Coding
"Eye 2"

Part:BBa_K1080011:Design

Designed by: Macquarie University   Group: iGEM13_Macquarie_Australia   (2013-09-22)

ChlI2


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 523
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Incorporated sequence overlap for Gibson assembly and no GC rich region or restriction site in sequence.


ChlI2 Clone: DNA sequence from translation start site:

Translated the DNA sequence into a protein sequence using "Translate" at http://au.expasy.org/tools Then used the translated protein sequence to analyse the protein using "ProtParam" at http://au.expasy.org/tools

MPSTKAAKKP NFPFVKIQGQ EEMKLALLLN VVDPNIGGVL IMGDRGTAKS VAVRALVDML
PDIDVVEGDA FNSSPTDPKF MGPDTLQRFR NGEKLPTVRM RTPLVELPLG ATEDRICGTI
DIEKALTQGI KAYEPGLLAK ANRGILYVDE VNLLDDGLVD VVLDSSASGL NTVEREGVSI
VHPARFIMIG SGNPQEGELR PQLLDRFGMS VNVATLQDTK QRTQLVLDRL AYEADPDAFV
DSCKAEQTAL TDKLEAARQR LRSVKISEEL QILISDICSR LDVDGLRGDI VINRAAKALV
AFEGRTEVTT NDVERVISGC LNHRLRKDPL DPIDNGTKVA ILFKRMTDPE IMKREEEAKK

Please note the modified algorithm for extinction coefficient.

Number of amino acids: 393

Molecular weight: 42954.3

Theoretical pI: 5.78

Amino acid composition: Ala (A) 38 9.7% Arg (R) 30 7.6% Asn (N) 14 3.6% Asp (D) 32 8.1% Cys (C) 4 1.0% Gln (Q) 12 3.1% Glu (E) 28 7.1% Gly (G) 29 7.4% His (H) 2 0.5% Ile (I) 22 5.6% Leu (L) 41 10.4% Lys (K) 27 6.9% Met (M) 10 2.5% Phe (F) 10 2.5% Pro (P) 22 5.6% Ser (S) 16 4.1% Thr (T) 21 5.3% Trp (W) 1 0.3% Tyr (Y) 3 0.8% Val (V) 31 7.9% 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): 60 Total number of positively charged residues (Arg + Lys): 57

Atomic composition:

Carbon C 1875 Hydrogen H 3103 Nitrogen N 541 Oxygen O 580 Sulfur S 14

Formula: C1875H3103N541O580S14 Total number of atoms: 6113

Extinction coefficients:

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

Ext. coefficient 10220 Abs 0.1% (=1 g/l) 0.238, assuming all pairs of Cys residues form cystines


Ext. coefficient 9970 Abs 0.1% (=1 g/l) 0.232, 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 32.87 This classifies the protein as stable.


Aliphatic index: 95.06

Grand average of hydropathicity (GRAVY): -0.296