.. _`chapter:seq_objects`:

Sequence objects
================

Biological sequences are arguably the central object in Bioinformatics,
and in this chapter we’ll introduce the Biopython mechanism for dealing
with sequences, the ``Seq`` object.
Chapter :ref:`chapter:seq_annot` will introduce the
related ``SeqRecord`` object, which combines the sequence information
with any annotation, used again in
Chapter :ref:`chapter:seqio` for Sequence Input/Output.

Sequences are essentially strings of letters like ``AGTACACTGGT``, which
seems very natural since this is the most common way that sequences are
seen in biological file formats.

The most important difference between ``Seq`` objects and standard
Python strings is they have different methods. Although the ``Seq``
object supports many of the same methods as a plain string, its
``translate()`` method differs by doing biological translation, and
there are also additional biologically relevant methods like
``reverse_complement()``.

Sequences act like strings
--------------------------

In most ways, we can deal with Seq objects as if they were normal Python
strings, for example getting the length, or iterating over the elements:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> my_seq = Seq("GATCG")
   >>> for index, letter in enumerate(my_seq):
   ...     print("%i %s" % (index, letter))
   ...
   0 G
   1 A
   2 T
   3 C
   4 G
   >>> print(len(my_seq))
   5

You can access elements of the sequence in the same way as for strings
(but remember, Python counts from zero!):

.. cont-doctest

.. code:: pycon

   >>> print(my_seq[0])  # first letter
   G
   >>> print(my_seq[2])  # third letter
   T
   >>> print(my_seq[-1])  # last letter
   G

The ``Seq`` object has a ``.count()`` method, just like a string. Note
that this means that like a Python string, this gives a
*non-overlapping* count:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> "AAAA".count("AA")
   2
   >>> Seq("AAAA").count("AA")
   2

For some biological uses, you may actually want an overlapping count
(i.e. :math:`3` in this trivial example). When searching for single
letters, this makes no difference:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> my_seq = Seq("GATCGATGGGCCTATATAGGATCGAAAATCGC")
   >>> len(my_seq)
   32
   >>> my_seq.count("G")
   9
   >>> 100 * (my_seq.count("G") + my_seq.count("C")) / len(my_seq)
   46.875

While you could use the above snippet of code to calculate a GC%, note
that the ``Bio.SeqUtils`` module has several GC functions already built.
For example:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> from Bio.SeqUtils import gc_fraction
   >>> my_seq = Seq("GATCGATGGGCCTATATAGGATCGAAAATCGC")
   >>> gc_fraction(my_seq)
   0.46875

Note that using the ``Bio.SeqUtils.gc_fraction()`` function should
automatically cope with mixed case sequences and the ambiguous
nucleotide S which means G or C.

Also note that just like a normal Python string, the ``Seq`` object is
in some ways “read-only”. If you need to edit your sequence, for example
simulating a point mutation, look at the
Section :ref:`sec:mutable-seq` below which talks about the
``MutableSeq`` object.

Slicing a sequence
------------------

A more complicated example, let’s get a slice of the sequence:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> my_seq = Seq("GATCGATGGGCCTATATAGGATCGAAAATCGC")
   >>> my_seq[4:12]
   Seq('GATGGGCC')

Note that ‘Seq‘ objects follow the usual indexing conventions for Python
strings, with the first element of the sequence numbered 0. When you do
a slice the first item is included (i.e. 4 in this case) and the last is
excluded (12 in this case).

Also like a Python string, you can do slices with a start, stop and
*stride* (the step size, which defaults to one). For example, we can get
the first, second and third codon positions of this DNA sequence:

.. cont-doctest

.. code:: pycon

   >>> my_seq[0::3]
   Seq('GCTGTAGTAAG')
   >>> my_seq[1::3]
   Seq('AGGCATGCATC')
   >>> my_seq[2::3]
   Seq('TAGCTAAGAC')

Another stride trick you might have seen with a Python string is the use
of a -1 stride to reverse the string. You can do this with a ``Seq``
object too:

.. cont-doctest

.. code:: pycon

   >>> my_seq[::-1]
   Seq('CGCTAAAAGCTAGGATATATCCGGGTAGCTAG')

.. _`sec:seq-to-string`:

Turning Seq objects into strings
--------------------------------

If you really do just need a plain string, for example to write to a
file, or insert into a database, then this is very easy to get:

.. cont-doctest

.. code:: pycon

   >>> str(my_seq)
   'GATCGATGGGCCTATATAGGATCGAAAATCGC'

Since calling ``str()`` on a ``Seq`` object returns the full sequence as
a string, you often don’t actually have to do this conversion
explicitly. Python does this automatically in the print function:

.. cont-doctest

.. code:: pycon

   >>> print(my_seq)
   GATCGATGGGCCTATATAGGATCGAAAATCGC

You can also use the ``Seq`` object directly with a ``%s`` placeholder
when using the Python string formatting or interpolation operator
(``%``):

.. cont-doctest

.. code:: pycon

   >>> fasta_format_string = ">Name\n%s\n" % my_seq
   >>> print(fasta_format_string)
   >Name
   GATCGATGGGCCTATATAGGATCGAAAATCGC
   <BLANKLINE>

This line of code constructs a simple FASTA format record (without
worrying about line wrapping).
Section :ref:`sec:SeqRecord-format` describes a
neat way to get a FASTA formatted string from a ``SeqRecord`` object,
while the more general topic of reading and writing FASTA format
sequence files is covered in
Chapter :ref:`chapter:seqio`.

Concatenating or adding sequences
---------------------------------

Two ``Seq`` objects can be concatenated by adding them:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> seq1 = Seq("ACGT")
   >>> seq2 = Seq("AACCGG")
   >>> seq1 + seq2
   Seq('ACGTAACCGG')

Biopython does not check the sequence contents and will not raise an
exception if for example you concatenate a protein sequence and a DNA
sequence (which is likely a mistake):

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> protein_seq = Seq("EVRNAK")
   >>> dna_seq = Seq("ACGT")
   >>> protein_seq + dna_seq
   Seq('EVRNAKACGT')

You may often have many sequences to add together, which can be done
with a for loop like this:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> list_of_seqs = [Seq("ACGT"), Seq("AACC"), Seq("GGTT")]
   >>> concatenated = Seq("")
   >>> for s in list_of_seqs:
   ...     concatenated += s
   ...
   >>> concatenated
   Seq('ACGTAACCGGTT')

Like Python strings, Biopython ``Seq`` also has a ``.join`` method:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> contigs = [Seq("ATG"), Seq("ATCCCG"), Seq("TTGCA")]
   >>> spacer = Seq("N" * 10)
   >>> spacer.join(contigs)
   Seq('ATGNNNNNNNNNNATCCCGNNNNNNNNNNTTGCA')

Changing case
-------------

Python strings have very useful ``upper`` and ``lower`` methods for
changing the case. For example,

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> dna_seq = Seq("acgtACGT")
   >>> dna_seq
   Seq('acgtACGT')
   >>> dna_seq.upper()
   Seq('ACGTACGT')
   >>> dna_seq.lower()
   Seq('acgtacgt')

These are useful for doing case insensitive matching:

.. cont-doctest

.. code:: pycon

   >>> "GTAC" in dna_seq
   False
   >>> "GTAC" in dna_seq.upper()
   True

.. _`sec:seq-reverse-complement`:

Nucleotide sequences and (reverse) complements
----------------------------------------------

For nucleotide sequences, you can easily obtain the complement or
reverse complement of a ``Seq`` object using its built-in methods:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> my_seq = Seq("GATCGATGGGCCTATATAGGATCGAAAATCGC")
   >>> my_seq
   Seq('GATCGATGGGCCTATATAGGATCGAAAATCGC')
   >>> my_seq.complement()
   Seq('CTAGCTACCCGGATATATCCTAGCTTTTAGCG')
   >>> my_seq.reverse_complement()
   Seq('GCGATTTTCGATCCTATATAGGCCCATCGATC')

As mentioned earlier, an easy way to just reverse a ``Seq`` object (or a
Python string) is slice it with -1 step:

.. cont-doctest

.. code:: pycon

   >>> my_seq[::-1]
   Seq('CGCTAAAAGCTAGGATATATCCGGGTAGCTAG')

If you do accidentally end up trying to do something weird like taking
the (reverse) complement of a protein sequence, the results are
biologically meaningless:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> protein_seq = Seq("EVRNAK")
   >>> protein_seq.complement()
   Seq('EBYNTM')

Here the letter “E” is not a valid IUPAC ambiguity code for nucleotides,
so was not complemented. However, “V” means “A”, “C” or “G” and has
complement “B“, and so on.

The example in
Section :ref:`sec:SeqIO-reverse-complement`
combines the ``Seq`` object’s reverse complement method with
``Bio.SeqIO`` for sequence input/output.

Transcription
-------------

Before talking about transcription, I want to try to clarify the strand
issue. Consider the following (made up) stretch of double stranded DNA
which encodes a short peptide:

.. math::

   \begin{gathered}
       \text{DNA coding strand (aka Crick strand, strand } +1 \text{)} \\
       \text{5'} \qquad \texttt{ATGGCCATTGTAATGGGCCGCTGAAAGGGTGCCCGATAG} \qquad \text{3'} \\
       \texttt{|||||||||||||||||||||||||||||||||||||||} \\
       \text{3'} \qquad \texttt{TACCGGTAACATTACCCGGCGACTTTCCCACGGGCTATC} \qquad \text{5'} \\
       \text{DNA template strand (aka Watson strand, strand } -1 \text{)}
   \end{gathered}

Transcription of this DNA sequence produces the following RNA sequence:

.. math::

   \begin{gathered}
       \text{5'} \qquad \texttt{AUGGCCAUUGUAAUGGGCCGCUGAAAGGGUGCCCGAUAG} \qquad \text{3'} \\
       \text{Single-stranded messenger RNA}
   \end{gathered}

The actual biological transcription process works from the template
strand, doing a reverse complement (TCAG :math:`\rightarrow` CUGA) to
give the mRNA. However, in Biopython and bioinformatics in general, we
typically work directly with the coding strand because this means we can
get the mRNA sequence just by switching T :math:`\rightarrow` U.

Now let’s actually get down to doing a transcription in Biopython.
First, let’s create ``Seq`` objects for the coding and template DNA
strands:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> coding_dna = Seq("ATGGCCATTGTAATGGGCCGCTGAAAGGGTGCCCGATAG")
   >>> coding_dna
   Seq('ATGGCCATTGTAATGGGCCGCTGAAAGGGTGCCCGATAG')
   >>> template_dna = coding_dna.reverse_complement()
   >>> template_dna
   Seq('CTATCGGGCACCCTTTCAGCGGCCCATTACAATGGCCAT')

These should match the figure above - remember by convention nucleotide
sequences are normally read from the 5’ to 3’ direction, while in the
figure the template strand is shown reversed.

Now let’s transcribe the coding strand into the corresponding mRNA,
using the ``Seq`` object’s built-in ``transcribe`` method:

.. cont-doctest

.. code:: pycon

   >>> coding_dna
   Seq('ATGGCCATTGTAATGGGCCGCTGAAAGGGTGCCCGATAG')
   >>> messenger_rna = coding_dna.transcribe()
   >>> messenger_rna
   Seq('AUGGCCAUUGUAAUGGGCCGCUGAAAGGGUGCCCGAUAG')

As you can see, all this does is to replace T by U.

If you do want to do a true biological transcription starting with the
template strand, then this becomes a two-step process:

.. cont-doctest

.. code:: pycon

   >>> template_dna.reverse_complement().transcribe()
   Seq('AUGGCCAUUGUAAUGGGCCGCUGAAAGGGUGCCCGAUAG')

The ``Seq`` object also includes a back-transcription method for going
from the mRNA to the coding strand of the DNA. Again, this is a simple U
:math:`\rightarrow` T substitution:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> messenger_rna = Seq("AUGGCCAUUGUAAUGGGCCGCUGAAAGGGUGCCCGAUAG")
   >>> messenger_rna
   Seq('AUGGCCAUUGUAAUGGGCCGCUGAAAGGGUGCCCGAUAG')
   >>> messenger_rna.back_transcribe()
   Seq('ATGGCCATTGTAATGGGCCGCTGAAAGGGTGCCCGATAG')

*Note:* The ``Seq`` object’s ``transcribe`` and ``back_transcribe``
methods were added in Biopython 1.49. For older releases you would have
to use the ``Bio.Seq`` module’s functions instead, see
Section :ref:`sec:seq-module-functions`.

.. _`sec:translation`:

Translation
-----------

Sticking with the same example discussed in the transcription section
above, now let’s translate this mRNA into the corresponding protein
sequence - again taking advantage of one of the ``Seq`` object’s
biological methods:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> messenger_rna = Seq("AUGGCCAUUGUAAUGGGCCGCUGAAAGGGUGCCCGAUAG")
   >>> messenger_rna
   Seq('AUGGCCAUUGUAAUGGGCCGCUGAAAGGGUGCCCGAUAG')
   >>> messenger_rna.translate()
   Seq('MAIVMGR*KGAR*')

You can also translate directly from the coding strand DNA sequence:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> coding_dna = Seq("ATGGCCATTGTAATGGGCCGCTGAAAGGGTGCCCGATAG")
   >>> coding_dna
   Seq('ATGGCCATTGTAATGGGCCGCTGAAAGGGTGCCCGATAG')
   >>> coding_dna.translate()
   Seq('MAIVMGR*KGAR*')

You should notice in the above protein sequences that in addition to the
end stop character, there is an internal stop as well. This was a
deliberate choice of example, as it gives an excuse to talk about some
optional arguments, including different translation tables (Genetic
Codes).

The translation tables available in Biopython are based on those `from
the NCBI <https://www.ncbi.nlm.nih.gov/Taxonomy/Utils/wprintgc.cgi>`__
(see the next section of this tutorial). By default, translation will
use the *standard* genetic code (NCBI table id 1). Suppose we are
dealing with a mitochondrial sequence. We need to tell the translation
function to use the relevant genetic code instead:

.. cont-doctest

.. code:: pycon

   >>> coding_dna.translate(table="Vertebrate Mitochondrial")
   Seq('MAIVMGRWKGAR*')

You can also specify the table using the NCBI table number which is
shorter, and often included in the feature annotation of GenBank files:

.. cont-doctest

.. code:: pycon

   >>> coding_dna.translate(table=2)
   Seq('MAIVMGRWKGAR*')

Now, you may want to translate the nucleotides up to the first in frame
stop codon, and then stop (as happens in nature):

.. cont-doctest

.. code:: pycon

   >>> coding_dna.translate()
   Seq('MAIVMGR*KGAR*')
   >>> coding_dna.translate(to_stop=True)
   Seq('MAIVMGR')
   >>> coding_dna.translate(table=2)
   Seq('MAIVMGRWKGAR*')
   >>> coding_dna.translate(table=2, to_stop=True)
   Seq('MAIVMGRWKGAR')

Notice that when you use the ``to_stop`` argument, the stop codon itself
is not translated - and the stop symbol is not included at the end of
your protein sequence.

You can even specify the stop symbol if you don’t like the default
asterisk:

.. cont-doctest

.. code:: pycon

   >>> coding_dna.translate(table=2, stop_symbol="@")
   Seq('MAIVMGRWKGAR@')

Now, suppose you have a complete coding sequence CDS, which is to say a
nucleotide sequence (e.g. mRNA – after any splicing) which is a whole
number of codons (i.e. the length is a multiple of three), commences
with a start codon, ends with a stop codon, and has no internal in-frame
stop codons. In general, given a complete CDS, the default translate
method will do what you want (perhaps with the ``to_stop`` option).
However, what if your sequence uses a non-standard start codon? This
happens a lot in bacteria – for example the gene yaaX in *E. coli*
K12:

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> gene = Seq(
   ...     "GTGAAAAAGATGCAATCTATCGTACTCGCACTTTCCCTGGTTCTGGTCGCTCCCATGGCA"
   ...     "GCACAGGCTGCGGAAATTACGTTAGTCCCGTCAGTAAAATTACAGATAGGCGATCGTGAT"
   ...     "AATCGTGGCTATTACTGGGATGGAGGTCACTGGCGCGACCACGGCTGGTGGAAACAACAT"
   ...     "TATGAATGGCGAGGCAATCGCTGGCACCTACACGGACCGCCGCCACCGCCGCGCCACCAT"
   ...     "AAGAAAGCTCCTCATGATCATCACGGCGGTCATGGTCCAGGCAAACATCACCGCTAA"
   ... )
   >>> gene.translate(table="Bacterial")
   Seq('VKKMQSIVLALSLVLVAPMAAQAAEITLVPSVKLQIGDRDNRGYYWDGGHWRDH...HR*',
   ProteinAlpabet())
   >>> gene.translate(table="Bacterial", to_stop=True)
   Seq('VKKMQSIVLALSLVLVAPMAAQAAEITLVPSVKLQIGDRDNRGYYWDGGHWRDH...HHR')

In the bacterial genetic code ``GTG`` is a valid start codon, and while
it does *normally* encode Valine, if used as a start codon it should be
translated as methionine. This happens if you tell Biopython your
sequence is a complete CDS:

.. code:: pycon

   >>> gene.translate(table="Bacterial", cds=True)
   Seq('MKKMQSIVLALSLVLVAPMAAQAAEITLVPSVKLQIGDRDNRGYYWDGGHWRDH...HHR')

In addition to telling Biopython to translate an alternative start codon
as methionine, using this option also makes sure your sequence really is
a valid CDS (you’ll get an exception if not).

The example in Section :ref:`sec:SeqIO-translate`
combines the ``Seq`` object’s translate method with ``Bio.SeqIO`` for
sequence input/output.

Translation Tables
------------------

In the previous sections we talked about the ``Seq`` object translation
method (and mentioned the equivalent function in the ``Bio.Seq`` module
– see Section :ref:`sec:seq-module-functions`). Internally these
use codon table objects derived from the NCBI information at
ftp://ftp.ncbi.nlm.nih.gov/entrez/misc/data/gc.prt, also shown on
https://www.ncbi.nlm.nih.gov/Taxonomy/Utils/wprintgc.cgi in a much more
readable layout.

As before, let’s just focus on two choices: the Standard translation
table, and the translation table for Vertebrate Mitochondrial DNA.

.. doctest

.. code:: pycon

   >>> from Bio.Data import CodonTable
   >>> standard_table = CodonTable.unambiguous_dna_by_name["Standard"]
   >>> mito_table = CodonTable.unambiguous_dna_by_name["Vertebrate Mitochondrial"]

Alternatively, these tables are labeled with ID numbers 1 and 2,
respectively:

.. cont-doctest

.. code:: pycon

   >>> from Bio.Data import CodonTable
   >>> standard_table = CodonTable.unambiguous_dna_by_id[1]
   >>> mito_table = CodonTable.unambiguous_dna_by_id[2]

You can compare the actual tables visually by printing them:

.. code:: pycon

   >>> print(standard_table)
   Table 1 Standard, SGC0

     |  T      |  C      |  A      |  G      |
   --+---------+---------+---------+---------+--
   T | TTT F   | TCT S   | TAT Y   | TGT C   | T
   T | TTC F   | TCC S   | TAC Y   | TGC C   | C
   T | TTA L   | TCA S   | TAA Stop| TGA Stop| A
   T | TTG L(s)| TCG S   | TAG Stop| TGG W   | G
   --+---------+---------+---------+---------+--
   C | CTT L   | CCT P   | CAT H   | CGT R   | T
   C | CTC L   | CCC P   | CAC H   | CGC R   | C
   C | CTA L   | CCA P   | CAA Q   | CGA R   | A
   C | CTG L(s)| CCG P   | CAG Q   | CGG R   | G
   --+---------+---------+---------+---------+--
   A | ATT I   | ACT T   | AAT N   | AGT S   | T
   A | ATC I   | ACC T   | AAC N   | AGC S   | C
   A | ATA I   | ACA T   | AAA K   | AGA R   | A
   A | ATG M(s)| ACG T   | AAG K   | AGG R   | G
   --+---------+---------+---------+---------+--
   G | GTT V   | GCT A   | GAT D   | GGT G   | T
   G | GTC V   | GCC A   | GAC D   | GGC G   | C
   G | GTA V   | GCA A   | GAA E   | GGA G   | A
   G | GTG V   | GCG A   | GAG E   | GGG G   | G
   --+---------+---------+---------+---------+--

and:

.. code:: pycon

   >>> print(mito_table)
   Table 2 Vertebrate Mitochondrial, SGC1

     |  T      |  C      |  A      |  G      |
   --+---------+---------+---------+---------+--
   T | TTT F   | TCT S   | TAT Y   | TGT C   | T
   T | TTC F   | TCC S   | TAC Y   | TGC C   | C
   T | TTA L   | TCA S   | TAA Stop| TGA W   | A
   T | TTG L   | TCG S   | TAG Stop| TGG W   | G
   --+---------+---------+---------+---------+--
   C | CTT L   | CCT P   | CAT H   | CGT R   | T
   C | CTC L   | CCC P   | CAC H   | CGC R   | C
   C | CTA L   | CCA P   | CAA Q   | CGA R   | A
   C | CTG L   | CCG P   | CAG Q   | CGG R   | G
   --+---------+---------+---------+---------+--
   A | ATT I(s)| ACT T   | AAT N   | AGT S   | T
   A | ATC I(s)| ACC T   | AAC N   | AGC S   | C
   A | ATA M(s)| ACA T   | AAA K   | AGA Stop| A
   A | ATG M(s)| ACG T   | AAG K   | AGG Stop| G
   --+---------+---------+---------+---------+--
   G | GTT V   | GCT A   | GAT D   | GGT G   | T
   G | GTC V   | GCC A   | GAC D   | GGC G   | C
   G | GTA V   | GCA A   | GAA E   | GGA G   | A
   G | GTG V(s)| GCG A   | GAG E   | GGG G   | G
   --+---------+---------+---------+---------+--

You may find these following properties useful – for example if you are
trying to do your own gene finding:

.. cont-doctest

.. code:: pycon

   >>> mito_table.stop_codons
   ['TAA', 'TAG', 'AGA', 'AGG']
   >>> mito_table.start_codons
   ['ATT', 'ATC', 'ATA', 'ATG', 'GTG']
   >>> mito_table.forward_table["ACG"]
   'T'

.. _`sec:seq-comparison`:

Comparing Seq objects
---------------------

Sequence comparison is actually a very complicated topic, and there is
no easy way to decide if two sequences are equal. The basic problem is
the meaning of the letters in a sequence are context dependent - the
letter “A” could be part of a DNA, RNA or protein sequence. Biopython
can track the molecule type, so comparing two ``Seq`` objects could mean
considering this too.

Should a DNA fragment “ACG” and an RNA fragment “ACG” be equal? What
about the peptide “ACG”? Or the Python string “ACG”? In everyday use,
your sequences will generally all be the same type of (all DNA, all RNA,
or all protein). Well, as of Biopython 1.65, sequence comparison only
looks at the sequence and compares like the Python string.

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> seq1 = Seq("ACGT")
   >>> "ACGT" == seq1
   True
   >>> seq1 == "ACGT"
   True

As an extension to this, using sequence objects as keys in a Python
dictionary is equivalent to using the sequence as a plain string for the
key. See also Section :ref:`sec:seq-to-string`.

Sequences with unknown sequence contents
----------------------------------------

In some cases, the length of a sequence may be known but not the actual
letters constituting it. For example, GenBank and EMBL files may
represent a genomic DNA sequence only by its config information, without
specifying the sequence contents explicitly. Such sequences can be
represented by creating a ``Seq`` object with the argument ``None``,
followed by the sequence length:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> unknown_seq = Seq(None, 10)

The ``Seq`` object thus created has a well-defined length. Any attempt
to access the sequence contents, however, will raise an
``UndefinedSequenceError``:

.. cont-doctest

.. code:: pycon

   >>> unknown_seq
   Seq(None, length=10)
   >>> len(unknown_seq)
   10
   >>> print(unknown_seq)
   Traceback (most recent call last):
   ...
   Bio.Seq.UndefinedSequenceError: Sequence content is undefined
   >>>

.. _`sec:partial-seq`:

Sequences with partially defined sequence contents
--------------------------------------------------

Sometimes the sequence contents is defined for parts of the sequence
only, and undefined elsewhere. For example, the following excerpt of a
MAF (Multiple Alignment Format) file shows an alignment of human, chimp,
macaque, mouse, rat, dog, and opossum genome sequences:

.. code:: text

   s hg38.chr7     117512683 36 + 159345973 TTGAAAACCTGAATGTGAGAGTCAGTCAAGGATAGT
   s panTro4.chr7  119000876 36 + 161824586 TTGAAAACCTGAATGTGAGAGTCACTCAAGGATAGT
   s rheMac3.chr3  156330991 36 + 198365852 CTGAAATCCTGAATGTGAGAGTCAATCAAGGATGGT
   s mm10.chr6      18207101 36 + 149736546 CTGAAAACCTAAGTAGGAGAATCAACTAAGGATAAT
   s rn5.chr4       42326848 36 + 248343840 CTGAAAACCTAAGTAGGAGAGACAGTTAAAGATAAT
   s canFam3.chr14  56325207 36 +  60966679 TTGAAAAACTGATTATTAGAGTCAATTAAGGATAGT
   s monDom5.chr8  173163865 36 + 312544902 TTAAGAAACTGGAAATGAGGGTTGAATGACAAACTT

In each row, the first number indicates the starting position (in
zero-based coordinates) of the aligned sequence on the chromosome,
followed by the size of the aligned sequence, the strand, the size of
the full chromosome, and the aligned sequence.

A ``Seq`` object representing such a partially defined sequence can be
created using a dictionary for the ``data`` argument, where the keys are
the starting coordinates of the known sequence segments, and the values
are the corresponding sequence contents. For example, for the first
sequence we would use

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> seq = Seq({117512683: "TTGAAAACCTGAATGTGAGAGTCAGTCAAGGATAGT"}, length=159345973)

Extracting a subsequence from a partially define sequence may return a
fully defined sequence, an undefined sequence, or a partially defined
sequence, depending on the coordinates:

.. cont-doctest

.. code:: pycon

   >>> seq[1000:1020]
   Seq(None, length=20)
   >>> seq[117512690:117512700]
   Seq('CCTGAATGTG')
   >>> seq[117512670:117512690]
   Seq({13: 'TTGAAAA'}, length=20)
   >>> seq[117512700:]
   Seq({0: 'AGAGTCAGTCAAGGATAGT'}, length=41833273)

Partially defined sequences can also be created by appending sequences,
if at least one of the sequences is partially or fully undefined:

.. cont-doctest

.. code:: pycon

   >>> seq = Seq("ACGT")
   >>> undefined_seq = Seq(None, length=10)
   >>> seq + undefined_seq + seq
   Seq({0: 'ACGT', 14: 'ACGT'}, length=18)

.. _`sec:mutable-seq`:

MutableSeq objects
------------------

Just like the normal Python string, the ``Seq`` object is “read only”,
or in Python terminology, immutable. Apart from wanting the ``Seq``
object to act like a string, this is also a useful default since in many
biological applications you want to ensure you are not changing your
sequence data:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> my_seq = Seq("GCCATTGTAATGGGCCGCTGAAAGGGTGCCCGA")

Observe what happens if you try to edit the sequence:

.. cont-doctest

.. code:: pycon

   >>> my_seq[5] = "G"
   Traceback (most recent call last):
   ...
   TypeError: 'Seq' object does not support item assignment

However, you can convert it into a mutable sequence (a ``MutableSeq``
object) and do pretty much anything you want with it:

.. cont-doctest

.. code:: pycon

   >>> from Bio.Seq import MutableSeq
   >>> mutable_seq = MutableSeq(my_seq)
   >>> mutable_seq
   MutableSeq('GCCATTGTAATGGGCCGCTGAAAGGGTGCCCGA')

Alternatively, you can create a ``MutableSeq`` object directly from a
string:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import MutableSeq
   >>> mutable_seq = MutableSeq("GCCATTGTAATGGGCCGCTGAAAGGGTGCCCGA")

Either way will give you a sequence object which can be changed:

.. cont-doctest

.. code:: pycon

   >>> mutable_seq
   MutableSeq('GCCATTGTAATGGGCCGCTGAAAGGGTGCCCGA')
   >>> mutable_seq[5] = "C"
   >>> mutable_seq
   MutableSeq('GCCATCGTAATGGGCCGCTGAAAGGGTGCCCGA')
   >>> mutable_seq.remove("T")
   >>> mutable_seq
   MutableSeq('GCCACGTAATGGGCCGCTGAAAGGGTGCCCGA')
   >>> mutable_seq.reverse()
   >>> mutable_seq
   MutableSeq('AGCCCGTGGGAAAGTCGCCGGGTAATGCACCG')

Note that the ``MutableSeq`` object’s ``reverse()`` method, like the
``reverse()`` method of a Python list, reverses the sequence in place.

An important technical difference between mutable and immutable objects
in Python means that you can’t use a ``MutableSeq`` object as a
dictionary key, but you can use a Python string or a ``Seq`` object in
this way.

Once you have finished editing your a ``MutableSeq`` object, it’s easy
to get back to a read-only ``Seq`` object should you need to:

.. cont-doctest

.. code:: pycon

   >>> from Bio.Seq import Seq
   >>> new_seq = Seq(mutable_seq)
   >>> new_seq
   Seq('AGCCCGTGGGAAAGTCGCCGGGTAATGCACCG')

You can also get a string from a ``MutableSeq`` object just like from a
``Seq`` object (Section :ref:`sec:seq-to-string`).

Finding subsequences
--------------------

Sequence objects have ``find``, ``rfind``, ``index``, and ``rindex``
methods that perform the same function as the corresponding methods
on plain string objects. The only difference is that the subsequence
can be a string (``str``), ``bytes``, ``bytearray``, ``Seq``, or
``MutableSeq`` object:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import Seq, MutableSeq
   >>> seq = Seq("GCCATTGTAATGGGCCGCTGAAAGGGTGCCCGA")
   >>> seq.index("ATGGGCCGC")
   9
   >>> seq.index(b"ATGGGCCGC")
   9
   >>> seq.index(bytearray(b"ATGGGCCGC"))
   9
   >>> seq.index(Seq("ATGGGCCGC"))
   9
   >>> seq.index(MutableSeq("ATGGGCCGC"))
   9

A ``ValueError`` is raised if the subsequence is not found:

.. cont-doctest

.. code:: pycon

   >>> seq.index("ACTG")  # doctest:+ELLIPSIS
   Traceback (most recent call last):
   ...
   ValueError: ...

while the ``find`` method returns -1 if the subsequence is not found:

.. cont-doctest

.. code:: pycon

   >>> seq.find("ACTG")
   -1

The methods ``rfind`` and ``rindex`` search for the subsequence starting
from the right hand side of the sequence:

.. cont-doctest

.. code:: pycon

   >>> seq.find("CC")
   1
   >>> seq.rfind("CC")
   29

Use the ``search`` method to search for multiple subsequences at the same
time. This method returns an iterator:

.. cont-doctest

.. code:: pycon

   >>> for index, sub in seq.search(["CC", "GGG", "CC"]):
   ...     print(index, sub)
   ...
   1 CC
   11 GGG
   14 CC
   23 GGG
   28 CC
   29 CC

The ``search`` method also takes plain strings, ``bytes``, ``bytearray``,
``Seq``, and ``MutableSeq`` objects as subsequences; identical subsequences
are reported only once, as in the example above.

.. _`sec:seq-module-functions`:

Working with strings directly
-----------------------------

To close this chapter, for those you who *really* don’t want to use the
sequence objects (or who prefer a functional programming style to an
object orientated one), there are module level functions in ``Bio.Seq``
will accept plain Python strings, ``Seq`` objects or ``MutableSeq``
objects:

.. doctest

.. code:: pycon

   >>> from Bio.Seq import reverse_complement, transcribe, back_transcribe, translate
   >>> my_string = "GCTGTTATGGGTCGTTGGAAGGGTGGTCGTGCTGCTGGTTAG"
   >>> reverse_complement(my_string)
   'CTAACCAGCAGCACGACCACCCTTCCAACGACCCATAACAGC'
   >>> transcribe(my_string)
   'GCUGUUAUGGGUCGUUGGAAGGGUGGUCGUGCUGCUGGUUAG'
   >>> back_transcribe(my_string)
   'GCTGTTATGGGTCGTTGGAAGGGTGGTCGTGCTGCTGGTTAG'
   >>> translate(my_string)
   'AVMGRWKGGRAAG*'

You are, however, encouraged to work with ``Seq`` objects by default.
