Coding
"The Gun"

Part:BBa_K1080003:Design

Designed by: Macquarie University   Group: iGEM13_Macquarie_Australia   (2013-09-12)

GUN4


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 719
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 404
  • 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.

GUN4 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

Ribbon structure of protein

http://pfam.sanger.ac.uk/structure/getimage?id=1y6i


Amino acid sequence

MAMRVTVAAG KLDSVSLFGG DTASLMGGSQ TVEKKKSGKE AVMEVQLSST AGIDYTVLRD
HLANGEFREA EDETRALLIK LAGPEAVKRN WVYFTEVKNI SVTDFQTLDN LWKASSNNKF
GYSVQKEIWV QNQKRWPKFF KQIDWTQGEN NNYRKWPMEF IYSMDAPRGH LPLTNALRGT
QLFQAIMEHP AFEKSSTAKT LDQKAAEAAG RTQSLF

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

Number of amino acids: 216

Molecular weight: 24350.6

Theoretical pI: 8.98

Amino acid composition:

Ala(A) 21 ( 9.7% )
Arg(R) 10 ( 4.6% )
Asn(N) 11 ( 5.1% )
Asp(D) 10 ( 4.6% )
Cys(C) 00 ( 0.0% )
Gln(Q) 12 ( 5.6% )
Glu(E) 15 ( 6.9% )
Gly(G) 14 ( 6.5% )
His(H) 03 ( 1.4% )
Ile(I) 07 ( 3.2% )
Leu(L) 17 ( 7.9% )
Lys(K) 18 ( 8.3% )
Met(M) 07 ( 3.2% )
Phe(F) 11 ( 5.1% )
Pro(P) 06 ( 2.8% )
Ser(S) 15 ( 6.9% )
Thr(T) 15 ( 6.9% )
Trp(W) 06 ( 2.8% )
Tyr(Y) 05 ( 2.3% )
Val(V) 13 ( 6.0% )
Pyl(O) 00 ( 0.0% )
Sec(U) 00 ( 0.0% ) 

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


Total number of negatively charged residues (Asp + Glu): 25 Total number of positively charged residues (Arg + Lys): 28

Atomic composition:

Carbon C 1085 Hydrogen H 1693 Nitrogen N 299 Oxygen O 325 Sulfur S 7

Formula: C1085H1693N299O325S7 Total number of atoms: 3409

Extinction coefficients:

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

Ext. coefficient 40450 Abs 0.1% (=1 g/l) 1.661

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 22.44 This classifies the protein as stable.


Aliphatic index: 70.51

Grand average of hydropathicity (GRAVY): -0.508

Source

Chlamydomonas reinhardtii

References