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Entrez Programming Utilities Help [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2010-.

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Entrez Direct Examples

, PhD.

Author Information

Created: ; Last Update: August 2, 2022.

Estimated reading time: 16 minutes

Some early EDirect examples required post-processing with shell script:

  start=$(( start + 1 ))
stop=$(( stop + 1 ))

or awk commands:

  awk -F '\t' -v 'OFS=\t' '{print $1, $2+1, $3+1}'

to increment 0-based sequence coordinates so that the proper region was retrieved:

  efetch -db nuccore -id $accn -format fasta -seq_start $start -seq_stop $stop

This led to -element derivatives that could do simple adjustments within an xtract command:

  -element ChrAccVer -inc ChrStart -1-based ChrStop

and to efetch arguments that could take 0-based coordinates directly:

  efetch -db nuccore -id $accn -format fasta -chr_start $start -chr_stop $stop

These helped eliminate the need for scripts to perform otherwise trivial modifications on extracted data.

More recent work allows easier after-the-fact numeric manipulation using the filter-columns:

  filter-columns '10 <= $2 && $2 <= 30'

and print-columns:

  print-columns '$1, $2+1, $3-1, "\042" $4 "\042", tolower($5), total += $2'

scripts, which accept column designators in an argument and pass them to internal awk commands. The NF (number of fields) and NR (current record number) built-in variables can also be used.


Author Frequency

Who are the most prolific authors on rattlesnake phospholipase?

  esearch -db pubmed -query \
"crotalid venoms [MAJR] AND phospholipase [TIAB]" |
efetch -format xml |
xtract -pattern PubmedArticle \
-block Author -sep " " -tab "\n" -element LastName,Initials |

Placing author names on separate lines allows sort-uniq-count-rank to produce a frequency table:

  87    Lomonte B
77 Gutiérrez JM
64 Soares AM
53 Marangoni S
43 Giglio JR
39 Bon C

Publication Distribution

When were the most papers about Legionnaires disease published?

  esearch -db pubmed -query "legionnaires disease [TITL]" |
efetch -format docsum |
xtract -pattern DocumentSummary -element PubDate |
cut -c 1-4 |

In this case sort-uniq-count-rank reports the number of selected papers per year:

  173    1979
102 1980
96 1978
92 1981
66 1983

Using ‑year "PubDate/*" on a PubmedArticle record takes the first four-digit number it encounters, and the result does not need to be trimmed:

  esearch -db pubmed -query "legionnaires disease [TITL]" |
efetch -format xml |
xtract -pattern PubmedArticle -year "PubDate/*" |

Treatment Locations

What is the geographic distribution of sepsis treatment studies?

  esearch -db pubmed -query \
"sepsis/therapy [MESH] AND geographic locations [MESH]" |
efetch -format xml |
xtract -pattern PubmedArticle \
-block MeshHeading -if DescriptorName@Type -equals Geographic \
-tab "\n" -element DescriptorName |

This returns the number of articles ranked by country or region:

  660    United States
250 Spain
182 Germany
168 India
155 Taiwan
139 Japan
126 China
124 France
123 Europe

Note that England and United Kingdom will appear as two separate entries.

Indexed Date Fields

What date fields are indexed for PubMed?

  einfo -db pubmed |
xtract -pattern Field \
-if IsDate -equals Y -and IsHidden -equals N \
-pfx "[" -sfx "]" -element Name \
-pfx "" -sfx "" -element FullName |
sort -k 2f | expand

Indexed dates are shown with field abbreviations and descriptive names:

  [CDAT]  Date - Completion
[CRDT] Date - Create
[EDAT] Date - Entrez
[MHDA] Date - MeSH
[MDAT] Date - Modification
[PDAT] Date - Publication

Digital Object Identifiers

How are digital object identifiers obtained from PubMed articles?

  esearch -db pubmed -query "Rowley JD [AUTH]" |
efetch -format xml |
xtract -head '<html><body>' -tail '</body></html>' \
-pattern PubmedArticle -PMID MedlineCitation/PMID \
-block ArticleId -if @IdType -equals doi \
-tab "" -pfx '<p><a href="' -sfx '">' -doi ArticleId \
-tab "\n" -pfx '' -sfx '</a></p>' -element "&PMID" |
transmute -format

The ‑doi command extracts the DOIs and constructs URL references. The ‑pfx and ‑sfx arguments used here enclose each PMID with a clickable link to its DOI:

  <?xml version="1.0" encoding="UTF-8" ?>
<!DOCTYPE html>
<a href="https://0-doi-org.brum.beds.ac.uk/10.1038%2Fleu.2013.340">24496283</a>
<a href="https://0-doi-org.brum.beds.ac.uk/10.1073%2Fpnas.1310656110">23818607</a>

Reference Formatting

How were references in the main EDirect documentation page formatted?

  efetch -db pubmed -format docsum \
-id 26931183 31738401 31600197 29718389 14728215 29490008 8743683 31114887 23175613 |

Relevant document summary fields were collected with:

  xtract -set Set -rec Rec -pattern DocumentSummary \
-sep ", " -sfx "." -NM Name -clr \
-sfx "." -SRC Source -clr \
-pfx ":" -PG Pages -clr \
-pfx "(PMID " -sfx ".)" -ID Id -clr \
-group ArticleId -if IdType -equals doi \
-pfx "https://0-doi-org.brum.beds.ac.uk/" -sfx "." -DOI Value -PG "(.)" -clr \
-group DocumentSummary \
-wrp Name -element "&NM" \
-wrp Title -element Title \
-wrp Source -element "&SRC" \
-wrp Year -year PubDate \
-wrp Pages -element "&PG" \
-wrp DOI -element "&DOI" \
-wrp Id -element "&ID" |

This generated intermediate XML with partial formatting:

<Name>Wei CH, Allot A, Leaman R, Lu Z.</Name>
<Title>PubTator central: automated concept ... full text articles.</Title>
<Source>Nucleic Acids Res.</Source>
<Id>(PMID 31114887.)</Id>

References were printed with:

  xtract -pattern Rec -deq "\n\n" -tab " " -sep "" \
-element Name Title Source Year,Pages DOI Id

followed by manual editing of hyphenated initials:

Wei C-H, Allot A, Leaman R, Lu Z. PubTator central: automated concept
annotation for biomedical full text articles. Nucleic Acids Res. 2019.
https://0-doi-org.brum.beds.ac.uk/10.1093/nar/gkz389. (PMID 31114887.)

Using xtract ‑reg and ‑exp regular expressions to guide xtract ‑replace:

  efetch -db pubmed -id 31114887 -format xml |
xtract -pattern PubmedArticle -block Author -deq "\n" \
-element LastName -reg "[a-z .]" -exp "" -replace ForeName

can generate hyphenated initials directly from the ForeName field:


by removing lower-case letters, spaces, and periods.


Coding Sequences

What are the coding sequences in the Escherichia coli lac operon?

  efetch -db nuccore -id J01636.1 -format gbc |
xtract -insd CDS gene sub_sequence

Sequence under a feature location is obtained with the ‑insd "sub_sequence" argument:


Untranslated Region Sequences

What are the 5' and 3' UTR sequences for lycopene cyclase mRNAs?

  SubSeq() {

xtract -pattern INSDSeq -ACC INSDSeq_accession-version -SEQ INSDSeq_sequence \
-group INSDFeature -if INSDFeature_key -equals CDS -PRD "(-)" \
-block INSDQualifier -if INSDQualifier_name -equals product \
-PRD INSDQualifier_value \
-block INSDFeature -pfc "\n" -element "&ACC" -rst \
-first INSDInterval_from -last INSDInterval_to -element "&PRD" "&SEQ"

UTRs() {

while IFS=$'\t' read acc fst lst prd seq

if [ $fst -gt 1 ]
echo -e ">$acc 5'UTR: 1..$((fst-1)) $prd"
echo "${seq:1:$((fst-2))}" | fold -w 50
echo -e ">$acc NO 5'UTR"
if [ $lst -lt ${#seq} ]
echo -e ">$acc 3'UTR: $((lst+1))..${#seq} $prd"
echo "${seq:$lst}" | fold -w 50
echo -e ">$acc NO 3'UTR"

esearch -db nuccore -query " [ECNO]" |
efilter -molecule mrna -feature cds -source refseq |
efetch -format gbc |
SubSeq | UTRs

Sequences before the start codon and after the stop codon are obtained with Unix substring commands:

  >NM_001324787.1 NO 5'UTR
>NM_001324787.1 NO 3'UTR
>NM_001324979.1 5'UTR: 1..262 lycopene beta cyclase, chloroplastic/chromoplastic
>NM_001324979.1 3'UTR: 1760..1782 lycopene beta cyclase, chloroplastic/chromoplastic

5-Column Feature Table

How can you generate a 5-column feature table from GenBank format?

  XtoT() {

xtract -pattern INSDSeq -pfx ">Feature " \
-first INSDSeqid,INSDSeq_accession-version \
-group INSDFeature -FKEY INSDFeature_key \
-block INSDInterval -deq "\n" \
-element INSDInterval_from INSDInterval_to \
INSDInterval_point INSDInterval_point \
"&FKEY" -FKEY "()" \
-block INSDQualifier -deq "\n\t\t\t" \
-element INSDQualifier_name INSDQualifier_value

efetch -db nuccore -id U54469 -format gb |
transmute -g2x |

Exploring the INSDSeq XML hierarchy with a ‑pattern {sequence} ‑group {feature} ‑block {qualifier} construct, and adding proper indentation, can produce feature table submission format:

  >Feature U54469.1
1 2881 source
organism Drosophila melanogaster
mol_type genomic DNA
db_xref taxon:7227
chromosome 3
map 67A8-B2
80 2881 gene
gene eIF4E
80 224 mRNA
892 1458
1550 1920
1986 2085
2317 2404
2466 2881
gene eIF4E
product eukaryotic initiation factor 4E-I

An xml2tbl script containing this xtract command is now included with EDirect.

WGS Components

How can you get FASTA format for all components of a WGS project?

  GenerateWGSAccessions() {

gb=$( efetch -db nuccore -id "$1" -format gbc < /dev/null )
ln=$( echo "$gb" | xtract -pattern INSDSeq -element INSDSeq_length )
echo "$gb" |
xtract -pattern INSDSeq -element INSDSeq_locus |
sed -e 's/010*/01/g'
seq -f "${pf}%07.0f" 1 "$ln"

GenerateWGSAccessions "JABRPF000000000" |

returns the expanded list of accessions from the WGS master:


Piping those accessions to:

  efetch -db nuccore -format fasta

retrieves the individual components in FASTA format:

  >JABRPF010000001.1 Enterococcus faecalis strain G109-2 contig00001, ...


Amino Acid Composition

What is the amino acid composition of human titin?

  #!/bin/bash -norc

AAComp() {

abbrev=( Ala Asx Cys Asp Glu Phe Gly His Ile \
Xle Lys Leu Met Asn Pyl Pro Gln Arg \
Ser Thr Sec Val Trp Xxx Tyr Glx )

tr A-Z a-z |
sed 's/[^a-z]//g' |
fold -w 1 |
sort-uniq-count |
while read num lttr
idx=$(printf %i "'$lttr'")
echo -e "${abbrev[$ofs]}\t$num"
done |

efetch -db protein -id Q8WZ42 -format gpc |
xtract -pattern INSDSeq -element INSDSeq_sequence |

This produces a table of residue counts using the three-letter amino acid abbreviations:

  Ala    2084
Arg 1640
Asn 1111
Asp 1720
Cys 513
Gln 942
Glu 3193
Gly 2066
His 478
Ile 2062
Leu 2117
Lys 2943
Met 398
Phe 908
Pro 2517
Ser 2463
Thr 2546
Trp 466
Tyr 999
Val 3184

Longest Sequences

What are the longest known insulin precursor molecules?

  esearch -db protein -query "insulin [PROT]" |
efetch -format docsum |
xtract -pattern DocumentSummary -element Caption Slen Title |
grep -v receptor | sort -k 2,2nr | head -n 5 | cut -f 1 |
xargs -n 1 sh -c 'efetch -db protein -id "$0" -format gp > "$0".gpf'

Post-processing excludes the longer "insulin-like receptor" sequences, sorts by sequence length, and saves the GenPept results to individual files named by their sequence accessions, using the right angle bracket (">") Unix output redirection character:


Archaea Enzyme

Which archaebacteria have chloramphenicol acetyltransferase?

  esearch -db protein -organism archaea \
-query "chloramphenicol acetyltransferase [PROT]" |
efetch -format gpc |
xtract -pattern INSDSeq -element INSDSeq_organism INSDSeq_definition |
grep -i chloramphenicol | grep -v MULTISPECIES |
cut -f 1 | sort -f | uniq -i

Filtering on the definition line produces a list of archaeal organism names:

  Euryarchaeota archaeon
Methanobrevibacter arboriphilus
Methanobrevibacter gottschalkii
Methanobrevibacter millerae
Methanobrevibacter oralis


Gene Counts

How many genes are on each human chromosome?

  for chr in {1..22} X Y MT
esearch -db gene -query "Homo sapiens [ORGN] AND $chr [CHR]" |
efilter -status alive -type coding |
efetch -format docsum |
xtract -pattern DocumentSummary -NAME Name \
-block GenomicInfoType -if ChrLoc -equals "$chr" \
-tab "\n" -element ChrLoc,"&NAME" |
sort | uniq | cut -f 1 |
sort-uniq-count-rank |
reorder-columns 2 1

This returns a count of unique protein-coding genes per chromosome:

  1     2011
2 1224
3 1046
4 742
5 854
6 1015
7 911
8 666
9 763
10 718
11 1279
12 1012
13 327
14 601
15 585
16 835
17 1147
18 266
19 1391
20 528
21 232
22 429
X 839
Y 63
MT 13

The range construct cannot be used for Roman numerals, so the equivalent query on Saccharomyces cerevisiae would need to explicitly list all chromosomes, including the mitochondrion:


Plastid genes can be selected with "source plastid [PROP]" or ‑location plastid.

Chromosome Locations

Where are mammalian calmodulin genes located?

  esearch -db gene -query "calmodulin * [PFN] AND mammalia [ORGN]" |
efetch -format docsum |
xtract -pattern DocumentSummary \
-def "-" -element Id Name MapLocation ScientificName

The ‑def command adds a dash to prevent missing data from shifting columns in the table:

  801       CALM1     14q32.11     Homo sapiens
808 CALM3 19q13.32 Homo sapiens
805 CALM2 2p21 Homo sapiens
24242 Calm1 6q32 Rattus norvegicus
12313 Calm1 12 E Mus musculus
50663 Calm2 6q12 Rattus norvegicus
24244 Calm3 1q21 Rattus norvegicus
12315 Calm3 7 9.15 cM Mus musculus
12314 Calm2 17 E4 Mus musculus
80796 Calm4 13 A1 Mus musculus
617095 CALM1 - Bos taurus
396838 CALM3 - Sus scrofa
520277 CALM3 - Bos taurus
364774 Calml5 17q12.2 Rattus norvegicus

Exon Counts

How many exons are in each dystrophin transcript variant?

  esearch -db gene -query "DMD [GENE] AND human [ORGN]" |
efetch -format docsum |
xtract -pattern DocumentSummary -block GenomicInfoType \
-tab "\n" -element ChrAccVer,ChrStart,ChrStop |
xargs -n 3 sh -c 'efetch -db nuccore -format gbc \
-id "$0" -chr_start "$1" -chr_stop "$2"' |

This retrieves an INSDSeq XML subset record for the indicated gene region, which contains a number of alternatively-spliced dystrophin mRNA and CDS features.

Data extraction computes the number of intervals for each mRNA location (individual exons plus non-adjacent UTRs), and obtains the transcript sequence accession, transcript length, and product name from qualifiers:

  xtract -insd complete mRNA "#INSDInterval" \
transcript_id "%transcription" product |

Final filtering and sorting:

  grep -i dystrophin |
sed 's/dystrophin, transcript variant //g' |
sort -k 2,2nr -k 4,4nr

results in a table of exon counts and transcript lengths:

  NC_000023.11    79    NM_004010.3       14083    Dp427p2
NC_000023.11 79 NM_004009.3 14000 Dp427p1
NC_000023.11 79 NM_004006.3 13992 Dp427m
NC_000023.11 79 NM_000109.4 13854 Dp427c
NC_000023.11 78 XM_006724468.2 13923 X1
NC_000023.11 78 XM_017029328.1 13916 X4
NC_000023.11 78 XM_006724469.3 13897 X2
NC_000023.11 77 XM_006724470.3 13875 X3

Upstream Sequences

What sequences are upstream of phenylalanine hydroxylase genes?

  esearch -db nuccore -query "U49897 [ACCN]" |
elink -target gene |
elink -target homologene |
elink -target gene |
efetch -format docsum |
xtract -pattern DocumentSummary -if GenomicInfoType -element Id \
-block GenomicInfoType -element ChrAccVer -1-based ChrStart ChrStop |

This produces a table with 1-based sequence coordinates:

  5053       NC_000012.12    102958441    102836889
18478 NC_000076.7 87357657 87419999
38871 NT_037436.4 7760453 7763166
24616 NC_005106.4 28066639 28129772
378962 NC_007115.7 17409367 17391680

Then, given a shell script named "upstream.sh":

  #!/bin/bash -norc

if [ -n "$1" ]

while read id accn start stop
if [ $start -eq 0 ] || [ $stop -eq 0 ] || [ $start -eq $stop ]
echo "Skipping $id due to ambiguous coordinates"
if [ $start -gt $stop ]
stop=$(( start + bases ))
start=$(( start + 1 ))
stop=$(( start - 1 ))
start=$(( start - bases ))
rslt=$( efetch -db nuccore -id $accn -format fasta \
-seq_start $start -seq_stop $stop -strand $strand < /dev/null )
echo "$rslt"

the data lines can be piped through:

  upstream.sh 500

to extract and print the 500 nucleotides immediately upstream of each gene:

  >NC_000012.12:c102958941-102958442 Homo sapiens chromosome 12, GRCh38.p13 ...


Complete Genomes

What complete genomes are available for Escherichia coli?

  esearch -db assembly -query \
"Escherichia coli [ORGN] AND representative [PROP]" |
elink -target nuccore -name assembly_nuccore_refseq |
efetch -format docsum |
xtract -pattern DocumentSummary -element AccessionVersion Slen Title |
sed 's/,.*//' |
sort-table -k 2,2nr

This search finds genomic assemblies and sorts the results by sequence length, allowing complete genomes to be easily distinguished from smaller plasmids:

  NC_002695.2    5498578    Escherichia coli O157:H7 str. Sakai DNA
NC_000913.3 4641652 Escherichia coli str. K-12 substr. MG1655
NC_002128.1 92721 Escherichia coli O157:H7 str. Sakai plasmid pO157
NC_002127.1 3306 Escherichia coli O157:H7 str. Sakai plasmid pOSAK1

The sed command removes text (e.g., complete genome, complete sequence, primary assembly) after a comma.

A similar query for humans, additionally filtering out scaffolds, contigs, and plasmids:

  esearch -db assembly -query "Homo sapiens [ORGN] AND representative [PROP]" |
elink -target nuccore -name assembly_nuccore_refseq |
efetch -format docsum |
xtract -pattern DocumentSummary -element AccessionVersion Slen Title |
sed 's/,.*//' |
grep -v scaffold | grep -v contig | grep -v plasmid | grep -v patch |

returns the assembled chromosome and mitochondrial sequence records:

  NC_000001.11    248956422    Homo sapiens chromosome 1
NC_000002.12 242193529 Homo sapiens chromosome 2
NC_000003.12 198295559 Homo sapiens chromosome 3
NC_000004.12 190214555 Homo sapiens chromosome 4
NC_000005.10 181538259 Homo sapiens chromosome 5
NC_000006.12 170805979 Homo sapiens chromosome 6
NC_000007.14 159345973 Homo sapiens chromosome 7
NC_000008.11 145138636 Homo sapiens chromosome 8
NC_000009.12 138394717 Homo sapiens chromosome 9
NC_000010.11 133797422 Homo sapiens chromosome 10
NC_000011.10 135086622 Homo sapiens chromosome 11
NC_000012.12 133275309 Homo sapiens chromosome 12
NC_000013.11 114364328 Homo sapiens chromosome 13
NC_000014.9 107043718 Homo sapiens chromosome 14
NC_000015.10 101991189 Homo sapiens chromosome 15
NC_000016.10 90338345 Homo sapiens chromosome 16
NC_000017.11 83257441 Homo sapiens chromosome 17
NC_000018.10 80373285 Homo sapiens chromosome 18
NC_000019.10 58617616 Homo sapiens chromosome 19
NC_000020.11 64444167 Homo sapiens chromosome 20
NC_000021.9 46709983 Homo sapiens chromosome 21
NC_000022.11 50818468 Homo sapiens chromosome 22
NC_000023.11 156040895 Homo sapiens chromosome X
NC_000024.10 57227415 Homo sapiens chromosome Y
NC_012920.1 16569 Homo sapiens mitochondrion

This process can be automated to loop through a list of specified organisms:

  for org in \
"Agrobacterium tumefaciens" \
"Bacillus anthracis" \
"Escherichia coli" \
"Neisseria gonorrhoeae" \
"Pseudomonas aeruginosa" \
"Shigella flexneri" \
"Streptococcus pneumoniae"
esearch -db assembly -query "$org [ORGN]" |
efilter -query "representative [PROP]" |
elink -target nuccore -name assembly_nuccore_refseq |
efetch -format docsum |
xtract -pattern DocumentSummary -element AccessionVersion Slen Title |
sed 's/,.*//' |
grep -v -i -e scaffold -e contig -e plasmid -e sequence -e patch |
sort-table -k 2,2nr

which generates:

  NC_011985.1    4005130    Agrobacterium radiobacter K84 chromosome 1
NC_011983.1 2650913 Agrobacterium radiobacter K84 chromosome 2
NC_005945.1 5228663 Bacillus anthracis str. Sterne chromosome
NC_003997.3 5227293 Bacillus anthracis str. Ames chromosome
NC_002695.1 5498450 Escherichia coli O157:H7 str. Sakai chromosome
NC_018658.1 5273097 Escherichia coli O104:H4 str. 2011C-3493 ...
NC_011751.1 5202090 Escherichia coli UMN026 chromosome
NC_011750.1 5132068 Escherichia coli IAI39 chromosome
NC_017634.1 4747819 Escherichia coli O83:H1 str. NRG 857C chromosome
NC_000913.3 4641652 Escherichia coli str. K-12 substr. MG1655
NC_002946.2 2153922 Neisseria gonorrhoeae FA 1090 chromosome
NC_002516.2 6264404 Pseudomonas aeruginosa PAO1 chromosome
NC_004337.2 4607202 Shigella flexneri 2a str. 301 chromosome
NC_003028.3 2160842 Streptococcus pneumoniae TIGR4 chromosome
NC_003098.1 2038615 Streptococcus pneumoniae R6 chromosome


SNP Data Table

How can you obtain a tab-delimited table of SNP attributes from a document summary?

  efetch -db snp -id 11549407 -format docsum |

The snp2tbl script extracts fields from HGVS data and prints the individual values for further processing:

  rs11549407    NC_000011.10    5226773    G    A    Genomic    Substitution    HBB
rs11549407 NC_000011.10 5226773 G C Genomic Substitution HBB
rs11549407 NC_000011.10 5226773 G T Genomic Substitution HBB
rs11549407 NG_000007.3 70841 C A Genomic Substitution HBB
rs11549407 NG_000007.3 70841 C G Genomic Substitution HBB
rs11549407 NG_000007.3 70841 C T Genomic Substitution HBB
rs11549407 NM_000518.5 167 C A Coding Substitution HBB
rs11549407 NM_000518.5 167 C G Coding Substitution HBB
rs11549407 NM_000518.5 167 C T Coding Substitution HBB
rs11549407 NP_000509.1 39 Q * Protein Termination HBB
rs11549407 NP_000509.1 39 Q E Protein Missense HBB
rs11549407 NP_000509.1 39 Q K Protein Missense HBB

Columns are SNP identifier, accession.version, offset from start of sequence, letters to delete, letters to insert, sequence class, variant type, and gene name. The internal steps (snp2hgvs, hgvs2spdi, and spdi2tbl) are described below.

Amino Acid Substitutions

What are the missense products of green-sensitive opsin?

  esearch -db gene -query "OPN1MW [PREF] AND human [ORGN]" |
elink -target snp | efilter -class missense |
efetch -format docsum |

SNP document summaries contain HGVS data of the form:

  NC_000023.11:g.154193517C>A, ... ,NP_000504.1:p.Ala285Val

This can be parsed by an xtract ‑hgvs command within the snp2hgvs script:

  snp2hgvs |

into a structured representation of nucleotide and amino acid replacements:


where the original 1-based HGVS positions are converted to 0-based in the XML.

Passing those results through the hgvs2spdi script:

  hgvs2spdi |

converts CDS-relative offsets to sequence-relative positions:


Piping this through the spdi2tbl script:

  spdi2tbl |

completes the final step of the snp2tbl process to generate a SNP Data Table. Filtering that through:

  grep Protein | grep Missense | cut -f 1-5

results in a table of amino acid substitutions sorted by accession, position, and residue:

  rs1238141906    NP_000504.1    40    E    K
rs1189783086 NP_000504.1 42 P L
rs1257135801 NP_000504.1 45 H Y
rs1284438666 NP_000504.1 63 V I
rs1223726997 NP_000504.1 64 I T

Those rows are processed in a while loop that caches the current sequence data:

  while read rsid accn ofs del ins
if [ "$accn" != "$last" ]
seq=$( efetch -db protein -id "$accn" -format gpc < /dev/null |
xtract -pattern INSDSeq -lower INSDSeq_sequence )
pos=$((ofs + 1))
echo ">$rsid [$accn $ins@$pos]"
echo "$seq" |
transmute -replace -offset "$ofs" -delete "$del" -insert "$ins" -lower |
fold -w 50

and uses transmute ‑replace to generate modified FASTA with substituted residues in upper case:

  >rs1238141906 [NP_000504.1 K@41]

SNP-Modified Product Pairs

How can you match codon modifications with amino acid substitutions?

  efetch -db snp -id 11549407 -format docsum |
snp2tbl |

For SNPs that have different substitutions at the same position, the tbl2prod script translates coding sequences (after nucleotide modification), and sorts them with protein sequences (after residue replacement), to produce adjacent matching CDS/protein pairs:


The "+" sign indicates the unmodified "wild-type" nucleotide or amino acid.


Structural Similarity

What archaea structures are similar to snake venom phospholipase?

  esearch -db structure -query "crotalus [ORGN] AND phospholipase A2" |
elink -related |
efilter -query "archaea [ORGN]" |
efetch -format docsum |
xtract -pattern DocumentSummary \
-if PdbClass -equals Hydrolase \
-element PdbDescr |
sort -f | uniq -i

Structure neighbors use geometric comparison to find proteins that are too divergent to be detected by sequence similarity with a BLAST search:

  Crystal Structure Of Autoprocessed Form Of Tk-Subtilisin
Crystal Structure Of Ca2 Site Mutant Of Pro-S324a
Crystal Structure Of Ca3 Site Mutant Of Pro-S324a


Taxonomic Lineage

What are the major taxonomic lineage nodes for humans?

  efetch -db taxonomy -id 9606 -format xml |
xtract -pattern Taxon -first TaxId -tab "\n" -element ScientificName \
-block "**/Taxon" \
-if Rank -is-not "no rank" -and Rank -is-not clade \
-tab "\n" -element Rank,ScientificName

This uses the double star / child construct to recursively explore the data hierarchy:

  9606            Homo sapiens
superkingdom Eukaryota
kingdom Metazoa
phylum Chordata
subphylum Craniata
superclass Sarcopterygii
class Mammalia
superorder Euarchontoglires
order Primates


Installation of EDirect on Cloud

To install the EDirect software, open a terminal window and execute the following command:

  sh -c "$(curl -fsSL ftp://ftp.ncbi.nlm.nih.gov/entrez/entrezdirect/install-edirect.sh)"

At the end of installation, answer "y" to have the script run the PATH update command to edit your configuration file, so that EDirect programs can be run in subsequent terminal sessions.

One installation is complete, run:

  export PATH=${PATH}:${HOME}/edirect

to set the PATH for the current terminal session.

Downloading BLAST Software

Obtain the Magic-BLAST software if it is not already installed:

  download-ncbi-software magic-blast

Preparing Chromosome Files

Retrieve the sequence of human chromosome 7 with:

  efetch -db nuccore -id NC_000007 -format fasta -immediate > NC_000007.fsa

Download protein-coding genes on human chromosome 7 in document summary format, and extract the gene ranges into a tab-delimited table:

  esearch -db gene -query "Homo sapiens [ORGN] AND 7 [CHR]" |
efilter -status alive -type coding | efetch -format docsum |
gene2range "7" > NC_000007.gen

SRA Gene Analysis

Run a magicblast search of SRR6314034 against chromosome 7:

  magicblast -sra SRR6314034 -subject NC_000007.fsa -outfmt asn |
asn2xml > SRR6314034.xml

Passing the alignment details through the blst2tkns script:

  cat SRR6314034.xml |
blst2tkns |

tokenizes the alignment parts:

  index          1
score 6268
start 33088037
stop 33167397
strand plus
match 12
mismatch 1
genomic-ins 1

Piping these to the split-at-intron script:

  split-at-intron |

detects large genomic insertions:

  6268    1    plus    33088037..33091003,33163798..33167397
4910 2 plus 33089462..33091003,33163799..33167397
1814 3 plus 33164965..33167397

These are filtered by minimum score with:

  filter-columns '$1 > 1000' |

The fuse-ranges script then merges overlapping alignments:

  fuse-ranges |

into a minimal set of extended ranges:

  plus     33088037    33091003    2967
plus 33163798 33167398 3601
plus 33272248 33272278 31
plus 33394238 33394551 314
minus 33088037 33091004 2968
minus 33163798 33167398 3601
minus 33255886 33256012 127
minus 33394326 33394551 226

Running each range through the find-in-gene script:

  while read std min max len
cat NC_000007.gen |
find-in-gene "$std" "$min" "$max"
done |
sort -f | uniq -i

returns the names of gene(s) that overlap the aligned segments:



PubChem in Entrez

Entrez can search on the complete synonym of a compound:

  esearch -db pccompound -query "catechol [CSYN]" |
efetch -format uid

to return a small number of closely matching compound identifiers (CIDs):


Entrez document summaries for a compound:

  efetch -db pccompound -id 289 -format docsum |
xtract -pattern DocumentSummary -element MolecularFormula IsomericSmiles

contains general descriptive information fields for the compound:

  C6H6O2    C1=CC=C(C(=C1)O)O

Power User Gateway (PUG) REST Query Form

PubChem also supports a RESTful service for more advanced queries. Nquire provides a -pugrest URL shortcut. The base form of a search request is is:

  nquire -pugrest [compound|substance|assay] [input] [operation] [output]

Searches can use the name of a compound:

  nquire -pugrest compound name catechol cids TXT

to obtain the best matching compound identifier:


The CID can be used as the input key to obtain a title and description:

  nquire -pugrest compound cid 289 description XML

or to retrieve a much more detailed record:

  nquire -pugrest compound cid 289 record XML |

that includes the canonical or isomeric SMILES codes:

  xtract -pattern PC-InfoData \
-if PC-Urn_label -equals SMILES -and PC-Urn_name -equals Isomeric \
-element PC-InfoData_value_sval

PubChem Chemical Identifiers

Certain identifier types require POST arguments to encode special symbols:

  nquire -pugrest compound smiles description XML \
-smiles "C1=CC=C(C(=C1)O)O" |
xtract -pattern InformationList -element Title Description

This returns the chemical name and description:

  Catechol    Catechol is a benzenediol comprising...

Other identifier key types that are encoded in separate arguments are shown below:

  nquire -pugrest compound inchi synonyms TXT \
-inchi "1S/C6H6O2/c7-5-3-1-2-4-6(5)8/h1-4,7-8H"

nquire -pugrest compound inchikey cids JSON \

nquire -pugrest compound/fastsubstructure/smarts/cids/XML \
-smarts "[#7]-[#6]-1=[#6]-[#6](C#C)=[#6](-[#6]-[#8])-[#6]=[#6]-1"

(Nquire -inchi will supply the expected "InChI=" prefix if it is missing in the argument string.)

PUG-REST Asynchronous Queries

Some PUG-REST queries are computationally intensive and run asynchronously:

  nquire -pugrest compound/superstructure/cid/2244/XML |

The returned <ListKey> token is piped to an nquire -pugwait command, which polls the server until the results are available:

  nquire -pugwait

Identifiers are then downloaded and placed directly in an ENTREZ_DIRECT message:


Support for this form was added when EDirect was redesigned in 2020.

Bookshelf ID: NBK565821


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