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- EMDB-73644: AI-generated RNA-guided nuclease TAM-bound state -

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Basic information

Entry
Database: EMDB / ID: EMD-73644
TitleAI-generated RNA-guided nuclease TAM-bound state
Map data
Sample
  • Complex: AI-designed RNA-guided nuclease TAM-bound state
    • Protein or peptide: AI-designed RNA-guided nuclease
    • RNA: RNA (116-MER)
    • DNA: DNA (5'-D(*GP*GP*TP*TP*GP*AP*TP*C)-3')
    • DNA: DNA (5'-D(*AP*CP*AP*TP*CP*AP*AP*CP*C)-3')
KeywordsCRISPR / nuclease / AI / Evolution Scale Modeling / Inverse Folding / ESM-IF1 / TnpB / TAM / PAM / spacer / exonuclease / DNA binding protein-DNA complex / enzyme / ribonucleoprotein / RNA / RNA-guided nuclease / Cas12 / ISDra2 / ESM / RNA BINDING PROTEIN
Biological speciessynthetic construct (others) / Deinococcus radiodurans (radioresistant)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.8 Å
AuthorsSkopintsev P / Esain-Garcia I / Doudna J
Funding support United States, Switzerland, 2 items
OrganizationGrant numberCountry
National Science Foundation (NSF, United States)DGE 2334027 United States
Swiss National Science FoundationP500PB_214418 Switzerland
CitationJournal: To Be Published
Title: AI-generated RNA-guided nuclease TAM-bound state
Authors: Skopintsev P / Esain-Garcia I / Doudna J
History
DepositionOct 29, 2025-
Header (metadata) releaseJul 15, 2026-
Map releaseJul 15, 2026-
UpdateJul 15, 2026-
Current statusJul 15, 2026Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_73644.png

Downloads & links

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Map

FileDownload / File: emd_73644.map.gz / Format: CCP4 / Size: 83.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.85 Å/pix.
x 280 pix.
= 237.44 Å		0.85 Å/pix.
x 280 pix.
= 237.44 Å		0.85 Å/pix.
x 280 pix.
= 237.44 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.848 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.05
Minimum - Maximum-0.0017720968 - 1.6106251
Average (Standard dev.)0.00085021515 (±0.018604504)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions280280280
Spacing280280280
CellA=B=C: 237.44 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #2

Fileemd_73644_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #1

Fileemd_73644_half_map_2.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : AI-designed RNA-guided nuclease TAM-bound state

EntireName: AI-designed RNA-guided nuclease TAM-bound state
Components
  • Complex: AI-designed RNA-guided nuclease TAM-bound state
    • Protein or peptide: AI-designed RNA-guided nuclease
    • RNA: RNA (116-MER)
    • DNA: DNA (5'-D(*GP*GP*TP*TP*GP*AP*TP*C)-3')
    • DNA: DNA (5'-D(*AP*CP*AP*TP*CP*AP*AP*CP*C)-3')

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Supramolecule #1: AI-designed RNA-guided nuclease TAM-bound state

SupramoleculeName: AI-designed RNA-guided nuclease TAM-bound state / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Molecular weightTheoretical: 149 KDa

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Macromolecule #1: AI-designed RNA-guided nuclease

MacromoleculeName: AI-designed RNA-guided nuclease / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: Hydrolases; Acting on ester bonds
Source (natural)Organism: synthetic construct (others)
Molecular weightTheoretical: 46.649828 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MIHRKAFVVR LYPNARQEEL IERTLGSARF VYNHFLAERK AAYKESGKPL TYGQMSSLLT KLKQAPETAW LSEVDKFALQ NSLKNLERA FQNFFRTLKK KGEKVGFPKF RKKKTGESFR TQFTNNNIQI GKGKLKLPKL GWVKTKGQQE IKGKVLNVTV K RVHEGHYE ...String:
MIHRKAFVVR LYPNARQEEL IERTLGSARF VYNHFLAERK AAYKESGKPL TYGQMSSLLT KLKQAPETAW LSEVDKFALQ NSLKNLERA FQNFFRTLKK KGEKVGFPKF RKKKTGESFR TQFTNNNIQI GKGKLKLPKL GWVKTKGQQE IKGKVLNVTV K RVHEGHYE ASVLVEVEIP ELPEAPLLAA GVDLGIKNFA VVTDGEEFEK VENPKFLKKT EKRLARAQRR LSRRKKGSAR WE KARTRLA RIHKRIVNQR QDFLHKLTTG LVKTYQIIAV EDLNPKNMMK NHHLARAISD AAWGEFRRQL EYKAEWYGRT VAK VSRWFP SSQLCHVCGH KNPEVKDLSV RTWTCPSCGT THDRDENAAL NIRREALVAA GISDTLNAHG GYVRPASAGN GLRS ENHAT LVV

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Macromolecule #2: RNA (116-MER)

MacromoleculeName: RNA (116-MER) / type: rna / ID: 2 / Number of copies: 1
Source (natural)Organism: Deinococcus radiodurans (radioresistant)
Molecular weightTheoretical: 42.598207 KDa
SequenceString:
GGUGGCUGCG GGAAUCUCAG ACACCUUAAA CGCUCAUGGA GGCUAUGUCA GACCUGCUUC GGCGGGCAAU GGUCUGCGAA GUGAGAAUC ACGCGACUUU AGUCGUGUGA GGUUCAAGAG UCCCUUGGCG CCC

GENBANK: GENBANK: OV024757.1

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Macromolecule #3: DNA (5'-D(*GP*GP*TP*TP*GP*AP*TP*C)-3')

MacromoleculeName: DNA (5'-D(*GP*GP*TP*TP*GP*AP*TP*C)-3') / type: dna / ID: 3 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: synthetic construct (others)
Molecular weightTheoretical: 2.456639 KDa
SequenceString:
(DT)(DA)(DT)(DT)(DG)(DA)(DT)(DG)

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Macromolecule #4: DNA (5'-D(*AP*CP*AP*TP*CP*AP*AP*CP*C)-3')

MacromoleculeName: DNA (5'-D(*AP*CP*AP*TP*CP*AP*AP*CP*C)-3') / type: dna / ID: 4 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: synthetic construct (others)
Molecular weightTheoretical: 2.698812 KDa
SequenceString:
(DT)(DC)(DA)(DT)(DC)(DA)(DA)(DT)(DA)

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 8
GridModel: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: GOLD / Support film - topology: HOLEY
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 281 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeTFS KRIOS
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 50.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.8 µm
Sample stageCooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

CTF correctionSoftware - Name: cryoSPARC (ver. 4.3.0) / Software - details: PatchCTF / Type: PHASE FLIPPING AND AMPLITUDE CORRECTION
Startup modelType of model: OTHER / Details: Ab-initio map calculated in cryoSPARC
Final reconstructionResolution.type: BY AUTHOR / Resolution: 2.8 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 4.3.0) / Number images used: 389833
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 4.3.0)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 4.3.0)
Final 3D classificationSoftware - Name: cryoSPARC (ver. 4.3.0)

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Atomic model buiding 1

RefinementSpace: REAL / Protocol: OTHER
Output model

PDB-9yyg:
AI-generated RNA-guided nuclease TAM-bound state

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