| esim4

## Wiki

The master copies of EMBOSS documentation are available at http://emboss.open-bio.org/wiki/Appdocs on the EMBOSS Wiki.

Please help by correcting and extending the Wiki pages.

## Function

Align an mRNA to a genomic DNA sequence

## Description

The original sim4 program was developed by Florea et al. for aligning cdna/est sequence to genomic sequences.

The program is available for download from http://globin.cse.psu.edu/

## Algorithm

Florea L, Hartzell G, Zhang Z, Rubin GM, Miller W. "A computer program for aligning a cDNA sequence with a genomic DNA sequence." Genome Res 1998 Sep;8(9):967-74

## Usage

Here is a sample session with esim4

    % esim4
    Align an mRNA to a genomic DNA sequence
    Input nucleotide sequence: tembl:h45989
    Genomic sequence: tembl:z69719
    Sim4 program output file [h45989.esim4]:

Go to the input files for this example
Go to the output files for this example

## Command line arguments

    Align an mRNA to a genomic DNA sequence
    Version: EMBOSS:6.2.0

       Standard (Mandatory) qualifiers:
      [-asequence]         sequence   Nucleotide sequence filename and optional
                                      format, or reference (input USA)
      [-bsequence]         seqall     Genomic sequence
      [-outfile]           outfile    [*.esim4] Sim4 program output file

       Additional (Optional) qualifiers (* if not always prompted):
       -word               integer    [12] Sets the word size (W) for blast hits.
                                      Default value: 12. (Any integer value)
       -extend             integer    [12] Sets the word extension termination
                                      limit (X) for the blast-like stage of the
                                      algorithm. Default value: 12. (Any integer
                                      value)
       -cutoff             integer    [3] Sets the cutoff (E) in range [3,10].
                                      (Integer up to 10)
       -useramsp           toggle     [N] False: esim4 calculates mspA, True:
                                      value from mspA command line argument.
    *  -amsp               integer    [16] MSP score threshold (K) for the first
                                      stage of the algorithm. (If this option is
                                      not specified, the threshold is computed
                                      from the lengths of the sequences, using
                                      statistical criteria.) For example, a good
                                      value for genomic sequences in the range of
                                      a few hundred Kb is 16. To avoid spurious
                                      matches, however, a larger value may be
                                      needed for longer sequences. (Any integer
                                      value)
       -userbmsp           toggle     [N] False: esim4 calculates mspB, True:
                                      value from mspB command line argument.
    *  -bmsp               integer    [12] Sets the threshold for the MSP scores
                                      (C) when aligning the as-yet-unmatched
                                      fragments, during the second stage of the
                                      algorithm. By default, the smaller of the
                                      constant 12 and a statistics-based threshold
                                      is chosen. (Any integer value)
       -weight             integer    [0] Weight value (H) (undocumented). 0 uses
                                      a default, >0 is a value (Any integer value)
       -diagonal           integer    [10] Bound (K) for the diagonal distance
                                      within consecutive MSPs in an exon. (Any
                                      integer value)
       -strand             menu       [both] This determines the strand of the
                                      genome (R) with which the mRNA will be
                                      aligned. The default value is 'both', in
                                      which case both strands of the genome are
                                      attempted. The other allowed modes are
                                      'forward' and 'reverse'. (Values: both (Both
                                      strands); forward (Forward strand only);
                                      reverse (Reverse strand only))
       -format             integer    [0] Sets the output format (A). Exon
                                      endpoints only (format=0), exon endpoints
                                      and boundaries of the coding region (CDS) in
                                      the genomic sequence, when specified for
                                      the input mRNA (-format=5), alignment text
                                      (-format=1), alignment in lav-block format
                                      (-format=2), or both exon endpoints and
                                      alignment text (-format=3 or -format=4). If
                                      a reverse complement match is found,
                                      -format=0,1,2,3,5 will give its position in
                                      the plus strand of the longer sequence and
                                      the minus strand of the shorter sequence.
                                      -format=4 will give its position in the plus
                                      strand of the first sequence (mRNA) and the
                                      minus strand of the second sequence
                                      (genome), regardless of which sequence is
                                      longer. The -format=5 option can be used
                                      with the S command line option to specify
                                      the endpoints of the CDS in the mRNA, and
                                      produces output in the exons file format
                                      required by PipMaker. (Integer from 0 to 5)
       -cliptails          boolean    [N] Trim poly-A tails (P). Specifies whether
                                      or not the program should report the
                                      fragment of the alignment containing the
                                      poly-A tail (if found). By default
                                      (-nocliptails) the alignment is displayed as
                                      computed. When this feature is enabled
                                      (-cliptails), sim4 will remove the poly-A
                                      tails and all format options will produce
                                      additional lav alignment headers.
       -smallexons         boolean    [N] Requests an additional search for small
                                      marginal exons (N) (N=1) guided by the
                                      splice-site recognition signals. This option
                                      can be used when a high accuracy match is
                                      expected. The default value is N=0,
                                      specifying no additional search.
       -[no]ambiguity      boolean    [Y] Controls the set of characters allowed
                                      in the input sequences (B). By default
                                      (-ambiguity), ambiguity characters
                                      (ABCDGHKMNRSTVWXY) are allowed. By
                                      specifying -noambiguity, the set of
                                      acceptable characters is restricted to
                                      A,C,G,T,N and X only.
       -cdsregion          string     Allows the user to specify the endpoints of
                                      the CDS in the input mRNA (S), with the
                                      syntax: -cdsregion=n1..n2. This option is
                                      only available with the -format=5 flag,
                                      which produces output in the format required
                                      by PipMaker. Alternatively, the CDS
                                      coordinates could appear in a construct
                                      CDS=n1..n2 in the FastA header of the mRNA
                                      sequence. When the second file is an mRNA
                                      database, the command line specification for
                                      the CDS will apply to the first sequence in
                                      the file only. (Any string)
       -aoffset            integer    [0] Undocumented (f) - some sort of offset
                                      in first sequence. (Any integer value)
       -boffset            integer    [0] Undocumented (F) - some sort of offset
                                      in second sequence. (Any integer value)
       -toa                integer    [0] Undocumented (t)- offset end of first
                                      sequence? (Any integer value)
       -tob                integer    [0] Undocumented (T) - offset end of second
                                      sequence? (Any integer value)

       Advanced (Unprompted) qualifiers: (none)
       Associated qualifiers:

       "-asequence" associated qualifiers
       -sbegin1            integer    Start of the sequence to be used
       -send1              integer    End of the sequence to be used
       -sreverse1          boolean    Reverse (if DNA)
       -sask1              boolean    Ask for begin/end/reverse
       -snucleotide1       boolean    Sequence is nucleotide
       -sprotein1          boolean    Sequence is protein
       -slower1            boolean    Make lower case
       -supper1            boolean    Make upper case
       -sformat1           string     Input sequence format
       -sdbname1           string     Database name
       -sid1               string     Entryname
       -ufo1               string     UFO features
       -fformat1           string     Features format
       -fopenfile1         string     Features file name

       "-bsequence" associated qualifiers
       -sbegin2            integer    Start of each sequence to be used
       -send2              integer    End of each sequence to be used
       -sreverse2          boolean    Reverse (if DNA)
       -sask2              boolean    Ask for begin/end/reverse
       -snucleotide2       boolean    Sequence is nucleotide
       -sprotein2          boolean    Sequence is protein
       -slower2            boolean    Make lower case
       -supper2            boolean    Make upper case
       -sformat2           string     Input sequence format
       -sdbname2           string     Database name
       -sid2               string     Entryname
       -ufo2               string     UFO features
       -fformat2           string     Features format
       -fopenfile2         string     Features file name

       "-outfile" associated qualifiers
       -odirectory3        string     Output directory

       General qualifiers:
       -auto               boolean    Turn off prompts
       -stdout             boolean    Write first file to standard output
       -filter             boolean    Read first file from standard input, write
                                      first file to standard output
       -options            boolean    Prompt for standard and additional values
       -debug              boolean    Write debug output to program.dbg
       -verbose            boolean    Report some/full command line options
       -help               boolean    Report command line options and exit. More
                                      information on associated and general
                                      qualifiers can be found with -help -verbose
       -warning            boolean    Report warnings
       -error              boolean    Report errors
       -fatal              boolean    Report fatal errors
       -die                boolean    Report dying program messages
       -version            boolean    Report version number and exit

Qualifier | Type | Description | Allowed values | Default
--- | --- | --- | --- | ---
Standard (Mandatory) qualifiers
[-asequence] (Parameter 1) | sequence | Nucleotide sequence filename and optional format, or reference (input USA) | Readable sequence | Required
[-bsequence] (Parameter 2) | seqall | Genomic sequence | Readable sequence(s) | Required
[-outfile] (Parameter 3) | outfile | Sim4 program output file | Output file | <*>.esim4
Additional (Optional) qualifiers
-word | integer | Sets the word size (W) for blast hits. Default value: 12. | Any integer value | 12
-extend | integer | Sets the word extension termination limit (X) for the blast-like stage of the algorithm. Default value: 12. | Any integer value | 12
-cutoff | integer | Sets the cutoff (E) in range [3,10]. | Integer up to 10 | 3
-useramsp | toggle | False: esim4 calculates mspA, True: value from mspA command line argument. | Toggle value Yes/No | No

-amsp

integer

MSP score threshold (K) for the first stage of the algorithm. (If this option is not specified, the threshold is computed from the lengths of the sequences, using statistical criteria.) For example, a good value for genomic sequences in the range of a few hundred Kb is 16. To avoid spurious matches, however, a larger value may be needed for longer sequences.

Any integer value

16

-userbmsp | toggle | False: esim4 calculates mspB, True: value from mspB command line argument. | Toggle value Yes/No | No
-bmsp | integer | Sets the threshold for the MSP scores (C) when aligning the as-yet-unmatched fragments, during the second stage of the algorithm. By default, the smaller of the constant 12 and a statistics-based threshold is chosen. | Any integer value | 12
-weight | integer | Weight value (H) (undocumented). 0 uses a default, >0 is a value | Any integer value | 0
-diagonal | integer | Bound (K) for the diagonal distance within consecutive MSPs in an exon. | Any integer value | 10

-strand

list

This determines the strand of the genome (R) with which the mRNA will be aligned. The default value is 'both', in which case both strands of the genome are attempted. The other allowed modes are 'forward' and 'reverse'.

both | (Both strands)
forward | (Forward strand only)
reverse | (Reverse strand only)

both

-format

integer

Sets the output format (A). Exon endpoints only (format=0), exon endpoints and boundaries of the coding region (CDS) in the genomic sequence, when specified for the input mRNA (-format=5), alignment text (-format=1), alignment in lav-block format (-format=2), or both exon endpoints and alignment text (-format=3 or -format=4). If a reverse complement match is found, -format=0,1,2,3,5 will give its position in the plus strand of the longer sequence and the minus strand of the shorter sequence. -format=4 will give its position in the plus strand of the first sequence (mRNA) and the minus strand of the second sequence (genome), regardless of which sequence is longer. The -format=5 option can be used with the S command line option to specify the endpoints of the CDS in the mRNA, and produces output in the exons file format required by PipMaker.

Integer from 0 to 5

0

-cliptails

boolean

Trim poly-A tails (P). Specifies whether or not the program should report the fragment of the alignment containing the poly-A tail (if found). By default (-nocliptails) the alignment is displayed as computed. When this feature is enabled (-cliptails), sim4 will remove the poly-A tails and all format options will produce additional lav alignment headers.

Boolean value Yes/No

No

-smallexons | boolean | Requests an additional search for small marginal exons (N) (N=1) guided by the splice-site recognition signals. This option can be used when a high accuracy match is expected. The default value is N=0, specifying no additional search. | Boolean value Yes/No | No
-[no]ambiguity | boolean | Controls the set of characters allowed in the input sequences (B). By default (-ambiguity), ambiguity characters (ABCDGHKMNRSTVWXY) are allowed. By specifying -noambiguity, the set of acceptable characters is restricted to A,C,G,T,N and X only. | Boolean value Yes/No | Yes

-cdsregion

string

Allows the user to specify the endpoints of the CDS in the input mRNA (S), with the syntax: -cdsregion=n1..n2. This option is only available with the -format=5 flag, which produces output in the format required by PipMaker. Alternatively, the CDS coordinates could appear in a construct CDS=n1..n2 in the FastA header of the mRNA sequence. When the second file is an mRNA database, the command line specification for the CDS will apply to the first sequence in the file only.

Any string

-aoffset | integer | Undocumented (f) - some sort of offset in first sequence. | Any integer value | 0
-boffset | integer | Undocumented (F) - some sort of offset in second sequence. | Any integer value | 0
-toa | integer | Undocumented (t)- offset end of first sequence? | Any integer value | 0
-tob | integer | Undocumented (T) - offset end of second sequence? | Any integer value | 0
Advanced (Unprompted) qualifiers
(none)
Associated qualifiers
"-asequence" associated sequence qualifiers
-sbegin1 -sbegin_asequence | integer | Start of the sequence to be used | Any integer value | 0
-send1 -send_asequence | integer | End of the sequence to be used | Any integer value | 0
-sreverse1 -sreverse_asequence | boolean | Reverse (if DNA) | Boolean value Yes/No | N
-sask1 -sask_asequence | boolean | Ask for begin/end/reverse | Boolean value Yes/No | N
-snucleotide1 -snucleotide_asequence | boolean | Sequence is nucleotide | Boolean value Yes/No | N
-sprotein1 -sprotein_asequence | boolean | Sequence is protein | Boolean value Yes/No | N
-slower1 -slower_asequence | boolean | Make lower case | Boolean value Yes/No | N
-supper1 -supper_asequence | boolean | Make upper case | Boolean value Yes/No | N
-sformat1 -sformat_asequence | string | Input sequence format | Any string |
-sdbname1 -sdbname_asequence | string | Database name | Any string |
-sid1 -sid_asequence | string | Entryname | Any string |
-ufo1 -ufo_asequence | string | UFO features | Any string |
-fformat1 -fformat_asequence | string | Features format | Any string |
-fopenfile1 -fopenfile_asequence | string | Features file name | Any string |
"-bsequence" associated seqall qualifiers
-sbegin2 -sbegin_bsequence | integer | Start of each sequence to be used | Any integer value | 0
-send2 -send_bsequence | integer | End of each sequence to be used | Any integer value | 0
-sreverse2 -sreverse_bsequence | boolean | Reverse (if DNA) | Boolean value Yes/No | N
-sask2 -sask_bsequence | boolean | Ask for begin/end/reverse | Boolean value Yes/No | N
-snucleotide2 -snucleotide_bsequence | boolean | Sequence is nucleotide | Boolean value Yes/No | N
-sprotein2 -sprotein_bsequence | boolean | Sequence is protein | Boolean value Yes/No | N
-slower2 -slower_bsequence | boolean | Make lower case | Boolean value Yes/No | N
-supper2 -supper_bsequence | boolean | Make upper case | Boolean value Yes/No | N
-sformat2 -sformat_bsequence | string | Input sequence format | Any string |
-sdbname2 -sdbname_bsequence | string | Database name | Any string |
-sid2 -sid_bsequence | string | Entryname | Any string |
-ufo2 -ufo_bsequence | string | UFO features | Any string |
-fformat2 -fformat_bsequence | string | Features format | Any string |
-fopenfile2 -fopenfile_bsequence | string | Features file name | Any string |
"-outfile" associated outfile qualifiers
-odirectory3 -odirectory_outfile | string | Output directory | Any string |
General qualifiers
-auto | boolean | Turn off prompts | Boolean value Yes/No | N
-stdout | boolean | Write first file to standard output | Boolean value Yes/No | N
-filter | boolean | Read first file from standard input, write first file to standard output | Boolean value Yes/No | N
-options | boolean | Prompt for standard and additional values | Boolean value Yes/No | N
-debug | boolean | Write debug output to program.dbg | Boolean value Yes/No | N
-verbose | boolean | Report some/full command line options | Boolean value Yes/No | Y
-help | boolean | Report command line options and exit. More information on associated and general qualifiers can be found with -help -verbose | Boolean value Yes/No | N
-warning | boolean | Report warnings | Boolean value Yes/No | Y
-error | boolean | Report errors | Boolean value Yes/No | Y
-fatal | boolean | Report fatal errors | Boolean value Yes/No | Y
-die | boolean | Report dying program messages | Boolean value Yes/No | Y
-version | boolean | Report version number and exit | Boolean value Yes/No | N

## Input file format

esim4 reads any normal sequence USAs.

### Input files for usage example

'tembl:h45989' is a sequence entry in the example nucleic acid database 'tembl'

### Database entry: tembl:h45989

    ID   H45989; SV 1; linear; mRNA; EST; HUM; 495 BP.
    XX
    AC   H45989;
    XX
    DT   18-NOV-1995 (Rel. 45, Created)
    DT   04-MAR-2000 (Rel. 63, Last updated, Version 2)
    XX
    DE   yo13c02.s1 Soares adult brain N2b5HB55Y Homo sapiens cDNA clone
    DE   IMAGE:177794 3', mRNA sequence.
    XX
    KW   EST.
    XX
    OS   Homo sapiens (human)
    OC   Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
    OC   Eutheria; Euarchontoglires; Primates; Haplorrhini; Catarrhini; Hominidae;
    OC   Homo.
    XX
    RN   [1]
    RP   1-495
    RA   Hillier L., Clark N., Dubuque T., Elliston K., Hawkins M., Holman M.,
    RA   Hultman M., Kucaba T., Le M., Lennon G., Marra M., Parsons J., Rifkin L.,
    RA   Rohlfing T., Soares M., Tan F., Trevaskis E., Waterston R., Williamson A.,
    RA   Wohldmann P., Wilson R.;
    RT   "The WashU-Merck EST Project";
    RL   Unpublished.
    XX
    DR   GDB; 3839990.
    DR   GDB; 4193257.
    DR   ImaGenes; ENSEp780A0214D.
    DR   ImaGenes; ENSEp780A044Q.
    DR   ImaGenes; HU3_p972A0639D.
    DR   ImaGenes; HU3_p972B1110Q.
    DR   ImaGenes; HU3_p983A0639D.
    DR   ImaGenes; HU4_p940A0622D.
    DR   ImaGenes; IMAGp956A0431Q.
    DR   ImaGenes; IMAGp998F03326Q.
    DR   ImaGenes; RZPDp1096A101D.
    DR   ImaGenes; RZPDp1096A191Q.
    DR   ImaGenes; RZPDp200A0214D.
    DR   UNILIB; 555; 300.
    XX
    CC   On May 8, 1995 this sequence version replaced gi:800819.
    CC   Contact: Wilson RK
    CC   Washington University School of Medicine
    CC   4444 Forest Park Parkway, Box 8501, St. Louis, MO 63108
    CC   Tel: 314 286 1800
    CC   Fax: 314 286 1810
    CC   Email: est@watson.wustl.edu
    CC   Insert Size: 544
    CC   High quality sequence stops: 265
    CC   Source: IMAGE Consortium, LLNL
    CC   This clone is available royalty-free through LLNL ; contact the
    CC   IMAGE Consortium (info@image.llnl.gov) for further information.
    CC   Possible reversed clone: polyT not found
    CC   Insert Length: 544   Std Error: 0.00
    CC   Seq primer: SP6
    CC   High quality sequence stop: 265.
    XX
    FH   Key             Location/Qualifiers
    FH
    FT   source          1..495
    FT                   /organism="Homo sapiens"
    FT                   /lab_host="DH10B (ampicillin resistant)"
    FT                   /mol_type="mRNA"
    FT                   /sex="Male"
    FT                   /dev_stage="55-year old"
    FT                   /clone_lib="Soares adult brain N2b5HB55Y"
    FT                   /clone="IMAGE:177794"
    FT                   /note="Organ: brain; Vector: pT7T3D (Pharmacia) with a
    FT                   modified polylinker; Site_1: Not I; Site_2: Eco RI; 1st
    FT                   strand cDNA was primed with a Not I - oligo(dT) primer [5'
    FT                   TGTTACCAATCTGAAGTGGGAGCGGCCGCGCTTTTTTTTTTTTTTTTTTT 3'],
    FT                   double-stranded cDNA was size selected, ligated to Eco RI
    FT                   adapters (Pharmacia), digested with Not I and cloned into
    FT                   the Not I and Eco RI sites of a modified pT7T3 vector
    FT                   (Pharmacia). Library went through one round of
    FT                   normalization to a Cot = 53. Library constructed by Bento
    FT                   Soares and M.Fatima Bonaldo. The adult brain RNA was
    FT                   provided by Dr. Donald H. Gilden. Tissue was acquired 17-18
    FT                   hours after death which occurred in consequence of a
    FT                   ruptured aortic aneurysm. RNA was prepared from a pool of
    FT                   tissues representing the following areas of the brain:
    FT                   frontal, parietal, temporal and occipital cortex from the
    FT                   left and right hemispheres, subcortical white matter, basal
    FT                   ganglia, thalamus, cerebellum, midbrain, pons and medulla."
    FT                   /db_xref="taxon:9606"
    FT                   /db_xref="UNILIB:555"
    XX
    SQ   Sequence 495 BP; 73 A; 135 C; 169 G; 104 T; 14 other;
         ccggnaagct cancttggac caccgactct cgantgnntc gccgcgggag ccggntggan        60
         aacctgagcg ggactggnag aaggagcaga gggaggcagc acccggcgtg acggnagtgt       120
         gtggggcact caggccttcc gcagtgtcat ctgccacacg gaaggcacgg ccacgggcag       180
         gggggtctat gatcttctgc atgcccagct ggcatggccc cacgtagagt ggnntggcgt       240
         ctcggtgctg gtcagcgaca cgttgtcctg gctgggcagg tccagctccc ggaggacctg       300
         gggcttcagc ttcccgtagc gctggctgca gtgacggatg ctcttgcgct gccatttctg       360
         ggtgctgtca ctgtccttgc tcactccaaa ccagttcggc ggtccccctg cggatggtct       420
         gtgttgatgg acgtttgggc tttgcagcac cggccgccga gttcatggtn gggtnaagag       480
         atttgggttt tttcn                                                        495
    //

### Database entry: tembl:z69719

    ID   Z69719; SV 1; linear; genomic DNA; STD; HUM; 33760 BP.
    XX
    AC   Z69719;
    XX
    DT   26-FEB-1996 (Rel. 46, Created)
    DT   13-JAN-2009 (Rel. 99, Last updated, Version 7)
    XX
    DE   Human DNA sequence from clone XX-CNFG9 on chromosome 16
    XX
    KW   C16orf33; HTG; POLR3K; RHBDF1.
    XX
    OS   Homo sapiens (human)
    OC   Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
    OC   Eutheria; Euarchontoglires; Primates; Haplorrhini; Catarrhini; Hominidae;
    OC   Homo.
    XX
    RN   [1]
    RP   1-33760
    RA   Kershaw J.;
    RT   ;
    RL   Submitted (09-JAN-2009) to the EMBL/GenBank/DDBJ databases.
    RL   Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK.
    RL   E-mail enquiries: vega@sanger.ac.uk Clone requests: Geneservice
    RL   (http://www.geneservice.co.uk/) and BACPAC Resources
    RL   (http://bacpac.chori.org/)
    XX
    DR   EMBL-CON; GL000124.
    DR   EMBL-JOIN; Z69720.
    DR   GDB; 11502921.
    XX
    CC   -------------- Genome Center
    CC   Center: Wellcome Trust Sanger Institute
    CC   Center code: SC
    CC   Web site: http://www.sanger.ac.uk
    CC   Contact: vega@sanger.ac.uk
    CC   --------------
    CC
    CC   This sequence was finished as follows unless otherwise noted: all regions
    CC   were either double-stranded or sequenced with an alternate chemistry or
    CC   covered by high quality data (i.e., phred quality >= 30); an attempt was
    CC   made to resolve all sequencing problems, such as compressions and repeats;
    CC   all regions were covered by at least one subclone; and the assembly was
    CC   confirmed by restriction digest, except on the rare occasion of the clone
    CC   being a YAC.
    CC
    CC   The following abbreviations are used to associate primary accession
    CC   numbers given in the feature table with their source databases:
    CC   Em:, EMBL; Sw:, SWISSPROT; Tr:, TREMBL; Wp:, WORMPEP;
    CC   Information on the WORMPEP database can be found at
    CC   http://www.sanger.ac.uk/Projects/C_elegans/wormpep

      [Part of this file has been deleted for brevity]

         gagacagcag agtgctcagc tcatgaagga ggcaccagcc gccatgcctc tacatccagg     30840
         tctcctgggg ttcccacctc cacaaaaacc cccactgcta ggagtgcagg caggagggga     30900
         cctgagaacc gacagttata ggtcctgcgg gtgggcagtg ctgggtgttc tggtctgccc     30960
         cacccctgtg tgcctagatc cccatctggg cctcaagtgg gtgggattcc aaaggaagag     31020
         ccggagtagg cgtggggagg ggcaggccca ggctggacaa agagtctggc cagggagcgg     31080
         cacattgccc tcccagagac agtggctcag tgtccaggcc ttccccaggc gcacagtggg     31140
         ctcttgttcc cagaaagccc ctcgggggga tccaaacagt gtctccccca ccccgctgac     31200
         ccctcagtgt atggggaaac cgtggcccac ggaaggcctc actgcctggg gtcacacagc     31260
         atctgagtca ctgcagcagc ctcacagctg ccagcccagg cccagcccca tcaggagaca     31320
         cccaaagcca cagtgcatcc caggaccagc tgggggggct gcgggcagga ctctcgatga     31380
         ggctgaggga cgaggagggt caagggagcc actggcgcca tgcatgctga cgtcccctct     31440
         ggctgcctgc agagcctggt gtggaagggc tgagtggggg atggtggaga gtcctgttaa     31500
         ctcaggtttc tgctctgggg atgtctgggc acccatcaag ctggccgcgt gcacaggtgc     31560
         agggagagcc agaaagcagg agccgatgca gggaggccac tggggacagc ccaggctgat     31620
         gcttgggccc catgtgtctc caccacctac aaccctaagc aagcctcagc tttcccatct     31680
         ggaaatcagg ggtcacagca gtgcctggca cagtagcagc ggctgactcc atcacagggt     31740
         ggtgtagcct gtgggtactt ggcactctct gaggggcagg agctgggggg tgaaaggacc     31800
         ctagagcata tgcaacaaga gggcagccct ggggacacct ggggacagaa ccctccaaag     31860
         gtgtcgagtt tgggaagaga ctagagagaa gctctggcca gtccaggcat agacagtggc     31920
         cacagccagt ggagagctgc atcctcaggt gtgagcagca accacctctg tactcaggcc     31980
         tgccctgcac actcacagga ccatgctggc agggacaact ggcggcggag ttgactgcca     32040
         accccggggc cagaaccatc aagcctgggc tctgctccgc ccaaggaact gcctgctgcc     32100
         gaggtcagct ggagcaaggg gcctcacccc gggacacctt cccagacgtg tcctcagctc     32160
         acatgagcct catcccaggg ggatgtggct cctccagcat ccccacccac acgctgctct     32220
         ctgaccctca gtcttctgtt tgactcctaa tctgaagctc aatcctagat ctcccttgag     32280
         aagggggtca ccagctgtct ggcagcccag cctccaggtc ttctggatta atgaagggaa     32340
         agtcacctgg cctctctgcc ttgtctatta atggcatcat gctgagaatg atatttgcta     32400
         ggccctttgc aaaccccaaa gtgctcttca accctcccag tgaagcctct tcttttctgt     32460
         ggaagaaatg aggttcaggg tggagcaggg caggcctgag acctttgcag ggttctctcc     32520
         aggtccccag caggacagac tggcaccctg cctcccctca tcaccctaga caaggagaca     32580
         gaacaagagg ttccctgcta caggccatct gtgagggaag ccgccctagg gcctgtagac     32640
         acaggaatcc ctgaggacct gacctgtgag ggtagtgcac aaaggggcca gcacttggca     32700
         ggaggggggg gggcactgcc ccaaggctca gctagcaaat gtggcacagg ggtcaccaga     32760
         gctaaacccc tgactcagtt gggtctgaca ggggctgaca tggcagacac acccaggaat     32820
         caggggacac caagtgcagc tcagggcacc tgtccaggcc acacagtcag aaaggggatg     32880
         gcagcaagga cttagctaca ctagattctg ggggtaaact gcctggtatg ctggtcactg     32940
         ctagtcccca gtctggagtc tagctgggtc tcaggagtta ggcgaaaaca ccctccccag     33000
         gctgcaggtg ggagaggccc acatcccctg cacacgtctg gccagaggac agatgggcag     33060
         cccagtcacc agtcagagcc ctccagaggt gtccctgact gaccctacac acatgcaccc     33120
         aggtgcccag gcacccttgg gctcagcaac cctgcaaccc cctcccagga cccaccagaa     33180
         gcaggatagg actagagagg ccacaggagg gaaaccaagt cagagcagaa atggcttcgg     33240
         tcctcagcag cctggctcag cttcctcaaa ccagatcctg actgatcaca ctggtctgtc     33300
         taacccctgg gaggggtcct ctgtatccat cttacagata aggaaactga ggctcagaga     33360
         agcccatcac tgcctaaggt cccagggcct ataagggagc tcaaagcctt gggccaggtc     33420
         tgcccaggag ctgcagtgga agggaccctg tctgcagacc cccagaagac aaggcagacc     33480
         acctgggttc ttcagccttg tggctgtgga cggctgtcag acccttctaa gaccccttgc     33540
         cacctgctcc atcaggggca tctcagttga agaaggaagg actcaccccc aaaatcgtcc     33600
         aactcagaaa aaaaggcaga agccaaggaa tccaatcact gggcaaaatg tgatcctggc     33660
         acagacactg aggtggggga actggagccg gtgtggcgga ggccctcaca gccaagagca     33720
         actgggggtg ccctgggcag ggactgtagc tgggaagatc                           33760
    //

## Output file format

esim4 outputs a graph to the specified graphics device. outputs a report format file. The default format is ...

### Output files for usage example

### File: h45989.esim4

    seq1 = H45989, 495 bp
    seq2 = Z69719 (Z69719), 33760 bp

    1-193  (25686-25874)   92% <-
    194-407  (26279-26492)   98% <-
    408-487  (27391-27469)   88%

## Data files

## Notes

None.

## References

None.

## Warnings

None.

## Diagnostic Error Messages

None.

## Exit status

It always exits with status 0.

## Known bugs

None.

## See also

Program name | Description
--- | ---
est2genome | Align EST sequences to genomic DNA sequence
needle | Needleman-Wunsch global alignment of two sequences
needleall | Many-to-many pairwise alignments of two sequence sets
stretcher | Needleman-Wunsch rapid global alignment of two sequences

## Author(s)

This program is an EMBOSS conversion of a program written by Liliana Florea as part of the SIM4 package.

Although we take every care to ensure that the results of the EMBOSS version are identical to those from the original package, we recommend that you check your inputs give the same results in both versions before publication.

Please report all bugs in the EMBOSS version to the EMBOSS bug team, not to the original author.

## History

## Target users

This program is intended to be used by everyone and everything, from naive users to embedded scripts.