Coding
"Eye 1"

Part:BBa_K1080000:Design

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


ChlI1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 231
    Illegal BglII site found at 887
    Illegal BglII site found at 1082
    Illegal BamHI site found at 279
  • 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

ChlI1 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

No Biobrick restriction sites (EcoRI, XbaI, SpeI or PstI).

Amino Acid Sequence

MAATEVKAAE GRTEKELGQA RPIFPFTAIV GQDEMKLALI LNVIDPKIGG VMIMGDRGTG KSTTIRALAD LLPEMQVVAN DPFNSDPTDP ELMSEEVRNR
VKAGEQLPVS SKKIPMVDLP LGATEDRVCG TIDIEKALTE GVKAFEPGLL AKANRGILYV DEVNLLDDHL VDVLLDSAAS GWNTVEREGI SISHPARFIL
VGSGNPEEGE LRPQLLDRFG MHAQIGTVKD PRLRVQIVSQ RSTFDENPAA FRKDYEAGQM ALTQRIVDAR KLLKQGEVNY DFRVKISQIC SDLNVDGIRG
DIVTNRAAKA LAAFEGRTEV TPEDIYRVIP LCLRHRLRKD PLAEIDDGDR VREIFKQVFG ME

References and documentation are available. Please note the modified algorithm for extinction coefficient.

Number of amino acids: 362

Molecular weight: 39952.7

Theoretical pI: 5.11

Amino acid composition: Ala (A) 31 8.6% Arg (R) 28 7.7% Asn (N) 12 3.3% Asp (D) 29 8.0% Cys (C) 3 0.8% Gln (Q) 13 3.6% Glu (E) 29 8.0% Gly (G) 28 7.7% His (H) 4 1.1% Ile (I) 25 6.9% Leu (L) 34 9.4% Lys (K) 19 5.2% Met (M) 10 2.8% Phe (F) 12 3.3% Pro (P) 19 5.2% Ser (S) 14 3.9% Thr (T) 17 4.7% Trp (W) 1 0.3% Tyr (Y) 4 1.1% Val (V) 30 8.3% 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): 58 Total number of positively charged residues (Arg + Lys): 47

Atomic composition:

Carbon C 1752 Hydrogen H 2857 Nitrogen N 499 Oxygen O 539 Sulfur S 13

Formula: C1752H2857N499O539S13 Total number of atoms: 5660

Extinction coefficients:

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

Ext. coefficient 11585 Abs 0.1% (=1 g/l) 0.290, assuming all pairs of Cys residues form cystines


Ext. coefficient 11460 Abs 0.1% (=1 g/l) 0.287, 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 29.73 This classifies the protein as stable.


Aliphatic index: 96.16

Grand average of hydropathicity (GRAVY): -0.250

Source

Chlamydomonas reinhardtii

References