EMDB-13539: S. cerevisiae replisome-SCF(Dia2) complex bound to double-strande...

Basic information

• Entry: Database: EMDB / ID: EMD-13539
• Title: S. cerevisiae replisome-SCF(Dia2) complex bound to double-stranded DNA (conformation II)
• Map data: Composite cryo-EM density map for the budding yeast CMG-Csm3-Tof1-Mrc1-Ctf4-PolE-SCF(Dia2) complex on double-stranded DNA (conformation II), produced using the Phenix combine_focused_maps program

Sample

• Complex: Budding yeast replisome on double-stranded DNA engaged with SCF(Dia2) (conformation II)
  • Complex: DNA
    • DNA: x 2 types
  • Complex: Replisome
    • Protein or peptide: x 16 types
  • Protein or peptide: x 2 types
• Ligand: x 3 types

• Keywords: Genome stability / DNA replication / Ubiquitination / termination / replisome / cryo-EM / CMG / SCF(Dia2) / REPLICATION

Function / homology

Function:

RAVE complex / Iron uptake and transport / CBF3 complex / regulation of transcription by galactose / regulation of sulfur amino acid metabolic process / cellular response to methylmercury / establishment of sister chromatid cohesion / vacuolar proton-transporting V-type ATPase complex assembly / mitotic DNA replication termination / DNA-templated DNA replication maintenance of fidelity / gene conversion / septin ring assembly / maintenance of DNA repeat elements / Unwinding of DNA / invasive growth in response to glucose limitation / DNA replication initiation / Cul8-RING ubiquitin ligase complex / replication fork arrest / meiotic chromosome segregation / : / protein-containing complex disassembly / epsilon DNA polymerase complex / DNA strand elongation involved in mitotic DNA replication / GINS complex / MCM core complex / Assembly of the pre-replicative complex / Switching of origins to a post-replicative state / MCM complex binding / mitotic DNA replication preinitiation complex assembly / nuclear DNA replication / premeiotic DNA replication / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / SUMO binding / regulation of exit from mitosis / nucleotide-excision repair, DNA gap filling / mitotic DNA replication / Activation of the pre-replicative complex / kinetochore assembly / DNA replication checkpoint signaling / CMG complex / establishment of mitotic sister chromatid cohesion / nuclear pre-replicative complex / DNA replication proofreading / Termination of translesion DNA synthesis / Activation of ATR in response to replication stress / DNA replication preinitiation complex / single-stranded DNA 3'-5' DNA exonuclease activity / MCM complex / vacuolar acidification / replication fork protection complex / positive regulation of D-glucose transmembrane transport / mitotic DNA replication checkpoint signaling / double-strand break repair via break-induced replication / single-stranded DNA helicase activity / protein neddylation / mitotic DNA replication initiation / mitotic intra-S DNA damage checkpoint signaling / regulation of metabolic process / mitochondrial fusion / silent mating-type cassette heterochromatin formation / regulation of DNA-templated DNA replication initiation / exit from mitosis / Hydrolases; Acting on ester bonds; Exodeoxyribonucleases producing 5'-phosphomonoesters / SCF ubiquitin ligase complex / mitotic sister chromatid cohesion / SCF-dependent proteasomal ubiquitin-dependent protein catabolic process / nuclear chromosome / FBXL7 down-regulates AURKA during mitotic entry and in early mitosis / DNA strand elongation involved in DNA replication / Orc1 removal from chromatin / leading strand elongation / replication fork processing / nuclear replication fork / regulation of DNA replication / cullin family protein binding / Antigen processing: Ubiquitination & Proteasome degradation / Dual incision in TC-NER / DNA replication origin binding / DNA replication initiation / regulation of protein-containing complex assembly / error-prone translesion synthesis / subtelomeric heterochromatin formation / negative regulation of cytoplasmic translation / base-excision repair, gap-filling / endomembrane system / regulation of mitotic cell cycle / replication fork / meiotic cell cycle / helicase activity / transcription elongation by RNA polymerase II / base-excision repair / kinetochore / double-strand break repair via nonhomologous end joining / DNA-templated DNA replication / G1/S transition of mitotic cell cycle / heterochromatin formation / nucleosome assembly / G2/M transition of mitotic cell cycle / double-strand break repair / mitotic cell cycle

Domain/homology:

F-box domain / Chromosome segregation in meiosis protein 3 / TIPIN/Csm3/Swi3 / Replication Fork Protection Component Swi3 / DNA polymerase epsilon, subunit B / Timeless, N-terminal / Timeless / Timeless protein / : / DNA polymerase alpha-binding protein Ctf4, C-terminal domain / Minichromosome loss protein Mcl1, middle region / Minichromosome loss protein, Mcl1, middle region / Leucine-rich repeat, cysteine-containing subtype / Leucine-rich repeat - CC (cysteine-containing) subfamily / : / : / DNA polymerase epsilon catalytic subunit A, thumb domain / Zinc finger domain of DNA polymerase-epsilon / DNA polymerase epsilon, catalytic subunit A, C-terminal / DNA polymerase epsilon catalytic subunit / Domain of unknown function (DUF1744) / DUF1744 / CDC45 family / DNA replication complex GINS protein Psf2 / CDC45 / A Receptor for Ubiquitination Targets / GINS complex, subunit Psf1 / GINS complex, subunit Psf3 / DNA replication complex GINS protein SLD5, C-terminal / GINS, helical bundle-like domain superfamily / GINS complex, subunit Psf3 superfamily / GINS complex protein Sld5, alpha-helical domain / : / DNA replication complex GINS protein SLD5 C-terminus / MCM3 winged helix domain / GINS complex subunit Sld5 / GINS subunit, domain A / GINS complex protein helical bundle domain / F-box domain profile. / MCM4, winged helix domain / F-box-like domain superfamily / SKP1 component, dimerisation / S-phase kinase-associated protein 1 / SKP1-like, dimerisation domain superfamily / Skp1 family, dimerisation domain / DNA polymerase alpha/delta/epsilon, subunit B / DNA polymerase alpha/epsilon subunit B / F-box domain / : / MCM5, C-terminal domain / DNA replication licensing factor Mcm5 / MCM3-like, winged helix domain / 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. / MCM N-terminal domain / MCM N-terminal domain / MCM OB domain / MCM OB domain / Mini-chromosome maintenance protein / MCM, AAA-lid domain / MCM P-loop domain / MCM AAA-lid domain / MCM family domain profile. / minichromosome maintenance proteins / MCM domain / DNA polymerase family B, thumb domain / S-phase kinase-associated protein 1-like / SKP1 component, POZ domain / Skp1 family, tetramerisation domain / Found in Skp1 protein family / DNA polymerase family B, exonuclease domain / DNA-directed DNA polymerase, family B, exonuclease domain / DNA polymerase, palm domain superfamily / DNA polymerase type-B family / DNA-directed DNA polymerase, family B / TPR repeat region circular profile. / Leucine-rich repeat profile. / SKP1/BTB/POZ domain superfamily / Leucine-rich repeat / Leucine-rich repeat domain superfamily / Tetratricopeptide-like helical domain superfamily / Ribonuclease H superfamily / Ribonuclease H-like superfamily / WD40-repeat-containing domain superfamily / Winged helix-like DNA-binding domain superfamily / WD40/YVTN repeat-like-containing domain superfamily / Nucleic acid-binding, OB-fold / ATPases associated with a variety of cellular activities / AAA+ ATPase domain / DNA/RNA polymerase superfamily / P-loop containing nucleoside triphosphate hydrolase

Component:

DNA replication complex GINS protein PSF3 / DNA polymerase epsilon catalytic subunit A / DNA replication licensing factor MCM3 / DNA polymerase epsilon subunit B / DNA replication licensing factor MCM2 / Minichromosome maintenance protein 5 / DNA replication licensing factor MCM4 / DNA replication licensing factor MCM7 / DNA replication complex GINS protein PSF2 / Suppressor of kinetochore protein 1 / DNA replication licensing factor MCM6 / Topoisomerase 1-associated factor 1 / DNA polymerase alpha-binding protein / DNA replication complex GINS protein SLD5 / Chromosome segregation in meiosis protein 3 / Cell division control protein 45 / Protein DIA2 / DNA replication complex GINS protein PSF1

• Biological species: DNA molecule (others) / Saccharomyces cerevisiae (brewer's yeast) / Synthetic construct (others)
• Method: single particle reconstruction / cryo EM / Resolution: 3.2 Å
• Authors: Jenkyn-Bedford M / Yeeles JTP

Funding support

United Kingdom, 3 items

Organization	Grant number	Country
Medical Research Council (MRC, United Kingdom)	MC_UP_1201/12	United Kingdom
Medical Research Council (MRC, United Kingdom)	MC_UU_12016/13	United Kingdom
Wellcome Trust	204678/Z/16/Z	United Kingdom

• Citation: Journal: Nature / Year: 2021; Title: A conserved mechanism for regulating replisome disassembly in eukaryotes.; Authors: Michael Jenkyn-Bedford / Morgan L Jones / Yasemin Baris / Karim P M Labib / Giuseppe Cannone / Joseph T P Yeeles / Tom D Deegan / ; Abstract: Replisome disassembly is the final step of eukaryotic DNA replication and is triggered by ubiquitylation of the CDC45-MCM-GINS (CMG) replicative helicase. Despite being driven by evolutionarily diverse E3 ubiquitin ligases in different eukaryotes (SCF in budding yeast, CUL2 in metazoa), replisome disassembly is governed by a common regulatory principle, in which ubiquitylation of CMG is suppressed before replication termination, to prevent replication fork collapse. Recent evidence suggests that this suppression is mediated by replication fork DNA. However, it is unknown how SCF and CUL2 discriminate terminated from elongating replisomes, to selectively ubiquitylate CMG only after termination. Here we used cryo-electron microscopy to solve high-resolution structures of budding yeast and human replisome-E3 ligase assemblies. Our structures show that the leucine-rich repeat domains of Dia2 and LRR1 are structurally distinct, but bind to a common site on CMG, including the MCM3 and MCM5 zinc-finger domains. The LRR-MCM interaction is essential for replisome disassembly and, crucially, is occluded by the excluded DNA strand at replication forks, establishing the structural basis for the suppression of CMG ubiquitylation before termination. Our results elucidate a conserved mechanism for the regulation of replisome disassembly in eukaryotes, and reveal a previously unanticipated role for DNA in preserving replisome integrity.

History

Deposition	Sep 2, 2021	-
Header (metadata) release	Nov 10, 2021	-
Map release	Nov 10, 2021	-
Update	Jul 17, 2024	-
Current status	Jul 17, 2024	Processing site: PDBe / Status: Released

Map

• File: Download / File: emd_13539.map.gz / Format: CCP4 / Size: 202.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
• Annotation: Composite cryo-EM density map for the budding yeast CMG-Csm3-Tof1-Mrc1-Ctf4-PolE-SCF(Dia2) complex on double-stranded DNA (conformation II), produced using the Phenix combine_focused_maps program
• Voxel size: X=Y=Z: 1.06 Å

Density

Contour Level	By AUTHOR: 6.82 / Movie #1: 6.82
Minimum - Maximum	-32.137005000000002 - 63.837997000000001
Average (Standard dev.)	0.037581224 (±1.5334363)

• Symmetry: Space group: 1

Details

EMDB XML:

Map geometry	Axis order Z Y X Origin 0 0 0 Dimensions 376 376 376 Spacing 376 376 376
Cell	A=B=C: 398.55997 Å α=β=γ: 90.0 °

CCP4 map header:

mode	Image stored as Reals
Å/pix. X/Y/Z	1.06	1.06	1.06
M x/y/z	376	376	376
origin x/y/z	0.000	0.000	0.000
length x/y/z	398.560	398.560	398.560
α/β/γ	90.000	90.000	90.000
start NX/NY/NZ	0	0	0
NX/NY/NZ	376	376	376
MAP C/R/S	3	2	1
start NC/NR/NS	0	0	0
NC/NR/NS	376	376	376
D min/max/mean	-32.137	63.838	0.038

Supplemental data

Sample components

Entire : Budding yeast replisome on double-stranded DNA engaged with SCF(D...

• Entire: Name: Budding yeast replisome on double-stranded DNA engaged with SCF(Dia2) (conformation II)

Components

• Complex: Budding yeast replisome on double-stranded DNA engaged with SCF(Dia2) (conformation II)
  • Complex: DNA
    • DNA: Leading strand template DNA
    • DNA: Lagging strand template DNA
  • Complex: Replisome
    • 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
    • Protein or peptide: DNA replication complex GINS protein PSF1
    • Protein or peptide: DNA replication complex GINS protein PSF2
    • Protein or peptide: DNA replication complex GINS protein PSF3
    • Protein or peptide: DNA replication complex GINS protein SLD5
    • Protein or peptide: Cell division control protein 45,Cell division control protein 45
    • Protein or peptide: DNA polymerase alpha-binding protein
    • Protein or peptide: DNA polymerase epsilon catalytic subunit A
    • Protein or peptide: DNA polymerase epsilon subunit B
    • Protein or peptide: Topoisomerase 1-associated factor 1
    • Protein or peptide: Chromosome segregation in meiosis protein 3
  • Protein or peptide: Suppressor of kinetochore protein 1
  • Protein or peptide: Protein DIA2
• Ligand: PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER
• Ligand: MAGNESIUM ION
• Ligand: ZINC ION

Supramolecule #1: Budding yeast replisome on double-stranded DNA engaged with SCF(D...

• Supramolecule: Name: Budding yeast replisome on double-stranded DNA engaged with SCF(Dia2) (conformation II); type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#20; Details: Complex reconstituted in vitro from purified proteins and DNA

Supramolecule #2: DNA

• Supramolecule: Name: DNA / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #13-#14
• Source (natural): Organism: DNA molecule (others)

Supramolecule #3: Replisome

• Supramolecule: Name: Replisome / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #1-#12, #17-#20
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)

Macromolecule #1: DNA replication licensing factor MCM2

• Macromolecule: Name: DNA replication licensing factor MCM2 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 98.911539 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

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 #2: DNA replication licensing factor MCM3

• Macromolecule: Name: DNA replication licensing factor MCM3 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 111.987562 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

MKRRWKKNFI AVSAANRFKK ISSSGALENL YFQGEAPVME GSTGFDGDAT TFFAPDAVFG DRVRRFQEFL DTFTSYRDSV RSIQVYNSN NAANYNDDQD DADERDLLGD DDGDDLEKEK KAASSTSLNI LPHRIIISLD DLREFDRSFW SGILVEPAYF I PPAEKALT DLADSMDDVP HPNASAVSSR HPWKLSFKGS FGAHALSPRT LTAQHLNKLV SVEGIVTKTS LVRPKLIRSV HY AAKTGRF HYRDYTDATT TLTTRIPTPA IYPTEDTEGN KLTTEYGYST FIDHQRITVQ EMPEMAPAGQ LPRSIDVILD DDL VDKTKP GDRVNVVGVF KSLGAGGMNQ SNSNTLIGFK TLILGNTVYP LHARSTGVAA RQMLTDFDIR NINKLSKKKD IFDI LSQSL APSIYGHDHI KKAILLMLMG GVEKNLENGS HLRGDINILM VGDPSTAKSQ LLRFVLNTAS LAIATTGRGS SGVGL TAAV TTDRETGERR LEAGAMVLAD RGVVCIDEFD KMTDVDRVAI HEVMEQQTVT IAKAGIHTTL NARCSVIAAA NPVFGQ YDV NRDPHQNIAL PDSLLSRFDL LFVVTDDINE IRDRSISEHV LRTHRYLPPG YLEGEPVRER LNLSLAVGED ADINPEE HS NSGAGVENEG EDDEDHVFEK FNPLLQAGAK LAKNKGNYNG TEIPKLVTIP FLRKYVQYAK ERVIPQLTQE AINVIVKN Y TDLRNDDNTK KSPITARTLE TLIRLATAHA KVRLSKTVNK VDAKVAANLL RFALLGEDIG NDIDEEESEY EEALSKRSP QKSPKKRQRV RQPASNSGSP IKSTPRRSTA SSVNATPSSA RRILRFQDDE QNAGEDDNDI MSPLPADEEA ELQRRLQLGL RVSPRRREH LHAPEEGSSG PLTEVGTPRL PNVSSAGQDD EQQQSVISFD NVEPGTISTG RLSLISGIIA RLMQTEIFEE E SYPVASLF ERINEELPEE EKFSAQEYLA GLKIMSDRNN LMVADDKVWR V

UniProtKB: DNA replication licensing factor MCM3

Macromolecule #3: DNA replication licensing factor MCM4

• Macromolecule: Name: DNA replication licensing factor MCM4 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 105.138375 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

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 #4: Minichromosome maintenance protein 5

• Macromolecule: Name: Minichromosome maintenance protein 5 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 86.505734 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

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 #5: DNA replication licensing factor MCM6

• Macromolecule: Name: DNA replication licensing factor MCM6 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 113.110211 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

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 #6: DNA replication licensing factor MCM7

• Macromolecule: Name: DNA replication licensing factor MCM7 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA helicase
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 95.049875 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

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 #7: DNA replication complex GINS protein PSF1

• Macromolecule: Name: DNA replication complex GINS protein PSF1 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 24.230576 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

MYGDLGNKLV LEAKRTKQLY ARSNQDVNLP MYHEDIIRNI LKEVSNLRKN TEYLKEQQQL GMLDDKVAKC QYFVTLLCME RNKRCLLAY QRLRTDILDS MAWNNNGLDL MSSITFSQQD TNNLSHQEQE YLKEYCDLIT DLKSGDLVDI DLSGSLVPPS D VFIDVRVL KDAGEIQTEY GVFNLIKDSQ FFVRQSDVER LIQQGYLQKI

UniProtKB: DNA replication complex GINS protein PSF1

Macromolecule #8: DNA replication complex GINS protein PSF2

• Macromolecule: Name: DNA replication complex GINS protein PSF2 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 25.096807 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

MSLPAHLQQT FSPEEIQFIV ENEPIKIFPR ITTRQKIRGD DRGTGNHTRW QLITTDDKAL NNMVAMRSTE VVLWIALLLK QQSKCSIVA PQWLTTKELD RKIQYEKTHP DRFSELPWNW LVLARILFNK AKDDFHDPIH ELRGKIQDLR EIRQIKVLKG L KYLNESHL QLDNLSLLEI NELRPFITEI MDKLREIHTA SLTAGTENDE EEFNI

UniProtKB: DNA replication complex GINS protein PSF2

Macromolecule #9: DNA replication complex GINS protein PSF3

• Macromolecule: Name: DNA replication complex GINS protein PSF3 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 22.00416 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

MGYYDIDDVL ADGTEFPCKF QYDIPGLGYL ENNPGRPITK NTKLNLPLWL ARILAIVGGD EALVDEEPVP FVELLPPDMF STKVMNAIK TDPVALDLHS INSHFFSLAI KWIMLFSEKE LANVVSELLL QRAQELNHHA SSLSIDLNAD STGKNSANTN I ATSTFLLK LEEMEKEIYK KSHESYKDTK RWMFKK

UniProtKB: DNA replication complex GINS protein PSF3

Macromolecule #10: DNA replication complex GINS protein SLD5

• Macromolecule: Name: DNA replication complex GINS protein SLD5 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 33.983617 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

MDINIDDILA ELDKETTAVD STKITQGSSS TTHRDANTIV GSSLDLNDKT QIYVSPQQDF SDLMKSWKNE RCSPELLPYP HQLMKRLLN RISMQSQLIE NISMGFLDMQ NASNANPPMP NESKLPLLCM ETELERLKFV IRSYIRCRLS KIDKFSLYLR Q LNEDENSL ISLTDLLSKD EIKYHDTHSL IWLKLVNDSI LKYMPEELQA INDTEGSVNM IDEPDWNKFV FIHVNGPPDG KW NEDPLLQ ENEFGKPCYT VTIPDLKEEV ELTIGSIYVM RYEVIRDLLR DDKVALI

UniProtKB: DNA replication complex GINS protein SLD5

Macromolecule #11: Cell division control protein 45,Cell division control protein 45

• Macromolecule: Name: Cell division control protein 45,Cell division control protein 45; type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 75.154703 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

MYYGISQFSE AYNKILRNSS SHSSCQLVIF VSCLNIDALC ATKMLSLLFK KQLVQSQIVP IFGYSELRRH YSQLDDNINS LLLVGFGGV IDLEAFLEID PQEYVIDTDE KSGEQSFRRD IYVLDAHRPW NLDNIFGSQI IQCFDDGTVD DTLGEQKEAY Y KLLELDEE SGDDELSGDE NDNNGGDDEA TDADEVTDED YKDDDGDYKD DDETISNKRG NSSIGPNDLS KRKQRKKQIH EY EGVLEEY YSQGTTVVNS ISAQIYSLLS AIGETNLSNL WLNILGTTSL DIAYAQVYNR LYPLLQDEVK RLTPSSRNSV KTP DTLTLN IQPDYYLFLL RHSSLYDSFY YSNYVNAKLS LWNENGKKRL HKMFARMGIP LSTAQETWLY MDHSIKRELG IIFD KNLDR YGLQDIIRDG FVRTLGYRGS ISASEFVEAL TALLEVGNST DKDSVKINND NNDDTDGEEE EDNSAQKLTN LRKRW VSNF WLSWDALDDR KVELLNRGIQ LAQDLQRAIF NTGVAILEKK LIKHLRIYRL CVLQDGPDLD LYRNPLTLLR LGNWLI ECC AESEDKQLLP MVLASIDENT DTYLVAGLTP RYPRGLDTIH TKKPILNNFS MAFQQITAET DAKVRIDNFE SSIIEIR RE DLSPFLEKLT LSGLL

UniProtKB: Cell division control protein 45, Cell division control protein 45

Macromolecule #12: DNA polymerase alpha-binding protein

• Macromolecule: Name: DNA polymerase alpha-binding protein / type: protein_or_peptide / ID: 12 / Number of copies: 3 / Enantiomer: LEVO
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 108.610148 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

MKRRWKKNFI AVSAANRFKK ISSSGALENL YFQGEMVSVI DKLVFDFGGK TLVSLAPDNN TLCVANKNGL TKILKTNNPE EEPETLDSS KLVSSIKCYS NSHFLMTTMQ GDALRYNIDS SQEELLARFA LPLRDCCVIH SGKMAVFGGD DLELILLELD D ETHKKHAI KIDEQVSQIS YNSQMNILAV SMINGKVQIF SLTSTIPNKV HELNDYIVAN SYDDTHRDKI LSNMMDDIDK DN DNDLSET ADPDENNVAD PEFCAANRIC TRVAWHPKGL HFALPCADDT VKIFSIKGYS LQKTLSTNLS STKAHFIDLQ FDP LRGTYI AAVDLNNKLT VWNWETSEIH YTREFKRKIT NIAWKIQADS KTLDLVLGTW SGSIAIVQNL AESVVSNIPD QSVA ESSTK HGLFVDSESD LENLEGNDDI NKSDKLFSDI TQEANAEDVF TQTHDGPSGL SEKRKYNFED EEDFIDDDDG AGYIS GKKP HNEHSYSRVH KTHSFPISLA NTGKFRYMPF SPAGTPFGFT DRRYLTMNEV GYVSTVKNSE QYSITVSFFD VGRFRE YHF EDLFGYDLCF LNEKGTLFGQ SKTGQIQYRP HDSIHSNWTK IIPLQAGERI TSVAATPVRV IVGTSLGYFR SFNQFGV PF AVEKTSPIVA LTAQNYRVFS VHYSQFHGLS YSLSELGTSS KRYYKRECPL PMSLPNINSD MKKDANLDYY NFNPMGIK S LFFSSYGDPC IFGSDNTLLL LSKWRSPEES KWLPILDSNM EIWKMSGGKE TTDIHVWPLA LAYDTLNCIL VKGKHIWPE FPLPLPSEME IRMPVFVKSK LLEENKAILN KKNEIGADTE AEEGEEDKEI QIPVSMAAEE EYLRSKVLSE LLTDTLENDG EMYGNENEV LAALNGAYDK ALLRLFASAC SDQNVEKALS LAHELKQDRA LTAAVKISER AELPSLVKKI NNIREARYEQ Q LK

UniProtKB: DNA polymerase alpha-binding protein

Macromolecule #15: Suppressor of kinetochore protein 1

• Macromolecule: Name: Suppressor of kinetochore protein 1 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 22.35727 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

MVTSNVVLVS GEGERFTVDK KIAERSLLLK NYLNDMHDSN LQNNSDSESD SDSETNHKSK DNNNGDDDDE DDDEIVMPVP NVRSSVLQK VIEWAEHHRD SNFPDEDDDD SRKSAPVDSW DREFLKVDQE MLYEIILAAN YLNIKPLLDA GCKVVAEMIR G RSPEEIRR TFNIVNDFTP EEEAAIRREN EWAEDR

UniProtKB: Suppressor of kinetochore protein 1

Macromolecule #16: Protein DIA2

• Macromolecule: Name: Protein DIA2 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 85.368844 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

GAGMSSPGNS GVAIDSTVLK AIELGTRLFK SGEYLQAKRI FTNALRVCDS YSQEQIMRIR NAYQLDTARP DNKRLYHPRY IKILDNICA CYEKLNDLKS CLDVSQRLLK LEPGNIKCYI RCTRTLIKLK DWKRAYKTCS RGLQLCNNDS NHLRQQKQFI K NNMVQKQD GKRSYIDPLE ETKIAKKKKN NNVLESLPKK KIKGSTKKTD LVGNLPIEIL PIIFQRFTTK ELVTLSLVCN KW RDKILYH LDCFQEFNLA PINFKNFVKF MDFLQQNFTR TYRKYILSQV KVSSRITSEE LRITQLLFSK MPKCINIERL ILS MPTLTT TQIFKLMVRG GTDFFTRLLE LSLMITYRPD KQHELEILQT CPLLKKIELI FVNSLVPIFD GNNSVGRDGS FNVM ARHTN MQISTADNDE QGIVEEKVIY SELEKITLIC DKKKIKNFPL CRALLRGQFP LLQKLTITGV TFPMNNQDIM NFQWL LNFP DLKELWIEDN DNCELSKFLQ LLKFSNVWKN LEKLTFRENK LYPIVNLDED QPVTNDDEVP SMLFYKENLQ NLEKLD LMG TSISGSALTR LCEQEYLDGR KLRSLNIGNC PNIQFPNNHA HTARMILDVN AVLKRLSKLE EINLSHLSSL NDSTMKS FI INVPFLENLK RLDISHNFEI TGISIYEFLK KFQMDHDNEA GGQPLAYLNI DGCSQVSHIT VNMIRAQNLV TQVDCVYE R DVWRKFGINS YSYS

UniProtKB: Protein DIA2

Macromolecule #17: DNA polymerase epsilon catalytic subunit A

• Macromolecule: Name: DNA polymerase epsilon catalytic subunit A / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed DNA polymerase
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 255.890422 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

MMFGKKKNNG GSSTARYSAG NKYNTLSNNY ALSAQQLLNA SKIDDIDSMM GFERYVPPQY NGRFDAKDID QIPGRVGWLT NMHATLVSQ ETLSSGSNGG GNSNDGERVT TNQGISGVDF YFLDEEGGSF KSTVVYDPYF FIACNDESRV NDVEELVKKY L ESCLKSLQ IIRKEDLTMD NHLLGLQKTL IKLSFVNSNQ LFEARKLLRP ILQDNANNNV QRNIYNVAAN GSEKVDAKHL IE DIREYDV PYHVRVSIDK DIRVGKWYKV TQQGFIEDTR KIAFADPVVM AFAIATTKPP LKFPDSAVDQ IMMISYMIDG EGF LITNRE IISEDIEDFE YTPKPEYPGF FTIFNENDEV ALLQRFFEHI RDVRPTVIST FNGDFFDWPF IHNRSKIHGL DMFD EIGFA PDAEGEYKSS YCSHMDCFRW VKRDSYLPQG SQGLKAVTQS KLGYNPIELD PELMTPYAFE KPQHLSEYSV SDAVA TYYL YMKYVHPFIF SLCTIIPLNP DETLRKGTGT LCEMLLMVQA YQHNILLPNK HTDPIERFYD GHLLESETYV GGHVES LEA GVFRSDLKNE FKIDPSAIDE LLQELPEALK FSVEVENKSS VDKVTNFEEI KNQITQKLLE LKENNIRNEL PLIYHVD VA SMYPNIMTTN RLQPDSIKAE RDCASCDFNR PGKTCARKLK WAWRGEFFPS KMDEYNMIKR ALQNETFPNK NKFSKKKV L TFDELSYADQ VIHIKKRLTE YSRKVYHRVK VSEIVEREAI VCQRENPFYV DTVKSFRDRR YEFKGLAKTW KGNLSKIDP SDKHARDEAK KMIVLYDSLQ LAHKVILNSF YGYVMRKGSR WYSMEMAGIT CLTGATIIQM ARALVERVGR PLELDTDGIW CILPKSFPE TYFFTLENGK KLYLSYPCSM LNYRVHQKFT NHQYQELKDP LNYIYETHSE NTIFFEVDGP YKAMILPSSK E EGKGIKKR YAVFNEDGSL AELKGFELKR RGELQLIKNF QSDIFKVFLE GDTLEGCYSA VASVCNRWLD VLDSHGLMLE DE DLVSLIC ENRSMSKTLK EYEGQKSTSI TTARRLGDFL GEDMVKDKGL QCKYIISSKP FNAPVTERAI PVAIFSADIP IKR SFLRRW TLDPSLEDLD IRTIIDWGYY RERLGSAIQK IITIPAALQG VSNPVPRVEH PDWLKRKIAT KEDKFKQTSL TKFF SKTKN VPTMGKIKDI EDLFEPTVEE DNAKIKIART TKKKAVSKRK RNQLTNEEDP LVLPSEIPSM DEDYVGWLNY QKIKW KIQA RDRKRRDQLF GNTNSSRERS ALGSMIRKQA ESYANSTWEV LQYKDSGEPG VLEVFVTING KVQNITFHIP KTIYMK FKS QTMPLQKIKN CLIEKSSASL PNNPKTSNPA GGQLFKITLP ESVFLEEKEN CTSIFNDENV LGVFEGTITP HQRAIMD LG ASVTFRSKAM GALGKGIQQG FEMKDLSMAE NERYLSGFSM DIGYLLHFPT SIGYEFFSLF KSWGDTITIL VLKPSNQA Q EINASSLGQI YKQMFEKKKG KIETYSYLVD IKEDINFEFV YFTDISKLYR RLSQETTKLK EERGLQFLLL LQSPFITKL LGTIRLLNQM PIVKLSLNEV LLPQLNWQPT LLKKLVNHVL SSGSWISHLI KLSQYSNIPI CNLRLDSMDY IIDVLYARKL KKENIVLWW NEKAPLPDHG GIQNDFDLNT SWIMNDSEFP KINNSGVYDN VVLDVGVDNL TVNTILTSAL INDAEGSDLV N NNMGIDDK DAVINSPSEF VHDAFSNDAL NVLRGMLKEW WDEALKENST ADLLVNSLAS WVQNPNAKLF DGLLRYHVHN LT KKALLQL VNEFSALGST IVYADRNQIL IKTNKYSPEN CYAYSQYMMK AVRTNPMFSY LDLNIKRYWD LLIWMDKFNF SGL ACIEIE EKENQDYTAV SQWQLKKFLS PIYQPEFEDW MMIILDSMLK TKQSYLKLNS GTQRPTQIVN VKKQDKEDSV ENSL NGFSH LFSKPLMKRV KKLFKNQQEF ILDPQYEADY VIPVLPGSHL NVKNPLLELV KSLCHVMLLS KSTILEIRTL RKELL KIFE LREFAKVAEF KDPSLSLVVP DFLCEYCFFI SDIDFCKAAP ESIFSCVRCH KAFNQVLLQE HLIQKLRSDI ESYLIQ DLR CSRCHKVKRD YMSAHCPCAG AWEGTLPRES IVQKLNVFKQ VAKYYGFDIL LSCIADLTI

UniProtKB: DNA polymerase epsilon catalytic subunit A

Macromolecule #18: DNA polymerase epsilon subunit B

• Macromolecule: Name: DNA polymerase epsilon subunit B / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 78.425852 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

MFGSGNVLPV KIQPPLLRPL AYRVLSRKYG LSIKSDGLSA LAEFVGTNIG ANWRQGPATI KFLEQFAAVW KQQERGLFID QSGVKEVIQ EMKEREKVEW SHEHPIQHEE NILGRTDDDE NNSDDEMPIA ADSSLQNVSL SSPMRQPTER DEYKQPFKPE S SKALDWRD YFKVINASQQ QRFSYNPHKM QFIFVPNKKQ NGLGGIAGFL PDIEDKVQMF LTRYYLTNDR VMRNENFQNS DM FNPLSSM VSLQNELSNT NRQQQSSSMS ITPIKNLLGR DAQNFLLLGL LNKNFKGNWS LEDPSGSVEI DISQTIPTQG HYY VPGCMV LVEGIYYSVG NKFHVTSMTL PPGERREITL ETIGNLDLLG IHGISNNNFI ARLDKDLKIR LHLLEKELTD HKFV ILGAN LFLDDLKIMT ALSKILQKLN DDPPTLLIWQ GSFTSVPVFA SMSSRNISSS TQFKNNFDAL ATLLSRFDNL TENTT MIFI PGPNDLWGSM VSLGASGTLP QDPIPSAFTK KINKVCKNVV WSSNPTRIAY LSQEIVIFRD DLSGRFKRHR LEFPFN ESE DVYTENDNMM SKDTDIVPID ELVKEPDQLP QKVQETRKLV KTILDQGHLS PFLDSLRPIS WDLDHTLTLC PIPSTMV LC DTTSAQFDLT YNGCKVINPG SFIHNRRARY MEYVPSSKKT IQEEIYI

UniProtKB: DNA polymerase epsilon subunit B

Macromolecule #19: Topoisomerase 1-associated factor 1

• Macromolecule: Name: Topoisomerase 1-associated factor 1 / type: protein_or_peptide / ID: 19 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 141.296875 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

MSADLQQGTT NAADFSLTVL RARIALLATA IGGPDYTSQI DPPPYKLGDD CLACLKDLKR WFKLVDDQQK RWDVAMAVAE YRILTDDLL PILIDWENKC SLAAKLAKNN PDHEEFRNKA YYDKIALNCL QLLVLMTWPL IVTEQSSSNQ ITLYGELKKH Q LVYKKTIL SMESGKVLRA AIRLALDVIK IDRLSRTPRD NMVLKLVLNF FRNVIAIEPG EFTINTKKSM PKKGITSIDT LP PNVSMDD ISLNTVISSF HKNKVFGFLL TLTSSLSKEF DQDFINIPLL EIMFYFTKDV NQELLFPRQF ETGTHSKVVN KNE SSSANN IVTSAGFELS KLLQKEHQMR KNVIKHTSAR HSRFGGLLSI QTPDKTRLTV SGSQALVDEK IALQKLDDSK KWNK RIIKK HQSVAAEGLP NSLLNSQTGK AIFFTESNGK HFKEFINNFI DSGFNILLHS VTNYFTTEQD RMVTLEQVEY LLFFA WFVK YQLLRSKIDN SADIKQVSEA LKEVTFILVS SLLRSAYDLK NWTVTHAGMI AFNELLNLVS RTKAAQEEDS TDIEFI VSR LFSDERIQLL SNLPKIGSKY SLQFMKSCIE LTHSVLKVLE QYSDDKTLVI EGKSRRQKKF NISEGDITKL IEEENVD RD EALDILTSSL RSIEVNFQKV QANYMTEPVI ETYINFLERF RELEDDSIKK VFSFFHRVFV QAKEQALLFR FDLIILLR E MLSPDGLDRM SRSRKYVSQF SDYFLARLKK RLKKSPAWFV GLLFPPLHNS EVGFYQRYGE YNVLNNESMY AAPASQFKP IPDEEALPPS ILLDMKYGVL VSTLLDDGKT ELLDQLLKHI THTLDIFKSW LTVNVNAGKE TVNPPNEYFT LTGVLNNDPI FKDKDYRAL LLLIGYSIPR KINEPCFLPG TVEVSDLTVS CELVKKYLST PFETPNGLPS SSYLLRVRSE KDSFSHNEQD G WEGDDDYD YNDPYIVPDD QILSKSDAAY FKDLDNNASD KLKGTKFSKG IARSKKKDKR KRRKGEAKTN LPMFGDQDDE RP QTVRERH GVFSKEFISD SEDDEDLMNP IFFENETYMR WLLDKNNGQL TEDRYIQFAK FAAERMNNGG VVTGDYTSLF GGS IPSIES IRATESSSFA PDKSLISLAS HVASEMSIFD VNNNNNNQLS DDDVNSESRN SLGSSQPSNS QNMFQSEVYS RKES TKRSL EASAADESDE DEEAIRLFGK KSRVVLSQGD SDD

UniProtKB: Topoisomerase 1-associated factor 1

Macromolecule #20: Chromosome segregation in meiosis protein 3

• Macromolecule: Name: Chromosome segregation in meiosis protein 3 / type: protein_or_peptide / ID: 20 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 36.588758 KDa
• Recombinant expression: Organism: Saccharomyces cerevisiae (brewer's yeast)

Sequence

String:

GEMDQDFDSL LLGFNDSDSV QKDPTVPNGL DGSVVDPTIA DPTAITARKR RPQVKLTAEK LLSDKGLPYV LKNAHKRIRI SSKKNSYDN LSNIIQFYQL WAHELFPKAK FKDFMKICQT VGKTDPVLRE YRVSLFRDEM GMSFDVGTRE TGQDLERQSP M VEEHVTSA EERPIVADSF AQDKRNVNNV DYDNDEDDDI YHLSYRNRRG RVLDERGNNE TVLNNVVPPK EDLDALLKTF RV QGPVGLE ENEKKLLLGW LDAHRKMEKG SMTEEDVQLI QSLEEWEMND IEGQHTHYDL LPGGDEFGVD QDELDAMKEM GF

UniProtKB: Chromosome segregation in meiosis protein 3

Macromolecule #13: Leading strand template DNA

• Macromolecule: Name: Leading strand template DNA / type: dna / ID: 13 / Number of copies: 1 / Classification: DNA
• Source (natural): Organism: Synthetic construct (others)
• Molecular weight: Theoretical: 37.738676 KDa

Sequence

String:

(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG) (DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG) (DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG) (DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG) (DG) (DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG) (DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG) (DG)(DG) (DG)(DG)(DG)(DG)(DG)(DG)(DG) (DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG) (DG)(DG)(DG) (DG)(DG)(DG)(DG)(DG)(DG) (DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG) (DG)(DG)(DG)(DG) (DG)(DG)(DT)(DT)(DT) (DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)(DG)

Macromolecule #14: Lagging strand template DNA

• Macromolecule: Name: Lagging strand template DNA / type: dna / ID: 14 / Number of copies: 1 / Classification: DNA
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 35.259238 KDa

Sequence

String:

(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC) (DA)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC) (DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC) (DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC) (DC) (DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC) (DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC) (DC)(DC) (DC)(DC)(DC)(DC)(DC)(DC)(DC) (DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC) (DC)(DC)(DC) (DC)(DC)(DC)(DC)(DC)(DC) (DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC) (DC)(DC)(DC)(DC) (DC)(DC)(DC)(DC)(DC) (DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC)(DC) (DC)(DC)(DC)(DC)(DC) (DC)(DC)

Macromolecule #21: PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER

• Macromolecule: Name: PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER / type: ligand / ID: 21 / Number of copies: 2 / Formula: ANP
• Molecular weight: Theoretical: 506.196 Da

Chemical component information

ChemComp-ANP:
PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER / AMP-PNP, energy-carrying molecule analogue*YM

Macromolecule #22: MAGNESIUM ION

• Macromolecule: Name: MAGNESIUM ION / type: ligand / ID: 22 / Number of copies: 2 / Formula: MG
• Molecular weight: Theoretical: 24.305 Da

Macromolecule #23: ZINC ION

• Macromolecule: Name: ZINC ION / type: ligand / ID: 23 / Number of copies: 7 / Formula: ZN
• Molecular weight: Theoretical: 65.409 Da

Experimental details

Structure determination

• Method: cryo EM
• Processing: single particle reconstruction
• Aggregation state: particle

Sample preparation

• Buffer: pH: 7.6
• Grid: Model: Quantifoil R2/2 / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 5 sec. / Details: 15 mA
• Vitrification: Cryogen name: ETHANE / Details: Manual plunger.

Electron microscopy

• Microscope: FEI TITAN KRIOS
• Specialist optics: Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
• Image recording: Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number real images: 12730 / Average exposure time: 4.0 sec. / Average electron dose: 38.8 e/Ų
• Electron beam: Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
• Electron optics: Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.2 µm / Nominal defocus min: 0.4 µm / Nominal magnification: 81000
• Sample stage: Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
• Experimental equipment: Model: Titan Krios / Image courtesy: FEI Company

Image processing

• Particle selection: Number selected: 2160000
• Startup model: Type of model: OTHER
• Final reconstruction: Resolution.type: BY AUTHOR / Resolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3); Details: Composite map produced from individual cryo-EM density maps (resolutions 3.2 - 4.0 A) using Phenix combine_focused_maps. See publication for details.; Number images used: 369254
• Initial angle assignment: Type: ANGULAR RECONSTITUTION / Software - Name: RELION (ver. 3)
• Final angle assignment: Type: ANGULAR RECONSTITUTION / Software - Name: RELION (ver. 3)