Coding

Part:BBa_K1159102

Designed by: Dong-Jiunn Jeffery TRUONG   Group: iGEM13_TU-Munich   (2013-09-14)

C-terminal half of TEV Protease S219V mutant (for Split-TEV-Protease applications) in RFC[25]

C-terminal half of the TEV Protease (autolysis resistant S219V mutant) can be used for Split-TEV-Protease applications where autolysis of the reconstituted TEV Protease is not wanted. The necessary N-terminal half can be found under BBa_K1159101. This part is flanked by RFC[25] pre- and suffix for further protein fusions.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 310

Protein data table for BioBrick BBa_ automatically created by the BioBrick-AutoAnnotator version 1.0
Nucleotide sequence in RFC 25, so ATGGCCGGC and ACCGGT were added (in italics) to the 5' and 3' ends: (underlined part encodes the protein)
 ATGGCCGGCAAGAGCATG ... CTCATGAATACCGGT
 ORF from nucleotide position -8 to 360 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)

101 
MAGKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFIVGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHK
VFMVKPEEPFQPVKEATQLMNTG*
Sequence features: (with their position in the amino acid sequence, see the list of supported features)
None of the supported features appeared in the sequence
Amino acid composition:
Ala (A)5 (4.1%)
Arg (R)2 (1.6%)
Asn (N)8 (6.5%)
Asp (D)5 (4.1%)
Cys (C)2 (1.6%)
Gln (Q)7 (5.7%)
Glu (E)5 (4.1%)
Gly (G)10 (8.1%)
His (H)3 (2.4%)
Ile (I)4 (3.3%)
Leu (L)6 (4.9%)
Lys (K)7 (5.7%)
Met (M)6 (4.9%)
Phe (F)8 (6.5%)
Pro (P)6 (4.9%)
Ser (S)14 (11.4%)
Thr (T)10 (8.1%)
Trp (W)5 (4.1%)
Tyr (Y)1 (0.8%)
Val (V)9 (7.3%)
Amino acid counting
Total number:123
Positively charged (Arg+Lys):9 (7.3%)
Negatively charged (Asp+Glu):10 (8.1%)
Aromatic (Phe+His+Try+Tyr):17 (13.8%)
Biochemical parameters
Atomic composition:C608H918N162O184S8
Molecular mass [Da]:13697.4
Theoretical pI:6.25
Extinction coefficient at 280 nm [M-1 cm-1]:28990 / 29115 (all Cys red/ox)
Plot for hydrophobicity, charge, predicted secondary structure, solvent accessability, transmembrane helices and disulfid bridges 
Codon usage
Organism:E. coliB. subtilisS. cerevisiaeA. thalianaP. patensMammals
Codon quality (CAI):good (0.75)good (0.73)good (0.68)good (0.75)excellent (0.80)good (0.70)
Alignments (obtained from PredictProtein.org)
SwissProt:P04517 (98% identity on 119 AAs), P13529 (47% identity on 119 AAs)
TrEML:Q000U0 (97% identity on 119 AAs), G9IZP9 (95% identity on 119 AAs)
PDB: -
Predictions (obtained from PredictProtein.org)
Subcellular Localization (reliability in brackets)
Archaea:secreted (100%)
Bacteria:secreted (76%)
Eukarya:cytosol (17%)
Gene Ontology (reliability in brackets)
Molecular Function Ontology: -
Biological Process Ontology:GO:0006508 (41%), GO:0019079 (41%)
 
Predicted features:
Disulfid bridges: -
Transmembrane helices: -
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Categories
Parameters
None