Berkeley Structural Genomics Center

Target info for 1073B

This target is gi number 4981923 from Thermotoga maritima.

This gene has the annotation "peptide chain release factor RF-1".

The target was selected as part of Igor's sets.

Available sections:


Experimental status of 1073B

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
SelectedDonejmc2001-08-24
ClonedDoneabwaight2001-08-24
Expression testedDoneabwaight2001-08-24
Solubility testedSabwaight2001-08-24
PurifiedDoneabwaight2001-08-24
CrystallizedDoneabwaight2001-08-24
Diffraction quality crystalsDoneabwaight2001-08-24
Phasing diffraction dataDoneabwaight2001-08-24
Traceable mapDoneabwaight2001-08-24
Crystal structureDonedhshin2003-07-28
In PDB1RQ0dhshin2003-12-15
Biochemical functionDonedhshin2003-01-23

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Potential MP Homologues of 1073B

MP genelog10(PSIBLAST E)% identical% coverage (of MP gene)
MP475 / MPN361-113.0045.7897.08

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1073B DNA sequence

GTGAAAGAAAAGAAAAAAGAGATCGAAAAGCTCCTCGCCAGGCCAGATCTGACTCCCGAACAGATGAAGAACTACGGGAT
GGAATACGCAAAGATAGAAGAAATAGAGAACATCACAAACAGAATAAAAGAAACTCAAGAGTTCATCGAACTTTTGAGAG
AAGAAGGAGAAAATGAACTGGAGATAGAAAAGTACGAGAAAGAACTTGACCAGCTCTATCAAGAACTGCTCTTTCTTCTT
TCTCCAGAAGCGAGCGATAAAGCAATCGTAGAAATCAGACCAGGCACGGGTGGAGAAGAAGCCGCTCTCTTCGCGCGGGA
TCTGTTCAGAATGTACACCAGGTACGCAGAAAGAAAAGGCTGGAACCTCGAAGTCGCTGAAATCCACGAAACGGATCTCG
GTGGAATAAGGGAAGTGGTCTTCTTTGTGAAGGGAAAGAACGCTTACGGTATTTTGAAGTACGAAAGCGGTGTCCATAGA
GTCCAGAGAGTTCCTGTTACAGAGTCTGGAGGGAGAATCCATACGTCCACCGCTACCGTTGCCGTTCTTCCAGAGATTGA
AGAAAAGGATATAGAAATCAGGCCGGAAGACTTGAAGATAGAAACTTTCAGGGCATCGGGACACGGTGGACAGTACGTCA
ACAAGACCGAATCCGCTGTGAGGATCACACATCTTCCAACGGGAATAGTGGTCTCGTGTCAGAACGAAAGATCTCAGTAT
CAGAACAAACAAACCGCTCTGAGGATCCTCAGAGCAAGACTCTACCAGCTACAGAAGGAACAGAAAGAAAGGGAGATCTC
GCAGAAGAGAAAATCACAGATAGGAACTGGTGAGAGAAGCGAAAAAATCAGAACTTACAACTTCCCCCAGAACAGAGTTA
CGGATCATAGAATAAACTACACTTCTTATAGACTGCAGGAGATCCTCGATGGAGACCTGGACGAAATCATATCGAAGCTT
ATAGAACACGACATAGAAAACAACCTAGAGGAAGTTTTAGGAATTGGAGCCTCAGTCGAAGAAAAGTAA

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1073B Codon Usage

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

UUU5.8(   2)  UCU11.7(   4)  UAU8.7(   3)  UGU2.9(   1)  
UUC17.5(   6)  UCC5.8(   2)  UAC32.1(  11)  UGC0.0(   0)  
UUA2.9(   1)  UCA5.8(   2)  UAA2.9(   1)  UGA0.0(   0)  
UUG8.7(   3)  UCG11.7(   4)  UAG0.0(   0)  UGG2.9(   1)  

CUU20.4(   7)  CCU2.9(   1)  CAU11.7(   4)  CGU0.0(   0)  
CUC29.2(  10)  CCC5.8(   2)  CAC8.7(   3)  CGC0.0(   0)  
CUA5.8(   2)  CCA14.6(   5)  CAA8.7(   3)  CGA0.0(   0)  
CUG20.4(   7)  CCG2.9(   1)  CAG40.8(  14)  CGG2.9(   1)  

AUU8.7(   3)  ACU17.5(   6)  AAU2.9(   1)  AGU0.0(   0)  
AUC40.8(  14)  ACC14.6(   5)  AAC37.9(  13)  AGC8.7(   3)  
AUA37.9(  13)  ACA8.7(   3)  AAA32.1(  11)  AGA49.6(  17)  
AUG8.7(   3)  ACG14.6(   5)  AAG43.7(  15)  AGG23.3(   8)  

GUU17.5(   6)  GCU17.5(   6)  GAU20.4(   7)  GGU17.5(   6)  
GUC20.4(   7)  GCC11.7(   4)  GAC14.6(   5)  GGC5.8(   2)  
GUA2.9(   1)  GCA14.6(   5)  GAA119.5(  41)  GGA35.0(  12)  
GUG14.6(   5)  GCG5.8(   2)  GAG32.1(  11)  GGG5.8(   2)  

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1073B Potential Cloning Problems

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

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

Key:

gtgaaagaaaagaaaaaagagatcgaaaagctcctcgccaggccAGATCTgactcccgaa
                                            4               
cagatgaagaactacgggatggaatacgcaaagatagaagaaatagagaacatcacaaac
                                                            
agaataaaagaaactcaagagttcatcgaacttttgagagaagaaggagaaaatgaactg
                                                            
gagatagaaaagtacgagaaagaacttgaccagctctatcaagaactgctctttcttctt
                                                            
tctccagaagcgagcgataaagcaatcgtagaaatcagaccaggcacgggtggagaagaa
                                                            
gccgctctcttcgcgcgggatctgttcagaatgtacaccaggtacgcagaaagaaaaggc
                                                            
tggaacctcgaagtcgctgaaatccacgaaacggatctcggtggaataagggaagtggtc
                                                            
ttctttgtgaagggaaagaacgcttacggtattttgaagtacgaaagcggtgtccataga
                                                            
gtccagagagttcctgttacagagtctggagggagaatccatacgtccaccgctaccgtt
                                                            
gccgttcttccagagattgaagaaaaggatatagaaatcaggccggaagacttgaagata
                                                            
gaaactttcagggcatcgggacacggtggacagtacgtcaacaagaccgaatccgctgtg
                                                            
aggatcacacatcttccaacgggaatagtggtctcgtgtcagaacgaaAGATCTcagtat
                                                4           
cagaacaaacaaaccgctctgaGGATCCtcagagcaagactctaccagctacagaaggaa
                      2                                     
cagaaagaaagggAGATCTcgcagaagagaaaatcacagataggaactggtgagagaagc
             4                                              
gaaaaaatcagaacttacaacttcccccagaacagagttacggatcatagaataaactac
                                                            
acttcttatagactgcaggagatcctcgatggagacctggacgaaatcatatcgaagctt
                                                            
atagaacacgacatagaaaacaacctagaggaagttttaggaattggagcctcagtcgaa
                                                            
gaaaagtaa
         

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1073B 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
1073B-R160.90CATATGAAAGAAAAGAAAAAAGAGATCGAA
1073B-R262.20TTACTTTTCTTCGACTGAGGCTCC
1073B-R1063.40GGCGGTGGTGGCGGCATGAAAGAAAAGAAAAAAGAGATCGAAAAG
1073B-R1163.60GTTCTTCTCCTTTGCGCCCCTACTTTTCTTCGACTGAGGCTCCA

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1073B Translation

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

gtgaaagaaaagaaaaaagagatcgaaaagctcctcgccaggccagatctgactcccgaa
 V  K  E  K  K  K  E  I  E  K  L  L  A  R  P  D  L  T  P  E 
cagatgaagaactacgggatggaatacgcaaagatagaagaaatagagaacatcacaaac
 Q  M  K  N  Y  G  M  E  Y  A  K  I  E  E  I  E  N  I  T  N 
agaataaaagaaactcaagagttcatcgaacttttgagagaagaaggagaaaatgaactg
 R  I  K  E  T  Q  E  F  I  E  L  L  R  E  E  G  E  N  E  L 
gagatagaaaagtacgagaaagaacttgaccagctctatcaagaactgctctttcttctt
 E  I  E  K  Y  E  K  E  L  D  Q  L  Y  Q  E  L  L  F  L  L 
tctccagaagcgagcgataaagcaatcgtagaaatcagaccaggcacgggtggagaagaa
 S  P  E  A  S  D  K  A  I  V  E  I  R  P  G  T  G  G  E  E 
gccgctctcttcgcgcgggatctgttcagaatgtacaccaggtacgcagaaagaaaaggc
 A  A  L  F  A  R  D  L  F  R  M  Y  T  R  Y  A  E  R  K  G 
tggaacctcgaagtcgctgaaatccacgaaacggatctcggtggaataagggaagtggtc
 W  N  L  E  V  A  E  I  H  E  T  D  L  G  G  I  R  E  V  V 
ttctttgtgaagggaaagaacgcttacggtattttgaagtacgaaagcggtgtccataga
 F  F  V  K  G  K  N  A  Y  G  I  L  K  Y  E  S  G  V  H  R 
gtccagagagttcctgttacagagtctggagggagaatccatacgtccaccgctaccgtt
 V  Q  R  V  P  V  T  E  S  G  G  R  I  H  T  S  T  A  T  V 
gccgttcttccagagattgaagaaaaggatatagaaatcaggccggaagacttgaagata
 A  V  L  P  E  I  E  E  K  D  I  E  I  R  P  E  D  L  K  I 
gaaactttcagggcatcgggacacggtggacagtacgtcaacaagaccgaatccgctgtg
 E  T  F  R  A  S  G  H  G  G  Q  Y  V  N  K  T  E  S  A  V 
aggatcacacatcttccaacgggaatagtggtctcgtgtcagaacgaaagatctcagtat
 R  I  T  H  L  P  T  G  I  V  V  S  C  Q  N  E  R  S  Q  Y 
cagaacaaacaaaccgctctgaggatcctcagagcaagactctaccagctacagaaggaa
 Q  N  K  Q  T  A  L  R  I  L  R  A  R  L  Y  Q  L  Q  K  E 
cagaaagaaagggagatctcgcagaagagaaaatcacagataggaactggtgagagaagc
 Q  K  E  R  E  I  S  Q  K  R  K  S  Q  I  G  T  G  E  R  S 
gaaaaaatcagaacttacaacttcccccagaacagagttacggatcatagaataaactac
 E  K  I  R  T  Y  N  F  P  Q  N  R  V  T  D  H  R  I  N  Y 
acttcttatagactgcaggagatcctcgatggagacctggacgaaatcatatcgaagctt
 T  S  Y  R  L  Q  E  I  L  D  G  D  L  D  E  I  I  S  K  L 
atagaacacgacatagaaaacaacctagaggaagttttaggaattggagcctcagtcgaa
 I  E  H  D  I  E  N  N  L  E  E  V  L  G  I  G  A  S  V  E 
gaaaagtaa
 E  K  * 

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1073B AA Sequence

MKEKKKEIEKLLARPDLTPEQMKNYGMEYAKIEEIENITNRIKETQEFIELLREEGENELEIEKYEKELDQLYQELLFLL
SPEASDKAIVEIRPGTGGEEAALFARDLFRMYTRYAERKGWNLEVAEIHETDLGGIREVVFFVKGKNAYGILKYESGVHR
VQRVPVTESGGRIHTSTATVAVLPEIEEKDIEIRPEDLKIETFRASGHGGQYVNKTESAVRITHLPTGIVVSCQNERSQY
QNKQTALRILRARLYQLQKEQKEREISQKRKSQIGTGERSEKIRTYNFPQNRVTDHRINYTSYRLQEILDGDLDEIISKL
IEHDIENNLEEVLGIGASVEEK

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1073B Protein Parameters

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

Number of amino acids: 342

Molecular weight: 39656.8

Theoretical pI: 5.35

Amino acid composition:

Ala (A)  17	  5.0%
Arg (R)  26	  7.6%
Asn (N)  14	  4.1%
Asp (D)  12	  3.5%
Cys (C)   1	  0.3%
Gln (Q)  17	  5.0%
Glu (E)  52	 15.2%
Gly (G)  22	  6.4%
His (H)   7	  2.0%
Ile (I)  30	  8.8%
Leu (L)  30	  8.8%
Lys (K)  26	  7.6%
Met (M)   4	  1.2%
Phe (F)   8	  2.3%
Pro (P)   9	  2.6%
Ser (S)  15	  4.4%
Thr (T)  19	  5.6%
Trp (W)   1	  0.3%
Tyr (Y)  14	  4.1%
Val (V)  18	  5.3%

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

Total number of negatively charged residues (Asp + Glu): 64
Total number of positively charged residues (Arg + Lys): 52

Atomic composition:

Carbon      C	      1746
Hydrogen    H	      2812
Nitrogen    N	       492
Oxygen      O	       550
Sulfur      S	         5

Formula: C1746H2812N492O550S5
Total number of atoms: 5605

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 .

The first table lists values computed assuming ALL Cys 
residues appear as half cystines, whereas the second table 
assumes that NONE do. 

                      276     278     279     280     282
                       nm      nm      nm      nm      nm
Ext. coefficient    25700   25200   24490   23610   22400
Abs 0.1% (=1 g/l)   0.648   0.635   0.618   0.595   0.565



                      276     278     279     280     282
                       nm      nm      nm      nm      nm
Ext. coefficient    25700   25200   24490   23610   22400
Abs 0.1% (=1 g/l)   0.648   0.635   0.618   0.595   0.565


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 57.38
This classifies the protein as unstable.



Aliphatic index: 88.65

Grand average of hydropathicity (GRAVY): -0.740

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