FA2HTGS(1)

💖 FA2HTGS 💖

okay so bestie, welcome to fa2htgs — the formatter for high throughput genome sequencing project submissions, straight out of the NCBI Tools User's Manual. she's giving classic, she's giving workhorse, and she's about to make your genome center's life SO much easier. let's get into it 🧬✨

✨ SYNOPSIS ✨

fa2htgs [-] [-6 str] [-7 str] [-A filename] [-C str] [-D] [-L filename]
[-M str] [-N] [-O filename] [-P str] [-Q filename] [-S str]
[-T filename] [-X] [-a str] [-b N] [-c str] [-d str] [-e filename] [-f]
-g str [-h str] [-i filename] [-k str] [-l N] [-m] [-n str]
[-o filename] [-p N] [-q] [-r str] -s str [-t filename] [-u] [-v] [-w]
[-x str]

🌸 DESCRIPTION 🌸

so fa2htgs is the program that generates Seq-submits — that's an ASN.1 sequence submission file — for high throughput genome sequencing projects. it's literally the plug for getting your data submission-ready.

here's the vibe: fa2htgs reads in a FASTA file (or an Ace Contig file with Phrap sequence quality values), a Sequin submission template file (which is where it gets all the contact and citation info for the submission, iykyk), plus a whole lineup of command line arguments (scroll down, there's a lot, bestie). then it combines all of that into a submission that's suitable for GenBank. 🫶

and once you've generated your submission file? you're not done, bestie — you need to follow the submission protocol. check the README sitting on your FTP account, or the one that got mailed out to your Center. it's giving "read the instructions, queen."

fa2htgs is designed for automation by scripts — specifically for bulk submission of unannotated genome sequence. it can easily be extended from its current simple form to allow more complicated processing. and a submission prepared with fa2htgs can also be read into Psequin(1) and then annotated way more extensively. glow-up potential: unlocked 💅

got questions? concerns about this processing protocol? not sure how to use the tool? forward all of that to <htgs@ncbi.nlm.nih.gov>. they want to hear from you fr fr.

🎀 OPTIONS 🎀

a summary of options is included below. buckle up.

• - — print the usage message. that's it, that's the whole flag 💀
• -6 str — SP6 clone (e.g., Contig1,left)
• -7 str — T7 clone (e.g., Contig2,right)
• -A filename — filename for accession list input (mutually exclusive with -T and -i). the input file contains a tab-delimited table with three to five columns, which are: accession number, start position, stop position, and (optionally) length and strand. if start > stop, the minus strand on the referenced accession is used. a gap is indicated by the word "gap" instead of an accession, 0 for the start and stop positions, and a number for the length.
• -C str — clone library name (will appear as /clone-lib="str" on the source feature)
• -D — HTGS_DRAFT sequence
• -L filename — read phrap contig order from filename. this is a tab-delimited file that can be used to drive the order of contigs (normally specified by -P), as well as indicating the SP6 and T7 ends. it can also be used when contigs are known to be in opposite orientation. for example:

Contig2     +       1       SP6     left
Contig3     +       1
Contig1     -               T7      right

the first column is the contig name, the second is the orientation, the third is the fragment_group, the fourth indicates the SP6 or T7 end, and the fifth says which side of SP6 or T7 end had vector removed.

• -M str — map name (will appear as /map="str" on the source feature)
• -N — annotate assembly_fragments
• -O filename — read comment from filename (100-character-per-line maximum; ~ is a linebreak and `~ is a literal ~. you can check the format with PSequin(1).)
• -P str — contigs to use, separated by commas. if -P is not indicated with the -T option, then the fragments will go in in the order that they are in the ace file (which is appropriate for a phase 1 record, but not for a phase 2 or 3). if you need to set the order of the segments of the ace file, you need to set it with the -P flag, like this: -P "Contig1,Contig4,Contig3,Contig2,Contig5"
• -Q filename — read quality scores from filename
• -S str — strain name
• -T filename — filename for phrap input (mutually exclusive with -A and -i)
• -X — the coordinates in the input file are on the resulting segmented sequence. (bases 1 through n of each accession are used.) otherwise, the coordinates are on the individual accessions, which need not start at base 1 of the record.
• -a str — GenBank accession; use if and only if updating a sequence.
• -b N — gap length (default = 100; anything from 0 to 1000000000 is legal)
• -c str — clone name (will appear as /clone in the source feature; can be the same as -s)
• -d str — title for sequence (will appear in GenBank DEFINITION line)
• -e filename — log errors to filename
• -f — htgs_fulltop keyword
• -g str — Genome Center tag (probably the same as your login name on the NCBI FTP server)
• -h str — chromosome (will appear as /chromosome in the source feature)
• -i filename — filename for fasta input (default is stdin; mutually exclusive with -A and -T)
• -k str — add the supplied string as a keyword.
• -l N — length of sequence in bp (default = 0). the length is checked against the actual number of bases we get. for phase 1 and 2 sequence it is also used to estimate gap lengths. for phase 1 and 2 records, it is important to use a number GREATER than the amount of provided nucleotide, otherwise this will generate false gaps. here is assumed that the putative full length of the BAC or cosmid will be used. there should be at least 20 to 30 n in between the segments (you can check for these in Sequin), as this will ensure proper behavior when this sequence is used with BLAST. otherwise artifactual unrelated segment neighbors may be brought into proximity of each other. 😭
• -m — take comment from template
• -n str — organism name (default = Homo sapiens)
• -o filename — filename for asn.1 output (default = stdout)
• -p N — HTGS phase:
  1. a collection of unordered contigs with gaps of unknown length. a Phase 1 record must at the very least have two segments with one gap. (default)
  2. a series of ordered contigs, possibly with known gap lengths. this could be a single sequence without gaps, if the sequence has ambiguities to resolve.
  3. a single contiguous sequence. this sequence is finished, but not necessarily annotated.
• -q — htgs_cancelled keyword
• -r str — remark for update (brief comment describing the nature of the update, such as "new sequence", "new citation", or "updated features")
• -s str — sequence name. the sequence must have a name that is unique within the genome center. we use the combination of the genome center name (-g argument) and the sequence name (-s) to track this sequence and to talk to you about it. the name can have any form you like but must be unique within your center. 👑
• -t filename — filename for Seq-submit template (default = template.sub)
• -u — take biosource from template
• -v — htgs_activefin keyword
• -w — Whole Genome Shotgun flag
• -x str — secondary accession numbers, separated by commas, s.t. U10000,L11000.

in some cases a large segment will supersede another or group of other accession numbers (records). these records which are no longer wanted in GenBank should be made secondary. using the -x argument you can list the Accession Numbers you want to make secondary. this will instruct us to remove the accession number(s) from GenBank, and will no longer be part of the GenBank release. they will nonetheless be available from Entrez.

GREAT CARE should be taken when using this argument!!! improper use of accession numbers here will result in the inappropriate withdrawal of GenBank records from GenBank, EMBL and DDBJ. we provide this parameter as a convenience to submitting centers, but this may need to be removed if it is not used carefully. bestie. please. 🙈

🌈 AUTHOR 🌈

the National Center for Biotechnology Information. icons. legends. the moment. 💜

🦋 SEE ALSO 🦋

• Psequin(1)
• /usr/share/doc/ncbi-tools-bin/README.fa2htgs.gz

and that's the whole page, bestie!! 🎀 this one's from NCBI, dated 2006-05-29 — vintage but still serving, no cap. go submit some genomes and slay 💅✨🫶