BESTORF_nuc_CODING, BESTORF_nuc_CDS_with_M, BESTORF_aa, BESTORF_aa_CODING, BESTORF_aa_CDS_with_M  = find_good_ORF_criteria_3(bash_fasta, bash_codeUniversel)   ### DEF 4 - PART 2 - ### 

    ## a ## OUTPUT BESTORF_nuc
    if BESTORF_nuc != {}:
        count_file_with_CDS = count_file_with_CDS +1
        OUT1 = open("%s/%s" %(Path_OUT1,file), "w")

            OUT1.write("%s\n" %fasta_name)
            OUT1.write("%s\n" %seq)
        OUT1.close()
    else:
        count_file_without_CDS = count_file_without_CDS + 1
        
   
    ## b ## OUTPUT BESTORF_nuc_CODING  ===> THE MOST INTERESTING!!!
    if BESTORF_aa != {}:
        OUT2 = open("%s/%s" %(Path_OUT2,file), "w")
        for fasta_name in BESTORF_aa.keys():
            seq = BESTORF_aa[fasta_name]
            OUT2.write("%s\n" %fasta_name)
            OUT2.write("%s\n" %seq)
        OUT2.close()    

    ## c ## OUTPUT BESTORF_aa
    if BESTORF_nuc_CODING != {}:
        OUT3 = open("%s/%s" %(Path_OUT3,file), "w")
        for fasta_name in BESTORF_nuc_CODING.keys():

        OUT4 = open("%s/%s" %(Path_OUT4,file), "w")
        for fasta_name in BESTORF_aa_CODING.keys():
            seq = BESTORF_aa_CODING[fasta_name]
            OUT4.write("%s\n" %fasta_name)
            OUT4.write("%s\n" %seq)
        OUT4.close()    

    ## e ## OUTPUT BESTORF_nuc_CDS_with_M
    if BESTORF_nuc_CDS_with_M != {}:
        count_file_with_CDS_plus_M = count_file_with_CDS_plus_M + 1
        OUT5 = open("%s/%s" %(Path_OUT5,file), "w")
        for fasta_name in BESTORF_nuc_CDS_with_M.keys():
            seq = BESTORF_nuc_CDS_with_M[fasta_name]
            OUT5.write("%s\n" %fasta_name)
            OUT5.write("%s\n" %seq)
        OUT5.close()    

    ## f ## OUTPUT BESTORF_aa_CDS_with_M
    if BESTORF_aa_CDS_with_M != {}:
        OUT6 = open("%s/%s" %(Path_OUT6,file), "w")
        for fasta_name in BESTORF_aa_CDS_with_M.keys():
            seq = BESTORF_aa_CDS_with_M[fasta_name]
            OUT6.write("%s\n" %fasta_name)
            OUT6.write("%s\n" %seq)
        OUT6.close()

    os.system("rm -rf %s" %file)    

## Print 
print "*************** CDS detection ***************"
print "\nFiles processed: %d" %count_file_processed
print "\tFiles with CDS: %d" %count_file_with_CDS
print "\t\tFiles with CDS plus M (codon start): %d" %count_file_with_CDS_plus_M
print "\tFiles without CDS: %d \n" %count_file_without_CDS
print ""

## Zipfile
f_bestORF_nuc = zipfile.ZipFile("ORF_Search_bestORF_nuc.zip", "w")
f_bestORF_aa = zipfile.ZipFile("ORF_Search_bestORF_aa.zip", "w")
f_CDS_nuc = zipfile.ZipFile("ORF_Search_CDS_nuc.zip", "w")
f_CDS_aa = zipfile.ZipFile("ORF_Search_CDS_aa.zip", "w")
f_CDSM_nuc = zipfile.ZipFile("ORF_Search_CDSM_nuc.zip", "w")
f_CDSM_aa = zipfile.ZipFile("ORF_Search_CDSM_aa.zip", "w")

os.chdir("%s" %Path_OUT1)
folder = os.listdir("./")
for i in folder :
    f_bestORF_nuc.write("./%s" %i)

os.chdir("../%s" %Path_OUT2)
folder = os.listdir("./")
for i in folder :
    f_bestORF_aa.write("./%s" %i)

os.chdir("../%s" %Path_OUT3)
folder = os.listdir("./")
for i in folder :
    f_CDS_nuc.write("./%s" %i)

os.chdir("../%s" %Path_OUT4)
folder = os.listdir("./")
for i in folder :
    f_CDS_aa.write("./%s" %i)

os.chdir("../%s" %Path_OUT5)
folder = os.listdir("./")
for i in folder :
    f_CDSM_nuc.write("./%s" %i)

os.chdir("../%s" %Path_OUT6)
folder = os.listdir("./")
for i in folder :
    f_CDSM_aa.write("./%s" %i)

2:0d2f72caea10

    return(bash_codeUniversel)
###########################################################


######################################################################################################################
##### DEF 3 : Test if the sequence is a multiple of 3, and if not correct the sequence to become a multiple of 3 #####
######################################################################################################################
### WEAKNESS OF THAT APPROACH = I remove extra base(s) at the end of the sequence ==> I can lost a codon, when I test ORF (as I will decay the ORF)
def multiple3(seq):
    leng = len(seq)
    modulo = leng%3
    if modulo == 0:   # the results of dividing leng per 3 is an integer
        new_seq = seq
    elif modulo == 1:    # means 1 extra nc (nucleotid) needs to be removed (the remaining of modulo indicate the part which is non-dividable per 3)
        new_seq = seq[:-1]  # remove the last nc
    elif modulo == 2:  # means 2 extra nc (nucleotid) needs to be removed (the remaining of modulo indicate the part which is non-dividable per 3)
        new_seq = seq[:-2]  # remove the 2 last nc
    len1 = len(new_seq)
    return(new_seq, modulo)
##########################################################


#############################

        base1 = string.capitalize(base1)
        base2 = seq_dna[i+1]
        base2 = string.capitalize(base2)
        base3 = seq_dna[i+2]
        base3 = string.capitalize(base3)

        codon = base1+base2+base3
        codon = string.replace(codon, "T", "U")

        if codon in bash_codeUniversel.keys():
            aa = bash_codeUniversel[codon]
            seq_aa = seq_aa + aa
        else:
            seq_aa = seq_aa +"?"    ### Take account for gap "-" and "N"
        i = i + 3

    return(seq_aa)
##########################################################


###### DEF 4 - Part 2 - ######
##############################
def find_good_ORF_criteria_3(bash_aligned_nc_seq, bash_codeUniversel):

    ## 1 ## Get the list of aligned aa seq for the 3 ORF:
    bash_of_aligned_aa_seq_3ORF = {}
    bash_of_aligned_nuc_seq_3ORF = {}
    BEST_LONGUEST_SUBSEQUENCE_LIST_POSITION = []
    for fasta_name in bash_aligned_nc_seq.keys():
        ## 1.1. ## Get the raw sequence
        sequence_nc = bash_aligned_nc_seq[fasta_name]

        ## 1.2. ## Check whether the sequence is multiple of 3, and correct it if not:
        new_sequence_nc, modulo = multiple3(sequence_nc)                                  ### DEF 3 ###

        ## 1.3. ## Get the 3 ORFs (nuc) for each sequence
        seq_nuc_ORF1 = new_sequence_nc
        seq_nuc_ORF2 = new_sequence_nc[1:-2]
        seq_nuc_ORF3 = new_sequence_nc[2:-1]
        seq_reversed=ReverseComplement2(seq_nuc_ORF1)
        seq_nuc_ORF4=seq_reversed
        seq_nuc_ORF5=seq_reversed[1:-2]
        seq_nuc_ORF6=seq_reversed[2:-1]

        LIST_6_ORF_nuc = [seq_nuc_ORF1, seq_nuc_ORF2, seq_nuc_ORF3,seq_nuc_ORF4,seq_nuc_ORF5,seq_nuc_ORF6]
        bash_of_aligned_nuc_seq_3ORF[fasta_name] = LIST_6_ORF_nuc     ### For each seq of the multialignment => give the 6 ORFs (in nuc)

        ## 1.4. ## Get the 3 ORFs (aa) for each sequence
        seq_prot_ORF1 = simply_get_ORF(seq_nuc_ORF1,bash_codeUniversel)                ### DEF 4 - Part 1 - ##
        seq_prot_ORF2 = simply_get_ORF(seq_nuc_ORF2,bash_codeUniversel)                ### DEF 4 - Part 1 - ##
        seq_prot_ORF3 = simply_get_ORF(seq_nuc_ORF3,bash_codeUniversel)                ### DEF 4 - Part 1 - ##
        seq_prot_ORF4 = simply_get_ORF(seq_nuc_ORF4,bash_codeUniversel)                ### DEF 4 - Part 1 - ##
        seq_prot_ORF5 = simply_get_ORF(seq_nuc_ORF5,bash_codeUniversel)                ### DEF 4 - Part 1 - ##
        seq_prot_ORF6 = simply_get_ORF(seq_nuc_ORF6,bash_codeUniversel)                ### DEF 4 - Part 1 - ##

        LIST_6_ORF_aa = [seq_prot_ORF1, seq_prot_ORF2, seq_prot_ORF3,seq_prot_ORF4,seq_prot_ORF5,seq_prot_ORF6]
        bash_of_aligned_aa_seq_3ORF[fasta_name] = LIST_6_ORF_aa     ### For each seq of the multialignment => give the 6 ORFs (in aa)

    ## 2 ## Test for the best ORF (Get the longuest segment in the alignment with no codon stop ... for each ORF ... the longuest should give the ORF)
    BEST_MAX = 0
    for i in [0,1,2,3,4,5]:   ### Test the 6 ORFs
        ORF_Aligned_aa = []
        ORF_Aligned_nuc = []


        ## 2.1 ## Get the alignment of sequence for a given ORF
        ## Compare the 1rst ORF between all sequence => list them in ORF_Aligned_aa // them do the same for the second ORF, and them the 3rd
        for fasta_name in bash_of_aligned_aa_seq_3ORF.keys():
            ORFsequence = bash_of_aligned_aa_seq_3ORF[fasta_name][i]
            aa_length = len(ORFsequence)
            ORF_Aligned_aa.append(ORFsequence)   ### List of all sequences in the ORF nb "i" =

        n = i+1

        for fasta_name in bash_of_aligned_nuc_seq_3ORF.keys():
            ORFsequence = bash_of_aligned_nuc_seq_3ORF[fasta_name][i]
            nuc_length = len(ORFsequence)
            ORF_Aligned_nuc.append(ORFsequence)   ### List of all sequences in the ORF nb "i" =

        ## 2.2 ## Get the list of sublist of positions whithout codon stop in the alignment
        ## For each ORF, now we have the list of sequences available (i.e. THE ALIGNMENT IN A GIVEN ORF)
        ## Next step is to get the longuest subsequence whithout stop
        ## We will explore the presence of stop "*" in each column of the alignment, and get the positions of the segments between the positions with "*"
        MAX_LENGTH = 0
        LONGUEST_SEGMENT_UNSTOPPED = ""
        j = 0 # Start from first position in alignment
        List_of_List_subsequences = []
        List_positions_subsequence = []
        while j < aa_length:
                column = []
                for seq in ORF_Aligned_aa:
                    column.append(seq[j])
                j = j+1
                if "*" in column:
                    List_of_List_subsequences.append(List_positions_subsequence) ## Add previous list of positions
                    List_positions_subsequence = []                              ## Re-initialyse list of positions
                else:
                    List_positions_subsequence.append(j)

        ## 2.3 ## Among all the sublists (separated by column with codon stop "*"), get the longuest one (BETTER SEGMENT for a given ORF)
        LONGUEST_SUBSEQUENCE_LIST_POSITION = []
        MAX=0
        for sublist in List_of_List_subsequences:
            if len(sublist) > MAX and len(sublist) > MINIMAL_CDS_LENGTH:
                MAX = len(sublist)
                LONGUEST_SUBSEQUENCE_LIST_POSITION = sublist

        ## 2.4. ## Test if the longuest subsequence start exactly at the beginning of the original sequence (i.e. means the ORF maybe truncated)
        if LONGUEST_SUBSEQUENCE_LIST_POSITION != []:
            if LONGUEST_SUBSEQUENCE_LIST_POSITION[0] == 0:
                CDS_maybe_truncated = 1
            else:
                CDS_maybe_truncated = 0
        else:
            CDS_maybe_truncated = 0


        ## 2.5 ## Test if this BETTER SEGMENT for a given ORF, is the better than the one for the other ORF (GET THE BEST ORF)
        ## Test whether it is the better ORF
        if MAX > BEST_MAX:
            BEST_MAX = MAX

        ## 4 ## Get the corresponding position (START/END of BEST CODING SEGMENT) for nucleotides alignment
        pos_MIN_nuc = pos_MIN_aa * 3
        pos_MAX_nuc = pos_MAX_aa * 3

        BESTORF_bash_aligned_nc_seq = {}
        BESTORF_bash_aligned_nc_seq_CODING = {}
        for fasta_name in bash_aligned_nc_seq.keys():
            seq = bash_of_aligned_nuc_seq_3ORF[fasta_name][index_BEST_ORF]
            seq_coding = seq[pos_MIN_nuc:pos_MAX_nuc]
            BESTORF_bash_aligned_nc_seq[fasta_name] = seq
            BESTORF_bash_aligned_nc_seq_CODING[fasta_name] = seq_coding

    ### Check whether their is a "M" or not, and if at least 1 "M" is present, that it is not in the last 50 aa
    ###########################################################################################################

    BESTORF_bash_of_aligned_aa_seq_CDS_with_M = {}
    BESTORF_bash_of_aligned_nuc_seq_CDS_with_M = {}

    Ortho = 0
    for fasta_name in BESTORF_bash_of_aligned_aa_seq_CODING.keys():
        seq_aa = BESTORF_bash_of_aligned_aa_seq_CODING[fasta_name]
        Ortho = detect_Methionine(seq_aa, Ortho)                                ### DEF6 ###

    if Ortho == 1:
        BESTORF_bash_of_aligned_aa_seq_CDS_with_M = BESTORF_bash_of_aligned_aa_seq_CODING
        BESTORF_bash_of_aligned_nuc_seq_CDS_with_M = BESTORF_bash_aligned_nc_seq_CODING

    ## CASE 2: in case the CDS is truncated, so the "M" is maybe missing:
    if Ortho == 0 and CDS_maybe_truncated == 1:
        BESTORF_bash_of_aligned_aa_seq_CDS_with_M = BESTORF_bash_of_aligned_aa_seq_CODING
        BESTORF_bash_of_aligned_nuc_seq_CDS_with_M = BESTORF_bash_aligned_nc_seq_CODING

    ## CASE 3: CDS not truncated AND no "M" found in good position (i.e. before the last 50 aa):
        ## => the 2 bash "CDS_with_M" are left empty ("{}")

    return(BESTORF_bash_aligned_nc_seq,  BESTORF_bash_aligned_nc_seq_CODING, BESTORF_bash_of_aligned_nuc_seq_CDS_with_M, BESTORF_bash_of_aligned_aa_seq, BESTORF_bash_of_aligned_aa_seq_CODING, BESTORF_bash_of_aligned_aa_seq_CDS_with_M)
##########################################################


##################################################################################################
###### DEF 5 : Detect all indices corresponding to all occurance of a substring in a string ######

    return listindex
######################################################


############################################################
###### DEF 6 : Detect if methionin in the aa sequence ######
############################################################
def detect_Methionine(seq_aa, Ortho):

    ln = len(seq_aa)
    nbre = sys.argv[2]
    CUTOFF_Last_50aa = ln - MINIMAL_CDS_LENGTH
    #Ortho = 0  ## means orthologs not found

    ## Find all indices of occurances of "M" in a string of aa
    list_indices = allindices(seq_aa, "M")                  ### DEF5 ###

    ## If some "M" are present, find whether the first "M" found is not in the 50 last aa (indice < CUTOFF_Last_50aa) ==> in this case: maybenot a CDS
    if list_indices != []:
        first_M = list_indices[0]
        if first_M < CUTOFF_Last_50aa:
            Ortho = 1  ## means orthologs found

    return(Ortho)
###################################

############################################################
###### DEF 7 : Reverse complement DNA sequence ######
###### Reference: http://crazyhottommy.blogspot.fr/2013/10/python-code-for-getting-reverse.html
############################################################


def ReverseComplement2(seq):

#######################
##### RUN RUN RUN #####
#######################
import string, os, time, re, zipfile, sys

infiles = sys.argv[1]
MINIMAL_CDS_LENGTH = int(sys.argv[3])  ## in aa number

## INPUT / OUTPUT
list_file = str.split(infiles,",")

### Get Universal Code
F2 = open(sys.argv[2], 'r')
bash_codeUniversel = code_universel(F2)  ### DEF2 ###
F2.close()

os.mkdir("04_BEST_ORF_nuc")
Path_OUT1 = "04_BEST_ORF_nuc"
os.mkdir("04_BEST_ORF_aa")
Path_OUT2 = "04_BEST_ORF_aa"

Path_OUT5 = "06_CDS_with_M_nuc"
os.mkdir("06_CDS_with_M_aa")
Path_OUT6 = "06_CDS_with_M_aa"




### Get the Bash corresponding to an alignment file in fasta format
count_file_processed = 0
count_file_with_CDS = 0
count_file_without_CDS = 0
count_file_with_CDS_plus_M = 0

for file in list_file:
    count_file_processed = count_file_processed + 1
    fasta_file_path = "./%s" %file
    bash_fasta = dico(fasta_file_path)   ### DEF 1 ###
    BESTORF_nuc, BESTORF_nuc_CODING, BESTORF_nuc_CDS_with_M, BESTORF_aa, BESTORF_aa_CODING, BESTORF_aa_CDS_with_M  = find_good_ORF_criteria_3(bash_fasta, bash_codeUniversel)   ### DEF 4 - PART 2 - ###

    ## a ## OUTPUT BESTORF_nuc
    if BESTORF_nuc != {}:
        count_file_with_CDS = count_file_with_CDS +1
        OUT1 = open("%s/%s" %(Path_OUT1,file), "w")

            OUT1.write("%s\n" %fasta_name)
            OUT1.write("%s\n" %seq)
        OUT1.close()
    else:
        count_file_without_CDS = count_file_without_CDS + 1


    ## b ## OUTPUT BESTORF_nuc_CODING  ===> THE MOST INTERESTING!!!
    if BESTORF_aa != {}:
        OUT2 = open("%s/%s" %(Path_OUT2,file), "w")
        for fasta_name in BESTORF_aa.keys():
            seq = BESTORF_aa[fasta_name]
            OUT2.write("%s\n" %fasta_name)
            OUT2.write("%s\n" %seq)
        OUT2.close()

    ## c ## OUTPUT BESTORF_aa
    if BESTORF_nuc_CODING != {}:
        OUT3 = open("%s/%s" %(Path_OUT3,file), "w")
        for fasta_name in BESTORF_nuc_CODING.keys():

        OUT4 = open("%s/%s" %(Path_OUT4,file), "w")
        for fasta_name in BESTORF_aa_CODING.keys():
            seq = BESTORF_aa_CODING[fasta_name]
            OUT4.write("%s\n" %fasta_name)
            OUT4.write("%s\n" %seq)
        OUT4.close()

    ## e ## OUTPUT BESTORF_nuc_CDS_with_M
    if BESTORF_nuc_CDS_with_M != {}:
        count_file_with_CDS_plus_M = count_file_with_CDS_plus_M + 1
        OUT5 = open("%s/%s" %(Path_OUT5,file), "w")
        for fasta_name in BESTORF_nuc_CDS_with_M.keys():
            seq = BESTORF_nuc_CDS_with_M[fasta_name]
            OUT5.write("%s\n" %fasta_name)
            OUT5.write("%s\n" %seq)
        OUT5.close()

    ## f ## OUTPUT BESTORF_aa_CDS_with_M
    if BESTORF_aa_CDS_with_M != {}:
        OUT6 = open("%s/%s" %(Path_OUT6,file), "w")
        for fasta_name in BESTORF_aa_CDS_with_M.keys():
            seq = BESTORF_aa_CDS_with_M[fasta_name]
            OUT6.write("%s\n" %fasta_name)
            OUT6.write("%s\n" %seq)
        OUT6.close()

    os.system("rm -rf %s" %file)

## Print
print "*************** CDS detection ***************"
print "\nFiles processed: %d" %count_file_processed
print "\tFiles with CDS: %d" %count_file_with_CDS
print "\t\tFiles with CDS plus M (codon start): %d" %count_file_with_CDS_plus_M
print "\tFiles without CDS: %d \n" %count_file_without_CDS
print ""