Preparing a Sequence Listing for the USPTO is a formal process governed by 37 CFR 1.821–1.825 and requires strict adherence to format, content, and electronic submission rules. The goal is to present nucleotide and amino acid sequence data in a standardized, computer-readable way that supports examination and database use.
Regulatory basis and scope
The USPTO sequence listing requirements are grounded in the U.S. Code of Federal Regulations, specifically 37 CFR §§1.821–1.825. These rules mandate standard symbols, standard formatting, and submission of the listing in computer-readable form for all patent applications that disclose qualifying biological sequences.
A sequence listing is required for:
All unbranched nucleotide sequences with 10 or more bases.
All unbranched, non-D amino acid sequences with 4 or more residues.
This requirement applies to every qualifying sequence mentioned in the application, including unclaimed sequences such as PCR primers.
File format and electronic submission
The sequence listing must be prepared as an ASCII text file, not as a word-processor or PDF file. The file must be compatible with common operating systems such as MS-DOS, MS-Windows, Unix, and Macintosh, using ASCII Carriage Return plus ASCII Line Feed as line terminators and continuous pagination (no hard page breaks).
Key electronic requirements include:
Computer-readable form (typically on CD or other accepted medium).
No page numbers within the listing file.
A labeled medium indicating applicant name, invention title, date of recording, and operating system used.
The USPTO “PatentIn” software is available to help applicants generate compliant listings in the correct format.
Content and organization of the listing
A compliant sequence listing must contain the actual nucleotide and/or amino acid sequences and a defined set of “numeric identifiers” that capture bibliographic, technical, and sequence-specific information. Items marked “M” (mandatory) must be present, and the identifiers must appear in a specified order.
Core mandatory identifiers include:
<110> Applicant (up to about ten names, one per line).
<120> Title of the invention.
<130> File reference (when filed before assignment or application number).
<140>, <141>, <150>, <151> for current and prior application numbers and filing dates, when applicable.
<160> Total number of SEQ ID NOs in the listing.
<210>–<213> for each sequence: SEQ ID NO, length, type (DNA/RNA/PRT), and organism.
Feature and annotation requirements
Biological features and special sequence characteristics are captured using identifiers <220>–<223> when certain conditions are met. These become mandatory if:
“n” bases, “Xaa” residues, or modified/unusual L-amino acids or bases are present.
The organism is designated “Artificial Sequence” or “Unknown”.
The molecule is a combined DNA/RNA sequence.
Within this feature section:
<221> Name/Key provides a feature identifier, preferably from WIPO ST.25 tables.
<222> Location specifies positions (e.g., from/to) within the sequence.
<223> Other Information allows brief explanatory text, limited to four lines.
Publication and sequence details
Optional identifiers <300>–<313> permit inclusion of related publication and database information, such as authors, journal details, database accession numbers, and patent document citations. These fields support cross-referencing to prior publications or database entries but are not mandatory.
The actual sequence is presented under identifier <400>, which must directly follow the corresponding SEQ ID NO line. SEQ ID NO:1 should be reserved for the most important claimed sequence, and sequences intended to be claimed are typically listed first to highlight their significance in the application.