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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
MGRDLYDDDD KDHPFTMAMR VTVAAGKLDS VSLFGGDTAS LMGGSQTVEK KKSGKEAVME
VQLSSTAGID YTVLRDHLAN GEFREAEDET RALLIKLAGP EAVKRNWVYF TEVKNISVTD
FQTLDNLWKA SSNNKFGYSV QKEIWVQNQK RWPKFFKQID WTQGENNNYR KWPMEFIYSM
DAPRGHLPLT NALRGTQLFQ AIMEHPAFEK SSTAKTLDQK AAEAAGRTQS LF
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Number of amino acids: 232
Molecular weight: 26272.6
Theoretical pI: 6.45
Amino acid composition:
Ala (A) 21 9.1% Arg (R) 11 4.7% Asn (N) 11 4.7% Asp (D) 16 6.9% Cys (C) 0 0.0% Gln (Q) 12 5.2% Glu (E) 15 6.5% Gly (G) 15 6.5% His (H) 4 1.7% Ile (I) 7 3.0% Leu (L) 18 7.8% Lys (K) 19 8.2% Met (M) 8 3.4% Phe (F) 12 5.2% Pro (P) 7 3.0% Ser (S) 15 6.5% Thr (T) 16 6.9% Trp (W) 6 2.6% Tyr (Y) 6 2.6% Val (V) 13 5.6% 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): 30
Atomic composition:
Carbon C 1167 Hydrogen H 1809 Nitrogen N 321 Oxygen O 355 Sulfur S 8
Formula: C1167H1809N321O355S8 Total number of atoms: 3660
Extinction coefficients:
Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.
Ext. coefficient 41940 Abs 0.1% (=1 g/l) 1.596
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 22.94 This classifies the protein as stable.
Aliphatic index: 67.33
Grand average of hydropathicity (GRAVY): -0.594
Source
Chlamydomonas reinhardtii