Part:BBa_K1080003:Design
GUN4
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 719
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 404
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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
No Biobrick sites: EcoRI, PstI, SpeI or XbaI
Amino acid sequence
MRGSHHHHHH GMASMTGGQQ MGRDLYDDDD KDHPFTMAMR VTVAAGKLDS VSLFGGDTAS
LMGGSQTVEK KKSGKEAVME VQLSSTAGID YTVLRDHLAN GEFREAEDET RALLIKLAGP
EAVKRNWVYF TEVKNISVTD FQTLDNLWKA SSNNKFGYSV QKEIWVQNQK RWPKFFKQID
WTQGENNNYR KWPMEFIYSM DAPRGHLPLT NALRGTQLFQ AIMEHPAFEK SSTAKTLDQK
AAEAAGRTQS LF
Number of amino acids: 252
Molecular weight: 28476.0
Theoretical pI: 7.18
Amino acid composition:
Ala (A) 22 8.7% Arg (R) 12 4.8% Asn (N) 11 4.4% Asp (D) 16 6.3% Cys (C) 0 0.0% Gln (Q) 14 5.6% Glu (E) 15 6.0% Gly (G) 19 7.5% His (H) 10 4.0% Ile (I) 7 2.8% Leu (L) 18 7.1% Lys (K) 19 7.5% Met (M) 11 4.4% Phe (F) 12 4.8% Pro (P) 7 2.8% Ser (S) 17 6.7% Thr (T) 17 6.7% Trp (W) 6 2.4% Tyr (Y) 6 2.4% Val (V) 13 5.2%
Asx (B) 0 0.0% Glx (Z) 0 0.0% Xaa (X) 0 0.0%
Total number of negatively charged residues (Asp + Glu): 31 Total number of positively charged residues (Arg + Lys): 31
Atomic composition:
Carbon C 1255 Hydrogen H 1940 Nitrogen N 358 Oxygen O 380 Sulfur S 11
Formula: C1255H1940N358O380S11 Total number of atoms: 3944
Extinction coefficients:
Extinction coefficients are in units of M-1 cm-1, at 280 nm.
Ext. coefficient 41940 Abs 0.1% (=1 g/l) 1.473, assuming ALL Cys residues appear as half cystines
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 22.16 This classifies the protein as stable.
Aliphatic index: 62.38
Grand average of hydropathicity (GRAVY): -0.655
Source
Chlamydomonas reinhardtii