Berkeley Structural Genomics Center

Target info for 1944B

This target is gi number 16128119 from Escherichia coli K12.

This gene has the annotation "putative carbonic anhdrase (EC 4.2.1.1)".

The target was selected as part of CompIII targets.

Available sections:


Experimental status of 1944B

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
SelectedDonejmc2004-04-26
ClonedDonedhshin2004-04-27
Expression testedDonedhshin2004-04-27
Solubility testedSdhshin2004-04-27
PurifiedDonedhshin2004-04-27
CrystallizedDonedhshin2004-04-27
Diffraction quality crystalsDonedhshin2004-04-27
Native diffraction dataDonedhshin2004-04-27
Phasing diffraction dataDonedhshin2004-04-27
Traceable mapDonedhshin2004-04-27
Crystal structureDonedhshin2004-04-27
In PDB1T75dhshin2004-05-10

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Potential MP Homologues of 1944B

No MP homologues identified for this gene.

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1944B DNA sequence

ATGAAAGACATAGATACACTCATCAGCAACAATGCACTATGGTCAAAAATGCTGGTGGAAGAGGATCCCGGGTTTTTTGA
GAAACTGGCACAAGCGCAAAAACCGCGCTTTCTATGGATTGGATGTTCCGACAGTCGCGTTCCTGCAGAACGTTTAACCG
GTCTTGAGCCGGGCGAACTCTTTGTTCACCGTAATGTTGCTAACCTGGTCATTCACACTGACCTGAACTGCCTTTCCGTG
GTTCAGTATGCAGTGGATGTACTCGAAGTTGAACACATTATTATCTGTGGCCACTACGGTTGCGGCGGCGTACAAGCCGC
AGTTGAAAACCCGGAACTGGGGCTTATCAACAACTGGCTGCTGCATATCCGCGATATCTGGTTCAAACATAGCTCATTGC
TCGGCGAAATGCCGCAAGAGCGCCGTCTGGATACCTTGTGTGAACTGAACGTCATGGAACAGGTGTATAACCTGGGCCAC
TCCACCATTATGCAATCAGCGTGGAAACGCGGGCAGAAAGTTACCATTCACGGCTGGGCCTACGGCATTCACGACGGCTT
GCTGCGTGATCTGGATGTTACCGCCACCAACCGCGAAACCCTTGAGCAACGTTACCGTCACGGGATTTCCAACCTCAAGC
TGAAACACGCCAACCACAAATAA

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1944B Codon Usage

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

UUU18.1(   4)  UCU0.0(   0)  UAU9.0(   2)  UGU13.6(   3)  
UUC4.5(   1)  UCC18.1(   4)  UAC13.6(   3)  UGC9.0(   2)  
UUA4.5(   1)  UCA13.6(   3)  UAA4.5(   1)  UGA0.0(   0)  
UUG13.6(   3)  UCG0.0(   0)  UAG0.0(   0)  UGG27.1(   6)  

CUU18.1(   4)  CCU4.5(   1)  CAU9.0(   2)  CGU27.1(   6)  
CUC22.6(   5)  CCC4.5(   1)  CAC45.2(  10)  CGC27.1(   6)  
CUA9.0(   2)  CCA0.0(   0)  CAA27.1(   6)  CGA0.0(   0)  
CUG58.8(  13)  CCG18.1(   4)  CAG13.6(   3)  CGG0.0(   0)  

AUU36.2(   8)  ACU4.5(   1)  AAU9.0(   2)  AGU4.5(   1)  
AUC22.6(   5)  ACC31.7(   7)  AAC49.8(  11)  AGC9.0(   2)  
AUA4.5(   1)  ACA4.5(   1)  AAA40.7(   9)  AGA0.0(   0)  
AUG22.6(   5)  ACG0.0(   0)  AAG4.5(   1)  AGG0.0(   0)  

GUU36.2(   8)  GCU4.5(   1)  GAU31.7(   7)  GGU9.0(   2)  
GUC9.0(   2)  GCC18.1(   4)  GAC18.1(   4)  GGC40.7(   9)  
GUA9.0(   2)  GCA22.6(   5)  GAA49.8(  11)  GGA4.5(   1)  
GUG18.1(   4)  GCG9.0(   2)  GAG22.6(   5)  GGG18.1(   4)  

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1944B 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:

atgaaagacatagatacactcatcagcaacaatgcactatggtcaaaaatgctggtggaa
                                                            
gaGGATCCcgggttttttgagaaactggcacaagcgcaaaaaccgcgctttctatggatt
  2                                                         
ggatgttccgacagtcgcgttcctgcagaacgtttaaccggtcttgagccgggcgaactc
                                                            
tttgttcaccgtaatgttgctaacctggtcattcacactgacctgaactgcctttccgtg
                                                            
gttcagtatgcagtggatgtactcgaagttgaacacattattatctgtggccactacggt
                                                            
tgcggcggcgtacaagccgcagttgaaaacccggaactggggcttatcaacaactggctg
                                                            
ctgcatatccgcgatatctggttcaaacatagctcattgctcggcgaaatgccgcaagag
                                                            
cgccgtctggataccttgtgtgaactgaacgtcatggaacaggtgtataacctgggccac
                                                            
tccaccattatgcaatcagcgtggaaacgcgggcagaaagttaccattcacggctgggcc
                                                            
tacggcattcacgacggcttgctgcgtgatctggatgttaccgccaccaaccgcgaaacc
                                                            
cttgagcaacgttaccgtcacgggatttccaacctcaagctgaaacacgccaaccacaaa
                                                            
taa
   

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1944B 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
1944B-R1063.00GGCGGTGGTGGCGGCATGATGAAAGACATAGATACACTCATCAGCAAC
1944B-R1164.40GTTCTTCTCCTTTGCGCCCCTATTATTTGTGGTTGGCGTGTTTCA

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1944B Translation

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

atgaaagacatagatacactcatcagcaacaatgcactatggtcaaaaatgctggtggaa
 M  K  D  I  D  T  L  I  S  N  N  A  L  W  S  K  M  L  V  E 
gaggatcccgggttttttgagaaactggcacaagcgcaaaaaccgcgctttctatggatt
 E  D  P  G  F  F  E  K  L  A  Q  A  Q  K  P  R  F  L  W  I 
ggatgttccgacagtcgcgttcctgcagaacgtttaaccggtcttgagccgggcgaactc
 G  C  S  D  S  R  V  P  A  E  R  L  T  G  L  E  P  G  E  L 
tttgttcaccgtaatgttgctaacctggtcattcacactgacctgaactgcctttccgtg
 F  V  H  R  N  V  A  N  L  V  I  H  T  D  L  N  C  L  S  V 
gttcagtatgcagtggatgtactcgaagttgaacacattattatctgtggccactacggt
 V  Q  Y  A  V  D  V  L  E  V  E  H  I  I  I  C  G  H  Y  G 
tgcggcggcgtacaagccgcagttgaaaacccggaactggggcttatcaacaactggctg
 C  G  G  V  Q  A  A  V  E  N  P  E  L  G  L  I  N  N  W  L 
ctgcatatccgcgatatctggttcaaacatagctcattgctcggcgaaatgccgcaagag
 L  H  I  R  D  I  W  F  K  H  S  S  L  L  G  E  M  P  Q  E 
cgccgtctggataccttgtgtgaactgaacgtcatggaacaggtgtataacctgggccac
 R  R  L  D  T  L  C  E  L  N  V  M  E  Q  V  Y  N  L  G  H 
tccaccattatgcaatcagcgtggaaacgcgggcagaaagttaccattcacggctgggcc
 S  T  I  M  Q  S  A  W  K  R  G  Q  K  V  T  I  H  G  W  A 
tacggcattcacgacggcttgctgcgtgatctggatgttaccgccaccaaccgcgaaacc
 Y  G  I  H  D  G  L  L  R  D  L  D  V  T  A  T  N  R  E  T 
cttgagcaacgttaccgtcacgggatttccaacctcaagctgaaacacgccaaccacaaa
 L  E  Q  R  Y  R  H  G  I  S  N  L  K  L  K  H  A  N  H  K 
taa
 * 

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1944B AA Sequence

MKDIDTLISNNALWSKMLVEEDPGFFEKLAQAQKPRFLWIGCSDSRVPAERLTGLEPGELFVHRNVANLVIHTDLNCLSV
VQYAVDVLEVEHIIICGHYGCGGVQAAVENPELGLINNWLLHIRDIWFKHSSLLGEMPQERRLDTLCELNVMEQVYNLGH
STIMQSAWKRGQKVTIHGWAYGIHDGLLRDLDVTATNRETLEQRYRHGISNLKLKHANHK

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1944B Protein Parameters

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

Number of amino acids: 220

Molecular weight: 25096.7

Theoretical pI: 6.16

Amino acid composition:

Ala (A)  12	  5.5%
Arg (R)  12	  5.5%
Asn (N)  13	  5.9%
Asp (D)  11	  5.0%
Cys (C)   5	  2.3%
Gln (Q)   9	  4.1%
Glu (E)  16	  7.3%
Gly (G)  16	  7.3%
His (H)  12	  5.5%
Ile (I)  14	  6.4%
Leu (L)  28	 12.7%
Lys (K)  10	  4.5%
Met (M)   5	  2.3%
Phe (F)   5	  2.3%
Pro (P)   6	  2.7%
Ser (S)  10	  4.5%
Thr (T)   9	  4.1%
Trp (W)   6	  2.7%
Tyr (Y)   5	  2.3%
Val (V)  16	  7.3%

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

Total number of negatively charged residues (Asp + Glu): 27
Total number of positively charged residues (Arg + Lys): 22

Atomic composition:

Carbon      C	      1117
Hydrogen    H	      1756
Nitrogen    N	       318
Oxygen      O	       321
Sulfur      S	        10

Formula: C1117H1756N318O321S10
Total number of atoms: 3522

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    39940   40854   40925   40780   39840
Abs 0.1% (=1 g/l)   1.591   1.628   1.631   1.625   1.587



                      276     278     279     280     282
                       nm      nm      nm      nm      nm
Ext. coefficient    39650   40600   40685   40540   39600
Abs 0.1% (=1 g/l)   1.580   1.618   1.621   1.615   1.578


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 32.94
This classifies the protein as stable.



Aliphatic index: 101.00

Grand average of hydropathicity (GRAVY): -0.231

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