Starting phenix.real_space_refine (version: 1.21rc1) on Mon May 8 07:05:07 2023 by dcliebschner =============================================================================== Processing files: ------------------------------------------------------------------------------- Found model, /net/marbles/raid1/dorothee/rerefine/cryoem/data_no_H/8saq_40275/05_2023/8saq_40275_neut.pdb Found real_map, /net/marbles/raid1/dorothee/rerefine/cryoem/data_no_H/8saq_40275/05_2023/8saq_40275.map Processing PHIL parameters: ------------------------------------------------------------------------------- Adding command-line PHIL: ------------------------- refinement.macro_cycles=10 scattering_table=electron resolution=3.9 write_initial_geo_file=False Final processed PHIL parameters: ------------------------------------------------------------------------------- data_manager { real_map_files = "/net/marbles/raid1/dorothee/rerefine/cryoem/data_no_H/8saq_40275/05_2023/8saq_40275.map" default_real_map = "/net/marbles/raid1/dorothee/rerefine/cryoem/data_no_H/8saq_40275/05_2023/8saq_40275.map" model { file = "/net/marbles/raid1/dorothee/rerefine/cryoem/data_no_H/8saq_40275/05_2023/8saq_40275_neut.pdb" } default_model = "/net/marbles/raid1/dorothee/rerefine/cryoem/data_no_H/8saq_40275/05_2023/8saq_40275_neut.pdb" } resolution = 3.9 write_initial_geo_file = False refinement { macro_cycles = 10 } Starting job =============================================================================== ------------------------------------------------------------------------------- Citation: ********* Afonine PV, Poon BK, Read RJ, Sobolev OV, Terwilliger TC, Urzhumtsev A, Adams PD. (2018) Real-space refinement in PHENIX for cryo-EM and crystallography. Acta Cryst. D74:531-544. Validating inputs ------------------------------------------------------------------------------- Processing inputs ***************** Set random seed Set to: 0 Set model cs if undefined Decide on map wrapping Map wrapping is set to: False Normalize map: mean=0, sd=1 Input map: mean= 0.002 sd= 0.020 Set stop_for_unknowns flag Set to: True Assert model is a single copy model Assert all atoms have isotropic ADPs Construct map_model_manager Extract box with map and model Check model and map are aligned Set scattering table Set to: electron Number of scattering types: 4 Type Number sf(0) Gaussians S 142 5.16 5 C 11892 2.51 5 N 3201 2.21 5 O 3711 1.98 5 sf(0) = scattering factor at diffraction angle 0. Process input model Symmetric amino acids flipped Residue "A ASP 113": "OD1" <-> "OD2" Residue "A GLU 190": "OE1" <-> "OE2" Residue "A GLU 362": "OE1" <-> "OE2" Residue "A GLU 492": "OE1" <-> "OE2" Residue "C ASP 107": "OD1" <-> "OD2" Residue "D TYR 625": "CD1" <-> "CD2" "CE1" <-> "CE2" Residue "E GLU 47": "OE1" <-> "OE2" Residue "E GLU 83": "OE1" <-> "OE2" Residue "E PHE 85": "CD1" <-> "CD2" "CE1" <-> "CE2" Residue "E GLU 106": "OE1" <-> "OE2" Residue "E ASP 113": "OD1" <-> "OD2" Residue "E GLU 190": "OE1" <-> "OE2" Residue "E GLU 211": "OE1" <-> "OE2" Residue "E GLU 231": "OE1" <-> "OE2" Residue "E TYR 435": "CD1" <-> "CD2" "CE1" <-> "CE2" Residue "F GLU 584": "OE1" <-> "OE2" Residue "F GLU 654": "OE1" <-> "OE2" Residue "H ASP 107": "OD1" <-> "OD2" Residue "H TYR 111": "CD1" <-> "CD2" "CE1" <-> "CE2" Residue "I TYR 271": "CD1" <-> "CD2" "CE1" <-> "CE2" Residue "K GLU 106": "OE1" <-> "OE2" Residue "K GLU 213": "OE1" <-> "OE2" Residue "K GLU 233": "OE1" <-> "OE2" Residue "K GLU 364": "OE1" <-> "OE2" Residue "K TYR 437": "CD1" <-> "CD2" "CE1" <-> "CE2" Residue "L GLU 586": "OE1" <-> "OE2" Residue "M TYR 94": "CD1" <-> "CD2" "CE1" <-> "CE2" Time to flip residues: 0.05s Monomer Library directory: "/net/cci-filer2/raid1/xp/phenix/phenix-1.21rc1-4953/modules/chem_data/mon_lib" Total number of atoms: 18946 Number of models: 1 Model: "" Number of chains: 21 Chain: "A" Number of atoms: 3367 Number of conformers: 1 Conformer: "" Number of residues, atoms: 426, 3367 Classifications: {'peptide': 426} Link IDs: {'CIS': 1, 'PTRANS': 22, 'TRANS': 402} Chain breaks: 4 Chain: "B" Number of atoms: 1008 Number of conformers: 1 Conformer: "" Number of residues, atoms: 127, 1008 Classifications: {'peptide': 127} Modifications used: {'COO': 1} Link IDs: {'PTRANS': 1, 'TRANS': 125} Chain breaks: 1 Chain: "C" Number of atoms: 996 Number of conformers: 1 Conformer: "" Number of residues, atoms: 126, 996 Classifications: {'peptide': 126} Link IDs: {'PTRANS': 5, 'TRANS': 120} Chain: "D" Number of atoms: 796 Number of conformers: 1 Conformer: "" Number of residues, atoms: 109, 796 Classifications: {'peptide': 109} Link IDs: {'PTRANS': 5, 'TRANS': 103} Chain: "E" Number of atoms: 3367 Number of conformers: 1 Conformer: "" Number of residues, atoms: 426, 3367 Classifications: {'peptide': 426} Link IDs: {'CIS': 1, 'PTRANS': 22, 'TRANS': 402} Chain breaks: 4 Chain: "F" Number of atoms: 1008 Number of conformers: 1 Conformer: "" Number of residues, atoms: 127, 1008 Classifications: {'peptide': 127} Modifications used: {'COO': 1} Link IDs: {'PTRANS': 1, 'TRANS': 125} Chain breaks: 1 Chain: "H" Number of atoms: 996 Number of conformers: 1 Conformer: "" Number of residues, atoms: 126, 996 Classifications: {'peptide': 126} Link IDs: {'PTRANS': 5, 'TRANS': 120} Chain: "I" Number of atoms: 796 Number of conformers: 1 Conformer: "" Number of residues, atoms: 109, 796 Classifications: {'peptide': 109} Link IDs: {'PTRANS': 5, 'TRANS': 103} Chain: "K" Number of atoms: 3379 Number of conformers: 1 Conformer: "" Number of residues, atoms: 428, 3379 Classifications: {'peptide': 428} Link IDs: {'CIS': 1, 'PTRANS': 22, 'TRANS': 404} Chain breaks: 4 Chain: "L" Number of atoms: 991 Number of conformers: 1 Conformer: "" Number of residues, atoms: 125, 991 Classifications: {'peptide': 125} Link IDs: {'PTRANS': 1, 'TRANS': 123} Chain breaks: 1 Chain: "M" Number of atoms: 996 Number of conformers: 1 Conformer: "" Number of residues, atoms: 126, 996 Classifications: {'peptide': 126} Link IDs: {'PTRANS': 5, 'TRANS': 120} Chain: "N" Number of atoms: 796 Number of conformers: 1 Conformer: "" Number of residues, atoms: 109, 796 Classifications: {'peptide': 109} Link IDs: {'PTRANS': 5, 'TRANS': 103} Chain: "G" Number of atoms: 39 Number of conformers: 1 Conformer: "" Number of residues, atoms: 3, 39 Unusual residues: {'BMA': 1, 'NAG': 2} Classifications: {'undetermined': 3} Link IDs: {None: 2} Unresolved non-hydrogen bonds: 3 Unresolved non-hydrogen angles: 6 Unresolved non-hydrogen chiralities: 3 Chain: "J" Number of atoms: 83 Number of conformers: 1 Conformer: "" Number of residues, atoms: 7, 83 Unusual residues: {'BMA': 1, 'MAN': 4, 'NAG': 2} Classifications: {'undetermined': 7} Link IDs: {None: 6} Unresolved non-hydrogen bonds: 7 Unresolved non-hydrogen angles: 14 Unresolved non-hydrogen chiralities: 7 Chain: "Q" Number of atoms: 39 Number of conformers: 1 Conformer: "" Number of residues, atoms: 3, 39 Unusual residues: {'BMA': 1, 'NAG': 2} Classifications: {'undetermined': 3} Link IDs: {None: 2} Unresolved non-hydrogen bonds: 3 Unresolved non-hydrogen angles: 6 Unresolved non-hydrogen chiralities: 3 Chain: "R" Number of atoms: 83 Number of conformers: 1 Conformer: "" Number of residues, atoms: 7, 83 Unusual residues: {'BMA': 1, 'MAN': 4, 'NAG': 2} Classifications: {'undetermined': 7} Link IDs: {None: 6} Unresolved non-hydrogen bonds: 7 Unresolved non-hydrogen angles: 14 Unresolved non-hydrogen chiralities: 7 Chain: "U" Number of atoms: 39 Number of conformers: 1 Conformer: "" Number of residues, atoms: 3, 39 Unusual residues: {'BMA': 1, 'NAG': 2} Classifications: {'undetermined': 3} Link IDs: {None: 2} Unresolved non-hydrogen bonds: 3 Unresolved non-hydrogen angles: 6 Unresolved non-hydrogen chiralities: 3 Chain: "V" Number of atoms: 83 Number of conformers: 1 Conformer: "" Number of residues, atoms: 7, 83 Unusual residues: {'BMA': 1, 'MAN': 4, 'NAG': 2} Classifications: {'undetermined': 7} Link IDs: {None: 6} Unresolved non-hydrogen bonds: 7 Unresolved non-hydrogen angles: 14 Unresolved non-hydrogen chiralities: 7 Chain: "A" Number of atoms: 28 Number of conformers: 1 Conformer: "" Number of residues, atoms: 2, 28 Unusual residues: {'NAG': 2} Classifications: {'undetermined': 2} Link IDs: {None: 1} Unresolved non-hydrogen bonds: 2 Unresolved non-hydrogen angles: 4 Unresolved non-hydrogen chiralities: 2 Chain: "E" Number of atoms: 28 Number of conformers: 1 Conformer: "" Number of residues, atoms: 2, 28 Unusual residues: {'NAG': 2} Classifications: {'undetermined': 2} Link IDs: {None: 1} Unresolved non-hydrogen bonds: 2 Unresolved non-hydrogen angles: 4 Unresolved non-hydrogen chiralities: 2 Chain: "K" Number of atoms: 28 Number of conformers: 1 Conformer: "" Number of residues, atoms: 2, 28 Unusual residues: {'NAG': 2} Classifications: {'undetermined': 2} Link IDs: {None: 1} Unresolved non-hydrogen bonds: 2 Unresolved non-hydrogen angles: 4 Unresolved non-hydrogen chiralities: 2 Time building chain proxies: 10.00, per 1000 atoms: 0.53 Number of scatterers: 18946 At special positions: 0 Unit cell: (155.636, 155.636, 137.514, 90, 90, 90) Space group: P 1 (No. 1) Number of sites at special positions: 0 Number of scattering types: 4 Type Number sf(0) S 142 16.00 O 3711 8.00 N 3201 7.00 C 11892 6.00 sf(0) = scattering factor at diffraction angle 0. Number of disulfides: simple=39, symmetry=0 Simple disulfide: pdb=" SG CYS A 54 " - pdb=" SG CYS A 74 " distance=2.04 Simple disulfide: pdb=" SG CYS A 119 " - pdb=" SG CYS A 205 " distance=2.04 Simple disulfide: pdb=" SG CYS A 126 " - pdb=" SG CYS A 196 " distance=2.03 Simple disulfide: pdb=" SG CYS A 131 " - pdb=" SG CYS A 157 " distance=2.03 Simple disulfide: pdb=" SG CYS A 218 " - pdb=" SG CYS A 247 " distance=2.04 Simple disulfide: pdb=" SG CYS A 228 " - pdb=" SG CYS A 239 " distance=2.03 Simple disulfide: pdb=" SG CYS A 296 " - pdb=" SG CYS A 331 " distance=2.03 Simple disulfide: pdb=" SG CYS A 378 " - pdb=" SG CYS A 445 " distance=2.03 Simple disulfide: pdb=" SG CYS A 385 " - pdb=" SG CYS A 418 " distance=2.03 Simple disulfide: pdb=" SG CYS A 501 " - pdb=" SG CYS B 581 " distance=2.04 Simple disulfide: pdb=" SG CYS B 574 " - pdb=" SG CYS B 580 " distance=2.03 Simple disulfide: pdb=" SG CYS C 22 " - pdb=" SG CYS C 96 " distance=2.03 Simple disulfide: pdb=" SG CYS D 541 " - pdb=" SG CYS D 626 " distance=2.03 Simple disulfide: pdb=" SG CYS E 54 " - pdb=" SG CYS E 74 " distance=2.03 Simple disulfide: pdb=" SG CYS E 119 " - pdb=" SG CYS E 205 " distance=2.03 Simple disulfide: pdb=" SG CYS E 126 " - pdb=" SG CYS E 196 " distance=2.03 Simple disulfide: pdb=" SG CYS E 131 " - pdb=" SG CYS E 157 " distance=2.03 Simple disulfide: pdb=" SG CYS E 218 " - pdb=" SG CYS E 247 " distance=2.03 Simple disulfide: pdb=" SG CYS E 228 " - pdb=" SG CYS E 239 " distance=2.04 Simple disulfide: pdb=" SG CYS E 296 " - pdb=" SG CYS E 331 " distance=2.03 Simple disulfide: pdb=" SG CYS E 378 " - pdb=" SG CYS E 445 " distance=2.03 Simple disulfide: pdb=" SG CYS E 385 " - pdb=" SG CYS E 418 " distance=2.03 Simple disulfide: pdb=" SG CYS E 501 " - pdb=" SG CYS F 605 " distance=2.04 Simple disulfide: pdb=" SG CYS F 598 " - pdb=" SG CYS F 604 " distance=2.03 Simple disulfide: pdb=" SG CYS H 22 " - pdb=" SG CYS H 96 " distance=2.03 Simple disulfide: pdb=" SG CYS I 252 " - pdb=" SG CYS I 320 " distance=2.03 Simple disulfide: pdb=" SG CYS K 54 " - pdb=" SG CYS K 74 " distance=2.03 Simple disulfide: pdb=" SG CYS K 119 " - pdb=" SG CYS K 207 " distance=2.03 Simple disulfide: pdb=" SG CYS K 126 " - pdb=" SG CYS K 198 " distance=2.03 Simple disulfide: pdb=" SG CYS K 131 " - pdb=" SG CYS K 159 " distance=2.03 Simple disulfide: pdb=" SG CYS K 220 " - pdb=" SG CYS K 249 " distance=2.05 Simple disulfide: pdb=" SG CYS K 230 " - pdb=" SG CYS K 241 " distance=2.03 Simple disulfide: pdb=" SG CYS K 298 " - pdb=" SG CYS K 333 " distance=2.03 Simple disulfide: pdb=" SG CYS K 380 " - pdb=" SG CYS K 447 " distance=2.03 Simple disulfide: pdb=" SG CYS K 387 " - pdb=" SG CYS K 420 " distance=2.03 Simple disulfide: pdb=" SG CYS K 503 " - pdb=" SG CYS L 607 " distance=2.04 Simple disulfide: pdb=" SG CYS L 600 " - pdb=" SG CYS L 606 " distance=2.03 Simple disulfide: pdb=" SG CYS M 22 " - pdb=" SG CYS M 96 " distance=2.03 Simple disulfide: pdb=" SG CYS N 252 " - pdb=" SG CYS N 320 " distance=2.03 Automatic linking Parameters for automatic linking Linking & cutoffs Metal : Auto - 3.50 Amino acid : False - 1.90 Carbohydrate : True - 1.99 Ligands : True - 1.99 Small molecules : False - 1.98 Amino acid - RNA/DNA : False Number of custom bonds: simple=0, symmetry=0 Links applied ALPHA1-2 " MAN J 4 " - " MAN J 5 " " MAN J 5 " - " MAN J 6 " " MAN R 4 " - " MAN R 5 " " MAN R 5 " - " MAN R 6 " " MAN V 4 " - " MAN V 5 " " MAN V 5 " - " MAN V 6 " ALPHA1-3 " BMA J 3 " - " MAN J 4 " " BMA R 3 " - " MAN R 4 " " BMA V 3 " - " MAN V 4 " ALPHA1-6 " BMA J 3 " - " MAN J 7 " " BMA R 3 " - " MAN R 7 " " BMA V 3 " - " MAN V 7 " BETA1-4 " NAG G 1 " - " NAG G 2 " " NAG G 2 " - " BMA G 3 " " NAG J 1 " - " NAG J 2 " " NAG J 2 " - " BMA J 3 " " NAG Q 1 " - " NAG Q 2 " " NAG Q 2 " - " BMA Q 3 " " NAG R 1 " - " NAG R 2 " " NAG R 2 " - " BMA R 3 " " NAG U 1 " - " NAG U 2 " " NAG U 2 " - " BMA U 3 " " NAG V 1 " - " NAG V 2 " " NAG V 2 " - " BMA V 3 " NAG-ASN " NAG A 601 " - " ASN A 156 " " NAG A 602 " - " ASN A 442 " " NAG E 601 " - " ASN E 156 " " NAG E 602 " - " ASN E 442 " " NAG G 1 " - " ASN A 301 " " NAG J 1 " - " ASN A 332 " " NAG K 601 " - " ASN K 158 " " NAG K 602 " - " ASN K 444 " " NAG Q 1 " - " ASN E 301 " " NAG R 1 " - " ASN E 332 " " NAG U 1 " - " ASN K 303 " " NAG V 1 " - " ASN K 334 " Time building additional restraints: 7.92 Conformation dependent library (CDL) restraints added in 2.9 seconds 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. Adding C-beta torsion restraints... Number of C-beta restraints generated: 4360 Finding SS restraints... Secondary structure from input PDB file: 55 helices and 44 sheets defined 21.4% alpha, 29.1% beta 0 base pairs and 0 stacking pairs defined. Time for finding SS restraints: 1.29 Creating SS restraints... Processing helix chain 'A' and resid 60 through 64 Processing helix chain 'A' and resid 99 through 117 Processing helix chain 'A' and resid 122 through 126 Processing helix chain 'A' and resid 195 through 198 Processing helix chain 'A' and resid 334 through 353 removed outlier: 4.147A pdb=" N LYS A 351 " --> pdb=" O LYS A 347 " (cutoff:3.500A) removed outlier: 4.359A pdb=" N HIS A 352 " --> pdb=" O GLU A 348 " (cutoff:3.500A) removed outlier: 3.650A pdb=" N PHE A 353 " --> pdb=" O LEU A 349 " (cutoff:3.500A) Processing helix chain 'A' and resid 368 through 373 Processing helix chain 'A' and resid 377 through 381 removed outlier: 3.615A pdb=" N GLY A 380 " --> pdb=" O ASN A 377 " (cutoff:3.500A) removed outlier: 3.741A pdb=" N GLU A 381 " --> pdb=" O CYS A 378 " (cutoff:3.500A) No H-bonds generated for 'chain 'A' and resid 377 through 381' Processing helix chain 'A' and resid 475 through 480 removed outlier: 3.960A pdb=" N TRP A 479 " --> pdb=" O MET A 475 " (cutoff:3.500A) Processing helix chain 'B' and resid 505 through 510 Processing helix chain 'B' and resid 514 through 519 Processing helix chain 'B' and resid 545 through 572 removed outlier: 4.104A pdb=" N ILE B 549 " --> pdb=" O THR B 545 " (cutoff:3.500A) removed outlier: 4.326A pdb=" N ALA B 554 " --> pdb=" O LYS B 550 " (cutoff:3.500A) removed outlier: 3.689A pdb=" N TRP B 572 " --> pdb=" O LEU B 568 " (cutoff:3.500A) Processing helix chain 'B' and resid 587 through 591 Processing helix chain 'B' and resid 597 through 602 removed outlier: 3.836A pdb=" N MET B 602 " --> pdb=" O ILE B 598 " (cutoff:3.500A) Processing helix chain 'B' and resid 603 through 612 Processing helix chain 'B' and resid 614 through 639 removed outlier: 3.776A pdb=" N GLU B 624 " --> pdb=" O GLY B 620 " (cutoff:3.500A) removed outlier: 3.738A pdb=" N GLN B 629 " --> pdb=" O SER B 625 " (cutoff:3.500A) removed outlier: 5.051A pdb=" N ASN B 632 " --> pdb=" O GLN B 628 " (cutoff:3.500A) removed outlier: 3.774A pdb=" N GLU B 633 " --> pdb=" O GLN B 629 " (cutoff:3.500A) removed outlier: 4.412A pdb=" N LEU B 637 " --> pdb=" O GLU B 633 " (cutoff:3.500A) Processing helix chain 'C' and resid 28 through 30 No H-bonds generated for 'chain 'C' and resid 28 through 30' Processing helix chain 'C' and resid 61 through 65 Processing helix chain 'C' and resid 87 through 91 Processing helix chain 'D' and resid 617 through 621 Processing helix chain 'E' and resid 60 through 64 Processing helix chain 'E' and resid 99 through 117 Processing helix chain 'E' and resid 122 through 126 Processing helix chain 'E' and resid 194 through 199 removed outlier: 4.095A pdb=" N THR E 198 " --> pdb=" O ILE E 194 " (cutoff:3.500A) Processing helix chain 'E' and resid 334 through 350 removed outlier: 3.587A pdb=" N GLN E 350 " --> pdb=" O GLY E 346 " (cutoff:3.500A) Processing helix chain 'E' and resid 368 through 373 Processing helix chain 'F' and resid 529 through 534 Processing helix chain 'F' and resid 538 through 543 Processing helix chain 'F' and resid 569 through 596 removed outlier: 3.898A pdb=" N ILE F 573 " --> pdb=" O THR F 569 " (cutoff:3.500A) removed outlier: 4.160A pdb=" N ALA F 578 " --> pdb=" O LYS F 574 " (cutoff:3.500A) removed outlier: 3.696A pdb=" N TRP F 596 " --> pdb=" O LEU F 592 " (cutoff:3.500A) Processing helix chain 'F' and resid 611 through 615 Processing helix chain 'F' and resid 621 through 626 Processing helix chain 'F' and resid 627 through 636 Processing helix chain 'F' and resid 638 through 650 removed outlier: 3.653A pdb=" N ILE F 642 " --> pdb=" O TYR F 638 " (cutoff:3.500A) removed outlier: 3.584A pdb=" N GLU F 648 " --> pdb=" O GLY F 644 " (cutoff:3.500A) Processing helix chain 'F' and resid 650 through 662 removed outlier: 4.037A pdb=" N ASN F 656 " --> pdb=" O GLN F 652 " (cutoff:3.500A) removed outlier: 3.913A pdb=" N GLU F 657 " --> pdb=" O GLN F 653 " (cutoff:3.500A) Processing helix chain 'H' and resid 28 through 30 No H-bonds generated for 'chain 'H' and resid 28 through 30' Processing helix chain 'H' and resid 61 through 65 removed outlier: 3.565A pdb=" N GLN H 65 " --> pdb=" O ARG H 62 " (cutoff:3.500A) Processing helix chain 'H' and resid 87 through 91 Processing helix chain 'I' and resid 257 through 262 removed outlier: 4.775A pdb=" N HIS I 262 " --> pdb=" O ASP I 258 " (cutoff:3.500A) Processing helix chain 'I' and resid 311 through 315 Processing helix chain 'K' and resid 60 through 64 Processing helix chain 'K' and resid 99 through 117 Processing helix chain 'K' and resid 122 through 126 Processing helix chain 'K' and resid 197 through 200 Processing helix chain 'K' and resid 336 through 356 removed outlier: 4.028A pdb=" N LYS K 353 " --> pdb=" O LYS K 349 " (cutoff:3.500A) removed outlier: 4.147A pdb=" N HIS K 354 " --> pdb=" O GLU K 350 " (cutoff:3.500A) removed outlier: 3.628A pdb=" N PHE K 356 " --> pdb=" O LEU K 351 " (cutoff:3.500A) Processing helix chain 'K' and resid 370 through 375 Processing helix chain 'K' and resid 477 through 483 removed outlier: 3.904A pdb=" N TRP K 481 " --> pdb=" O MET K 477 " (cutoff:3.500A) removed outlier: 3.847A pdb=" N SER K 483 " --> pdb=" O ASP K 479 " (cutoff:3.500A) Processing helix chain 'L' and resid 531 through 536 Processing helix chain 'L' and resid 540 through 545 removed outlier: 3.642A pdb=" N ASN L 545 " --> pdb=" O VAL L 541 " (cutoff:3.500A) Processing helix chain 'L' and resid 571 through 598 removed outlier: 3.742A pdb=" N ILE L 575 " --> pdb=" O THR L 571 " (cutoff:3.500A) removed outlier: 3.699A pdb=" N ALA L 580 " --> pdb=" O LYS L 576 " (cutoff:3.500A) removed outlier: 3.652A pdb=" N TRP L 598 " --> pdb=" O LEU L 594 " (cutoff:3.500A) Processing helix chain 'L' and resid 613 through 617 Processing helix chain 'L' and resid 623 through 627 removed outlier: 3.685A pdb=" N ASN L 627 " --> pdb=" O ILE L 624 " (cutoff:3.500A) Processing helix chain 'L' and resid 629 through 638 Processing helix chain 'L' and resid 640 through 662 removed outlier: 3.750A pdb=" N ILE L 644 " --> pdb=" O TYR L 640 " (cutoff:3.500A) removed outlier: 3.583A pdb=" N GLU L 650 " --> pdb=" O GLY L 646 " (cutoff:3.500A) removed outlier: 4.176A pdb=" N ASN L 653 " --> pdb=" O GLU L 649 " (cutoff:3.500A) removed outlier: 4.051A pdb=" N GLN L 655 " --> pdb=" O SER L 651 " (cutoff:3.500A) Processing helix chain 'M' and resid 28 through 30 No H-bonds generated for 'chain 'M' and resid 28 through 30' Processing helix chain 'M' and resid 87 through 91 Processing helix chain 'N' and resid 311 through 315 removed outlier: 3.540A pdb=" N GLU N 315 " --> pdb=" O ALA N 312 " (cutoff:3.500A) Processing sheet with id=AA1, first strand: chain 'A' and resid 494 through 499 removed outlier: 5.218A pdb=" N VAL B 584 " --> pdb=" O VAL A 36 " (cutoff:3.500A) removed outlier: 7.281A pdb=" N VAL A 38 " --> pdb=" O THR B 582 " (cutoff:3.500A) removed outlier: 7.593A pdb=" N THR B 582 " --> pdb=" O VAL A 38 " (cutoff:3.500A) removed outlier: 8.123A pdb=" N TYR A 40 " --> pdb=" O CYS B 580 " (cutoff:3.500A) removed outlier: 8.300A pdb=" N CYS B 580 " --> pdb=" O TYR A 40 " (cutoff:3.500A) Processing sheet with id=AA2, first strand: chain 'A' and resid 45 through 47 removed outlier: 3.815A pdb=" N ILE A 225 " --> pdb=" O VAL A 245 " (cutoff:3.500A) Processing sheet with id=AA3, first strand: chain 'A' and resid 53 through 56 Processing sheet with id=AA4, first strand: chain 'A' and resid 91 through 94 removed outlier: 3.621A pdb=" N GLU A 91 " --> pdb=" O CYS A 239 " (cutoff:3.500A) removed outlier: 3.863A pdb=" N PHE A 93 " --> pdb=" O GLY A 237 " (cutoff:3.500A) removed outlier: 4.024A pdb=" N GLY A 237 " --> pdb=" O PHE A 93 " (cutoff:3.500A) Processing sheet with id=AA5, first strand: chain 'A' and resid 169 through 177 Processing sheet with id=AA6, first strand: chain 'A' and resid 260 through 261 removed outlier: 3.994A pdb=" N GLY A 451 " --> pdb=" O LEU A 260 " (cutoff:3.500A) removed outlier: 7.281A pdb=" N GLY A 451 " --> pdb=" O THR A 290 " (cutoff:3.500A) removed outlier: 10.904A pdb=" N THR A 290 " --> pdb=" O GLY A 451 " (cutoff:3.500A) removed outlier: 12.273A pdb=" N LEU A 453 " --> pdb=" O LEU A 288 " (cutoff:3.500A) removed outlier: 12.458A pdb=" N LEU A 288 " --> pdb=" O LEU A 453 " (cutoff:3.500A) removed outlier: 11.295A pdb=" N THR A 455 " --> pdb=" O VAL A 286 " (cutoff:3.500A) removed outlier: 11.354A pdb=" N VAL A 286 " --> pdb=" O THR A 455 " (cutoff:3.500A) removed outlier: 4.416A pdb=" N PHE A 382 " --> pdb=" O LYS A 421 " (cutoff:3.500A) Processing sheet with id=AA7, first strand: chain 'A' and resid 271 through 273 removed outlier: 3.779A pdb=" N ARG A 273 " --> pdb=" O ILE A 285 " (cutoff:3.500A) removed outlier: 11.354A pdb=" N VAL A 286 " --> pdb=" O THR A 455 " (cutoff:3.500A) removed outlier: 11.295A pdb=" N THR A 455 " --> pdb=" O VAL A 286 " (cutoff:3.500A) removed outlier: 12.458A pdb=" N LEU A 288 " --> pdb=" O LEU A 453 " (cutoff:3.500A) removed outlier: 12.273A pdb=" N LEU A 453 " --> pdb=" O LEU A 288 " (cutoff:3.500A) removed outlier: 10.904A pdb=" N THR A 290 " --> pdb=" O GLY A 451 " (cutoff:3.500A) removed outlier: 7.281A pdb=" N GLY A 451 " --> pdb=" O THR A 290 " (cutoff:3.500A) removed outlier: 6.674A pdb=" N LYS A 360 " --> pdb=" O PHE A 468 " (cutoff:3.500A) removed outlier: 3.635A pdb=" N TYR A 361 " --> pdb=" O GLY A 393 " (cutoff:3.500A) removed outlier: 3.919A pdb=" N GLY A 393 " --> pdb=" O TYR A 361 " (cutoff:3.500A) Processing sheet with id=AA8, first strand: chain 'A' and resid 304 through 308 Processing sheet with id=AA9, first strand: chain 'A' and resid 423 through 425 Processing sheet with id=AB1, first strand: chain 'C' and resid 11 through 12 removed outlier: 4.070A pdb=" N LYS C 12 " --> pdb=" O THR C 124 " (cutoff:3.500A) No H-bonds generated for sheet with id=AB1 Processing sheet with id=AB2, first strand: chain 'C' and resid 18 through 22 removed outlier: 3.651A pdb=" N VAL C 18 " --> pdb=" O LEU C 83 " (cutoff:3.500A) Processing sheet with id=AB3, first strand: chain 'C' and resid 45 through 51 removed outlier: 6.765A pdb=" N TRP C 36 " --> pdb=" O MET C 48 " (cutoff:3.500A) removed outlier: 4.375A pdb=" N TRP C 50 " --> pdb=" O ILE C 34 " (cutoff:3.500A) removed outlier: 6.478A pdb=" N ILE C 34 " --> pdb=" O TRP C 50 " (cutoff:3.500A) removed outlier: 5.206A pdb=" N TYR C 33 " --> pdb=" O GLY C 99 " (cutoff:3.500A) Processing sheet with id=AB4, first strand: chain 'D' and resid 528 through 531 removed outlier: 3.570A pdb=" N ALA D 622 " --> pdb=" O VAL D 643 " (cutoff:3.500A) Processing sheet with id=AB5, first strand: chain 'D' and resid 528 through 531 removed outlier: 3.570A pdb=" N ALA D 622 " --> pdb=" O VAL D 643 " (cutoff:3.500A) Processing sheet with id=AB6, first strand: chain 'D' and resid 536 through 542 Processing sheet with id=AB7, first strand: chain 'E' and resid 495 through 499 removed outlier: 5.380A pdb=" N VAL F 608 " --> pdb=" O VAL E 36 " (cutoff:3.500A) removed outlier: 7.578A pdb=" N VAL E 38 " --> pdb=" O THR F 606 " (cutoff:3.500A) removed outlier: 7.605A pdb=" N THR F 606 " --> pdb=" O VAL E 38 " (cutoff:3.500A) Processing sheet with id=AB8, first strand: chain 'E' and resid 45 through 47 removed outlier: 3.517A pdb=" N ILE E 225 " --> pdb=" O VAL E 245 " (cutoff:3.500A) Processing sheet with id=AB9, first strand: chain 'E' and resid 53 through 56 removed outlier: 3.730A pdb=" N CYS E 218 " --> pdb=" O PHE E 53 " (cutoff:3.500A) Processing sheet with id=AC1, first strand: chain 'E' and resid 91 through 93 removed outlier: 3.934A pdb=" N GLU E 91 " --> pdb=" O CYS E 239 " (cutoff:3.500A) removed outlier: 3.860A pdb=" N PHE E 93 " --> pdb=" O GLY E 237 " (cutoff:3.500A) removed outlier: 4.152A pdb=" N GLY E 237 " --> pdb=" O PHE E 93 " (cutoff:3.500A) Processing sheet with id=AC2, first strand: chain 'E' and resid 169 through 177 Processing sheet with id=AC3, first strand: chain 'E' and resid 202 through 203 removed outlier: 6.739A pdb=" N THR E 202 " --> pdb=" O TYR E 435 " (cutoff:3.500A) Processing sheet with id=AC4, first strand: chain 'E' and resid 259 through 261 removed outlier: 3.792A pdb=" N GLY E 451 " --> pdb=" O LEU E 260 " (cutoff:3.500A) removed outlier: 7.148A pdb=" N GLY E 451 " --> pdb=" O THR E 290 " (cutoff:3.500A) removed outlier: 10.784A pdb=" N THR E 290 " --> pdb=" O GLY E 451 " (cutoff:3.500A) removed outlier: 12.224A pdb=" N LEU E 453 " --> pdb=" O LEU E 288 " (cutoff:3.500A) removed outlier: 12.158A pdb=" N LEU E 288 " --> pdb=" O LEU E 453 " (cutoff:3.500A) removed outlier: 11.327A pdb=" N THR E 455 " --> pdb=" O VAL E 286 " (cutoff:3.500A) removed outlier: 11.639A pdb=" N VAL E 286 " --> pdb=" O THR E 455 " (cutoff:3.500A) removed outlier: 4.412A pdb=" N PHE E 382 " --> pdb=" O LYS E 421 " (cutoff:3.500A) Processing sheet with id=AC5, first strand: chain 'E' and resid 271 through 273 removed outlier: 11.639A pdb=" N VAL E 286 " --> pdb=" O THR E 455 " (cutoff:3.500A) removed outlier: 11.327A pdb=" N THR E 455 " --> pdb=" O VAL E 286 " (cutoff:3.500A) removed outlier: 12.158A pdb=" N LEU E 288 " --> pdb=" O LEU E 453 " (cutoff:3.500A) removed outlier: 12.224A pdb=" N LEU E 453 " --> pdb=" O LEU E 288 " (cutoff:3.500A) removed outlier: 10.784A pdb=" N THR E 290 " --> pdb=" O GLY E 451 " (cutoff:3.500A) removed outlier: 7.148A pdb=" N GLY E 451 " --> pdb=" O THR E 290 " (cutoff:3.500A) removed outlier: 6.857A pdb=" N LYS E 360 " --> pdb=" O PHE E 468 " (cutoff:3.500A) removed outlier: 3.698A pdb=" N TYR E 361 " --> pdb=" O GLY E 393 " (cutoff:3.500A) removed outlier: 3.939A pdb=" N GLY E 393 " --> pdb=" O TYR E 361 " (cutoff:3.500A) Processing sheet with id=AC6, first strand: chain 'E' and resid 304 through 308 Processing sheet with id=AC7, first strand: chain 'H' and resid 11 through 12 removed outlier: 3.889A pdb=" N LYS H 12 " --> pdb=" O THR H 124 " (cutoff:3.500A) No H-bonds generated for sheet with id=AC7 Processing sheet with id=AC8, first strand: chain 'H' and resid 21 through 22 Processing sheet with id=AC9, first strand: chain 'H' and resid 45 through 51 removed outlier: 5.354A pdb=" N TRP H 47 " --> pdb=" O ARG H 38 " (cutoff:3.500A) removed outlier: 5.905A pdb=" N ARG H 38 " --> pdb=" O TRP H 47 " (cutoff:3.500A) removed outlier: 3.823A pdb=" N GLY H 49 " --> pdb=" O TRP H 36 " (cutoff:3.500A) removed outlier: 5.175A pdb=" N TYR H 33 " --> pdb=" O GLY H 99 " (cutoff:3.500A) removed outlier: 3.519A pdb=" N TYR H 94 " --> pdb=" O THR H 121 " (cutoff:3.500A) removed outlier: 3.588A pdb=" N THR H 121 " --> pdb=" O TYR H 94 " (cutoff:3.500A) Processing sheet with id=AD1, first strand: chain 'I' and resid 239 through 242 removed outlier: 3.595A pdb=" N ALA I 316 " --> pdb=" O VAL I 337 " (cutoff:3.500A) Processing sheet with id=AD2, first strand: chain 'I' and resid 239 through 242 removed outlier: 3.595A pdb=" N ALA I 316 " --> pdb=" O VAL I 337 " (cutoff:3.500A) Processing sheet with id=AD3, first strand: chain 'I' and resid 248 through 250 Processing sheet with id=AD4, first strand: chain 'K' and resid 496 through 501 removed outlier: 5.415A pdb=" N VAL L 610 " --> pdb=" O VAL K 36 " (cutoff:3.500A) removed outlier: 8.063A pdb=" N VAL K 38 " --> pdb=" O THR L 608 " (cutoff:3.500A) removed outlier: 7.701A pdb=" N THR L 608 " --> pdb=" O VAL K 38 " (cutoff:3.500A) removed outlier: 8.037A pdb=" N TYR K 40 " --> pdb=" O CYS L 606 " (cutoff:3.500A) removed outlier: 8.377A pdb=" N CYS L 606 " --> pdb=" O TYR K 40 " (cutoff:3.500A) Processing sheet with id=AD5, first strand: chain 'K' and resid 45 through 47 removed outlier: 3.629A pdb=" N ILE K 227 " --> pdb=" O VAL K 247 " (cutoff:3.500A) Processing sheet with id=AD6, first strand: chain 'K' and resid 92 through 93 removed outlier: 3.878A pdb=" N PHE K 93 " --> pdb=" O GLY K 239 " (cutoff:3.500A) removed outlier: 4.076A pdb=" N GLY K 239 " --> pdb=" O PHE K 93 " (cutoff:3.500A) No H-bonds generated for sheet with id=AD6 Processing sheet with id=AD7, first strand: chain 'K' and resid 171 through 179 removed outlier: 3.697A pdb=" N ASN K 158 " --> pdb=" O SER K 132 " (cutoff:3.500A) Processing sheet with id=AD8, first strand: chain 'K' and resid 261 through 263 removed outlier: 3.972A pdb=" N GLY K 453 " --> pdb=" O LEU K 262 " (cutoff:3.500A) removed outlier: 10.281A pdb=" N THR K 446 " --> pdb=" O ILE K 296 " (cutoff:3.500A) removed outlier: 11.690A pdb=" N ILE K 296 " --> pdb=" O THR K 446 " (cutoff:3.500A) removed outlier: 10.192A pdb=" N LYS K 448 " --> pdb=" O VAL K 294 " (cutoff:3.500A) removed outlier: 11.025A pdb=" N VAL K 294 " --> pdb=" O LYS K 448 " (cutoff:3.500A) removed outlier: 9.777A pdb=" N ASN K 450 " --> pdb=" O THR K 292 " (cutoff:3.500A) removed outlier: 10.317A pdb=" N THR K 292 " --> pdb=" O ASN K 450 " (cutoff:3.500A) removed outlier: 7.286A pdb=" N THR K 452 " --> pdb=" O LEU K 290 " (cutoff:3.500A) removed outlier: 4.475A pdb=" N PHE K 384 " --> pdb=" O LYS K 423 " (cutoff:3.500A) Processing sheet with id=AD9, first strand: chain 'K' and resid 273 through 275 removed outlier: 7.286A pdb=" N THR K 452 " --> pdb=" O LEU K 290 " (cutoff:3.500A) removed outlier: 10.317A pdb=" N THR K 292 " --> pdb=" O ASN K 450 " (cutoff:3.500A) removed outlier: 9.777A pdb=" N ASN K 450 " --> pdb=" O THR K 292 " (cutoff:3.500A) removed outlier: 11.025A pdb=" N VAL K 294 " --> pdb=" O LYS K 448 " (cutoff:3.500A) removed outlier: 10.192A pdb=" N LYS K 448 " --> pdb=" O VAL K 294 " (cutoff:3.500A) removed outlier: 11.690A pdb=" N ILE K 296 " --> pdb=" O THR K 446 " (cutoff:3.500A) removed outlier: 10.281A pdb=" N THR K 446 " --> pdb=" O ILE K 296 " (cutoff:3.500A) removed outlier: 6.877A pdb=" N LYS K 362 " --> pdb=" O PHE K 470 " (cutoff:3.500A) removed outlier: 3.722A pdb=" N TYR K 363 " --> pdb=" O GLY K 412 " (cutoff:3.500A) removed outlier: 3.896A pdb=" N GLY K 412 " --> pdb=" O TYR K 363 " (cutoff:3.500A) Processing sheet with id=AE1, first strand: chain 'K' and resid 306 through 313 removed outlier: 4.085A pdb=" N GLN K 317 " --> pdb=" O ILE K 313 " (cutoff:3.500A) Processing sheet with id=AE2, first strand: chain 'K' and resid 425 through 426 Processing sheet with id=AE3, first strand: chain 'M' and resid 10 through 12 removed outlier: 4.198A pdb=" N THR M 124 " --> pdb=" O GLN M 10 " (cutoff:3.500A) removed outlier: 4.195A pdb=" N LYS M 12 " --> pdb=" O THR M 124 " (cutoff:3.500A) No H-bonds generated for sheet with id=AE3 Processing sheet with id=AE4, first strand: chain 'M' and resid 19 through 22 removed outlier: 3.521A pdb=" N CYS M 22 " --> pdb=" O ALA M 79 " (cutoff:3.500A) removed outlier: 3.597A pdb=" N ALA M 79 " --> pdb=" O CYS M 22 " (cutoff:3.500A) Processing sheet with id=AE5, first strand: chain 'M' and resid 46 through 51 removed outlier: 5.582A pdb=" N TRP M 47 " --> pdb=" O ARG M 38 " (cutoff:3.500A) removed outlier: 5.923A pdb=" N ARG M 38 " --> pdb=" O TRP M 47 " (cutoff:3.500A) removed outlier: 3.719A pdb=" N GLY M 49 " --> pdb=" O TRP M 36 " (cutoff:3.500A) removed outlier: 5.130A pdb=" N TYR M 33 " --> pdb=" O GLY M 99 " (cutoff:3.500A) Processing sheet with id=AE6, first strand: chain 'N' and resid 239 through 242 removed outlier: 3.782A pdb=" N THR N 338 " --> pdb=" O VAL N 240 " (cutoff:3.500A) removed outlier: 3.578A pdb=" N ALA N 316 " --> pdb=" O VAL N 337 " (cutoff:3.500A) Processing sheet with id=AE7, first strand: chain 'N' and resid 239 through 242 removed outlier: 3.782A pdb=" N THR N 338 " --> pdb=" O VAL N 240 " (cutoff:3.500A) removed outlier: 3.578A pdb=" N ALA N 316 " --> pdb=" O VAL N 337 " (cutoff:3.500A) Processing sheet with id=AE8, first strand: chain 'N' and resid 248 through 250 610 hydrogen bonds defined for protein. 1635 hydrogen bond angles defined for protein. Restraints generated for nucleic acids: 0 hydrogen bonds 0 hydrogen bond angles 0 basepair planarities 0 basepair parallelities 0 stacking parallelities Total time for adding SS restraints: 6.40 Time building geometry restraints manager: 8.48 seconds NOTE: a complete listing of the restraints can be obtained by requesting output of .geo file. Histogram of bond lengths: 1.22 - 1.36: 6147 1.36 - 1.50: 5381 1.50 - 1.65: 7623 1.65 - 1.79: 17 1.79 - 1.93: 177 Bond restraints: 19345 Sorted by residual: bond pdb=" CB CYS K 220 " pdb=" SG CYS K 220 " ideal model delta sigma weight residual 1.808 1.927 -0.119 3.30e-02 9.18e+02 1.30e+01 bond pdb=" CB CYS A 218 " pdb=" SG CYS A 218 " ideal model delta sigma weight residual 1.808 1.919 -0.111 3.30e-02 9.18e+02 1.14e+01 bond pdb=" N GLY F 521 " pdb=" CA GLY F 521 " ideal model delta sigma weight residual 1.451 1.492 -0.041 1.60e-02 3.91e+03 6.46e+00 bond pdb=" N GLY B 497 " pdb=" CA GLY B 497 " ideal model delta sigma weight residual 1.451 1.491 -0.040 1.60e-02 3.91e+03 6.15e+00 bond pdb=" C2 MAN R 5 " pdb=" O2 MAN R 5 " ideal model delta sigma weight residual 1.407 1.452 -0.045 2.00e-02 2.50e+03 5.05e+00 ... (remaining 19340 not shown) Histogram of bond angle deviations from ideal: 95.87 - 104.58: 307 104.58 - 113.28: 10753 113.28 - 121.99: 10730 121.99 - 130.70: 4351 130.70 - 139.40: 112 Bond angle restraints: 26253 Sorted by residual: angle pdb=" C VAL E 120 " pdb=" N LYS E 121 " pdb=" CA LYS E 121 " ideal model delta sigma weight residual 121.70 139.40 -17.70 1.80e+00 3.09e-01 9.67e+01 angle pdb=" C VAL K 120 " pdb=" N LYS K 121 " pdb=" CA LYS K 121 " ideal model delta sigma weight residual 121.70 138.38 -16.68 1.80e+00 3.09e-01 8.59e+01 angle pdb=" C VAL A 120 " pdb=" N LYS A 121 " pdb=" CA LYS A 121 " ideal model delta sigma weight residual 121.70 138.32 -16.62 1.80e+00 3.09e-01 8.53e+01 angle pdb=" CA CYS K 249 " pdb=" CB CYS K 249 " pdb=" SG CYS K 249 " ideal model delta sigma weight residual 114.40 127.41 -13.01 2.30e+00 1.89e-01 3.20e+01 angle pdb=" CA CYS A 247 " pdb=" CB CYS A 247 " pdb=" SG CYS A 247 " ideal model delta sigma weight residual 114.40 126.79 -12.39 2.30e+00 1.89e-01 2.90e+01 ... (remaining 26248 not shown) Histogram of dihedral angle deviations from ideal: 0.00 - 17.99: 10474 17.99 - 35.98: 725 35.98 - 53.97: 87 53.97 - 71.96: 32 71.96 - 89.95: 9 Dihedral angle restraints: 11327 sinusoidal: 4528 harmonic: 6799 Sorted by residual: dihedral pdb=" CB CYS K 220 " pdb=" SG CYS K 220 " pdb=" SG CYS K 249 " pdb=" CB CYS K 249 " ideal model delta sinusoidal sigma weight residual 93.00 25.25 67.75 1 1.00e+01 1.00e-02 5.97e+01 dihedral pdb=" CB CYS M 22 " pdb=" SG CYS M 22 " pdb=" SG CYS M 96 " pdb=" CB CYS M 96 " ideal model delta sinusoidal sigma weight residual 93.00 32.38 60.62 1 1.00e+01 1.00e-02 4.89e+01 dihedral pdb=" CB CYS A 218 " pdb=" SG CYS A 218 " pdb=" SG CYS A 247 " pdb=" CB CYS A 247 " ideal model delta sinusoidal sigma weight residual 93.00 34.18 58.82 1 1.00e+01 1.00e-02 4.63e+01 ... (remaining 11324 not shown) Histogram of chiral volume deviations from ideal: 0.000 - 0.057: 2411 0.057 - 0.114: 449 0.114 - 0.171: 153 0.171 - 0.228: 9 0.228 - 0.285: 6 Chirality restraints: 3028 Sorted by residual: chirality pdb=" C5 BMA U 3 " pdb=" C4 BMA U 3 " pdb=" C6 BMA U 3 " pdb=" O5 BMA U 3 " both_signs ideal model delta sigma weight residual False -2.27 -2.56 0.28 2.00e-01 2.50e+01 2.03e+00 chirality pdb=" C5 BMA Q 3 " pdb=" C4 BMA Q 3 " pdb=" C6 BMA Q 3 " pdb=" O5 BMA Q 3 " both_signs ideal model delta sigma weight residual False -2.27 -2.56 0.28 2.00e-01 2.50e+01 1.98e+00 chirality pdb=" C5 BMA G 3 " pdb=" C4 BMA G 3 " pdb=" C6 BMA G 3 " pdb=" O5 BMA G 3 " both_signs ideal model delta sigma weight residual False -2.27 -2.56 0.28 2.00e-01 2.50e+01 1.98e+00 ... (remaining 3025 not shown) Planarity restraints: 3301 Sorted by residual: delta sigma weight rms_deltas residual plane pdb=" CD ARG E 456 " 0.286 9.50e-02 1.11e+02 1.28e-01 1.09e+01 pdb=" NE ARG E 456 " -0.022 2.00e-02 2.50e+03 pdb=" CZ ARG E 456 " 0.013 2.00e-02 2.50e+03 pdb=" NH1 ARG E 456 " -0.008 2.00e-02 2.50e+03 pdb=" NH2 ARG E 456 " 0.005 2.00e-02 2.50e+03 delta sigma weight rms_deltas residual plane pdb=" CB HIS A 287 " 0.009 2.00e-02 2.50e+03 2.16e-02 7.00e+00 pdb=" CG HIS A 287 " -0.040 2.00e-02 2.50e+03 pdb=" ND1 HIS A 287 " 0.025 2.00e-02 2.50e+03 pdb=" CD2 HIS A 287 " 0.016 2.00e-02 2.50e+03 pdb=" CE1 HIS A 287 " -0.013 2.00e-02 2.50e+03 pdb=" NE2 HIS A 287 " 0.003 2.00e-02 2.50e+03 delta sigma weight rms_deltas residual plane pdb=" C SER D 532 " 0.045 5.00e-02 4.00e+02 6.53e-02 6.82e+00 pdb=" N PRO D 533 " -0.113 5.00e-02 4.00e+02 pdb=" CA PRO D 533 " 0.031 5.00e-02 4.00e+02 pdb=" CD PRO D 533 " 0.037 5.00e-02 4.00e+02 ... (remaining 3298 not shown) Histogram of nonbonded interaction distances: 1.98 - 2.57: 163 2.57 - 3.15: 15648 3.15 - 3.73: 26560 3.73 - 4.32: 37553 4.32 - 4.90: 63650 Nonbonded interactions: 143574 Sorted by model distance: nonbonded pdb=" O PRO N 237 " pdb=" OG1 THR N 335 " model vdw 1.985 2.440 nonbonded pdb=" O MET E 369 " pdb=" OG1 THR E 373 " model vdw 2.024 2.440 nonbonded pdb=" O MET A 369 " pdb=" OG1 THR A 373 " model vdw 2.025 2.440 nonbonded pdb=" O MET K 371 " pdb=" OG1 THR K 375 " model vdw 2.041 2.440 nonbonded pdb=" OG SER N 299 " pdb=" O THR N 302 " model vdw 2.064 2.440 ... (remaining 143569 not shown) NOTE: a complete listing of the restraints can be obtained by requesting output of .geo file. Find NCS groups from input model Found NCS groups: ncs_group { reference = chain 'A' selection = chain 'E' selection = (chain 'K' and (resid 31 through 152 or resid 155 through 522 or resid 601 throu \ gh 602)) } ncs_group { reference = (chain 'B' and resid 497 through 638) selection = (chain 'F' and resid 521 through 662) selection = chain 'L' } ncs_group { reference = chain 'C' selection = chain 'H' selection = chain 'M' } ncs_group { reference = chain 'D' selection = chain 'I' selection = chain 'N' } ncs_group { reference = chain 'G' selection = chain 'Q' selection = chain 'U' } ncs_group { reference = chain 'J' selection = chain 'R' selection = chain 'V' } Set up NCS constraints No NCS constraints will be used in refinement. Set refine NCS operators Adjust number of macro_cycles Number of macro_cycles: 10 Reset NCS operators Extract rigid body selections Check and reset occupancies Occupancies: min=1.00 max=1.00 mean=1.00 Load rotamer database and sin/cos tables Set ADP refinement strategy ADPs will be refined as group one per residue Make a string to write initial .geo file Internal consistency checks Time: Set random seed: 0.000 Set model cs if undefined: 0.000 Decide on map wrapping: 0.000 Normalize map: mean=0, sd=1: 0.740 Set stop_for_unknowns flag: 0.000 Assert model is a single copy model: 0.000 Assert all atoms have isotropic ADPs: 0.000 Construct map_model_manager: 0.010 Extract box with map and model: 3.490 Check model and map are aligned: 0.290 Set scattering table: 0.160 Process input model: 48.670 Find NCS groups from input model: 1.330 Set up NCS constraints: 0.190 Set refine NCS operators: 0.000 Adjust number of macro_cycles: 0.000 Reset NCS operators: 0.000 Extract rigid body selections: 0.000 Check and reset occupancies: 0.000 Load rotamer database and sin/cos tables:3.150 Set ADP refinement strategy: 0.000 Make a string to write initial .geo file:0.000 Internal consistency checks: 0.000 Total: 58.030 ------------------------------------------------------------------------------- Set refinement monitor ********************** ------------------------------------------------------------------------------- Setup refinement engine *********************** ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7494 moved from start: 0.0000 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd. rmsZ for bonds and angles. Bond : 0.005 0.119 19345 Z= 0.356 Angle : 0.885 17.703 26253 Z= 0.448 Chirality : 0.053 0.285 3028 Planarity : 0.006 0.128 3289 Dihedral : 12.508 89.947 6850 Min Nonbonded Distance : 1.985 Molprobity Statistics. All-atom Clashscore : 5.03 Ramachandran Plot: Outliers : 0.22 % Allowed : 7.75 % Favored : 92.03 % Rotamer Outliers : 0.05 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.13 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: -1.78 (0.18), residues: 2310 helix: -0.60 (0.27), residues: 381 sheet: -1.16 (0.19), residues: 666 loop : -1.33 (0.18), residues: 1263 *********************** REFINEMENT MACRO_CYCLE 1 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 393 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 1 poor density : 392 time to evaluate : 2.374 Fit side-chains revert: symmetry clash revert: symmetry clash revert: symmetry clash revert: symmetry clash revert: symmetry clash outliers start: 1 outliers final: 0 residues processed: 392 average time/residue: 0.3318 time to fit residues: 195.9125 Evaluate side-chains 286 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 0 poor density : 286 time to evaluate : 2.203 Switching outliers to nearest non-outliers outliers start: 0 outliers final: 0 residues processed: 0 time to fit residues: 2.9953 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Rotamers are restrained with sigma=5.00 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 228 random chunks: chunk 192 optimal weight: 1.9990 chunk 172 optimal weight: 8.9990 chunk 95 optimal weight: 9.9990 chunk 59 optimal weight: 8.9990 chunk 116 optimal weight: 0.9990 chunk 92 optimal weight: 0.3980 chunk 178 optimal weight: 8.9990 chunk 69 optimal weight: 10.0000 chunk 108 optimal weight: 1.9990 chunk 133 optimal weight: 7.9990 chunk 207 optimal weight: 9.9990 overall best weight: 2.6788 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... Flipped N/Q/H residues before XYZ refinement: A 249 HIS ** B 634 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** C 39 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** C 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** E 377 ASN H 6 GLN I 301 ASN K 72 HIS M 6 GLN ** M 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** Total number of N/Q/H flips: 6 ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7554 moved from start: 0.1396 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd. rmsZ for bonds and angles. Bond : 0.004 0.053 19345 Z= 0.249 Angle : 0.714 32.550 26253 Z= 0.352 Chirality : 0.047 0.294 3028 Planarity : 0.004 0.049 3289 Dihedral : 6.053 88.398 2537 Min Nonbonded Distance : 2.070 Molprobity Statistics. All-atom Clashscore : 7.42 Ramachandran Plot: Outliers : 0.17 % Allowed : 6.67 % Favored : 93.16 % Rotamer Outliers : 1.42 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.13 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: -1.45 (0.18), residues: 2310 helix: -0.03 (0.27), residues: 380 sheet: -0.98 (0.19), residues: 711 loop : -1.19 (0.19), residues: 1219 *********************** REFINEMENT MACRO_CYCLE 2 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 320 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 29 poor density : 291 time to evaluate : 2.105 Fit side-chains revert: symmetry clash revert: symmetry clash revert: symmetry clash outliers start: 29 outliers final: 18 residues processed: 302 average time/residue: 0.2907 time to fit residues: 137.0303 Evaluate side-chains 276 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 18 poor density : 258 time to evaluate : 2.266 Switching outliers to nearest non-outliers outliers start: 18 outliers final: 0 residues processed: 18 average time/residue: 0.1691 time to fit residues: 8.6587 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Rotamers are restrained with sigma=4.50 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 228 random chunks: chunk 115 optimal weight: 9.9990 chunk 64 optimal weight: 8.9990 chunk 172 optimal weight: 8.9990 chunk 140 optimal weight: 10.0000 chunk 57 optimal weight: 7.9990 chunk 207 optimal weight: 5.9990 chunk 224 optimal weight: 30.0000 chunk 184 optimal weight: 1.9990 chunk 205 optimal weight: 10.0000 chunk 70 optimal weight: 9.9990 chunk 166 optimal weight: 10.0000 overall best weight: 6.7990 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... Flipped N/Q/H residues before XYZ refinement: B 583 ASN ** C 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** E 66 HIS E 114 GLN E 377 ASN H 10 GLN ** H 46 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** H 54 GLN K 480 ASN M 10 GLN ** M 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** N 236 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** Total number of N/Q/H flips: 8 ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7710 moved from start: 0.2132 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd. rmsZ for bonds and angles. Bond : 0.008 0.065 19345 Z= 0.499 Angle : 0.868 36.948 26253 Z= 0.426 Chirality : 0.051 0.217 3028 Planarity : 0.006 0.057 3289 Dihedral : 6.708 88.842 2537 Min Nonbonded Distance : 2.031 Molprobity Statistics. All-atom Clashscore : 11.83 Ramachandran Plot: Outliers : 0.17 % Allowed : 9.31 % Favored : 90.52 % Rotamer Outliers : 2.73 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.13 % Twisted Proline : 0.00 % Twisted General : 0.04 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: -1.81 (0.18), residues: 2310 helix: -0.45 (0.26), residues: 395 sheet: -1.07 (0.22), residues: 599 loop : -1.50 (0.17), residues: 1316 *********************** REFINEMENT MACRO_CYCLE 3 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 342 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 56 poor density : 286 time to evaluate : 2.215 Fit side-chains revert: symmetry clash revert: symmetry clash outliers start: 56 outliers final: 41 residues processed: 324 average time/residue: 0.2916 time to fit residues: 147.2087 Evaluate side-chains 310 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 41 poor density : 269 time to evaluate : 2.048 Switching outliers to nearest non-outliers outliers start: 41 outliers final: 0 residues processed: 41 average time/residue: 0.1669 time to fit residues: 15.7921 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Rotamers are restrained with sigma=4.00 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 228 random chunks: chunk 204 optimal weight: 4.9990 chunk 155 optimal weight: 3.9990 chunk 107 optimal weight: 0.9980 chunk 22 optimal weight: 10.0000 chunk 98 optimal weight: 5.9990 chunk 139 optimal weight: 3.9990 chunk 208 optimal weight: 5.9990 chunk 220 optimal weight: 8.9990 chunk 108 optimal weight: 1.9990 chunk 197 optimal weight: 9.9990 chunk 59 optimal weight: 3.9990 overall best weight: 2.9988 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... Flipped N/Q/H residues before XYZ refinement: A 66 HIS C 3 GLN ** C 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** D 535 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** E 377 ASN H 3 GLN H 54 GLN ** M 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** M 46 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** Total number of N/Q/H flips: 5 ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7638 moved from start: 0.2240 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd. rmsZ for bonds and angles. Bond : 0.004 0.044 19345 Z= 0.262 Angle : 0.736 41.040 26253 Z= 0.348 Chirality : 0.046 0.217 3028 Planarity : 0.004 0.055 3289 Dihedral : 6.168 88.904 2537 Min Nonbonded Distance : 2.075 Molprobity Statistics. All-atom Clashscore : 9.99 Ramachandran Plot: Outliers : 0.13 % Allowed : 7.27 % Favored : 92.60 % Rotamer Outliers : 1.71 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.13 % Twisted Proline : 0.00 % Twisted General : 0.09 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: -1.53 (0.18), residues: 2310 helix: -0.03 (0.27), residues: 393 sheet: -0.86 (0.21), residues: 630 loop : -1.40 (0.18), residues: 1287 *********************** REFINEMENT MACRO_CYCLE 4 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 329 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 35 poor density : 294 time to evaluate : 2.257 Fit side-chains revert: symmetry clash revert: symmetry clash outliers start: 35 outliers final: 12 residues processed: 314 average time/residue: 0.3204 time to fit residues: 157.0330 Evaluate side-chains 275 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 12 poor density : 263 time to evaluate : 2.398 Switching outliers to nearest non-outliers outliers start: 12 outliers final: 0 residues processed: 12 average time/residue: 0.1857 time to fit residues: 7.2775 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Rotamers are restrained with sigma=3.50 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 228 random chunks: chunk 183 optimal weight: 7.9990 chunk 125 optimal weight: 2.9990 chunk 3 optimal weight: 7.9990 chunk 164 optimal weight: 8.9990 chunk 90 optimal weight: 10.0000 chunk 188 optimal weight: 0.0370 chunk 152 optimal weight: 10.0000 chunk 0 optimal weight: 10.0000 chunk 112 optimal weight: 3.9990 chunk 197 optimal weight: 9.9990 chunk 55 optimal weight: 0.8980 overall best weight: 3.1864 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... Flipped N/Q/H residues before XYZ refinement: A 114 GLN A 287 HIS ** C 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** D 535 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** E 377 ASN H 3 GLN H 10 GLN H 54 GLN M 39 GLN ** M 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** M 46 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** N 270 GLN Total number of N/Q/H flips: 8 ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7639 moved from start: 0.2415 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd. rmsZ for bonds and angles. Bond : 0.004 0.048 19345 Z= 0.267 Angle : 0.734 39.513 26253 Z= 0.345 Chirality : 0.046 0.235 3028 Planarity : 0.004 0.053 3289 Dihedral : 6.058 89.034 2537 Min Nonbonded Distance : 2.080 Molprobity Statistics. All-atom Clashscore : 10.23 Ramachandran Plot: Outliers : 0.13 % Allowed : 8.18 % Favored : 91.69 % Rotamer Outliers : 2.15 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.13 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: -1.52 (0.18), residues: 2310 helix: 0.09 (0.27), residues: 392 sheet: -0.81 (0.21), residues: 646 loop : -1.47 (0.18), residues: 1272 *********************** REFINEMENT MACRO_CYCLE 5 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 333 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 44 poor density : 289 time to evaluate : 2.268 Fit side-chains revert: symmetry clash revert: symmetry clash outliers start: 44 outliers final: 23 residues processed: 312 average time/residue: 0.3120 time to fit residues: 149.9211 Evaluate side-chains 289 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 23 poor density : 266 time to evaluate : 2.213 Switching outliers to nearest non-outliers outliers start: 23 outliers final: 0 residues processed: 23 average time/residue: 0.1731 time to fit residues: 10.4582 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Rotamers are restrained with sigma=3.00 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 228 random chunks: chunk 74 optimal weight: 10.0000 chunk 198 optimal weight: 4.9990 chunk 43 optimal weight: 10.0000 chunk 129 optimal weight: 5.9990 chunk 54 optimal weight: 1.9990 chunk 220 optimal weight: 7.9990 chunk 183 optimal weight: 5.9990 chunk 102 optimal weight: 10.0000 chunk 18 optimal weight: 4.9990 chunk 72 optimal weight: 30.0000 chunk 115 optimal weight: 10.0000 overall best weight: 4.7990 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... Flipped N/Q/H residues before XYZ refinement: ** A 478 ASN ** both conformations clash, **PLEASE CHECK MANUALLY** ** C 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** C 119 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** D 535 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** E 114 GLN E 377 ASN H 54 GLN M 39 GLN ** M 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** M 46 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** N 270 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** Total number of N/Q/H flips: 4 ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7682 moved from start: 0.2643 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd. rmsZ for bonds and angles. Bond : 0.006 0.047 19345 Z= 0.361 Angle : 0.776 36.477 26253 Z= 0.368 Chirality : 0.047 0.202 3028 Planarity : 0.005 0.057 3289 Dihedral : 6.238 89.039 2537 Min Nonbonded Distance : 2.057 Molprobity Statistics. All-atom Clashscore : 11.94 Ramachandran Plot: Outliers : 0.13 % Allowed : 9.09 % Favored : 90.78 % Rotamer Outliers : 2.15 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.13 % Twisted Proline : 0.00 % Twisted General : 0.09 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: -1.66 (0.18), residues: 2310 helix: -0.03 (0.27), residues: 392 sheet: -0.92 (0.21), residues: 607 loop : -1.53 (0.18), residues: 1311 *********************** REFINEMENT MACRO_CYCLE 6 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 333 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 44 poor density : 289 time to evaluate : 2.229 Fit side-chains revert: symmetry clash revert: symmetry clash revert: symmetry clash outliers start: 44 outliers final: 24 residues processed: 315 average time/residue: 0.3091 time to fit residues: 150.8866 Evaluate side-chains 295 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 24 poor density : 271 time to evaluate : 2.197 Switching outliers to nearest non-outliers outliers start: 24 outliers final: 0 residues processed: 24 average time/residue: 0.1705 time to fit residues: 10.9019 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Rotamers are restrained with sigma=2.50 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 228 random chunks: chunk 212 optimal weight: 9.9990 chunk 24 optimal weight: 0.9990 chunk 125 optimal weight: 0.9980 chunk 161 optimal weight: 7.9990 chunk 124 optimal weight: 0.6980 chunk 185 optimal weight: 10.0000 chunk 123 optimal weight: 2.9990 chunk 219 optimal weight: 20.0000 chunk 137 optimal weight: 0.9990 chunk 133 optimal weight: 6.9990 chunk 101 optimal weight: 7.9990 overall best weight: 1.3386 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... Flipped N/Q/H residues before XYZ refinement: ** A 478 ASN ** both conformations clash, **PLEASE CHECK MANUALLY** B 583 ASN ** C 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** C 119 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** D 535 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** E 377 ASN H 3 GLN ** I 270 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** K 332 HIS M 39 GLN ** M 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** M 46 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** N 246 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** N 270 GLN Total number of N/Q/H flips: 6 ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7594 moved from start: 0.2702 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd. rmsZ for bonds and angles. Bond : 0.003 0.053 19345 Z= 0.184 Angle : 0.724 42.168 26253 Z= 0.336 Chirality : 0.045 0.201 3028 Planarity : 0.004 0.053 3289 Dihedral : 5.828 89.116 2537 Min Nonbonded Distance : 2.103 Molprobity Statistics. All-atom Clashscore : 9.93 Ramachandran Plot: Outliers : 0.13 % Allowed : 6.45 % Favored : 93.42 % Rotamer Outliers : 1.17 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.13 % Twisted Proline : 0.00 % Twisted General : 0.00 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: -1.37 (0.18), residues: 2310 helix: 0.32 (0.27), residues: 392 sheet: -0.73 (0.21), residues: 653 loop : -1.40 (0.18), residues: 1265 *********************** REFINEMENT MACRO_CYCLE 7 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 317 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 24 poor density : 293 time to evaluate : 2.199 Fit side-chains revert: symmetry clash revert: symmetry clash revert: symmetry clash revert: symmetry clash outliers start: 24 outliers final: 8 residues processed: 305 average time/residue: 0.3105 time to fit residues: 145.1556 Evaluate side-chains 272 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 8 poor density : 264 time to evaluate : 2.221 Switching outliers to nearest non-outliers outliers start: 8 outliers final: 0 residues processed: 8 average time/residue: 0.1779 time to fit residues: 5.7032 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Rotamers are restrained with sigma=2.00 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 228 random chunks: chunk 135 optimal weight: 0.9980 chunk 87 optimal weight: 9.9990 chunk 131 optimal weight: 10.0000 chunk 66 optimal weight: 20.0000 chunk 43 optimal weight: 20.0000 chunk 42 optimal weight: 7.9990 chunk 139 optimal weight: 10.0000 chunk 149 optimal weight: 20.0000 chunk 108 optimal weight: 4.9990 chunk 20 optimal weight: 8.9990 chunk 172 optimal weight: 9.9990 overall best weight: 6.5988 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... Flipped N/Q/H residues before XYZ refinement: A 478 ASN ** C 119 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** D 535 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** D 607 ASN ** both conformations clash, **PLEASE CHECK MANUALLY** E 377 ASN M 39 GLN ** M 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** M 46 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** N 246 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** N 270 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** Total number of N/Q/H flips: 3 ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7725 moved from start: 0.2986 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd. rmsZ for bonds and angles. Bond : 0.007 0.060 19345 Z= 0.474 Angle : 0.866 43.570 26253 Z= 0.414 Chirality : 0.050 0.362 3028 Planarity : 0.005 0.060 3289 Dihedral : 6.481 89.081 2537 Min Nonbonded Distance : 2.016 Molprobity Statistics. All-atom Clashscore : 14.59 Ramachandran Plot: Outliers : 0.17 % Allowed : 10.04 % Favored : 89.78 % Rotamer Outliers : 1.95 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.13 % Twisted Proline : 0.00 % Twisted General : 0.13 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: -1.80 (0.18), residues: 2310 helix: -0.10 (0.27), residues: 386 sheet: -0.99 (0.21), residues: 608 loop : -1.66 (0.17), residues: 1316 *********************** REFINEMENT MACRO_CYCLE 8 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 319 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 40 poor density : 279 time to evaluate : 2.169 Fit side-chains revert: symmetry clash revert: symmetry clash revert: symmetry clash outliers start: 40 outliers final: 20 residues processed: 306 average time/residue: 0.3088 time to fit residues: 147.2647 Evaluate side-chains 282 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 20 poor density : 262 time to evaluate : 2.184 Switching outliers to nearest non-outliers outliers start: 20 outliers final: 0 residues processed: 20 average time/residue: 0.2062 time to fit residues: 10.1576 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Rotamers are restrained with sigma=1.50 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 228 random chunks: chunk 199 optimal weight: 1.9990 chunk 210 optimal weight: 2.9990 chunk 192 optimal weight: 6.9990 chunk 204 optimal weight: 5.9990 chunk 123 optimal weight: 0.9990 chunk 89 optimal weight: 0.1980 chunk 160 optimal weight: 10.0000 chunk 62 optimal weight: 6.9990 chunk 185 optimal weight: 3.9990 chunk 193 optimal weight: 0.8980 chunk 134 optimal weight: 2.9990 overall best weight: 1.4186 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... Flipped N/Q/H residues before XYZ refinement: A 315 GLN ** C 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** D 535 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** E 330 HIS E 377 ASN H 3 GLN I 301 ASN L 632 GLN ** M 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** M 46 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** M 63 ASN ** N 246 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** Total number of N/Q/H flips: 7 ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7604 moved from start: 0.3010 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd. rmsZ for bonds and angles. Bond : 0.003 0.054 19345 Z= 0.192 Angle : 0.757 41.310 26253 Z= 0.346 Chirality : 0.045 0.290 3028 Planarity : 0.004 0.055 3289 Dihedral : 5.914 89.040 2537 Min Nonbonded Distance : 2.082 Molprobity Statistics. All-atom Clashscore : 10.68 Ramachandran Plot: Outliers : 0.13 % Allowed : 6.41 % Favored : 93.46 % Rotamer Outliers : 0.93 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.13 % Twisted Proline : 0.00 % Twisted General : 0.09 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: -1.43 (0.18), residues: 2310 helix: 0.27 (0.27), residues: 392 sheet: -0.51 (0.21), residues: 609 loop : -1.58 (0.18), residues: 1309 *********************** REFINEMENT MACRO_CYCLE 9 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 312 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 19 poor density : 293 time to evaluate : 2.400 Fit side-chains revert: symmetry clash revert: symmetry clash revert: symmetry clash revert: symmetry clash outliers start: 19 outliers final: 11 residues processed: 300 average time/residue: 0.3117 time to fit residues: 144.9575 Evaluate side-chains 281 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 11 poor density : 270 time to evaluate : 2.255 Switching outliers to nearest non-outliers outliers start: 11 outliers final: 0 residues processed: 11 average time/residue: 0.1839 time to fit residues: 6.9950 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Rotamers are restrained with sigma=1.00 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 228 random chunks: chunk 216 optimal weight: 9.9990 chunk 132 optimal weight: 3.9990 chunk 102 optimal weight: 2.9990 chunk 150 optimal weight: 0.9990 chunk 227 optimal weight: 5.9990 chunk 209 optimal weight: 5.9990 chunk 180 optimal weight: 3.9990 chunk 18 optimal weight: 10.0000 chunk 139 optimal weight: 6.9990 chunk 110 optimal weight: 4.9990 chunk 143 optimal weight: 7.9990 overall best weight: 3.3990 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... Flipped N/Q/H residues before XYZ refinement: ** C 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** C 119 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** D 535 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** E 92 ASN E 377 ASN ** M 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** M 46 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** Total number of N/Q/H flips: 2 ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7645 moved from start: 0.3075 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd. rmsZ for bonds and angles. Bond : 0.004 0.046 19345 Z= 0.282 Angle : 0.771 39.507 26253 Z= 0.358 Chirality : 0.046 0.268 3028 Planarity : 0.004 0.055 3289 Dihedral : 5.980 89.136 2537 Min Nonbonded Distance : 2.062 Molprobity Statistics. All-atom Clashscore : 12.37 Ramachandran Plot: Outliers : 0.13 % Allowed : 8.83 % Favored : 91.04 % Rotamer Outliers : 0.20 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.13 % Twisted Proline : 0.00 % Twisted General : 0.09 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: -1.51 (0.18), residues: 2310 helix: 0.16 (0.27), residues: 393 sheet: -0.59 (0.21), residues: 618 loop : -1.61 (0.18), residues: 1299 ********************** REFINEMENT MACRO_CYCLE 10 OF 10 ************************ ------------------------------------------------------------------------------- Update Rama plot phi/psi targets (oldfield only) ************************************************ 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Re-set Ramachandran plot restraints *********************************** favored: oldfield allowed: oldfield outlier: oldfield 4620 Ramachandran restraints generated. 2310 Oldfield, 0 Emsley, 2310 emsley8k and 0 Phi/Psi/2. ------------------------------------------------------------------------------- Optimize residue side-chains **************************** Evaluate side-chains 281 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 4 poor density : 277 time to evaluate : 2.201 Fit side-chains revert: symmetry clash revert: symmetry clash revert: symmetry clash revert: symmetry clash revert: symmetry clash outliers start: 4 outliers final: 3 residues processed: 278 average time/residue: 0.3127 time to fit residues: 134.3056 Evaluate side-chains 272 residues out of total 2048 non-(ALA, GLY, PRO) need fitting. rotamer outliers: 3 poor density : 269 time to evaluate : 2.248 Switching outliers to nearest non-outliers outliers start: 3 outliers final: 0 residues processed: 3 average time/residue: 0.1887 time to fit residues: 4.1950 ------------------------------------------------------------------------------- Set rotamer restraints ********************** Rotamers are restrained with sigma=1.00 ------------------------------------------------------------------------------- XYZ refinement ************** Weight determination summary: number of chunks: 228 random chunks: chunk 192 optimal weight: 9.9990 chunk 55 optimal weight: 0.9990 chunk 166 optimal weight: 6.9990 chunk 26 optimal weight: 10.0000 chunk 50 optimal weight: 3.9990 chunk 181 optimal weight: 5.9990 chunk 75 optimal weight: 0.6980 chunk 186 optimal weight: 3.9990 chunk 22 optimal weight: 0.4980 chunk 33 optimal weight: 0.2980 chunk 159 optimal weight: 2.9990 overall best weight: 1.0984 ------------------------------------------------------------------------------- NQH flips ********* Analyzing N/Q/H residues for possible flip corrections... Flipped N/Q/H residues before XYZ refinement: A 330 HIS ** C 43 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** ** D 535 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** E 92 ASN E 377 ASN ** H 3 GLN ** both conformations clash, **PLEASE CHECK MANUALLY** L 593 GLN Total number of N/Q/H flips: 4 ------------------------------------------------------------------------------- ADP refinement ************** |-group b-factor refinement (macro cycle = 0; iterations = 0)-----------------| | r_work = 0.3829 r_free = 0.3829 target = 0.141879 restraints weight = None | |-----------------------------------------------------------------------------| |-group b-factor refinement (macro cycle = 1; iterations = 41)----------------| | r_work = 0.3529 r_free = 0.3529 target = 0.119151 restraints weight = 33157.864| |-----------------------------------------------------------------------------| |-group b-factor refinement (macro cycle = 2; iterations = 40)----------------| | r_work = 0.3568 r_free = 0.3568 target = 0.121952 restraints weight = 22634.573| |-----------------------------------------------------------------------------| |-group b-factor refinement (macro cycle = 3; iterations = 48)----------------| | r_work = 0.3595 r_free = 0.3595 target = 0.123872 restraints weight = 17369.632| |-----------------------------------------------------------------------------| r_work (final): 0.3601 ------------------------------------------------------------------------------- Occupancy refinement ******************** ------------------------------------------------------------------------------- Overall statistics ****************** model-to-map fit, CC_mask: 0.7721 moved from start: 0.3213 Geometry Restraints Library: GeoStd + Monomer Library + CDL v1.2 Deviations from Ideal Values - rmsd. rmsZ for bonds and angles. Bond : 0.003 0.053 19345 Z= 0.175 Angle : 0.726 38.114 26253 Z= 0.333 Chirality : 0.044 0.187 3028 Planarity : 0.004 0.051 3289 Dihedral : 5.603 89.406 2537 Min Nonbonded Distance : 2.101 Molprobity Statistics. All-atom Clashscore : 10.84 Ramachandran Plot: Outliers : 0.13 % Allowed : 6.36 % Favored : 93.51 % Rotamer Outliers : 0.29 % Cbeta Deviations : 0.00 % Peptide Plane: Cis-proline : 0.00 % Cis-general : 0.13 % Twisted Proline : 0.00 % Twisted General : 0.04 % Rama-Z (Ramachandran plot Z-score): Interpretation: bad |Rama-Z| > 3; suspicious 2 < |Rama-Z| < 3; good |Rama-Z| < 2. Scores for whole/helix/sheet/loop are scaled independently; therefore, the values are not related in a simple manner. whole: -1.33 (0.18), residues: 2310 helix: 0.40 (0.27), residues: 394 sheet: -0.48 (0.20), residues: 639 loop : -1.54 (0.18), residues: 1277 =============================================================================== Job complete usr+sys time: 3442.59 seconds wall clock time: 64 minutes 3.29 seconds (3843.29 seconds total)