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- EMDB-4980: Cryo-EM structure of an MCM loading intermediate -

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Basic information

Entry
Database: EMDB / ID: EMD-4980
TitleCryo-EM structure of an MCM loading intermediate
Map dataFull map - complete MCM-ORC (MO) origin licensing intermediate
Sample
  • Complex: The MCM-ORC (MO) loading intermediate
    • Complex: MCM-Orc6N lobe of the MCM-ORC (MO) origin licensing intermediate.
      • Protein or peptide: x 7 types
      • DNA: x 2 types
    • Complex: Orc1-5-Orc6C lobe of the MCM-ORC (MO) origin licensing intermediate
      • Protein or peptide: x 5 types
    • Complex: The MCM-ORC (MO) loading intermediate protein complex
    • Complex: DNA
  • Ligand: x 4 types
KeywordsDNA Replication / Origin licensing / MCM2-7 helicase / Origin Recognition Complex / REPLICATION
Function / homology
Function and homology information


CDC6 association with the ORC:origin complex / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / MCM complex binding / Assembly of the ORC complex at the origin of replication / nuclear DNA replication / premeiotic DNA replication ...CDC6 association with the ORC:origin complex / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / MCM complex binding / Assembly of the ORC complex at the origin of replication / nuclear DNA replication / premeiotic DNA replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / nuclear origin of replication recognition complex / mitotic DNA replication / Activation of the pre-replicative complex / nucleosome organization / CMG complex / nuclear pre-replicative complex / Activation of ATR in response to replication stress / DNA replication preinitiation complex / MCM complex / replication fork protection complex / mitotic DNA replication checkpoint signaling / double-strand break repair via break-induced replication / mitotic DNA replication initiation / single-stranded DNA helicase activity / regulation of DNA-templated DNA replication initiation / DNA strand elongation involved in DNA replication / silent mating-type cassette heterochromatin formation / DNA unwinding involved in DNA replication / Orc1 removal from chromatin / nuclear replication fork / regulation of DNA replication / DNA replication origin binding / nucleosome binding / DNA replication initiation / subtelomeric heterochromatin formation / DNA helicase activity / helicase activity / transcription elongation by RNA polymerase II / heterochromatin formation / single-stranded DNA binding / DNA helicase / chromosome, telomeric region / DNA damage response / chromatin binding / ATP hydrolysis activity / nucleoplasm / ATP binding / nucleus / metal ion binding / cytoplasm
Similarity search - Function
Origin recognition complex, subunit 6, fungi / : / Origin recognition complex subunit 1 C-terminal winged HTH domain / Origin recognition complex, subunit 6 / Origin recognition complex subunit 6 (ORC6) / Origin recognition complex subunit 4 / Origin recognition complex, subunit 3 / Origin recognition complex, subunit 5 / Origin recognition complex subunit 4, C-terminal / Origin recognition complex subunit 3, winged helix C-terminal ...Origin recognition complex, subunit 6, fungi / : / Origin recognition complex subunit 1 C-terminal winged HTH domain / Origin recognition complex, subunit 6 / Origin recognition complex subunit 6 (ORC6) / Origin recognition complex subunit 4 / Origin recognition complex, subunit 3 / Origin recognition complex, subunit 5 / Origin recognition complex subunit 4, C-terminal / Origin recognition complex subunit 3, winged helix C-terminal / Origin recognition complex subunit 3, N-terminal / : / : / Origin recognition complex (ORC) subunit 3 N-terminus / Origin recognition complex (ORC) subunit 4 C-terminus / Origin recognition complex (ORC) subunit 5 C-terminus / Origin recognition complex winged helix C-terminal / ORC5, lid domain / Orc1-like, AAA ATPase domain / Origin recognition complex subunit 2 / AAA ATPase domain / Origin recognition complex, subunit 2 / AAA lid domain / AAA lid domain / : / : / MCM3 winged helix domain / MCM4, winged helix domain / : / MCM5, C-terminal domain / DNA replication licensing factor Mcm5 / DNA replication licensing factor Mcm3 / Mini-chromosome maintenance complex protein 4 / DNA replication licensing factor Mcm6 / DNA replication licensing factor Mcm7 / Mcm6, C-terminal winged-helix domain / MCM6 C-terminal winged-helix domain / DNA replication licensing factor Mcm2 / Mini-chromosome maintenance protein 2 / Mini-chromosome maintenance, conserved site / MCM family signature. / Bromo adjacent homology domain / BAH domain / Bromo adjacent homology (BAH) domain / Bromo adjacent homology (BAH) domain superfamily / BAH domain profile. / MCM N-terminal domain / MCM N-terminal domain / MCM OB domain / Mini-chromosome maintenance protein / MCM, AAA-lid domain / MCM P-loop domain / MCM OB domain / MCM AAA-lid domain / MCM family domain profile. / minichromosome maintenance proteins / MCM domain / ATPase family associated with various cellular activities (AAA) / ATPase, AAA-type, core / EF-Hand 1, calcium-binding site / Winged helix-like DNA-binding domain superfamily / ATPases associated with a variety of cellular activities / AAA+ ATPase domain / Nucleic acid-binding, OB-fold / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
DNA replication licensing factor MCM3 / DNA replication licensing factor MCM2 / Minichromosome maintenance protein 5 / DNA replication licensing factor MCM4 / Origin recognition complex subunit 2 / DNA replication licensing factor MCM7 / Origin recognition complex subunit 6 / Origin recognition complex subunit 5 / DNA replication licensing factor MCM6 / Origin recognition complex subunit 1 ...DNA replication licensing factor MCM3 / DNA replication licensing factor MCM2 / Minichromosome maintenance protein 5 / DNA replication licensing factor MCM4 / Origin recognition complex subunit 2 / DNA replication licensing factor MCM7 / Origin recognition complex subunit 6 / Origin recognition complex subunit 5 / DNA replication licensing factor MCM6 / Origin recognition complex subunit 1 / Origin recognition complex subunit 3 / Origin recognition complex subunit 4
Similarity search - Component
Biological speciesSaccharomyces cerevisiae S288C (yeast) / Saccharomyces cerevisiae S288c (yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.4 Å
AuthorsMiller TCR / Locke J
Funding support United Kingdom, 2 items
OrganizationGrant numberCountry
The Francis Crick InstituteFC0010065 United Kingdom
European Research Council820102 United Kingdom
CitationJournal: Nature / Year: 2019
Title: Mechanism of head-to-head MCM double-hexamer formation revealed by cryo-EM.
Authors: Thomas C R Miller / Julia Locke / Julia F Greiwe / John F X Diffley / Alessandro Costa /
Abstract: In preparation for bidirectional DNA replication, the origin recognition complex (ORC) loads two hexameric MCM helicases to form a head-to-head double hexamer around DNA. The mechanism of MCM double- ...In preparation for bidirectional DNA replication, the origin recognition complex (ORC) loads two hexameric MCM helicases to form a head-to-head double hexamer around DNA. The mechanism of MCM double-hexamer formation is debated. Single-molecule experiments have suggested a sequential mechanism, in which the ORC-dependent loading of the first hexamer drives the recruitment of the second hexamer. By contrast, biochemical data have shown that two rings are loaded independently via the same ORC-mediated mechanism, at two inverted DNA sites. Here we visualize MCM loading using time-resolved electron microscopy, and identify intermediates in the formation of the double hexamer. We confirm that both hexamers are recruited via the same interaction that occurs between ORC and the C-terminal domains of the MCM helicases. Moreover, we identify the mechanism of coupled MCM loading. The loading of the first MCM hexamer around DNA creates a distinct interaction site, which promotes the engagement of ORC at the N-terminal homodimerization interface of MCM. In this configuration, ORC is poised to direct the recruitment of the second hexamer in an inverted orientation, which is suitable for the formation of the double hexamer. Our results therefore reconcile the two apparently contrasting models derived from single-molecule experiments and biochemical data.
History
DepositionMay 15, 2019-
Header (metadata) releaseNov 20, 2019-
Map releaseNov 20, 2019-
UpdateMay 22, 2024-
Current statusMay 22, 2024Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.016
  • Imaged by UCSF Chimera
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  • Surface view colored by radius
  • Surface level: 0.016
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-6rqc
  • Surface level: 0.016
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_4980.png

Downloads & links

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Map

FileDownload / File: emd_4980.map.gz / Format: CCP4 / Size: 149.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationFull map - complete MCM-ORC (MO) origin licensing intermediate
Projections & slices

Image control

Size
Brightness
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Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.38 Å/pix.
x 340 pix.
= 469.2 Å		1.38 Å/pix.
x 340 pix.
= 469.2 Å		1.38 Å/pix.
x 340 pix.
= 469.2 Å

Surface

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Images are generated by Spider.

Voxel sizeX=Y=Z: 1.38 Å
Density

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Histogram (log scale)
Contour LevelBy AUTHOR: 0.016 / Movie #1: 0.016
Minimum - Maximum-0.036754306 - 0.06955264
Average (Standard dev.)0.00020005555 (±0.0018424687)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions340340340
Spacing340340340
CellA=B=C: 469.2 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.381.381.38
M x/y/z340340340
origin x/y/z0.0000.0000.000
length x/y/z469.200469.200469.200
α/β/γ90.00090.00090.000
start NX/NY/NZ929262
NX/NY/NZ290290360
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS340340340
D min/max/mean-0.0370.0700.000

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Supplemental data

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Mask #1

Fileemd_4980_msk_1.map
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Mask #2

Fileemd_4980_msk_2.map
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Mask #3

Fileemd_4980_msk_3.map
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Additional map: Half map - MCM-Orc6N lobe of MCM-ORC (MO)...

Fileemd_4980_additional_1.map
AnnotationHalf map - MCM-Orc6N lobe of MCM-ORC (MO) origin licensing intermediate - Map from RELION multibody refinement of full MO complex.
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Additional map: Half map - MCM-Orc6N lobe of MCM-ORC (MO)...

Fileemd_4980_additional_2.map
AnnotationHalf map - MCM-Orc6N lobe of MCM-ORC (MO) origin licensing intermediate - Map from RELION multibody refinement of full MO complex.
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Additional map: Full map - MCM-Orc6N lobe of MCM-ORC (MO)...

Fileemd_4980_additional_3.map
AnnotationFull map - MCM-Orc6N lobe of MCM-ORC (MO) origin licensing intermediate, derived from RELION multibody refinement of full MO complex.
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Additional map: Half map - Orc1-5-Orc6C lobe of MCM-ORC (MO)...

Fileemd_4980_additional_4.map
AnnotationHalf map - Orc1-5-Orc6C lobe of MCM-ORC (MO) origin licensing intermediate, derived from RELION multibody refinement of full MO complex.
Projections & Slices
AxesZYX

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Additional map: Full map - Orc1-5-Orc6C lobe of MCM-ORC (MO)...

Fileemd_4980_additional_5.map
AnnotationFull map - Orc1-5-Orc6C lobe of MCM-ORC (MO) origin licensing intermediate, derived from RELION multibody refinement of full MO complex.
Projections & Slices
AxesZYX

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Additional map: Half map – Orc1-5-Orc6C lobe of MCM-ORC (MO)...

Fileemd_4980_additional_6.map
AnnotationHalf map – Orc1-5-Orc6C lobe of MCM-ORC (MO) origin licensing intermediate, derived from RELION multibody refinement of full MO complex.
Projections & Slices
AxesZYX

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Half map: Half map - complete MCM-ORC (MO) origin licensing intermediate

Fileemd_4980_half_map_1.map
AnnotationHalf map - complete MCM-ORC (MO) origin licensing intermediate
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AxesZYX

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Half map: Half map - complete MCM-ORC (MO) origin licensing intermediate

Fileemd_4980_half_map_2.map
AnnotationHalf map - complete MCM-ORC (MO) origin licensing intermediate
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Sample components

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Entire : The MCM-ORC (MO) loading intermediate

EntireName: The MCM-ORC (MO) loading intermediate
Components
  • Complex: The MCM-ORC (MO) loading intermediate
    • Complex: MCM-Orc6N lobe of the MCM-ORC (MO) origin licensing intermediate.
      • Protein or peptide: Origin recognition complex subunit 6
      • Protein or peptide: DNA replication licensing factor MCM2
      • Protein or peptide: DNA replication licensing factor MCM3
      • Protein or peptide: DNA replication licensing factor MCM4
      • Protein or peptide: Minichromosome maintenance protein 5
      • Protein or peptide: DNA replication licensing factor MCM6
      • Protein or peptide: DNA replication licensing factor MCM7
      • DNA: DNA (88-MER)
      • DNA: DNA (88-MER)
    • Complex: Orc1-5-Orc6C lobe of the MCM-ORC (MO) origin licensing intermediate
      • Protein or peptide: Origin recognition complex subunit 1
      • Protein or peptide: Origin recognition complex subunit 2
      • Protein or peptide: Origin recognition complex subunit 3
      • Protein or peptide: Origin recognition complex subunit 4
      • Protein or peptide: Origin recognition complex subunit 5
    • Complex: The MCM-ORC (MO) loading intermediate protein complex
    • Complex: DNA
  • Ligand: ADENOSINE-5'-TRIPHOSPHATE
  • Ligand: MAGNESIUM ION
  • Ligand: ADENOSINE-5'-DIPHOSPHATE
  • Ligand: ZINC ION

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Supramolecule #1: The MCM-ORC (MO) loading intermediate

SupramoleculeName: The MCM-ORC (MO) loading intermediate / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#14

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Supramolecule #2: MCM-Orc6N lobe of the MCM-ORC (MO) origin licensing intermediate.

SupramoleculeName: MCM-Orc6N lobe of the MCM-ORC (MO) origin licensing intermediate.
type: complex / ID: 2 / Parent: 1 / Macromolecule list: #6-#14 / Details: Map derived from multibody refinement in RELION-3.

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Supramolecule #3: Orc1-5-Orc6C lobe of the MCM-ORC (MO) origin licensing intermediate

SupramoleculeName: Orc1-5-Orc6C lobe of the MCM-ORC (MO) origin licensing intermediate
type: complex / ID: 3 / Parent: 1 / Macromolecule list: #1-#6, #13-#14 / Details: Map derived from multibody refinement in RELION-3.

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Supramolecule #4: The MCM-ORC (MO) loading intermediate protein complex

SupramoleculeName: The MCM-ORC (MO) loading intermediate protein complex / type: complex / ID: 4 / Parent: 1 / Macromolecule list: #1-#12 / Details: Recombinant complex
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)

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Supramolecule #5: DNA

SupramoleculeName: DNA / type: complex / ID: 5 / Parent: 1 / Macromolecule list: #13-#14 / Details: Synthetic DNA
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)

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Macromolecule #1: Origin recognition complex subunit 1

MacromoleculeName: Origin recognition complex subunit 1 / type: protein_or_peptide / ID: 1
Details: Sequence includes an N-terminal CBP-tag and TEV cleavage site.
Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 108.612922 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MKRRWKKNFI AVSAANRFKK ISSSGALENL YFQGEMAKTL KDLQGWEIIT TDEQGNIIDG GQKRLRRRGA KTEHYLKRSS DGIKLGRGD SVVMHNEAAG TYSVYMIQEL RLNTLNNVVE LWALTYLRWF EVNPLAHYRQ FNPDANILNR PLNYYNKLFS E TANKNELY ...String:
MKRRWKKNFI AVSAANRFKK ISSSGALENL YFQGEMAKTL KDLQGWEIIT TDEQGNIIDG GQKRLRRRGA KTEHYLKRSS DGIKLGRGD SVVMHNEAAG TYSVYMIQEL RLNTLNNVVE LWALTYLRWF EVNPLAHYRQ FNPDANILNR PLNYYNKLFS E TANKNELY LTAELAELQL FNFIRVANVM DGSKWEVLKG NVDPERDFTV RYICEPTGEK FVDINIEDVK AYIKKVEPRE AQ EYLKDLT LPSKKKEIKR GPQKKDKATQ TAQISDAETR ATDITDNEDG NEDESSDYES PSDIDVSEDM DSGEISADEL EEE EDEEED EDEEEKEARH TNSPRKRGRK IKLGKDDIDA SVQPPPKKRG RKPKDPSKPR QMLLISSCRA NNTPVIRKFT KKNV ARAKK KYTPFSKRFK SIAAIPDLTS LPEFYGNSSE LMASRFENKL KTTQKHQIVE TIFSKVKKQL NSSYVKEEIL KSANF QDYL PARENEFASI YLSAYSAIES DSATTIYVAG TPGVGKTLTV REVVKELLSS SAQREIPDFL YVEINGLKMV KPTDCY ETL WNKVSGERLT WAASMESLEF YFKRVPKNKK KTIVVLLDEL DAMVTKSQDI MYNFFNWTTY ENAKLIVIAV ANTMDLP ER QLGNKITSRI GFTRIMFTGY THEELKNIID LRLKGLNDSF FYVDTKTGNA ILIDAAGNDT TVKQTLPEDV RKVRLRMS A DAIEIASRKV ASVSGDARRA LKVCKRAAEI AEKHYMAKHG YGYDGKTVIE DENEEQIYDD EDKDLIESNK AKDDNDDDD DNDGVQTVHI THVMKALNET LNSHVITFMT RLSFTAKLFI YALLNLMKKN GSQEQELGDI VDEIKLLIEV NGSNKFVMEI AKTLFQQGS DNISEQLRII SWDFVLNQLL DAGILFKQTM KNDRICCVKL NISVEEAKRA MNEDETLRNL

UniProtKB: Origin recognition complex subunit 1

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Macromolecule #2: Origin recognition complex subunit 2

MacromoleculeName: Origin recognition complex subunit 2 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 71.34218 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MLNGEDFVEH NDILSSPAKS RNVTPKRVDP HGERQLRRIH SSKKNLLERI SLVGNERKNT SPDPALKPKT PSKAPRKRGR PRKIQEELT DRIKKDEKDT ISSKKKRKLD KDTSGNVNEE SKTSNNKQVM EKTGIKEKRE REKIQVATTT YEDNVTPQTD D NFVSNSPE ...String:
MLNGEDFVEH NDILSSPAKS RNVTPKRVDP HGERQLRRIH SSKKNLLERI SLVGNERKNT SPDPALKPKT PSKAPRKRGR PRKIQEELT DRIKKDEKDT ISSKKKRKLD KDTSGNVNEE SKTSNNKQVM EKTGIKEKRE REKIQVATTT YEDNVTPQTD D NFVSNSPE PPEPATPSKK SLTTNHDFTS PLKQIIMNNL KEYKDSTSPG KLTLSRNFTP TPVPKNKKLY QTSETKSASS FL DTFEGYF DQRKIVRTNA KSRHTMSMAP DVTREEFSLV SNFFNENFQK RPRQKLFEIQ KKMFPQYWFE LTQGFSLLFY GVG SKRNFL EEFAIDYLSP KIAYSQLAYE NELQQNKPVN SIPCLILNGY NPSCNYRDVF KEITDLLVPA ELTRSETKYW GNHV ILQIQ KMIDFYKNQP LDIKLILVVH NLDGPSIRKN TFQTMLSFLS VIRQIAIVAS TDHIYAPLLW DNMKAQNYNF VFHDI SNFE PSTVESTFQD VMKMGKSDTS SGAEGAKYVL QSLTVNSKKM YKLLIETQMQ NMGNLSANTG PKRGTQRTGV ELKLFN HLC AADFIASNEI ALRSMLREFI EHKMANITKN NSGMEIIWVP YTYAELEKLL KTVLNTL

UniProtKB: Origin recognition complex subunit 2

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Macromolecule #3: Origin recognition complex subunit 3

MacromoleculeName: Origin recognition complex subunit 3 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 72.161766 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MSDLNQSKKM NVSEFADAQR SHYTVYPSLP QSNKNDKHIP FVKLLSGKES EVNVEKRWEL YHQLHSHFHD QVDHIIDNIE ADLKAEISD LLYSETTQKR RCFNTIFLLG SDSTTKIELK DESSRYNVLI ELTPKESPNV RMMLRRSMYK LYSAADAEEH P TIKYEDIN ...String:
MSDLNQSKKM NVSEFADAQR SHYTVYPSLP QSNKNDKHIP FVKLLSGKES EVNVEKRWEL YHQLHSHFHD QVDHIIDNIE ADLKAEISD LLYSETTQKR RCFNTIFLLG SDSTTKIELK DESSRYNVLI ELTPKESPNV RMMLRRSMYK LYSAADAEEH P TIKYEDIN DEDGDFTEQN NDVSYDLSLV ENFKRLFGKD LAMVFNFKDV DSINFNTLDN FIILLKSAFK YDHVKISLIF NI NTNLSNI EKNLRQSTIR LLKRNYHKLD VSSNKGFKYG NQIFQSFLDT VDGKLNLSDR FVEFILSKMA NNTNHNLQLL TKM LDYSLM SYFFQNAFSV FIDPVNVDFL NDDYLKILSR CPTFMFFVEG LIKQHAPADE ILSLLTNKNR GLEEFFVEFL VREN PINGH AKFVARFLEE ELNITNFNLI ELYHNLLIGK LDSYLDRWSA CKEYKDRLHF EPIDTIFQEL FTLDNRSGLL TQSIF PSYK SNIEDNLLSW EQVLPSLDKE NYDTLSGDLD KIMAPVLGQL FKLYREANMT INIYDFYIAF RETLPKEEIL NFIRKD PSN TKLLELAETP DAFDKVALIL FMQAIFAFEN MGLIKFQSTK SYDLVEKCVW RGI

UniProtKB: Origin recognition complex subunit 3

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Macromolecule #4: Origin recognition complex subunit 4

MacromoleculeName: Origin recognition complex subunit 4 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 60.772152 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MTISEARLSP QVNLLPIKRH SNEEVEETAA ILKKRTIDNE KCKDSDPGFG SLQRRLLQQL YGTLPTDEKI IFTYLQDCQQ EIDRIIKQS IIQKESHSVI LVGPRQSYKT YLLDYELSLL QQSYKEQFIT IRLNGFIHSE QTAINGIATQ LEQQLQKIHG S EEKIDDTS ...String:
MTISEARLSP QVNLLPIKRH SNEEVEETAA ILKKRTIDNE KCKDSDPGFG SLQRRLLQQL YGTLPTDEKI IFTYLQDCQQ EIDRIIKQS IIQKESHSVI LVGPRQSYKT YLLDYELSLL QQSYKEQFIT IRLNGFIHSE QTAINGIATQ LEQQLQKIHG S EEKIDDTS LETISSGSLT EVFEKILLLL DSTTKTRNED SGEVDRESIT KITVVFIFDE IDTFAGPVRQ TLLYNLFDMV EH SRVPVCI FGCTTKLNIL EYLEKRVKSR FSQRVIYMPQ IQNLDDMVDA VRNLLTVRSE ISPWVSQWNE TLEKELSDPR SNL NRHIRM NFETFRSLPT LKNSIIPLVA TSKNFGSLCT AIKSCSFLDI YNKNQLSNNL TGRLQSLSDL ELAILISAAR VALR AKDGS FNFNLAYAEY EKMIKAINSR IPTVAPTTNV GTGQSTFSID NTIKLWLKKD VKNVWENLVQ LDFFTEKSAV GLRDN ATAA FYASNYQFQG TMIPFDLRSY QMQIILQELR RIIPKSNMYY SWTQL

UniProtKB: Origin recognition complex subunit 4

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Macromolecule #5: Origin recognition complex subunit 5

MacromoleculeName: Origin recognition complex subunit 5 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 55.347168 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MNVTTPEVAF REYQTNCLAS YISADPDITP SNLILQGYSG TGKTYTLKKY FNANPNLHAV WLEPVELVSW KPLLQAIART VQYKLKTLY PNIPTTDYDP LQVEEPFLLV KTLHNIFVQY ESLQEKTCLF LILDGFDSLQ DLDAALFNKY IKLNELLPKD S KINIKFIY ...String:
MNVTTPEVAF REYQTNCLAS YISADPDITP SNLILQGYSG TGKTYTLKKY FNANPNLHAV WLEPVELVSW KPLLQAIART VQYKLKTLY PNIPTTDYDP LQVEEPFLLV KTLHNIFVQY ESLQEKTCLF LILDGFDSLQ DLDAALFNKY IKLNELLPKD S KINIKFIY TMLETSFLQR YSTHCIPTVM FPRYNVDEVS TILVMSRCGE LMEDSCLRKR IIEEQITDCT DDQFQNVAAN FI HLIVQAF HSYTGNDIFA LNDLIDFKWP KYVSRITKEN IFEPLALYKS AIKLFLSTDD NLSENGQGES AITTNRDDLE NSQ TYDLSI ISKYLLIASY ICSYLEPRYD ASIFSRKTRI IQGRAAYGRR KKKEVNPRYL QPSLFAIERL LAIFQAIFPI QGKA ESGSL SALREESLMK ANIEVFQNLS ELHTLKLIAT TMNKNIDYLS PKVRWKVNVP WEIIKEISES VHFNISDYFS DIHE

UniProtKB: Origin recognition complex subunit 5

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Macromolecule #6: Origin recognition complex subunit 6

MacromoleculeName: Origin recognition complex subunit 6 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 50.369531 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MSMQQVQHCV AEVLRLDPQE KPDWSSGYLK KLTNATSILY NTSLNKVMLK QDEEVARCHI CAYIASQKMN EKHMPDLCYY IDSIPLEPK KAKHLMNLFR QSLSNSSPMK QFAWTPSPKK NKRSPVKNGG RFTSSDPKEL RNQLFGTPTK VRKSQNNDSF V IPELPPMQ ...String:
MSMQQVQHCV AEVLRLDPQE KPDWSSGYLK KLTNATSILY NTSLNKVMLK QDEEVARCHI CAYIASQKMN EKHMPDLCYY IDSIPLEPK KAKHLMNLFR QSLSNSSPMK QFAWTPSPKK NKRSPVKNGG RFTSSDPKEL RNQLFGTPTK VRKSQNNDSF V IPELPPMQ TNESPSITRR KLAFEEDEDE DEEEPGNDGL SLKSHSNKSI TGTRNVDSDE YENHESDPTS EEEPLGVQES RS GRTKQNK AVGKPQSELK TAKALRKRGR IPNSLLVKKY CKMTTEEIIR LCNDFELPRE VAYKIVDEYN INASRLVCPW QLV CGLVLN CTFIVFNERR RKDPRIDHFI VSKMCSLMLT SKVDDVIECV KLVKELIIGE KWFRDLQIRY DDFDGIRYDE IIFR KLGSM LQTTNILVTD DQYNIWKKRI EMDLALTEPL

UniProtKB: Origin recognition complex subunit 6

+
Macromolecule #7: DNA replication licensing factor MCM2

MacromoleculeName: DNA replication licensing factor MCM2 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 98.911539 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MSDNRRRRRE EDDSDSENEL PPSSPQQHFR GGMNPVSSPI GSPDMINPEG DDNEVDDVPD IDEVEEQMNE VDLMDDNMYE DYAADHNRD RYDPDQVDDR EQQELSLSER RRIDAQLNER DRLLRNVAYI DDEDEEQEGA AQLDEMGLPV QRRRRRRQYE D LENSDDDL ...String:
MSDNRRRRRE EDDSDSENEL PPSSPQQHFR GGMNPVSSPI GSPDMINPEG DDNEVDDVPD IDEVEEQMNE VDLMDDNMYE DYAADHNRD RYDPDQVDDR EQQELSLSER RRIDAQLNER DRLLRNVAYI DDEDEEQEGA AQLDEMGLPV QRRRRRRQYE D LENSDDDL LSDMDIDPLR EELTLESLSN VKANSYSEWI TQPNVSRTIA RELKSFLLEY TDETGRSVYG ARIRTLGEMN SE SLEVNYR HLAESKAILA LFLAKCPEEM LKIFDLVAME ATELHYPDYA RIHSEIHVRI SDFPTIYSLR ELRESNLSSL VRV TGVVTR RTGVFPQLKY VKFNCLKCGS ILGPFFQDSN EEIRISFCTN CKSKGPFRVN GEKTVYRNYQ RVTLQEAPGT VPPG RLPRH REVILLADLV DVSKPGEEVE VTGIYKNNYD GNLNAKNGFP VFATIIEANS IKRREGNTAN EGEEGLDVFS WTEEE EREF RKISRDRGII DKIISSMAPS IYGHRDIKTA VACSLFGGVP KNVNGKHSIR GDINVLLLGD PGTAKSQILK YVEKTA HRA VFATGQGASA VGLTASVRKD PITKEWTLEG GALVLADKGV CLIDEFDKMN DQDRTSIHEA MEQQSISISK AGIVTTL QA RCSIIAAANP NGGRYNSTLP LAQNVSLTEP ILSRFDILCV VRDLVDEEAD ERLATFVVDS HVRSHPENDE DREGEELK N NGESAIEQGE DEINEQLNAR QRRLQRQRKK EEEISPIPQE LLMKYIHYAR TKIYPKLHQM DMDKVSRVYA DLRRESIST GSFPITVRHL ESILRIAESF AKMRLSEFVS SYDLDRAIKV VVDSFVDAQK VSVRRQLRRS FAIYTLGH

UniProtKB: DNA replication licensing factor MCM2

+
Macromolecule #8: DNA replication licensing factor MCM3

MacromoleculeName: DNA replication licensing factor MCM3 / type: protein_or_peptide / ID: 8
Details: Sequence includes an N-terminal CBP-tag and TEV cleavage site.
Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 111.720242 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MKRRWKKNFI AVSAANRFKK ISSSGALENL YFQGEMEGST GFDGDATTFF APDAVFGDRV RRFQEFLDTF TSYRDSVRSI QVYNSNNAA NYNDDQDDAD ERDLLGDDDG DDLEKEKKAA SSTSLNILPH RIIISLDDLR EFDRSFWSGI LVEPAYFIPP A EKALTDLA ...String:
MKRRWKKNFI AVSAANRFKK ISSSGALENL YFQGEMEGST GFDGDATTFF APDAVFGDRV RRFQEFLDTF TSYRDSVRSI QVYNSNNAA NYNDDQDDAD ERDLLGDDDG DDLEKEKKAA SSTSLNILPH RIIISLDDLR EFDRSFWSGI LVEPAYFIPP A EKALTDLA DSMDDVPHPN ASAVSSRHPW KLSFKGSFGA HALSPRTLTA QHLNKLVSVE GIVTKTSLVR PKLIRSVHYA AK TGRFHYR DYTDATTTLT TRIPTPAIYP TEDTEGNKLT TEYGYSTFID HQRITVQEMP EMAPAGQLPR SIDVILDDDL VDK TKPGDR VNVVGVFKSL GAGGMNQSNS NTLIGFKTLI LGNTVYPLHA RSTGVAARQM LTDFDIRNIN KLSKKKDIFD ILSQ SLAPS IYGHDHIKKA ILLMLMGGVE KNLENGSHLR GDINILMVGD PSTAKSQLLR FVLNTASLAI ATTGRGSSGV GLTAA VTTD RETGERRLEA GAMVLADRGV VCIDEFDKMT DVDRVAIHEV MEQQTVTIAK AGIHTTLNAR CSVIAAANPV FGQYDV NRD PHQNIALPDS LLSRFDLLFV VTDDINEIRD RSISEHVLRT HRYLPPGYLE GEPVRERLNL SLAVGEDADI NPEEHSN SG AGVENEGEDD EDHVFEKFNP LLQAGAKLAK NKGNYNGTEI PKLVTIPFLR KYVQYAKERV IPQLTQEAIN VIVKNYTD L RNDDNTKKSP ITARTLETLI RLATAHAKVR LSKTVNKVDA KVAANLLRFA LLGEDIGNDI DEEESEYEEA LSKRSPQKS PKKRQRVRQP ASNSGSPIKS TPRRSTASSV NATPSSARRI LRFQDDEQNA GEDDNDIMSP LPADEEAELQ RRLQLGLRVS PRRREHLHA PEEGSSGPLT EVGTPRLPNV SSAGQDDEQQ QSVISFDNVE PGTISTGRLS LISGIIARLM QTEIFEEESY P VASLFERI NEELPEEEKF SAQEYLAGLK IMSDRNNLMV ADDKVWRV

UniProtKB: DNA replication licensing factor MCM3

+
Macromolecule #9: DNA replication licensing factor MCM4

MacromoleculeName: DNA replication licensing factor MCM4 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 105.138375 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MSQQSSSPTK EDNNSSSPVV PNPDSVPPQL SSPALFYSSS SSQGDIYGRN NSQNLSQGEG NIRAAIGSSP LNFPSSSQRQ NSDVFQSQG RQGRIRSSAS ASGRSRYHSD LRSDRALPTS SSSLGRNGQN RVHMRRNDIH TSDLSSPRRI VDFDTRSGVN T LDTSSSSA ...String:
MSQQSSSPTK EDNNSSSPVV PNPDSVPPQL SSPALFYSSS SSQGDIYGRN NSQNLSQGEG NIRAAIGSSP LNFPSSSQRQ NSDVFQSQG RQGRIRSSAS ASGRSRYHSD LRSDRALPTS SSSLGRNGQN RVHMRRNDIH TSDLSSPRRI VDFDTRSGVN T LDTSSSSA PPSEASEPLR IIWGTNVSIQ ECTTNFRNFL MSFKYKFRKI LDEREEFINN TTDEELYYIK QLNEMRELGT SN LNLDARN LLAYKQTEDL YHQLLNYPQE VISIMDQTIK DCMVSLIVDN NLDYDLDEIE TKFYKVRPYN VGSCKGMREL NPN DIDKLI NLKGLVLRST PVIPDMKVAF FKCNVCDHTM AVEIDRGVIQ EPARCERIDC NEPNSMSLIH NRCSFADKQV IKLQ ETPDF VPDGQTPHSI SLCVYDELVD SCRAGDRIEV TGTFRSIPIR ANSRQRVLKS LYKTYVDVVH VKKVSDKRLD VDTST IEQE LMQNKVDHNE VEEVRQITDQ DLAKIREVAA REDLYSLLAR SIAPSIYELE DVKKGILLQL FGGTNKTFTK GGRYRG DIN ILLCGDPSTS KSQILQYVHK ITPRGVYTSG KGSSAVGLTA YITRDVDTKQ LVLESGALVL SDGGVCCIDE FDKMSDS TR SVLHEVMEQQ TISIAKAGII TTLNARSSIL ASANPIGSRY NPNLPVTENI DLPPPLLSRF DLVYLVLDKV DEKNDREL A KHLTNLYLED KPEHISQDDV LPVEFLTMYI SYAKEHIHPI ITEAAKTELV RAYVGMRKMG DDSRSDEKRI TATTRQLES MIRLAEAHAK MKLKNVVELE DVQEAVRLIR SAIKDYATDP KTGKIDMNLV QTGKSVIQRK LQEDLSREIM NVLKDQASDS MSFNELIKQ INEHSQDRVE SSDIQEALSR LQQEDKVIVL GEGVRRSVRL NNRV

UniProtKB: DNA replication licensing factor MCM4

+
Macromolecule #10: Minichromosome maintenance protein 5

MacromoleculeName: Minichromosome maintenance protein 5 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 86.505734 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MSFDRPEIYS APVLQGESPN DDDNTEIIKS FKNFILEFRL DSQFIYRDQL RNNILVKNYS LTVNMEHLIG YNEDIYKKLS DEPSDIIPL FETAITQVAK RISILSRAQS ANNNDKDPEN TSMDTDSLLL NSLPTFQLIL NSNANQIPLR DLDSEHVSKI V RLSGIIIS ...String:
MSFDRPEIYS APVLQGESPN DDDNTEIIKS FKNFILEFRL DSQFIYRDQL RNNILVKNYS LTVNMEHLIG YNEDIYKKLS DEPSDIIPL FETAITQVAK RISILSRAQS ANNNDKDPEN TSMDTDSLLL NSLPTFQLIL NSNANQIPLR DLDSEHVSKI V RLSGIIIS TSVLSSRATY LSIMCRNCRH TTSITINNFN SITGNTVSLP RSCLSTIESE SSMANESNIG DESTKKNCGP DP YIIIHES SKFIDQQFLK LQEIPELVPV GEMPRNLTMT CDRYLTNKVI PGTRVTIVGI YSIYNSKNGA GSGRSGGGNG GSG VAIRTP YIKILGIQSD VETSSIWNSV TMFTEEEEEE FLQLSRNPKL YEILTNSIAP SIFGNEDIKK AIVCLLMGGS KKIL PDGMR LRGDINVLLL GDPGTAKSQL LKFVEKVSPI AVYTSGKGSS AAGLTASVQR DPMTREFYLE GGAMVLADGG VVCID EFDK MRDEDRVAIH EAMEQQTISI AKAGITTVLN SRTSVLAAAN PIYGRYDDLK SPGDNIDFQT TILSRFDMIF IVKDDH NEE RDISIANHVI NIHTGNANAM QNQQEENGSE ISIEKMKRYI TYCRLKCAPR LSPQAAEKLS SNFVTIRKQL LINELES TE RSSIPITIRQ LEAIIRITES LAKLELSPIA QERHVDEAIR LFQASTMDAA SQDPIGGLNQ ASGTSLSEIR RFEQELKR R LPIGWSTSYQ TLRREFVDTH RFSQLALDKA LYALEKHETI QLRHQGQNIY RSGV

UniProtKB: Minichromosome maintenance protein 5

+
Macromolecule #11: DNA replication licensing factor MCM6

MacromoleculeName: DNA replication licensing factor MCM6 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 113.110211 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MSSPFPADTP SSNRPSNSSP PPSSIGAGFG SSSGLDSQIG SRLHFPSSSQ PHVSNSQTGP FVNDSTQFSS QRLQTDGSAT NDMEGNEPA RSFKSRALNH VKKVDDVTGE KVREAFEQFL EDFSVQSTDT GEVEKVYRAQ IEFMKIYDLN TIYIDYQHLS M RENGALAM ...String:
MSSPFPADTP SSNRPSNSSP PPSSIGAGFG SSSGLDSQIG SRLHFPSSSQ PHVSNSQTGP FVNDSTQFSS QRLQTDGSAT NDMEGNEPA RSFKSRALNH VKKVDDVTGE KVREAFEQFL EDFSVQSTDT GEVEKVYRAQ IEFMKIYDLN TIYIDYQHLS M RENGALAM AISEQYYRFL PFLQKGLRRV VRKYAPELLN TSDSLKRSEG DEGQADEDEQ QDDDMNGSSL PRDSGSSAAP GN GTSAMAT RSITTSTSPE QTERVFQISF FNLPTVHRIR DIRSEKIGSL LSISGTVTRT SEVRPELYKA SFTCDMCRAI VDN VEQSFK YTEPTFCPNP SCENRAFWTL NVTRSRFLDW QKVRIQENAN EIPTGSMPRT LDVILRGDSV ERAKPGDRCK FTGV EIVVP DVTQLGLPGV KPSSTLDTRG ISKTTEGLNS GVTGLRSLGV RDLTYKISFL ACHVISIGSN IGASSPDANS NNRET ELQM AANLQANNVY QDNERDQEVF LNSLSSDEIN ELKEMVKDEH IYDKLVRSIA PAVFGHEAVK KGILLQMLGG VHKSTV EGI KLRGDINICV VGDPSTSKSQ FLKYVVGFAP RSVYTSGKAS SAAGLTAAVV RDEEGGDYTI EAGALMLADN GICCIDE FD KMDISDQVAI HEAMEQQTIS IAKAGIHATL NARTSILAAA NPVGGRYNRK LSLRGNLNMT APIMSRFDLF FVILDDCN E KIDTELASHI VDLHMKRDEA IEPPFSAEQL RRYIKYARTF KPILTKEARS YLVEKYKELR KDDAQGFSRS SYRITVRQL ESMIRLSEAI ARANCVDEIT PSFIAEAYDL LRQSIIRVDV DDVEMDEEFD NIESQSHAAS GNNDDNDDGT GSGVITSEPP ADIEEGQSE ATARPGTSEK KKTTVTYDKY VSMMNMIVRK IAEVDREGAE ELTAVDIVDW YLLQKENDLG SLAEYWEERR L AFKVIKRL VKDRILMEIH GTRHNLRDLE NEENENNKTV YVIHPNCEVL DQLEPQDSS

UniProtKB: DNA replication licensing factor MCM6

+
Macromolecule #12: DNA replication licensing factor MCM7

MacromoleculeName: DNA replication licensing factor MCM7 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 95.049875 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MSAALPSIQL PVDYNNLFNE ITDFLVTFKQ DTLSSDATRN ENEDENLDAE NIEQHLLEKG PKYMAMLQKV ANRELNSVII DLDDILQYQ NEKFLQGTQA DDLVSAIQQN ANHFTELFCR AIDNNMPLPT KEIDYKDDVL DVILNQRRLR NERMLSDRTN E IRSENLMD ...String:
MSAALPSIQL PVDYNNLFNE ITDFLVTFKQ DTLSSDATRN ENEDENLDAE NIEQHLLEKG PKYMAMLQKV ANRELNSVII DLDDILQYQ NEKFLQGTQA DDLVSAIQQN ANHFTELFCR AIDNNMPLPT KEIDYKDDVL DVILNQRRLR NERMLSDRTN E IRSENLMD TTMDPPSSMN DALREVVEDE TELFPPNLTR RYFLYFKPLS QNCARRYRKK AISSKPLSVR QIKGDFLGQL IT VRGIITR VSDVKPAVEV IAYTCDQCGY EVFQEVNSRT FTPLSECTSE ECSQNQTKGQ LFMSTRASKF SAFQECKIQE LSQ QVPVGH IPRSLNIHVN GTLVRSLSPG DIVDVTGIFL PAPYTGFKAL KAGLLTETYL EAQFVRQHKK KFASFSLTSD VEER VMELI TSGDVYNRLA KSIAPEIYGN LDVKKALLLL LVGGVDKRVG DGMKIRGDIN VCLMGDPGVA KSQLLKAICK ISPRG VYTT GKGSSGVGLT AAVMKDPVTD EMILEGGALV LADNGICCID EFDKMDESDR TAIHEVMEQQ TISISKAGIN TTLNAR TSI LAAANPLYGR YNPRLSPLDN INLPAALLSR FDILFLMLDI PSRDDDEKLA EHVTYVHMHN KQPDLDFTPV EPSKMRE YI AYAKTKRPVM SEAVNDYVVQ AYIRLRQDSK REMDSKFSFG QATPRTLLGI IRLSQALAKL RLADMVDIDD VEEALRLV R VSKESLYQET NKSKEDESPT TKIFTIIKKM LQETGKNTLS YENIVKTVRL RGFTMLQLSN CIQEYSYLNV WHLINEGNT LKFVDDGTMD TDQEDSLVST PKLAPQTTAS ANVSAQDSDI DLQDA

UniProtKB: DNA replication licensing factor MCM7

+
Macromolecule #13: DNA (88-MER)

MacromoleculeName: DNA (88-MER) / type: dna / ID: 13 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 27.116334 KDa
SequenceString: (DT)(DG)(DG)(DT)(DT)(DT)(DT)(DT)(DA)(DT) (DA)(DT)(DG)(DT)(DT)(DT)(DT)(DG)(DT)(DT) (DA)(DT)(DG)(DT)(DA)(DT)(DT)(DG)(DT) (DT)(DT)(DA)(DT)(DT)(DT)(DT)(DC)(DC)(DC) (DT) (DT)(DG)(DA)(DC)(DT)(DG) ...String:
(DT)(DG)(DG)(DT)(DT)(DT)(DT)(DT)(DA)(DT) (DA)(DT)(DG)(DT)(DT)(DT)(DT)(DG)(DT)(DT) (DA)(DT)(DG)(DT)(DA)(DT)(DT)(DG)(DT) (DT)(DT)(DA)(DT)(DT)(DT)(DT)(DC)(DC)(DC) (DT) (DT)(DG)(DA)(DC)(DT)(DG)(DA)(DC) (DT)(DG)(DA)(DC)(DT)(DG)(DA)(DC)(DT)(DG) (DA)(DC) (DT)(DG)(DA)(DC)(DT)(DG)(DA) (DC)(DT)(DG)(DA)(DC)(DT)(DG)(DA)(DC)(DT) (DG)(DA)(DC) (DT)(DG)(DT)(DA)(DT)(DA) (DT)(DA)

+
Macromolecule #14: DNA (88-MER)

MacromoleculeName: DNA (88-MER) / type: dna / ID: 14 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Saccharomyces cerevisiae S288c (yeast)
Molecular weightTheoretical: 27.154562 KDa
SequenceString: (DT)(DA)(DT)(DA)(DT)(DA)(DC)(DA)(DG)(DT) (DC)(DA)(DG)(DT)(DC)(DA)(DG)(DT)(DC)(DA) (DG)(DT)(DC)(DA)(DG)(DT)(DC)(DA)(DG) (DT)(DC)(DA)(DG)(DT)(DC)(DA)(DG)(DT)(DC) (DA) (DG)(DT)(DC)(DA)(DG)(DT) ...String:
(DT)(DA)(DT)(DA)(DT)(DA)(DC)(DA)(DG)(DT) (DC)(DA)(DG)(DT)(DC)(DA)(DG)(DT)(DC)(DA) (DG)(DT)(DC)(DA)(DG)(DT)(DC)(DA)(DG) (DT)(DC)(DA)(DG)(DT)(DC)(DA)(DG)(DT)(DC) (DA) (DG)(DT)(DC)(DA)(DG)(DT)(DC)(DA) (DA)(DG)(DG)(DG)(DA)(DA)(DA)(DA)(DT)(DA) (DA)(DA) (DC)(DA)(DA)(DT)(DA)(DC)(DA) (DT)(DA)(DA)(DC)(DA)(DA)(DA)(DA)(DC)(DA) (DT)(DA)(DT) (DA)(DA)(DA)(DA)(DA)(DC) (DC)(DA)

+
Macromolecule #15: ADENOSINE-5'-TRIPHOSPHATE

MacromoleculeName: ADENOSINE-5'-TRIPHOSPHATE / type: ligand / ID: 15 / Number of copies: 3 / Formula: ATP
Molecular weightTheoretical: 507.181 Da
Chemical component information

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM

+
Macromolecule #16: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 16 / Number of copies: 3 / Formula: MG
Molecular weightTheoretical: 24.305 Da

+
Macromolecule #17: ADENOSINE-5'-DIPHOSPHATE

MacromoleculeName: ADENOSINE-5'-DIPHOSPHATE / type: ligand / ID: 17 / Number of copies: 4 / Formula: ADP
Molecular weightTheoretical: 427.201 Da
Chemical component information

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

+
Macromolecule #18: ZINC ION

MacromoleculeName: ZINC ION / type: ligand / ID: 18 / Number of copies: 5 / Formula: ZN
Molecular weightTheoretical: 65.409 Da

-
Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

-
Sample preparation

BufferpH: 7.6
Component:
ConcentrationName
25.0 mMHEPES
10.0 mMMagnesium acetate
100.0 mMPotassium acetate
1.0 mMDTT
0.75 mMATP
GridMaterial: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: LACEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR
VitrificationCryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 288 K / Instrument: LEICA EM GP
Details: 10 second incubation, 3.5 seconds single side blotting..

-
Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Frames/image: 1-30 / Number grids imaged: 1 / Average electron dose: 1.68 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

+
Image processing

Startup modelType of model: NONE / Details: Ab initio model generated using cryoSPARC v1.6
Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 4.4 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3) / Number images used: 177637
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3)
Final 3D classificationNumber classes: 20 / Software - Name: RELION (ver. 3)
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial model
PDB IDChain

5zr1

source_name: PDB, initial_model_type: experimental model

6eyc

source_name: PDB, initial_model_type: experimental model

5bk4

source_name: PDB, initial_model_type: experimental model

6f0l

source_name: PDB, initial_model_type: experimental model
RefinementSpace: REAL / Protocol: RIGID BODY FIT
Output model

PDB-6rqc:
Cryo-EM structure of an MCM loading intermediate

+
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