README.md
In gschofl/biofiles: An Interface for GenBank/GenPept Flat Files
biofiles - an interface to GenBank/GenPept files in R
biofiles provides interfacing to GenBank/GenPept or Embl flat file records. It includes utilities for reading and writing GenBank files, and methods for interacting with annotation and sequence data.
Installation
Install the latest stable release of the biofiles package from CRAN:
install.packages("biofiles")
Install the development version from github using the devtools package.
# install.packages("devtools")
devtools::install_github("gschofl/biofiles")
Basic functionality
Let's download a small bacterial genome, Chlamydophila psittaci DC15 (GenBank acc. CP002806; GI 334693792) from NCBI.
# install.packages("reutils")
gb_file <- reutils::efetch("CP002806", "nuccore", rettype = "gbwithparts", retmode = "text")
gb_file
#> Object of class 'efetch'
#> LOCUS CP002806 1172182 bp DNA circular BCT 31-AUG-2017
#> DEFINITION Chlamydia psittaci 02DC15, complete genome.
#> ACCESSION CP002806
#> VERSION CP002806.1
#> DBLINK BioProject: PRJNA66481
#> BioSample: SAMN02603518
#> KEYWORDS .
#> SOURCE Chlamydia psittaci 02DC15
#> ORGANISM Chlamydia psittaci 02DC15
#> Bacteria; Chlamydiae; Chlamydiales; Chlamydiaceae;
#> Chlamydia/Chlamydophila group; Chlamydia.
#> REFERENCE 1 (bases 1 to 1172182)
#> ...
#> EFetch query using the 'nuccore' database.
#> Query url: 'https://eutils.ncbi.nlm.nih.gov/entrez/eutils/efetch.fcgi...'
#> Retrieval type: 'gbwithparts', retrieval mode: 'text'
Next, we parse the efetch object into a gbRecord instance.
rec <- biofiles::gbRecord(gb_file)
rec
#> An object of class 'gbRecord', with 2040 features
#> LOCUS CP002806 1172182 bp DNA circular BCT 31 ...
#> DEFINITION Chlamydia psittaci 02DC15, complete genome.
#> ACCESSION CP002806
#> VERSION CP002806.1 GI:NA
#> DBLINK Project: PRJNA66481
#> KEYWORDS .
#> SOURCE Chlamydia psittaci 02DC15
#> ORGANISM Chlamydia psittaci 02DC15
#> Bacteria; Chlamydiae; Chlamydiales; Chlamydiaceae;
#> Chlamydia/Chlamydophila group; Chlamydia.
#> REFERENCE 1 (bases 1 to 1172182)
#> AUTHORS Schofl,G., Voigt,A., Litsche,K., Sachse,K. and Saluz,H.P.
#> TITLE Complete Genome Sequences of Four Mammalian Isolates of
#> Chlamydophila psittaci
#> JOURNAL J. Bacteriol. 193 (16), 4258 (2011)
#> PUBMED 21705611
#> REFERENCE 2 (bases 1 to 1172182)
#> AUTHORS Schofl,G.
#> TITLE Direct Submission
#> JOURNAL Submitted (20-MAY-2011) Cell and Molecular Biology,
#> Leibniz Institute for Natural Product Research and
#> Infection Biology, Beutenbergstrasse 11a, Jena D-07745,
#> Germany
#> COMMENT Source DNA available from Dr. Konrad Sachse
#> (Konrad.Sachse@fli.bund.de) at the Institute for Molecular
#> Pathogenesis, Jena, Germany.
#> ORIGIN CAAAGTTTTAAACATGTTAACCACGTTGTTTTCCCTCTATAAACCGAGTTTTAAACAGT
#> ...
#> ATCTTGTGTTCACAAACATGCAAAATCTCGATTTATTTACAAACTGAACAATGTTTTAA
#> CONTIG
The summary function provides an overview over the object:
biofiles::summary(rec)
#> [[CP002806]]
#> 1172182 bp: Chlamydia psittaci 02DC15, complete genome.
#> Id Feature Location GeneId Product ...
#> 1 source 1..1172182 NA NA ...
#> 2 gene 149..1144 hemB NA ...
#> 3 CDS 149..1144 hemB delta-aminolevuli ...
#> 4 gene complement(1160..2578) NA NA ...
#> 5 CDS complement(1160..2578) NA Na(+)-translocati ...
#> 6 gene complement(2601..3035) NA NA ...
#> 7 CDS complement(2601..3035) NA conserved hypothe ...
#> ... ... ... ... ... ...
#> 2034 CDS 1169125..1170006 NA geranylgeranyl py ...
#> 2035 gene complement(1170021..1170094) NA NA ...
#> 2036 tRNA complement(1170021..1170094) NA tRNA-Pro ...
#> 2037 gene complement(1170106..1170360) NA NA ...
#> 2038 CDS complement(1170106..1170360) NA conserved hypothe ...
#> 2039 gene complement(1170443..1172035) NA NA ...
#> 2040 CDS complement(1170443..1172035) NA conserved hypothe ...
Various getter methods provide access to the data contained in a GenBank record; for instance:
biofiles::getAccession(rec)
#> [1] "CP002806"
biofiles::getGeneID(rec)
#> [1] "NA"
biofiles::getDefinition(rec)
#> [1] "Chlamydia psittaci 02DC15, complete genome."
biofiles::getOrganism(rec)
#> [1] "Chlamydia psittaci 02DC15"
biofiles::getSequence(rec)
#> A DNAStringSet instance of length 1
#> width seq names
#> [1] 1172182 CAAAGTTTTAAACATGTTAA...AAACTGAACAATGTTTTAA CP002806
biofiles::getReference(rec)
#> A 'gbReferenceList' instance:
#> REFERENCE 1 (bases 1 to 1172182)
#> AUTHORS Schofl,G., Voigt,A., Litsche,K., Sachse,K. and Saluz,H.P.
#> TITLE Complete Genome Sequences of Four Mammalian Isolates of
#> Chlamydophila psittaci
#> JOURNAL J. Bacteriol. 193 (16), 4258 (2011)
#> PUBMED 21705611
#> REFERENCE 2 (bases 1 to 1172182)
#> AUTHORS Schofl,G.
#> TITLE Direct Submission
#> JOURNAL Submitted (20-MAY-2011) Cell and Molecular Biology,
#> Leibniz Institute for Natural Product Research and
#> Infection Biology, Beutenbergstrasse 11a, Jena D-07745,
#> Germany
The function uniqueQualifs() provides an overview over the feature qualifiers used in a record:
biofiles::uniqueQualifs(rec)
#> [1] "organism" "mol_type" "strain"
#> [4] "db_xref" "gene" "locus_tag"
#> [7] "note" "codon_start" "transl_table"
#> [10] "product" "protein_id" "translation"
#> [13] "EC_number" "artificial_location"
Genbank Features
The important part of a GenBank record will generally be the list of annotions or features.
We can access the gbFeatureList of a gbRecord using getFeatures() or ft():
f <- biofiles::ft(rec)
f
#> 'gbFeatureTable' with 2040 features:
#> Feature: Location/Qualifiers:
#> source 1..1172182
#> /organism = "Chlamydia psittaci 02DC15"
#> /mol_type = "genomic DNA"
#> /strain = "02DC15"
#> /db_xref = "taxon:1112254"
#> ...
#> Feature: Location/Qualifiers:
#> CDS complement(1170443..1172035)
#> /locus_tag = "CPS0B_1080"
#> /note = "[F] COG1351 Predicted alternative thymidylate
#> synthase"
#> /codon_start = "1"
#> /transl_table = "11"
#> /product = "conserved hypothetical protein"
#> /protein_id = "AEG87987.1"
#> /translation = "MLSRDDEFSSEQRKSLSHFVTNLETNIFALKNLPEVVKGA
#> LFSKYSRSTLGLRSLLLKEFLEGEGGDFLDSSGVDFEVGIHKAADFYRRVLDGFG
#> DDSIGELGGAHLAIESVSMLAAKILEDARIGGSPLEKSSRYVYFDQKVKGEYLYY
#> RDPILMTSAFKDVFLGTCDFLFDTYADLIPKVRTYFEKIYPKESEVSQSAYTISL
#> RAKVLDCLRGLLPAATLTNLGFFGNGRFWQTLLHKIQGHNLTEIRQIGESSLTEL
#> MKIIPSFVSRAESHHHHHQAMLSYRQTLREQLTSLAEKFSGGSHPSKQTGVRLVY
#> GDPEGIYKVAAGFLFPYSEHTYEELINICKSMPREDLIRVLEAGSSSRENRRHKS
#> PRGLECLEFGFDITADFGAYRDLQRHRILTQERQLLTTNLGYHIPEQLLDTPMEK
#> DFREAMEKAEEAYNQISLEFPEEAQYVVPLAYNIRWFFHINGRALQWLCELRSQV
#> QGHENYRKIAIDMVKEVIRFDPVYESFFKFVDYSECDLGRIKQESRKRSF"
#> Seqinfo:
#> CP002806 1172182 DNA Chlamydia psittaci 02DC15, complete genome.
We can extract features either by numeric subsetting:
f[[1]]
#> Feature: Location/Qualifiers:
#> source 1..1172182
#> /organism = "Chlamydia psittaci 02DC15"
#> /mol_type = "genomic DNA"
#> /strain = "02DC15"
#> /db_xref = "taxon:1112254"
#> Seqinfo:
#> CP002806 1172182 DNA Chlamydia psittaci 02DC15, complete genome.
or we can subset by feature key:
f["CDS"]
#> 'gbFeatureTable' with 978 features:
#> Feature: Location/Qualifiers:
#> CDS 149..1144
#> /gene = "hemB"
#> /locus_tag = "CPS0B_0001"
#> /note = "PRIAM ID: PRI000415; [H] COG0113
#> Delta-aminolevulinic acid dehydratase"
#> /codon_start = "1"
#> /transl_table = "11"
#> /product = "delta-aminolevulinic acid dehydratase"
#> /protein_id = "AEG87010.1"
#> /translation = "MSSLALSRRPRRNRRTEAIRDLVSETSLLPQDFICPFFVK
#> EGKNIREEIESLTGVYRWSIDLLLKEIERLCSLGLRAVILFPVIPSHLKDAYGSY
#> SSNPKNILCKSIYEVKKAFPNLCVISDIALDPYTTHGHDGIIDRGEVLNDESVRI
#> FGNIATLHAEMGADVVAPSDMMDGRVAHIRSKLDQAGWTQTLILSYSVKYASALY
#> NPFRDALGSHLQSGDKRNYQMNPKNVLEALLECSLDEQEGADMLMIKPAGLYLDV
#> LHRVKNSTTLPLAAYQVSGEYAMIAAASTMGWLDREKIVYESLIAIKRAGADMII
#> SYATPLILEMIASSRV"
#> ...
#> Feature: Location/Qualifiers:
#> CDS complement(1170443..1172035)
#> /locus_tag = "CPS0B_1080"
#> /note = "[F] COG1351 Predicted alternative thymidylate
#> synthase"
#> /codon_start = "1"
#> /transl_table = "11"
#> /product = "conserved hypothetical protein"
#> /protein_id = "AEG87987.1"
#> /translation = "MLSRDDEFSSEQRKSLSHFVTNLETNIFALKNLPEVVKGA
#> LFSKYSRSTLGLRSLLLKEFLEGEGGDFLDSSGVDFEVGIHKAADFYRRVLDGFG
#> DDSIGELGGAHLAIESVSMLAAKILEDARIGGSPLEKSSRYVYFDQKVKGEYLYY
#> RDPILMTSAFKDVFLGTCDFLFDTYADLIPKVRTYFEKIYPKESEVSQSAYTISL
#> RAKVLDCLRGLLPAATLTNLGFFGNGRFWQTLLHKIQGHNLTEIRQIGESSLTEL
#> MKIIPSFVSRAESHHHHHQAMLSYRQTLREQLTSLAEKFSGGSHPSKQTGVRLVY
#> GDPEGIYKVAAGFLFPYSEHTYEELINICKSMPREDLIRVLEAGSSSRENRRHKS
#> PRGLECLEFGFDITADFGAYRDLQRHRILTQERQLLTTNLGYHIPEQLLDTPMEK
#> DFREAMEKAEEAYNQISLEFPEEAQYVVPLAYNIRWFFHINGRALQWLCELRSQV
#> QGHENYRKIAIDMVKEVIRFDPVYESFFKFVDYSECDLGRIKQESRKRSF"
#> Seqinfo:
#> CP002806 1172182 DNA Chlamydia psittaci 02DC15, complete genome.
A more versatile method to narrow down the list of features of interest is the function filter(). For instance, we can filter for all coding sequences (CDS) with the annotation "hypothetical" in the product qualifiers:
hypo <- biofiles::filter(rec, key = "CDS", product = "hypothetical")
biofiles::summary(hypo)
#> [[CP002806]]
#> 1172182 bp: Chlamydia psittaci 02DC15, complete genome.
#> Id Feature Location GeneId Product ...
#> 7 CDS complement(2601..3035) NA conserved hypothe ...
#> 31 CDS 20427..21713 NA conserved hypothe ...
#> 33 CDS 21822..23741 NA conserved hypothe ...
#> 35 CDS complement(23931..26531) NA conserved hypothe ...
#> 37 CDS 26689..29340 NA conserved hypothe ...
#> 39 CDS 29390..29620 NA conserved hypothe ...
#> 43 CDS complement(30347..30832) NA conserved hypothe ...
#> ... ... ... ... ... ...
#> 2002 CDS 1152648..1153925 NA conserved hypothe ...
#> 2008 CDS complement(1155986..1156711) NA conserved hypothe ...
#> 2010 CDS complement(1156746..1157474) NA conserved hypothe ...
#> 2024 CDS 1163308..1164144 NA conserved hypothe ...
#> 2028 CDS complement(1165683..1167527) NA conserved hypothe ...
#> 2038 CDS complement(1170106..1170360) NA conserved hypothe ...
#> 2040 CDS complement(1170443..1172035) NA conserved hypothe ...
or we can filter for all elongation factors,
elong <- biofiles::filter(rec, key = "CDS", product = "elongation factor")
biofiles::summary(elong)
#> [[CP002806]]
#> 1172182 bp: Chlamydia psittaci 02DC15, complete genome.
#> Id Feature Location GeneId Product ...
#> 9 CDS 3148..5301 greA transcription elong ...
#> 103 CDS complement(56119..56967) tsf translation elongat ...
#> 408 CDS 206213..208297 fusA translation elongat ...
#> 950 CDS complement(526454..527758) nusA transcription elong ...
#> 1126 CDS 623470..624027 NA elongation factor P ...
#> 1406 CDS complement(805145..806329) tuf translation elongat ...
#> 1750 CDS complement(997270..997842) efp translation elongat ...
now let's extract the sequence for all elongation factors, and using the tools from the Biostrings packages, translate them into protein sequences. Note, that in order to do so, we first get the gbFeatureTable from the gbRecord, as otherwise we'd just extract the complete sequence associated with the GenBank record.
dna <- biofiles::getSequence(biofiles::ft(elong))
dna
#> A DNAStringSet instance of length 7
#> width seq names
#> [1] 2154 GTGGACTATCTAGAAAAGTTG...AATCTATTTGGGATGCATAA lcl|CDS.9|gb|CP00...
#> [2] 849 TTAAGCTCCTATTTTCCATAA...TCCATAGAAAAGTTGCTCAT lcl|CDS.103|gb|CP...
#> [3] 2085 ATGAGTGATCAAGAATTCGAT...AAGAGATTGTTAAGAAGTAA lcl|CDS.408|gb|CP...
#> [4] 1305 TTAATCTTCAATTTTAGGTTT...GCTACAAGATCTTTATTCAT lcl|CDS.950|gb|CP...
#> [5] 558 ATGGTATTAAGTAGCCAGCTC...AATATATTCAACGCGTCTAA lcl|CDS.1126|gb|C...
#> [6] 1185 TTAGGCAATAATTTTTGAAAT...TGAAAAGTTTCTTTTGACAT lcl|CDS.1406|gb|C...
#> [7] 573 TTACTTGGAAACACGCGATTC...CTAGTGCTTACACGAACCAT lcl|CDS.1750|gb|C...
str <- biofiles::strand(elong)
dna_revcomp <- c(Biostrings::reverseComplement(dna[str == -1]), dna[str == 1])
aa <- Biostrings::translate(dna_revcomp)
names(aa) <- names(dna_revcomp)
aa
#> A AAStringSet instance of length 7
#> width seq names
#> [1] 283 MSNFSMETLKLLRQQTGVGLT...KTSGNSVEVKEFILWKIGA* lcl|CDS.103|gb|CP...
#> [2] 435 MNKDLVAIFDYMEKEKGIQRP...EQVSKYGEGKVDEKPKIED* lcl|CDS.950|gb|CP...
#> [3] 395 MSKETFQRTKPHINIGTIGHV...AIREGGRTIGAGTISKIIA* lcl|CDS.1406|gb|C...
#> [4] 191 MVRVSTSEFRVGLRIEIDGQP...GEVVKVDTRTGSYESRVSK* lcl|CDS.1750|gb|C...
#> [5] 718 VDYLEKLQVLIDEEQPSSFFN...LQFQGKKYKISRIQSIWDA* lcl|CDS.9|gb|CP00...
#> [6] 695 MSDQEFDLSKIRNIGIMAHID...EPAFFAKVPQKIQEEIVKK* lcl|CDS.408|gb|CP...
#> [7] 186 MVLSSQLSVGMFISTKDGLYK...EIGDIIKIDTRTCEYIQRV* lcl|CDS.1126|gb|C...
We can use the ranges() method to extract GRanges objects defined in the Bioconductor package GenomicRanges:
elong_ranges <- biofiles::ranges(elong, include = c("locus_tag", "protein_id", "product"))
elong_ranges
#> GRanges object with 7 ranges and 4 metadata columns:
#> seqnames ranges strand | key locus_tag
#> <Rle> <IRanges> <Rle> | <character> <character>
#> CPS0B_0004 CP002806 [ 3148, 5301] + | CDS CPS0B_0004
#> CPS0B_0056 CP002806 [ 56119, 56967] - | CDS CPS0B_0056
#> CPS0B_0219 CP002806 [206213, 208297] + | CDS CPS0B_0219
#> CPS0B_0509 CP002806 [526454, 527758] - | CDS CPS0B_0509
#> CPS0B_0600 CP002806 [623470, 624027] + | CDS CPS0B_0600
#> CPS0B_0751 CP002806 [805145, 806329] - | CDS CPS0B_0751
#> CPS0B_0928 CP002806 [997270, 997842] - | CDS CPS0B_0928
#> protein_id
#> <character>
#> CPS0B_0004 AEG87013.1
#> CPS0B_0056 AEG87055.1
#> CPS0B_0219 AEG87200.1
#> CPS0B_0509 AEG87466.1
#> CPS0B_0600 AEG87550.1
#> CPS0B_0751 AEG87684.1
#> CPS0B_0928 AEG87849.1
#> product
#> <character>
#> CPS0B_0004 transcription elongation factor GreA domain protein
#> CPS0B_0056 translation elongation factor Ts
#> CPS0B_0219 translation elongation factor G
#> CPS0B_0509 transcription elongation factor
#> CPS0B_0600 elongation factor P
#> CPS0B_0751 translation elongation factor Tu
#> CPS0B_0928 translation elongation factor P
#> -------
#> seqinfo: 1 sequence (1 circular) from Chlamydia psittaci 02DC15, complete genome. genome
gschofl/biofiles documentation built on Sept. 27, 2020, 12:08 a.m.
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biofiles - an interface to GenBank/GenPept files in R
biofiles provides interfacing to GenBank/GenPept or Embl flat file records. It includes utilities for reading and writing GenBank files, and methods for interacting with annotation and sequence data.
Installation
Install the latest stable release of the biofiles package from CRAN:
install.packages("biofiles")
Install the development version from github using the devtools package.
# install.packages("devtools")
devtools::install_github("gschofl/biofiles")
Basic functionality
Let's download a small bacterial genome, Chlamydophila psittaci DC15 (GenBank acc. CP002806; GI 334693792) from NCBI.
# install.packages("reutils")
gb_file <- reutils::efetch("CP002806", "nuccore", rettype = "gbwithparts", retmode = "text")
gb_file
#> Object of class 'efetch'
#> LOCUS CP002806 1172182 bp DNA circular BCT 31-AUG-2017
#> DEFINITION Chlamydia psittaci 02DC15, complete genome.
#> ACCESSION CP002806
#> VERSION CP002806.1
#> DBLINK BioProject: PRJNA66481
#> BioSample: SAMN02603518
#> KEYWORDS .
#> SOURCE Chlamydia psittaci 02DC15
#> ORGANISM Chlamydia psittaci 02DC15
#> Bacteria; Chlamydiae; Chlamydiales; Chlamydiaceae;
#> Chlamydia/Chlamydophila group; Chlamydia.
#> REFERENCE 1 (bases 1 to 1172182)
#> ...
#> EFetch query using the 'nuccore' database.
#> Query url: 'https://eutils.ncbi.nlm.nih.gov/entrez/eutils/efetch.fcgi...'
#> Retrieval type: 'gbwithparts', retrieval mode: 'text'
Next, we parse the efetch object into a gbRecord instance.
rec <- biofiles::gbRecord(gb_file)
rec
#> An object of class 'gbRecord', with 2040 features
#> LOCUS CP002806 1172182 bp DNA circular BCT 31 ...
#> DEFINITION Chlamydia psittaci 02DC15, complete genome.
#> ACCESSION CP002806
#> VERSION CP002806.1 GI:NA
#> DBLINK Project: PRJNA66481
#> KEYWORDS .
#> SOURCE Chlamydia psittaci 02DC15
#> ORGANISM Chlamydia psittaci 02DC15
#> Bacteria; Chlamydiae; Chlamydiales; Chlamydiaceae;
#> Chlamydia/Chlamydophila group; Chlamydia.
#> REFERENCE 1 (bases 1 to 1172182)
#> AUTHORS Schofl,G., Voigt,A., Litsche,K., Sachse,K. and Saluz,H.P.
#> TITLE Complete Genome Sequences of Four Mammalian Isolates of
#> Chlamydophila psittaci
#> JOURNAL J. Bacteriol. 193 (16), 4258 (2011)
#> PUBMED 21705611
#> REFERENCE 2 (bases 1 to 1172182)
#> AUTHORS Schofl,G.
#> TITLE Direct Submission
#> JOURNAL Submitted (20-MAY-2011) Cell and Molecular Biology,
#> Leibniz Institute for Natural Product Research and
#> Infection Biology, Beutenbergstrasse 11a, Jena D-07745,
#> Germany
#> COMMENT Source DNA available from Dr. Konrad Sachse
#> (Konrad.Sachse@fli.bund.de) at the Institute for Molecular
#> Pathogenesis, Jena, Germany.
#> ORIGIN CAAAGTTTTAAACATGTTAACCACGTTGTTTTCCCTCTATAAACCGAGTTTTAAACAGT
#> ...
#> ATCTTGTGTTCACAAACATGCAAAATCTCGATTTATTTACAAACTGAACAATGTTTTAA
#> CONTIG
The summary function provides an overview over the object:
biofiles::summary(rec)
#> [[CP002806]]
#> 1172182 bp: Chlamydia psittaci 02DC15, complete genome.
#> Id Feature Location GeneId Product ...
#> 1 source 1..1172182 NA NA ...
#> 2 gene 149..1144 hemB NA ...
#> 3 CDS 149..1144 hemB delta-aminolevuli ...
#> 4 gene complement(1160..2578) NA NA ...
#> 5 CDS complement(1160..2578) NA Na(+)-translocati ...
#> 6 gene complement(2601..3035) NA NA ...
#> 7 CDS complement(2601..3035) NA conserved hypothe ...
#> ... ... ... ... ... ...
#> 2034 CDS 1169125..1170006 NA geranylgeranyl py ...
#> 2035 gene complement(1170021..1170094) NA NA ...
#> 2036 tRNA complement(1170021..1170094) NA tRNA-Pro ...
#> 2037 gene complement(1170106..1170360) NA NA ...
#> 2038 CDS complement(1170106..1170360) NA conserved hypothe ...
#> 2039 gene complement(1170443..1172035) NA NA ...
#> 2040 CDS complement(1170443..1172035) NA conserved hypothe ...
Various getter methods provide access to the data contained in a GenBank record; for instance:
biofiles::getAccession(rec)
#> [1] "CP002806"
biofiles::getGeneID(rec)
#> [1] "NA"
biofiles::getDefinition(rec)
#> [1] "Chlamydia psittaci 02DC15, complete genome."
biofiles::getOrganism(rec)
#> [1] "Chlamydia psittaci 02DC15"
biofiles::getSequence(rec)
#> A DNAStringSet instance of length 1
#> width seq names
#> [1] 1172182 CAAAGTTTTAAACATGTTAA...AAACTGAACAATGTTTTAA CP002806
biofiles::getReference(rec)
#> A 'gbReferenceList' instance:
#> REFERENCE 1 (bases 1 to 1172182)
#> AUTHORS Schofl,G., Voigt,A., Litsche,K., Sachse,K. and Saluz,H.P.
#> TITLE Complete Genome Sequences of Four Mammalian Isolates of
#> Chlamydophila psittaci
#> JOURNAL J. Bacteriol. 193 (16), 4258 (2011)
#> PUBMED 21705611
#> REFERENCE 2 (bases 1 to 1172182)
#> AUTHORS Schofl,G.
#> TITLE Direct Submission
#> JOURNAL Submitted (20-MAY-2011) Cell and Molecular Biology,
#> Leibniz Institute for Natural Product Research and
#> Infection Biology, Beutenbergstrasse 11a, Jena D-07745,
#> Germany
The function uniqueQualifs() provides an overview over the feature qualifiers used in a record:
biofiles::uniqueQualifs(rec)
#> [1] "organism" "mol_type" "strain"
#> [4] "db_xref" "gene" "locus_tag"
#> [7] "note" "codon_start" "transl_table"
#> [10] "product" "protein_id" "translation"
#> [13] "EC_number" "artificial_location"
Genbank Features
The important part of a GenBank record will generally be the list of annotions or features.
We can access the gbFeatureList of a gbRecord using getFeatures() or ft():
f <- biofiles::ft(rec)
f
#> 'gbFeatureTable' with 2040 features:
#> Feature: Location/Qualifiers:
#> source 1..1172182
#> /organism = "Chlamydia psittaci 02DC15"
#> /mol_type = "genomic DNA"
#> /strain = "02DC15"
#> /db_xref = "taxon:1112254"
#> ...
#> Feature: Location/Qualifiers:
#> CDS complement(1170443..1172035)
#> /locus_tag = "CPS0B_1080"
#> /note = "[F] COG1351 Predicted alternative thymidylate
#> synthase"
#> /codon_start = "1"
#> /transl_table = "11"
#> /product = "conserved hypothetical protein"
#> /protein_id = "AEG87987.1"
#> /translation = "MLSRDDEFSSEQRKSLSHFVTNLETNIFALKNLPEVVKGA
#> LFSKYSRSTLGLRSLLLKEFLEGEGGDFLDSSGVDFEVGIHKAADFYRRVLDGFG
#> DDSIGELGGAHLAIESVSMLAAKILEDARIGGSPLEKSSRYVYFDQKVKGEYLYY
#> RDPILMTSAFKDVFLGTCDFLFDTYADLIPKVRTYFEKIYPKESEVSQSAYTISL
#> RAKVLDCLRGLLPAATLTNLGFFGNGRFWQTLLHKIQGHNLTEIRQIGESSLTEL
#> MKIIPSFVSRAESHHHHHQAMLSYRQTLREQLTSLAEKFSGGSHPSKQTGVRLVY
#> GDPEGIYKVAAGFLFPYSEHTYEELINICKSMPREDLIRVLEAGSSSRENRRHKS
#> PRGLECLEFGFDITADFGAYRDLQRHRILTQERQLLTTNLGYHIPEQLLDTPMEK
#> DFREAMEKAEEAYNQISLEFPEEAQYVVPLAYNIRWFFHINGRALQWLCELRSQV
#> QGHENYRKIAIDMVKEVIRFDPVYESFFKFVDYSECDLGRIKQESRKRSF"
#> Seqinfo:
#> CP002806 1172182 DNA Chlamydia psittaci 02DC15, complete genome.
We can extract features either by numeric subsetting:
f[[1]]
#> Feature: Location/Qualifiers:
#> source 1..1172182
#> /organism = "Chlamydia psittaci 02DC15"
#> /mol_type = "genomic DNA"
#> /strain = "02DC15"
#> /db_xref = "taxon:1112254"
#> Seqinfo:
#> CP002806 1172182 DNA Chlamydia psittaci 02DC15, complete genome.
or we can subset by feature key:
f["CDS"]
#> 'gbFeatureTable' with 978 features:
#> Feature: Location/Qualifiers:
#> CDS 149..1144
#> /gene = "hemB"
#> /locus_tag = "CPS0B_0001"
#> /note = "PRIAM ID: PRI000415; [H] COG0113
#> Delta-aminolevulinic acid dehydratase"
#> /codon_start = "1"
#> /transl_table = "11"
#> /product = "delta-aminolevulinic acid dehydratase"
#> /protein_id = "AEG87010.1"
#> /translation = "MSSLALSRRPRRNRRTEAIRDLVSETSLLPQDFICPFFVK
#> EGKNIREEIESLTGVYRWSIDLLLKEIERLCSLGLRAVILFPVIPSHLKDAYGSY
#> SSNPKNILCKSIYEVKKAFPNLCVISDIALDPYTTHGHDGIIDRGEVLNDESVRI
#> FGNIATLHAEMGADVVAPSDMMDGRVAHIRSKLDQAGWTQTLILSYSVKYASALY
#> NPFRDALGSHLQSGDKRNYQMNPKNVLEALLECSLDEQEGADMLMIKPAGLYLDV
#> LHRVKNSTTLPLAAYQVSGEYAMIAAASTMGWLDREKIVYESLIAIKRAGADMII
#> SYATPLILEMIASSRV"
#> ...
#> Feature: Location/Qualifiers:
#> CDS complement(1170443..1172035)
#> /locus_tag = "CPS0B_1080"
#> /note = "[F] COG1351 Predicted alternative thymidylate
#> synthase"
#> /codon_start = "1"
#> /transl_table = "11"
#> /product = "conserved hypothetical protein"
#> /protein_id = "AEG87987.1"
#> /translation = "MLSRDDEFSSEQRKSLSHFVTNLETNIFALKNLPEVVKGA
#> LFSKYSRSTLGLRSLLLKEFLEGEGGDFLDSSGVDFEVGIHKAADFYRRVLDGFG
#> DDSIGELGGAHLAIESVSMLAAKILEDARIGGSPLEKSSRYVYFDQKVKGEYLYY
#> RDPILMTSAFKDVFLGTCDFLFDTYADLIPKVRTYFEKIYPKESEVSQSAYTISL
#> RAKVLDCLRGLLPAATLTNLGFFGNGRFWQTLLHKIQGHNLTEIRQIGESSLTEL
#> MKIIPSFVSRAESHHHHHQAMLSYRQTLREQLTSLAEKFSGGSHPSKQTGVRLVY
#> GDPEGIYKVAAGFLFPYSEHTYEELINICKSMPREDLIRVLEAGSSSRENRRHKS
#> PRGLECLEFGFDITADFGAYRDLQRHRILTQERQLLTTNLGYHIPEQLLDTPMEK
#> DFREAMEKAEEAYNQISLEFPEEAQYVVPLAYNIRWFFHINGRALQWLCELRSQV
#> QGHENYRKIAIDMVKEVIRFDPVYESFFKFVDYSECDLGRIKQESRKRSF"
#> Seqinfo:
#> CP002806 1172182 DNA Chlamydia psittaci 02DC15, complete genome.
A more versatile method to narrow down the list of features of interest is the function filter(). For instance, we can filter for all coding sequences (CDS) with the annotation "hypothetical" in the product qualifiers:
hypo <- biofiles::filter(rec, key = "CDS", product = "hypothetical")
biofiles::summary(hypo)
#> [[CP002806]]
#> 1172182 bp: Chlamydia psittaci 02DC15, complete genome.
#> Id Feature Location GeneId Product ...
#> 7 CDS complement(2601..3035) NA conserved hypothe ...
#> 31 CDS 20427..21713 NA conserved hypothe ...
#> 33 CDS 21822..23741 NA conserved hypothe ...
#> 35 CDS complement(23931..26531) NA conserved hypothe ...
#> 37 CDS 26689..29340 NA conserved hypothe ...
#> 39 CDS 29390..29620 NA conserved hypothe ...
#> 43 CDS complement(30347..30832) NA conserved hypothe ...
#> ... ... ... ... ... ...
#> 2002 CDS 1152648..1153925 NA conserved hypothe ...
#> 2008 CDS complement(1155986..1156711) NA conserved hypothe ...
#> 2010 CDS complement(1156746..1157474) NA conserved hypothe ...
#> 2024 CDS 1163308..1164144 NA conserved hypothe ...
#> 2028 CDS complement(1165683..1167527) NA conserved hypothe ...
#> 2038 CDS complement(1170106..1170360) NA conserved hypothe ...
#> 2040 CDS complement(1170443..1172035) NA conserved hypothe ...
or we can filter for all elongation factors,
elong <- biofiles::filter(rec, key = "CDS", product = "elongation factor")
biofiles::summary(elong)
#> [[CP002806]]
#> 1172182 bp: Chlamydia psittaci 02DC15, complete genome.
#> Id Feature Location GeneId Product ...
#> 9 CDS 3148..5301 greA transcription elong ...
#> 103 CDS complement(56119..56967) tsf translation elongat ...
#> 408 CDS 206213..208297 fusA translation elongat ...
#> 950 CDS complement(526454..527758) nusA transcription elong ...
#> 1126 CDS 623470..624027 NA elongation factor P ...
#> 1406 CDS complement(805145..806329) tuf translation elongat ...
#> 1750 CDS complement(997270..997842) efp translation elongat ...
now let's extract the sequence for all elongation factors, and using the tools from the Biostrings packages, translate them into protein sequences. Note, that in order to do so, we first get the gbFeatureTable from the gbRecord, as otherwise we'd just extract the complete sequence associated with the GenBank record.
dna <- biofiles::getSequence(biofiles::ft(elong))
dna
#> A DNAStringSet instance of length 7
#> width seq names
#> [1] 2154 GTGGACTATCTAGAAAAGTTG...AATCTATTTGGGATGCATAA lcl|CDS.9|gb|CP00...
#> [2] 849 TTAAGCTCCTATTTTCCATAA...TCCATAGAAAAGTTGCTCAT lcl|CDS.103|gb|CP...
#> [3] 2085 ATGAGTGATCAAGAATTCGAT...AAGAGATTGTTAAGAAGTAA lcl|CDS.408|gb|CP...
#> [4] 1305 TTAATCTTCAATTTTAGGTTT...GCTACAAGATCTTTATTCAT lcl|CDS.950|gb|CP...
#> [5] 558 ATGGTATTAAGTAGCCAGCTC...AATATATTCAACGCGTCTAA lcl|CDS.1126|gb|C...
#> [6] 1185 TTAGGCAATAATTTTTGAAAT...TGAAAAGTTTCTTTTGACAT lcl|CDS.1406|gb|C...
#> [7] 573 TTACTTGGAAACACGCGATTC...CTAGTGCTTACACGAACCAT lcl|CDS.1750|gb|C...
str <- biofiles::strand(elong)
dna_revcomp <- c(Biostrings::reverseComplement(dna[str == -1]), dna[str == 1])
aa <- Biostrings::translate(dna_revcomp)
names(aa) <- names(dna_revcomp)
aa
#> A AAStringSet instance of length 7
#> width seq names
#> [1] 283 MSNFSMETLKLLRQQTGVGLT...KTSGNSVEVKEFILWKIGA* lcl|CDS.103|gb|CP...
#> [2] 435 MNKDLVAIFDYMEKEKGIQRP...EQVSKYGEGKVDEKPKIED* lcl|CDS.950|gb|CP...
#> [3] 395 MSKETFQRTKPHINIGTIGHV...AIREGGRTIGAGTISKIIA* lcl|CDS.1406|gb|C...
#> [4] 191 MVRVSTSEFRVGLRIEIDGQP...GEVVKVDTRTGSYESRVSK* lcl|CDS.1750|gb|C...
#> [5] 718 VDYLEKLQVLIDEEQPSSFFN...LQFQGKKYKISRIQSIWDA* lcl|CDS.9|gb|CP00...
#> [6] 695 MSDQEFDLSKIRNIGIMAHID...EPAFFAKVPQKIQEEIVKK* lcl|CDS.408|gb|CP...
#> [7] 186 MVLSSQLSVGMFISTKDGLYK...EIGDIIKIDTRTCEYIQRV* lcl|CDS.1126|gb|C...
We can use the ranges() method to extract GRanges objects defined in the Bioconductor package GenomicRanges:
elong_ranges <- biofiles::ranges(elong, include = c("locus_tag", "protein_id", "product"))
elong_ranges
#> GRanges object with 7 ranges and 4 metadata columns:
#> seqnames ranges strand | key locus_tag
#> <Rle> <IRanges> <Rle> | <character> <character>
#> CPS0B_0004 CP002806 [ 3148, 5301] + | CDS CPS0B_0004
#> CPS0B_0056 CP002806 [ 56119, 56967] - | CDS CPS0B_0056
#> CPS0B_0219 CP002806 [206213, 208297] + | CDS CPS0B_0219
#> CPS0B_0509 CP002806 [526454, 527758] - | CDS CPS0B_0509
#> CPS0B_0600 CP002806 [623470, 624027] + | CDS CPS0B_0600
#> CPS0B_0751 CP002806 [805145, 806329] - | CDS CPS0B_0751
#> CPS0B_0928 CP002806 [997270, 997842] - | CDS CPS0B_0928
#> protein_id
#> <character>
#> CPS0B_0004 AEG87013.1
#> CPS0B_0056 AEG87055.1
#> CPS0B_0219 AEG87200.1
#> CPS0B_0509 AEG87466.1
#> CPS0B_0600 AEG87550.1
#> CPS0B_0751 AEG87684.1
#> CPS0B_0928 AEG87849.1
#> product
#> <character>
#> CPS0B_0004 transcription elongation factor GreA domain protein
#> CPS0B_0056 translation elongation factor Ts
#> CPS0B_0219 translation elongation factor G
#> CPS0B_0509 transcription elongation factor
#> CPS0B_0600 elongation factor P
#> CPS0B_0751 translation elongation factor Tu
#> CPS0B_0928 translation elongation factor P
#> -------
#> seqinfo: 1 sequence (1 circular) from Chlamydia psittaci 02DC15, complete genome. genome
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