longtext: 7oo4-pdb

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HEADER    FLUORESCENT PROTEIN                     26-MAY-21   7OO4
TITLE     HALOTAG ENGINEERING FOR ENHANCED FLUOROGENICITY AND KINETICS WITH A
TITLE    2 STYRYLPYRIDINE DYE
COMPND    MOL_ID: 1;
COMPND   2 MOLECULE: HALOALKANE DEHALOGENASE;
COMPND   3 CHAIN: A;
COMPND   4 EC: 3.8.1.5;
COMPND   5 ENGINEERED: YES
SOURCE    MOL_ID: 1;
SOURCE   2 ORGANISM_SCIENTIFIC: RHODOCOCCUS SP.;
SOURCE   3 ORGANISM_TAXID: 1831;
SOURCE   4 GENE: DHAA;
SOURCE   5 EXPRESSION_SYSTEM: ESCHERICHIA COLI BL21(DE3);
SOURCE   6 EXPRESSION_SYSTEM_TAXID: 469008;
SOURCE   7 EXPRESSION_SYSTEM_PLASMID: PET30
KEYWDS    HALOTAG, FLUORESCENCE, IMAGING, MAMMALIAN CELLS, FLUORESCENT
KEYWDS   2 REPORTER, CHANNEL DYE, DIRECTED EVOLUTION, FLUORESCENT PROTEIN
EXPDTA    X-RAY DIFFRACTION
AUTHOR    A.STEIN,A.D.LIANG
REVDAT   1   21-JUL-21 7OO4    0
JRNL        AUTH   C.MIRO-VINYALS,A.STEIN,S.FISCHER,T.R.WARD,A.D.LIANG
JRNL        TITL   HALOTAG ENGINEERING FOR ENHANCED FLUOROGENICITY AND KINETICS
JRNL        TITL 2 WITH A STYRYLPYRIDINE DYE
JRNL        REF    TO BE PUBLISHED
JRNL        REFN
REMARK   2
REMARK   2 RESOLUTION.    2.10 ANGSTROMS.
REMARK   3
REMARK   3 REFINEMENT.
REMARK   3   PROGRAM     : REFMAC 5.8.0267
REMARK   3   AUTHORS     : MURSHUDOV,SKUBAK,LEBEDEV,PANNU,STEINER,
REMARK   3               : NICHOLLS,WINN,LONG,VAGIN
REMARK   3
REMARK   3    REFINEMENT TARGET : MAXIMUM LIKELIHOOD
REMARK   3
REMARK   3  DATA USED IN REFINEMENT.
REMARK   3   RESOLUTION RANGE HIGH (ANGSTROMS) : 2.10
REMARK   3   RESOLUTION RANGE LOW  (ANGSTROMS) : 43.05
REMARK   3   DATA CUTOFF            (SIGMA(F)) : 0.000
REMARK   3   COMPLETENESS FOR RANGE        (%) : 99.9
REMARK   3   NUMBER OF REFLECTIONS             : 17847
REMARK   3
REMARK   3  FIT TO DATA USED IN REFINEMENT.
REMARK   3   CROSS-VALIDATION METHOD          : THROUGHOUT
REMARK   3   FREE R VALUE TEST SET SELECTION  : RANDOM
REMARK   3   R VALUE     (WORKING + TEST SET) : 0.212
REMARK   3   R VALUE            (WORKING SET) : 0.210
REMARK   3   FREE R VALUE                     : 0.263
REMARK   3   FREE R VALUE TEST SET SIZE   (%) : 4.900
REMARK   3   FREE R VALUE TEST SET COUNT      : 914
REMARK   3
REMARK   3  FIT IN THE HIGHEST RESOLUTION BIN.
REMARK   3   TOTAL NUMBER OF BINS USED           : 20
REMARK   3   BIN RESOLUTION RANGE HIGH       (A) : 2.10
REMARK   3   BIN RESOLUTION RANGE LOW        (A) : 2.16
REMARK   3   REFLECTION IN BIN     (WORKING SET) : 1304
REMARK   3   BIN COMPLETENESS (WORKING+TEST) (%) : 100.0
REMARK   3   BIN R VALUE           (WORKING SET) : 0.3530
REMARK   3   BIN FREE R VALUE SET COUNT          : 69
REMARK   3   BIN FREE R VALUE                    : 0.3610
REMARK   3
REMARK   3  NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.
REMARK   3   PROTEIN ATOMS            : 2346
REMARK   3   NUCLEIC ACID ATOMS       : 0
REMARK   3   HETEROGEN ATOMS          : 25
REMARK   3   SOLVENT ATOMS            : 37
REMARK   3
REMARK   3  B VALUES.
REMARK   3   FROM WILSON PLOT           (A**2) : NULL
REMARK   3   MEAN B VALUE      (OVERALL, A**2) : 37.94
REMARK   3   OVERALL ANISOTROPIC B VALUE.
REMARK   3    B11 (A**2) : -0.21000
REMARK   3    B22 (A**2) : -4.49000
REMARK   3    B33 (A**2) : 4.71000
REMARK   3    B12 (A**2) : 0.00000
REMARK   3    B13 (A**2) : 0.00000
REMARK   3    B23 (A**2) : 0.00000
REMARK   3
REMARK   3  ESTIMATED OVERALL COORDINATE ERROR.
REMARK   3   ESU BASED ON R VALUE                            (A): 0.242
REMARK   3   ESU BASED ON FREE R VALUE                       (A): 0.207
REMARK   3   ESU BASED ON MAXIMUM LIKELIHOOD                 (A): 0.202
REMARK   3   ESU FOR B VALUES BASED ON MAXIMUM LIKELIHOOD (A**2): 8.329
REMARK   3
REMARK   3 CORRELATION COEFFICIENTS.
REMARK   3   CORRELATION COEFFICIENT FO-FC      : 0.954
REMARK   3   CORRELATION COEFFICIENT FO-FC FREE : 0.920
REMARK   3
REMARK   3  RMS DEVIATIONS FROM IDEAL VALUES        COUNT    RMS    WEIGHT
REMARK   3   BOND LENGTHS REFINED ATOMS        (A):  2451 ; 0.010 ; 0.013
REMARK   3   BOND LENGTHS OTHERS               (A):  2247 ; 0.001 ; 0.017
REMARK   3   BOND ANGLES REFINED ATOMS   (DEGREES):  3346 ; 1.673 ; 1.659
REMARK   3   BOND ANGLES OTHERS          (DEGREES):  5187 ; 1.309 ; 1.570
REMARK   3   TORSION ANGLES, PERIOD 1    (DEGREES):   291 ; 7.273 ; 5.000
REMARK   3   TORSION ANGLES, PERIOD 2    (DEGREES):   132 ;29.084 ;21.667
REMARK   3   TORSION ANGLES, PERIOD 3    (DEGREES):   369 ;16.292 ;15.000
REMARK   3   TORSION ANGLES, PERIOD 4    (DEGREES):    17 ;19.213 ;15.000
REMARK   3   CHIRAL-CENTER RESTRAINTS       (A**3):   300 ; 0.082 ; 0.200
REMARK   3   GENERAL PLANES REFINED ATOMS      (A):  2748 ; 0.009 ; 0.020
REMARK   3   GENERAL PLANES OTHERS             (A):   558 ; 0.001 ; 0.020
REMARK   3   NON-BONDED CONTACTS REFINED ATOMS (A):  NULL ;  NULL ;  NULL
REMARK   3   NON-BONDED CONTACTS OTHERS        (A):  NULL ;  NULL ;  NULL
REMARK   3   NON-BONDED TORSION REFINED ATOMS  (A):  NULL ;  NULL ;  NULL
REMARK   3   NON-BONDED TORSION OTHERS         (A):  NULL ;  NULL ;  NULL
REMARK   3   H-BOND (X...Y) REFINED ATOMS      (A):  NULL ;  NULL ;  NULL
REMARK   3   H-BOND (X...Y) OTHERS             (A):  NULL ;  NULL ;  NULL
REMARK   3   POTENTIAL METAL-ION REFINED ATOMS (A):  NULL ;  NULL ;  NULL
REMARK   3   POTENTIAL METAL-ION OTHERS        (A):  NULL ;  NULL ;  NULL
REMARK   3   SYMMETRY VDW REFINED ATOMS        (A):  NULL ;  NULL ;  NULL
REMARK   3   SYMMETRY VDW OTHERS               (A):  NULL ;  NULL ;  NULL
REMARK   3   SYMMETRY H-BOND REFINED ATOMS     (A):  NULL ;  NULL ;  NULL
REMARK   3   SYMMETRY H-BOND OTHERS            (A):  NULL ;  NULL ;  NULL
REMARK   3   SYMMETRY METAL-ION REFINED ATOMS  (A):  NULL ;  NULL ;  NULL
REMARK   3   SYMMETRY METAL-ION OTHERS         (A):  NULL ;  NULL ;  NULL
REMARK   3
REMARK   3  ISOTROPIC THERMAL FACTOR RESTRAINTS.     COUNT   RMS    WEIGHT
REMARK   3   MAIN-CHAIN BOND REFINED ATOMS  (A**2):  NULL ;  NULL ;  NULL
REMARK   3   MAIN-CHAIN BOND OTHER ATOMS    (A**2):  NULL ;  NULL ;  NULL
REMARK   3   MAIN-CHAIN ANGLE REFINED ATOMS (A**2):  NULL ;  NULL ;  NULL
REMARK   3   MAIN-CHAIN ANGLE OTHER ATOMS   (A**2):  NULL ;  NULL ;  NULL
REMARK   3   SIDE-CHAIN BOND REFINED ATOMS  (A**2):  NULL ;  NULL ;  NULL
REMARK   3   SIDE-CHAIN BOND OTHER ATOMS    (A**2):  NULL ;  NULL ;  NULL
REMARK   3   SIDE-CHAIN ANGLE REFINED ATOMS (A**2):  NULL ;  NULL ;  NULL
REMARK   3   SIDE-CHAIN ANGLE OTHER ATOMS   (A**2):  NULL ;  NULL ;  NULL
REMARK   3   LONG RANGE B REFINED ATOMS     (A**2):  NULL ;  NULL ;  NULL
REMARK   3   LONG RANGE B OTHER ATOMS       (A**2):  NULL ;  NULL ;  NULL
REMARK   3
REMARK   3 ANISOTROPIC THERMAL FACTOR RESTRAINTS.    COUNT   RMS   WEIGHT
REMARK   3   RIGID-BOND RESTRAINTS          (A**2):  NULL ;  NULL ;  NULL
REMARK   3   SPHERICITY; FREE ATOMS         (A**2):  NULL ;  NULL ;  NULL
REMARK   3   SPHERICITY; BONDED ATOMS       (A**2):  NULL ;  NULL ;  NULL
REMARK   3
REMARK   3  NCS RESTRAINTS STATISTICS
REMARK   3   NUMBER OF DIFFERENT NCS GROUPS : NULL
REMARK   3
REMARK   3  TLS DETAILS
REMARK   3   NUMBER OF TLS GROUPS  : NULL
REMARK   3
REMARK   3  BULK SOLVENT MODELLING.
REMARK   3   METHOD USED : MASK
REMARK   3   PARAMETERS FOR MASK CALCULATION
REMARK   3   VDW PROBE RADIUS   : 1.20
REMARK   3   ION PROBE RADIUS   : 0.80
REMARK   3   SHRINKAGE RADIUS   : 0.80
REMARK   3
REMARK   3  OTHER REFINEMENT REMARKS: HYDROGENS HAVE BEEN ADDED IN THE RIDING
REMARK   3  POSITIONS U VALUES : REFINED INDIVIDUALLY
REMARK   4
REMARK   4 7OO4 COMPLIES WITH FORMAT V. 3.30, 13-JUL-11
REMARK 100
REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBE ON 27-MAY-21.
REMARK 100 THE DEPOSITION ID IS D_1292116118.
REMARK 200
REMARK 200 EXPERIMENTAL DETAILS
REMARK 200  EXPERIMENT TYPE                : X-RAY DIFFRACTION
REMARK 200  DATE OF DATA COLLECTION        : 20-MAR-21
REMARK 200  TEMPERATURE           (KELVIN) : 100
REMARK 200  PH                             : 5.6
REMARK 200  NUMBER OF CRYSTALS USED        : 1
REMARK 200
REMARK 200  SYNCHROTRON              (Y/N) : Y
REMARK 200  RADIATION SOURCE               : SLS
REMARK 200  BEAMLINE                       : X06DA
REMARK 200  X-RAY GENERATOR MODEL          : NULL
REMARK 200  MONOCHROMATIC OR LAUE    (M/L) : M
REMARK 200  WAVELENGTH OR RANGE        (A) : 1.000031
REMARK 200  MONOCHROMATOR                  : NULL
REMARK 200  OPTICS                         : NULL
REMARK 200
REMARK 200  DETECTOR TYPE                  : PIXEL
REMARK 200  DETECTOR MANUFACTURER          : DECTRIS PILATUS 2M-F
REMARK 200  INTENSITY-INTEGRATION SOFTWARE : XDS
REMARK 200  DATA SCALING SOFTWARE          : AIMLESS
REMARK 200
REMARK 200  NUMBER OF UNIQUE REFLECTIONS   : 18774
REMARK 200  RESOLUTION RANGE HIGH      (A) : 2.100
REMARK 200  RESOLUTION RANGE LOW       (A) : 43.050
REMARK 200  REJECTION CRITERIA  (SIGMA(I)) : NULL
REMARK 200
REMARK 200 OVERALL.
REMARK 200  COMPLETENESS FOR RANGE     (%) : 99.9
REMARK 200  DATA REDUNDANCY                : 12.60
REMARK 200  R MERGE                    (I) : 0.12130
REMARK 200  R SYM                      (I) : NULL
REMARK 200   FOR THE DATA SET  : 15.5000
REMARK 200
REMARK 200 IN THE HIGHEST RESOLUTION SHELL.
REMARK 200  HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 2.10
REMARK 200  HIGHEST RESOLUTION SHELL, RANGE LOW  (A) : 2.18
REMARK 200  COMPLETENESS FOR SHELL     (%) : NULL
REMARK 200  DATA REDUNDANCY IN SHELL       : NULL
REMARK 200  R MERGE FOR SHELL          (I) : 0.95830
REMARK 200  R SYM FOR SHELL            (I) : NULL
REMARK 200   FOR SHELL         : 2.750
REMARK 200
REMARK 200 DIFFRACTION PROTOCOL: SINGLE WAVELENGTH
REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MOLECULAR REPLACEMENT
REMARK 200 SOFTWARE USED: PHASER
REMARK 200 STARTING MODEL: 5Y2X
REMARK 200
REMARK 200 REMARK: NULL
REMARK 280
REMARK 280 CRYSTAL
REMARK 280 SOLVENT CONTENT, VS   (%): 45.78
REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.27
REMARK 280
REMARK 280 CRYSTALLIZATION CONDITIONS: 0.1 M NA3 CIT 5.6 PH 20 %V/V 2-PROPOH
REMARK 280  20 %W/V PEG 4K, PH 5.6, VAPOR DIFFUSION, SITTING DROP,
REMARK 280  TEMPERATURE 293K
REMARK 290
REMARK 290 CRYSTALLOGRAPHIC SYMMETRY
REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 21 21 21
REMARK 290
REMARK 290      SYMOP   SYMMETRY
REMARK 290     NNNMMM   OPERATOR
REMARK 290       1555   X,Y,Z
REMARK 290       2555   -X+1/2,-Y,Z+1/2
REMARK 290       3555   -X,Y+1/2,-Z+1/2
REMARK 290       4555   X+1/2,-Y+1/2,-Z
REMARK 290
REMARK 290     WHERE NNN -> OPERATOR NUMBER
REMARK 290           MMM -> TRANSLATION VECTOR
REMARK 290
REMARK 290 CRYSTALLOGRAPHIC SYMMETRY TRANSFORMATIONS
REMARK 290 THE FOLLOWING TRANSFORMATIONS OPERATE ON THE ATOM/HETATM
REMARK 290 RECORDS IN THIS ENTRY TO PRODUCE CRYSTALLOGRAPHICALLY
REMARK 290 RELATED MOLECULES.
REMARK 290   SMTRY1   1  1.000000  0.000000  0.000000        0.00000
REMARK 290   SMTRY2   1  0.000000  1.000000  0.000000        0.00000
REMARK 290   SMTRY3   1  0.000000  0.000000  1.000000        0.00000
REMARK 290   SMTRY1   2 -1.000000  0.000000  0.000000       26.15950
REMARK 290   SMTRY2   2  0.000000 -1.000000  0.000000        0.00000
REMARK 290   SMTRY3   2  0.000000  0.000000  1.000000       43.01000
REMARK 290   SMTRY1   3 -1.000000  0.000000  0.000000        0.00000
REMARK 290   SMTRY2   3  0.000000  1.000000  0.000000       34.49300
REMARK 290   SMTRY3   3  0.000000  0.000000 -1.000000       43.01000
REMARK 290   SMTRY1   4  1.000000  0.000000  0.000000       26.15950
REMARK 290   SMTRY2   4  0.000000 -1.000000  0.000000       34.49300
REMARK 290   SMTRY3   4  0.000000  0.000000 -1.000000        0.00000
REMARK 290
REMARK 290 REMARK: NULL
REMARK 300
REMARK 300 BIOMOLECULE: 1
REMARK 300 SEE REMARK 350 FOR THE AUTHOR PROVIDED AND/OR PROGRAM
REMARK 300 GENERATED ASSEMBLY INFORMATION FOR THE STRUCTURE IN
REMARK 300 THIS ENTRY. THE REMARK MAY ALSO PROVIDE INFORMATION ON
REMARK 300 BURIED SURFACE AREA.
REMARK 350
REMARK 350 COORDINATES FOR A COMPLETE MULTIMER REPRESENTING THE KNOWN
REMARK 350 BIOLOGICALLY SIGNIFICANT OLIGOMERIZATION STATE OF THE
REMARK 350 MOLECULE CAN BE GENERATED BY APPLYING BIOMT TRANSFORMATIONS
REMARK 350 GIVEN BELOW.  BOTH NON-CRYSTALLOGRAPHIC AND
REMARK 350 CRYSTALLOGRAPHIC OPERATIONS ARE GIVEN.
REMARK 350
REMARK 350 BIOMOLECULE: 1
REMARK 350 AUTHOR DETERMINED BIOLOGICAL UNIT: MONOMERIC
REMARK 350 SOFTWARE DETERMINED QUATERNARY STRUCTURE: MONOMERIC
REMARK 350 SOFTWARE USED: PISA
REMARK 350 TOTAL BURIED SURFACE AREA: 130 ANGSTROM**2
REMARK 350 SURFACE AREA OF THE COMPLEX: 11660 ANGSTROM**2
REMARK 350 CHANGE IN SOLVENT FREE ENERGY: -13.0 KCAL/MOL
REMARK 350 APPLY THE FOLLOWING TO CHAINS: A
REMARK 350   BIOMT1   1  1.000000  0.000000  0.000000        0.00000
REMARK 350   BIOMT2   1  0.000000  1.000000  0.000000        0.00000
REMARK 350   BIOMT3   1  0.000000  0.000000  1.000000        0.00000
REMARK 465
REMARK 465 MISSING RESIDUES
REMARK 465 THE FOLLOWING RESIDUES WERE NOT LOCATED IN THE
REMARK 465 EXPERIMENT. (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN
REMARK 465 IDENTIFIER; SSSEQ=SEQUENCE NUMBER; I=INSERTION CODE.)
REMARK 465
REMARK 465   M RES C SSSEQI
REMARK 465     MET A     1
REMARK 465     ALA A     2
REMARK 465     HIS A   295
REMARK 465     HIS A   296
REMARK 465     HIS A   297
REMARK 465     HIS A   298
REMARK 465     HIS A   299
REMARK 465     HIS A   300
REMARK 500
REMARK 500 GEOMETRY AND STEREOCHEMISTRY
REMARK 500 SUBTOPIC: COVALENT BOND LENGTHS
REMARK 500
REMARK 500 THE STEREOCHEMICAL PARAMETERS OF THE FOLLOWING RESIDUES
REMARK 500 HAVE VALUES WHICH DEVIATE FROM EXPECTED VALUES BY MORE
REMARK 500 THAN 6*RMSD (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN
REMARK 500 IDENTIFIER; SSEQ=SEQUENCE NUMBER; I=INSERTION CODE).
REMARK 500
REMARK 500 STANDARD TABLE:
REMARK 500 FORMAT: (10X,I3,1X,2(A3,1X,A1,I4,A1,1X,A4,3X),1X,F6.3)
REMARK 500
REMARK 500 EXPECTED VALUES PROTEIN: ENGH AND HUBER, 1999
REMARK 500 EXPECTED VALUES NUCLEIC ACID: CLOWNEY ET AL 1996
REMARK 500
REMARK 500  M RES CSSEQI ATM1   RES CSSEQI ATM2   DEVIATION
REMARK 500    ASP A 106   CG    ASP A 106   OD1     0.152
REMARK 500
REMARK 500 REMARK: NULL
REMARK 500
REMARK 500 GEOMETRY AND STEREOCHEMISTRY
REMARK 500 SUBTOPIC: COVALENT BOND ANGLES
REMARK 500
REMARK 500 THE STEREOCHEMICAL PARAMETERS OF THE FOLLOWING RESIDUES
REMARK 500 HAVE VALUES WHICH DEVIATE FROM EXPECTED VALUES BY MORE
REMARK 500 THAN 6*RMSD (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN
REMARK 500 IDENTIFIER; SSEQ=SEQUENCE NUMBER; I=INSERTION CODE).
REMARK 500
REMARK 500 STANDARD TABLE:
REMARK 500 FORMAT: (10X,I3,1X,A3,1X,A1,I4,A1,3(1X,A4,2X),12X,F5.1)
REMARK 500
REMARK 500 EXPECTED VALUES PROTEIN: ENGH AND HUBER, 1999
REMARK 500 EXPECTED VALUES NUCLEIC ACID: CLOWNEY ET AL 1996
REMARK 500
REMARK 500  M RES CSSEQI ATM1   ATM2   ATM3
REMARK 500    ASP A 106   CB  -  CA  -  C   ANGL. DEV. =  12.0 DEGREES
REMARK 500
REMARK 500 REMARK: NULL
REMARK 500
REMARK 500 GEOMETRY AND STEREOCHEMISTRY
REMARK 500 SUBTOPIC: TORSION ANGLES
REMARK 500
REMARK 500 TORSION ANGLES OUTSIDE THE EXPECTED RAMACHANDRAN REGIONS:
REMARK 500 (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER;
REMARK 500 SSEQ=SEQUENCE NUMBER; I=INSERTION CODE).
REMARK 500
REMARK 500 STANDARD TABLE:
REMARK 500 FORMAT:(10X,I3,1X,A3,1X,A1,I4,A1,4X,F7.2,3X,F7.2)
REMARK 500
REMARK 500 EXPECTED VALUES: GJ KLEYWEGT AND TA JONES (1996). PHI/PSI-
REMARK 500 CHOLOGY: RAMACHANDRAN REVISITED. STRUCTURE 4, 1395 - 1400
REMARK 500
REMARK 500  M RES CSSEQI        PSI       PHI
REMARK 500    PRO A  42       50.71   -104.16
REMARK 500    ASP A  76       73.86   -100.48
REMARK 500    GLU A  98      -82.87    -92.49
REMARK 500    ASP A 106     -125.45     55.31
REMARK 500    ASN A 119       43.03   -142.76
REMARK 500    ARG A 153       48.59    -88.15
REMARK 500    ASP A 156      -66.84    -96.96
REMARK 500    VAL A 245      -69.81   -126.36
REMARK 500    LEU A 271     -105.67   -121.47
REMARK 500
REMARK 500 REMARK: NULL
DBREF  7OO4 A    3   293  UNP    P0A3G3   DHAA_RHOSO       3    293
SEQADV 7OO4 MET A    1  UNP  P0A3G3              INITIATING METHIONINE
SEQADV 7OO4 ALA A    2  UNP  P0A3G3              EXPRESSION TAG
SEQADV 7OO4 VAL A   47  UNP  P0A3G3    LEU    47 ENGINEERED MUTATION
SEQADV 7OO4 THR A   58  UNP  P0A3G3    SER    58 ENGINEERED MUTATION
SEQADV 7OO4 GLY A   78  UNP  P0A3G3    ASP    78 ENGINEERED MUTATION
SEQADV 7OO4 PHE A   87  UNP  P0A3G3    TYR    87 ENGINEERED MUTATION
SEQADV 7OO4 MET A   88  UNP  P0A3G3    LEU    88 ENGINEERED MUTATION
SEQADV 7OO4 PHE A  128  UNP  P0A3G3    CYS   128 ENGINEERED MUTATION
SEQADV 7OO4 THR A  155  UNP  P0A3G3    ALA   155 ENGINEERED MUTATION
SEQADV 7OO4 LYS A  160  UNP  P0A3G3    GLU   160 ENGINEERED MUTATION
SEQADV 7OO4 VAL A  167  UNP  P0A3G3    ALA   167 ENGINEERED MUTATION
SEQADV 7OO4 THR A  172  UNP  P0A3G3    ALA   172 ENGINEERED MUTATION
SEQADV 7OO4 MET A  175  UNP  P0A3G3    LYS   175 ENGINEERED MUTATION
SEQADV 7OO4 GLY A  176  UNP  P0A3G3    CYS   176 ENGINEERED MUTATION
SEQADV 7OO4 ASN A  195  UNP  P0A3G3    LYS   195 ENGINEERED MUTATION
SEQADV 7OO4 GLU A  224  UNP  P0A3G3    ALA   224 ENGINEERED MUTATION
SEQADV 7OO4 ASP A  227  UNP  P0A3G3    ASN   227 ENGINEERED MUTATION
SEQADV 7OO4 LYS A  257  UNP  P0A3G3    GLU   257 ENGINEERED MUTATION
SEQADV 7OO4 ALA A  264  UNP  P0A3G3    THR   264 ENGINEERED MUTATION
SEQADV 7OO4 ASN A  272  UNP  P0A3G3    HIS   272 ENGINEERED MUTATION
SEQADV 7OO4 LEU A  273  UNP  P0A3G3    TYR   273 ENGINEERED MUTATION
SEQADV 7OO4 SER A  291  UNP  P0A3G3    PRO   291 ENGINEERED MUTATION
SEQADV 7OO4 THR A  292  UNP  P0A3G3    ALA   292 ENGINEERED MUTATION
SEQADV 7OO4 GLU A  294  UNP  P0A3G3              EXPRESSION TAG
SEQADV 7OO4 HIS A  295  UNP  P0A3G3              EXPRESSION TAG
SEQADV 7OO4 HIS A  296  UNP  P0A3G3              EXPRESSION TAG
SEQADV 7OO4 HIS A  297  UNP  P0A3G3              EXPRESSION TAG
SEQADV 7OO4 HIS A  298  UNP  P0A3G3              EXPRESSION TAG
SEQADV 7OO4 HIS A  299  UNP  P0A3G3              EXPRESSION TAG
SEQADV 7OO4 HIS A  300  UNP  P0A3G3              EXPRESSION TAG
SEQRES   1 A  300  MET ALA GLU ILE GLY THR GLY PHE PRO PHE ASP PRO HIS
SEQRES   2 A  300  TYR VAL GLU VAL LEU GLY GLU ARG MET HIS TYR VAL ASP
SEQRES   3 A  300  VAL GLY PRO ARG ASP GLY THR PRO VAL LEU PHE LEU HIS
SEQRES   4 A  300  GLY ASN PRO THR SER SER TYR VAL TRP ARG ASN ILE ILE
SEQRES   5 A  300  PRO HIS VAL ALA PRO THR HIS ARG CYS ILE ALA PRO ASP
SEQRES   6 A  300  LEU ILE GLY MET GLY LYS SER ASP LYS PRO ASP LEU GLY
SEQRES   7 A  300  TYR PHE PHE ASP ASP HIS VAL ARG PHE MET ASP ALA PHE
SEQRES   8 A  300  ILE GLU ALA LEU GLY LEU GLU GLU VAL VAL LEU VAL ILE
SEQRES   9 A  300  HIS ASP TRP GLY SER ALA LEU GLY PHE HIS TRP ALA LYS
SEQRES  10 A  300  ARG ASN PRO GLU ARG VAL LYS GLY ILE ALA PHE MET GLU
SEQRES  11 A  300  PHE ILE ARG PRO ILE PRO THR TRP ASP GLU TRP PRO GLU
SEQRES  12 A  300  PHE ALA ARG GLU THR PHE GLN ALA PHE ARG THR THR ASP
SEQRES  13 A  300  VAL GLY ARG LYS LEU ILE ILE ASP GLN ASN VAL PHE ILE
SEQRES  14 A  300  GLU GLY THR LEU PRO MET GLY VAL VAL ARG PRO LEU THR
SEQRES  15 A  300  GLU VAL GLU MET ASP HIS TYR ARG GLU PRO PHE LEU ASN
SEQRES  16 A  300  PRO VAL ASP ARG GLU PRO LEU TRP ARG PHE PRO ASN GLU
SEQRES  17 A  300  LEU PRO ILE ALA GLY GLU PRO ALA ASN ILE VAL ALA LEU
SEQRES  18 A  300  VAL GLU GLU TYR MET ASP TRP LEU HIS GLN SER PRO VAL
SEQRES  19 A  300  PRO LYS LEU LEU PHE TRP GLY THR PRO GLY VAL LEU ILE
SEQRES  20 A  300  PRO PRO ALA GLU ALA ALA ARG LEU ALA LYS SER LEU PRO
SEQRES  21 A  300  ASN CYS LYS ALA VAL ASP ILE GLY PRO GLY LEU ASN LEU
SEQRES  22 A  300  LEU GLN GLU ASP ASN PRO ASP LEU ILE GLY SER GLU ILE
SEQRES  23 A  300  ALA ARG TRP LEU SER THR LEU GLU HIS HIS HIS HIS HIS
SEQRES  24 A  300  HIS
HET     CL  A 401       1
HET    VF2  A 402      24
HETNAM      CL CHLORIDE ION
HETNAM     VF2 4-[(E)-2-[1-(7-CHLORANYLHEPTYL)PYRIDIN-1-IUM-4-
HETNAM   2 VF2  YL]ETHENYL]-N,N-DIMETHYL-ANILINE
FORMUL   2   CL    CL 1-
FORMUL   3  VF2    C22 H30 CL N2 1+
FORMUL   4  HOH   *37(H2 O)
HELIX    1 AA1 SER A   44  ARG A   49  5                                   6
HELIX    2 AA2 ILE A   51  ALA A   56  1                                   6
HELIX    3 AA3 PHE A   80  LEU A   95  1                                  16
HELIX    4 AA4 ASP A  106  ASN A  119  1                                  14
HELIX    5 AA5 PRO A  142  PHE A  144  5                                   3
HELIX    6 AA6 ALA A  145  ARG A  153  1                                   9
HELIX    7 AA7 ASP A  156  ILE A  163  1                                   8
HELIX    8 AA8 ASN A  166  GLY A  171  1                                   6
HELIX    9 AA9 GLY A  171  GLY A  176  1                                   6
HELIX   10 AB1 THR A  182  GLU A  191  1                                  10
HELIX   11 AB2 PRO A  192  LEU A  194  5                                   3
HELIX   12 AB3 ASN A  195  ASP A  198  5                                   4
HELIX   13 AB4 ARG A  199  LEU A  209  1                                  11
HELIX   14 AB5 PRO A  215  SER A  232  1                                  18
HELIX   15 AB6 PRO A  248  LEU A  259  1                                  12
HELIX   16 AB7 LEU A  273  ASP A  277  5                                   5
HELIX   17 AB8 ASN A  278  THR A  292  1                                  15
SHEET    1 AA1 8 HIS A  13  VAL A  17  0
SHEET    2 AA1 8 GLU A  20  VAL A  27 -1  O  GLU A  20   N  VAL A  17
SHEET    3 AA1 8 CYS A  61  PRO A  64 -1  O  CYS A  61   N  VAL A  27
SHEET    4 AA1 8 VAL A  35  LEU A  38  1  N  VAL A  35   O  ILE A  62
SHEET    5 AA1 8 VAL A 100  HIS A 105  1  O  VAL A 101   N  LEU A  36
SHEET    6 AA1 8 VAL A 123  MET A 129  1  O  ALA A 127   N  LEU A 102
SHEET    7 AA1 8 LYS A 236  PRO A 243  1  O  LEU A 237   N  PHE A 128
SHEET    8 AA1 8 CYS A 262  GLY A 270  1  O  ILE A 267   N  TRP A 240
LINK         OD1 ASP A 106                 C20 VF2 A 402     1555   1555  1.44
CISPEP   1 ASN A   41    PRO A   42          0       -10.99
CISPEP   2 GLU A  214    PRO A  215          0         0.62
CISPEP   3 THR A  242    PRO A  243          0         5.03
CRYST1   52.319   68.986   86.020  90.00  90.00  90.00 P 21 21 21    4
ORIGX1      1.000000  0.000000  0.000000        0.00000
ORIGX2      0.000000  1.000000  0.000000        0.00000
ORIGX3      0.000000  0.000000  1.000000        0.00000
SCALE1      0.019114  0.000000  0.000000        0.00000
SCALE2      0.000000  0.014496  0.000000        0.00000
SCALE3      0.000000  0.000000  0.011625        0.00000
TER    2347      GLU A 294
MASTER      319    0    2   17    8    0    0    6 2408    1   25   24
END