Part:BBa_K2740019

CR1 hesA

CR1 hesA encodes a NAD/FAD-binding protein, HesA, which is involved in molybdopterin and thiamine biosynthesis. It plays an important role in nitrogen fixation and is responsive to the availability of molybdenum.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 286
  

Parameter of Protein

Number of amino acids: 254

Molecular weight: 27871.11

Theoretical pI: 5.42

Amino acid composition:
Ala (A)  25    9.8%
Arg (R)  19    7.5%
Asn (N)   8   3.1%
Asp (D)  12   4.7%
Cys (C)   8    3.1%
Gln (Q)   8    3.1%
Glu (E)  19    7.5%
Gly (G)  26  10.2%
His (H)   6    2.4%
Ile (I)   14     5.5%
Leu (L)  27  10.6%
Lys (K)   5    2.0%
Met (M)  13   5.1%
Phe (F)   3     1.2%
Pro (P)  12     4.7%
Ser (S)   9     3.5%
Thr (T)  13    5.1%
Trp (W)  3     1.2%
Tyr (Y)   9    3.5%
Val (V)  15    5.9%
Pyl (O)   0     0.0%
Sec (U)   0    0.0%

 (B)   0         0.0%
(Z)   0   0.0%
(X)   0         0.0%

 

Total number of negatively charged residues (Asp + Glu): 31
Total number of positively charged residues (Arg + Lys): 24

Atomic composition:

Carbon      C          1212
Hydrogen    H         1941
Nitrogen    N            347
Oxygen      O          364
Sulfur      S              21

Formula: C1212H1941N347O364S21
Total number of atoms: 3885

Extinction coefficients:

Extinction coefficients are in units of  M-1 cm-1, at 280 nm measured in water.

Ext. coefficient    30410
Abs 0.1% (=1 g/l)   1.091, assuming all pairs of Cys residues form cystines

 

Ext. coefficient    29910
Abs 0.1% (=1 g/l)   1.073, assuming all Cys residues are reduced

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 44.28
This classifies the protein as unstable.

 

Aliphatic index: 89.92

Grand average of hydropathicity (GRAVY): -0.088

Design Notes

Nitrogenase is a complex enzyme system consisting of nine protein components. Additionally, to maintain stoichiometry of these protein components is an essential requirement for nitrogenase biosynthesis and activity. However, there is only one copy of each structure gene present in the nif gene cluster. Therefore, cloning each of these nif genes and setting as independent part can facilitate the regulation of balancing expression ratios from the transcription and/or translation level(s) when they are heterogeneously expressed in non-diazotrophic hosts.