Berkeley Structural Genomics Center

Target info for 1325B

This target is gi number 15675166 from Streptococcus pyogenes.

This gene has the annotation "putative DNA-binding protein".

The target was selected as part of JMC's third set.

Available sections:


Experimental status of 1325B

Note: Dates prior to 10/24/01 may be inaccurate, due to conversion from a record-keeping system which tracked only the last experimental date, to a new system which keeps track of all dates. The user is the person who entered the data, not the person who did the experiment. This will also change in the future.

ExperimentResultUserExperiment Date
SelectedDonejmc2002-11-07
ClonedDoner_kim2003-02-11
Expression testedDoner_kim2003-02-11
Solubility testedSr_kim2003-02-11
PurifiedDonedewemmer2003-06-09
CrystallizedDonedhshin2003-09-12
Diffraction quality crystalsDonedhshin2003-11-12
Native diffraction dataDonedhshin2003-11-12
Phasing diffraction dataDonedhshin2003-11-12
Traceable mapDonedhshin2003-11-12
Crystal structureDonedhshin2003-12-02
NMR characterizationDonedewemmer2003-06-09
HSQCDonedewemmer2003-06-09
In PDB1S7Odhshin2005-01-19

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Potential MP Homologues of 1325B

MP genelog10(PSIBLAST E)% identical% coverage (of MP gene)
MP417 / MPN424-21.1527.3693.81

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1325B DNA sequence

ATGAACATTATGGAAATTGAAAAAACAAACAGGATGAATGCCCTTTTTGAATTTTATGCAGCTTTGCTAACAGATAAACA
AATGAATTATATTGAACTGTATTATGCCGATGATTATAGCTTAGCTGAGATTGCTGATGAATTTGGTGTCAGTCGTCAGG
CTGTCTATGATAATATTAAGCGTACGGAAAAAATTTTAGAGACTTATGAGATGAAACTTCATATGTATTCGGATTATGTT
GTTCGAAGTGAAATCTTTGACGACATGATTGCTCACTACCCTCATGATGAGTATTTGCAAGAAAAAATTTCCATTCTAAC
CAGTATTGACAATAGAGAGTAA

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1325B Codon Usage

(114 codons)
fields: [triplet] [frequency: per thousand] ([number])

UUU35.1(   4)  UCU0.0(   0)  UAU87.7(  10)  UGU0.0(   0)  
UUC0.0(   0)  UCC8.8(   1)  UAC8.8(   1)  UGC0.0(   0)  
UUA17.5(   2)  UCA0.0(   0)  UAA8.8(   1)  UGA0.0(   0)  
UUG17.5(   2)  UCG8.8(   1)  UAG0.0(   0)  UGG0.0(   0)  

CUU17.5(   2)  CCU8.8(   1)  CAU17.5(   2)  CGU17.5(   2)  
CUC0.0(   0)  CCC0.0(   0)  CAC8.8(   1)  CGC0.0(   0)  
CUA17.5(   2)  CCA0.0(   0)  CAA17.5(   2)  CGA8.8(   1)  
CUG8.8(   1)  CCG0.0(   0)  CAG8.8(   1)  CGG0.0(   0)  

AUU87.7(  10)  ACU8.8(   1)  AAU35.1(   4)  AGU26.3(   3)  
AUC8.8(   1)  ACC8.8(   1)  AAC17.5(   2)  AGC8.8(   1)  
AUA0.0(   0)  ACA17.5(   2)  AAA43.9(   5)  AGA8.8(   1)  
AUG61.4(   7)  ACG8.8(   1)  AAG8.8(   1)  AGG8.8(   1)  

GUU17.5(   2)  GCU43.9(   5)  GAU61.4(   7)  GGU8.8(   1)  
GUC17.5(   2)  GCC17.5(   2)  GAC26.3(   3)  GGC0.0(   0)  
GUA0.0(   0)  GCA8.8(   1)  GAA70.2(   8)  GGA0.0(   0)  
GUG0.0(   0)  GCG0.0(   0)  GAG43.9(   5)  GGG0.0(   0)  

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1325B Potential Cloning Problems

No UGA codons.
1 restriction site(s) found.

Details (problem DNA region is in upper case, with the key on the line below):

Key:

atgaacattatggaaattgaaaaaacaaacaggatgaatgccctttttgaattttatgca
                                                            
gctttgctaacagataaacaaatgaattatattgaactgtattatgccgatgattatagc
                                                            
ttagctgagattgctgatgaatttggtgtcagtcgtcaggctgtctatgataatattaag
                                                            
cgtacggaaaaaattttagagacttatgagatgaaacttCATATGtattcggattatgtt
                                       1                    
gttcgaagtgaaatctttgacgacatgattgctcactaccctcatgatgagtatttgcaa
                                                            
gaaaaaatttccattctaaccagtattgacaatagagagtaa
                                          

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1325B Automatically Selected Primers

Note: These were generated using Hisao's algorithm, and do not take into account possible mispriming, self-complementarity, or requirements for mutations anywhere in the sequences. The -R1 primers are for the N-terminal end, and begin with an NdeI restriction site (CATATG). The -R2 primers are for the C-terminal end, and begin with a BamHI site (GGATCC) if the target has no other BamHI sites. If the target does have a BamHI site, the -R2 primer wil begin with a BglII site (AGATCT), unless the target has no BglII sites. If the target has both sites, the design program gives up and doesn't prepend a restriction site to the -R2 primer!

Predicted Tm was obtained using the Tm determination tool at the Virtual Genome Center.

Primer IDPredicted Tmsequence
1325B-R160.80CATATGAACATTATGGAAATTGAAAAAACA
1325B-R262.30GGATCCTTACTCTCTATTGTCAATACTGGTTAGAATGG
1325B-R1063.30GGCGGTGGTGGCGGCATGAACATTATGGAAATTGAAAAAACAAAC
1325B-R1163.50GTTCTTCTCCTTTGCGCCCCTACTCTCTATTGTCAATACTGGTTAGAATGGAA

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1325B Translation

Note: This was generated using the standard codon usage table; UGA codons in MP/MP genes will show up as terminators ("*")

atgaacattatggaaattgaaaaaacaaacaggatgaatgccctttttgaattttatgca
 M  N  I  M  E  I  E  K  T  N  R  M  N  A  L  F  E  F  Y  A 
gctttgctaacagataaacaaatgaattatattgaactgtattatgccgatgattatagc
 A  L  L  T  D  K  Q  M  N  Y  I  E  L  Y  Y  A  D  D  Y  S 
ttagctgagattgctgatgaatttggtgtcagtcgtcaggctgtctatgataatattaag
 L  A  E  I  A  D  E  F  G  V  S  R  Q  A  V  Y  D  N  I  K 
cgtacggaaaaaattttagagacttatgagatgaaacttcatatgtattcggattatgtt
 R  T  E  K  I  L  E  T  Y  E  M  K  L  H  M  Y  S  D  Y  V 
gttcgaagtgaaatctttgacgacatgattgctcactaccctcatgatgagtatttgcaa
 V  R  S  E  I  F  D  D  M  I  A  H  Y  P  H  D  E  Y  L  Q 
gaaaaaatttccattctaaccagtattgacaatagagagtaa
 E  K  I  S  I  L  T  S  I  D  N  R  E  * 

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1325B AA Sequence

MNIMEIEKTNRMNALFEFYAALLTDKQMNYIELYYADDYSLAEIADEFGVSRQAVYDNIKRTEKILETYEMKLHMYSDYV
VRSEIFDDMIAHYPHDEYLQEKISILTSIDNRE

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1325B Protein Parameters

This information was obtained using the Protein Parameters tool on the ExPASy Molecular Biology Server.

Number of amino acids: 113

Molecular weight: 13590.3

Theoretical pI: 4.52

Amino acid composition:

Ala (A)   8	  7.1%
Arg (R)   5	  4.4%
Asn (N)   6	  5.3%
Asp (D)  10	  8.8%
Cys (C)   0	  0.0%
Gln (Q)   3	  2.7%
Glu (E)  13	 11.5%
Gly (G)   1	  0.9%
His (H)   3	  2.7%
Ile (I)  11	  9.7%
Leu (L)   9	  8.0%
Lys (K)   6	  5.3%
Met (M)   7	  6.2%
Phe (F)   4	  3.5%
Pro (P)   1	  0.9%
Ser (S)   6	  5.3%
Thr (T)   5	  4.4%
Trp (W)   0	  0.0%
Tyr (Y)  11	  9.7%
Val (V)   4	  3.5%

Asx (B)   0	  0.0%
Glx (Z)   0	  0.0%
Xaa (X)   0	  0.0%

Total number of negatively charged residues (Asp + Glu): 23
Total number of positively charged residues (Arg + Lys): 11

Atomic composition:

Carbon      C	       607
Hydrogen    H	       925
Nitrogen    N	       149
Oxygen      O	       191
Sulfur      S	         7

Formula: C607H925N149O191S7
Total number of atoms: 1879

Extinction coefficients:

Conditions: 6.0 M guanidium hydrochloride
            0.02 M phosphate buffer
            pH 6.5

Extinction coefficients are in units of  M-1 cm-1 .

                      276     278     279     280     282
                       nm      nm      nm      nm      nm
Ext. coefficient    15950   15400   14795   14080   13200
Abs 0.1% (=1 g/l)   1.174   1.133   1.089   1.036   0.971


Estimated half-life:

The N-terminal of the sequence considered is M (Met).

The estimated half-life is: 30 hours (mammalian reticulocytes, in vitro).
                            >20 hours (yeast, in vivo).
                            >10 hours (Escherichia coli, in vivo).


Instability index:

The instability index (II) is computed to be 64.02
This classifies the protein as unstable.



Aliphatic index: 86.37

Grand average of hydropathicity (GRAVY): -0.466

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