Coding

Part:BBa_K1159007

Designed by: Andreas-David BRUNNER   Group: iGEM13_TU-Munich   (2013-09-15)
Revision as of 14:14, 8 October 2013 by JohannaB (Talk | contribs)

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Secretory Laccase (IgKappa-SigP_Laccase) in RFC[25] N-Part

Laccase from Bacillus pumilus, which degrades a wide variety of phenolic and non-phenolic compounds, is fused with a N-terminal signal peptide for secretion (Ig Kappa signal peptide from Mus musculus) with also works in moss (Physcomitrella patens). This construct is flanked by RFC[25] N-part prefix and suffix. Note: This means only protein fusions to the C-terminus of this part is possible, adding promoters (typically RFC[10]) into the prefix or terminators/IRES (typically RFC[10]) into the suffix of this part is nevertheless possible.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 193
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Protein data table for BioBrick BBa_ automatically created by the BioBrick-AutoAnnotator version 1.0
Nucleotide sequence in RFC 25 N-Part using the stop codon in the suffix, so ACCGGT was added (in italics) to the 3' end: (underlined part encodes the protein)
 AACAATGGAAACC ... GACGAGCTGACCGGT
 ORF from nucleotide position 5 to 106 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)
1 METDTLLLWVLLLWVPGSTGDTGNLEKFVDELTG*
Sequence features: (with their position in the amino acid sequence, see the list of supported features)
RFC25 scar (shown in bold): 19 to 20, 22 to 23
Amino acid composition:
Ala (A)0 (0.0%)
Arg (R)0 (0.0%)
Asn (N)1 (2.9%)
Asp (D)3 (8.8%)
Cys (C)0 (0.0%)
Gln (Q)0 (0.0%)
Glu (E)3 (8.8%)
Gly (G)4 (11.8%)
His (H)0 (0.0%)
Ile (I)0 (0.0%)
Leu (L)8 (23.5%)
Lys (K)1 (2.9%)
Met (M)1 (2.9%)
Phe (F)1 (2.9%)
Pro (P)1 (2.9%)
Ser (S)1 (2.9%)
Thr (T)5 (14.7%)
Trp (W)2 (5.9%)
Tyr (Y)0 (0.0%)
Val (V)3 (8.8%)
Amino acid counting
Total number:34
Positively charged (Arg+Lys):1 (2.9%)
Negatively charged (Asp+Glu):6 (17.6%)
Aromatic (Phe+His+Try+Tyr):3 (8.8%)
Biochemical parameters
Atomic composition:C172H268N38O54S1
Molecular mass [Da]:3764.3
Theoretical pI:3.71
Extinction coefficient at 280 nm [M-1 cm-1]:11000 / 11000 (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 (0.67)good (0.66)good (0.62)good (0.75)good (0.74)good (0.66)
Alignments (obtained from PredictProtein.org)
   There were no alignments for this protein in the data base. The BLAST search was initialized and should be ready in a few hours.
Predictions (obtained from PredictProtein.org)
   There were no predictions for this protein in the data base. The prediction was initialized and should be ready in a few hours.
The BioBrick-AutoAnnotator was created by TU-Munich 2013 iGEM team. For more information please see the documentation.
If you have any questions, comments or suggestions, please leave us a comment.


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Categories
Parameters
None