Coding

Part:BBa_K1159011

Designed by: Andreas-David BRUNNER   Group: iGEM13_TU-Munich   (2013-05-17)

Secretory Laccase (SERK-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 (signal peptide from SERK Receptor of 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 226
  • 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)
 AACAATGCCTGGT ... GATATCATCACCGGT
 ORF from nucleotide position 5 to 1636 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)
1 
101 
201 
301 
401 
501 		MPGEVAWWRPLFLIALMPIGVLSNAEGDALNTTGNLEKFVDELPIPEVAKPVKKNPKQTYYEIAMEEVFLKVHRDLPPTKLWTYNGSLPGPTIHANRNEK
VKVKWMNKLPLKHFLPVDHTIHEGHHDEPEVKTVVHLHGGVTPASSDGYPEAWFSRDFEATGPFFEREVYEYPNHQQACTLWYHDHAMALTRLNVYAGLA
GFYLISDAFEKSLELPKGEYDIPLMIMDRTFQEDGALFYPSRPNNTPEDSDIPDPSIVPFFCGETILVNGKVWPYLEVEPRKYRFRILNASNTRTYELHL
DNDATILQIGSDGGFLPRPVHHQSFSIAPAERFDVIIDFSAYENKTITLKNKAGCGQEVNPETDANIMQFKVTRPLKGRAPKTLRPIFKPLPPLRPCRAD
KERTLTLTGTQDKYGRPILLLDNQFWNDPVTENPRLGSVEVWSIVNPTRGTHPIHLHLVQFRVIDRRPFDTEVYQSTGDIVYTGPNEAPPLHEQGYKDTI
QAHAGEVIRIIARFVPYSGRYVWHCHILEHEDYDMMRPMDIITG*
Sequence features: (with their position in the amino acid sequence, see the list of supported features)
RFC25 scar (shown in bold): 33 to 34, 161 to 162
Amino acid composition:
Ala (A)30 (5.5%)
Arg (R)30 (5.5%)
Asn (N)25 (4.6%)
Asp (D)32 (5.9%)
Cys (C)5 (0.9%)
Gln (Q)14 (2.6%)
Glu (E)39 (7.2%)
Gly (G)33 (6.1%)
His (H)23 (4.2%)
Ile (I)33 (6.1%)
Leu (L)46 (8.5%)
Lys (K)27 (5.0%)
Met (M)11 (2.0%)
Phe (F)25 (4.6%)
Pro (P)50 (9.2%)
Ser (S)19 (3.5%)
Thr (T)35 (6.4%)
Trp (W)10 (1.8%)
Tyr (Y)22 (4.0%)
Val (V)35 (6.4%)
Amino acid counting
Total number:544
Positively charged (Arg+Lys):57 (10.5%)
Negatively charged (Asp+Glu):71 (13.1%)
Aromatic (Phe+His+Try+Tyr):80 (14.7%)
Biochemical parameters
Atomic composition:C2828H4312N756O802S16
Molecular mass [Da]:62246.9
Theoretical pI:5.92
Extinction coefficient at 280 nm [M-1 cm-1]:87780 / 88093 (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 (0.72)good (0.75)good (0.68)good (0.75)good (0.78)good (0.67)
Alignments (obtained from PredictProtein.org)
SwissProt:P07788 (67% identity on 507 AAs), Q12737 (27% identity on 452 AAs)
TrEML:B4AIB1 (99% identity on 509 AAs), A8FAG9 (98% identity on 508 AAs)
PDB:1gsk (67% identity on 502 AAs), 1of0 (67% identity on 502 AAs)
Predictions (obtained from PredictProtein.org)
Subcellular Localization (reliability in brackets)
Archaea:cytosol (99%)
Bacteria:secreted (41%)
Eukarya:secreted (66%)
Gene Ontology (reliability in brackets)
Molecular Function Ontology:GO:0005507 (34%), GO:0005515 (23%)
Biological Process Ontology:GO:0055114 (37%), GO:0017000 (21%)
 
Predicted features:
Disulfid bridges: -
Transmembrane helices: -
The BioBrick-AutoAnnotator was created by TU-Munich 2013 iGEM team. For more information please see the documentation.
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Categories
Parameters
None