Part:BBa_K1159012

Secretory Catechol-2,3-dioxygenase (SERK-SigP_XylE) in RFC[25] N-Part

This part encodes for the enzyme catechol-2,3-dioxygenase (metapyrocatechase), which converts catechol to the bright yellow product 2-hydroxy-cis,cis-muconic semialdehyde. This part 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
  COMPATIBLE WITH RFC[21]
• 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 ... GTGCTGACCACCGGT  ORF from nucleotide position 5 to 1030 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below) 1  101  201  301  MPGEVAWWRPLFLIALMPIGVLSNAEGDALNTTGNKGVMRPGHVQLRVLDMSKALEHYVELLGLIEMDRDDQGRVYLKAWTEVDKFSLVLREADEPGMDF MGFKVVDEDALRQLERDLMAYGCAVEQLPAGELNSCGRRVRFQAPSGHHFELYADKEYTGKWGLNDVNPEAWPRDLKGMAAVRFDHALMYGDELPATYDL FTKVLGFYLAEQVLDENGTRVAQFLSLSTKAHDVAFIHHPEKGRLHHVSFHLETWEDLLRAADLISMTDTSIDIGPTRHGLTHGKTIYFFDPSGNRNEVF CGGDYNYPDHKPVTWTTDQLGKAIFYHDRILNERFMTVLTTG*
Sequence features: (with their position in the amino acid sequence, see the list of supported features) RFC25 scar (shown in bold):  33 to 34
Amino acid composition: Ala (A) 25 (7.3%) Arg (R) 20 (5.8%) Asn (N) 11 (3.2%) Asp (D) 28 (8.2%) Cys (C) 3 (0.9%) Gln (Q) 8 (2.3%) Glu (E) 23 (6.7%) Gly (G) 29 (8.5%) His (H) 15 (4.4%) Ile (I) 10 (2.9%) Leu (L) 39 (11.4%) Lys (K) 14 (4.1%) Met (M) 12 (3.5%) Phe (F) 17 (5.0%) Pro (P) 15 (4.4%) Ser (S) 11 (3.2%) Thr (T) 20 (5.8%) Trp (W) 7 (2.0%) Tyr (Y) 12 (3.5%) Val (V) 23 (6.7%)
Amino acid counting Total number: 342 Positively charged (Arg+Lys): 34 (9.9%) Negatively charged (Asp+Glu): 51 (14.9%) Aromatic (Phe+His+Try+Tyr): 51 (14.9%)	Biochemical parameters Atomic composition: C1742H2658N472O507S15 Molecular mass [Da]: 38806.0 Theoretical pI: 5.28 Extinction coefficient at 280 nm [M-1 cm-1]: 56380 / 56568 (all Cys red/ox)
Plot for hydrophobicity, charge, predicted secondary structure, solvent accessability, transmembrane helices and disulfid bridges
Codon usage Organism: E. coli B. subtilis S. cerevisiae A. thaliana P. patens Mammals Codon quality (CAI): good (0.73) good (0.70) acceptable (0.55) good (0.65) excellent (0.83) good (0.78)
Alignments (obtained from PredictProtein.org) SwissProt: P06622 (100% identity on 306 AAs), P27887 (94% identity on 306 AAs) TrEML: D2XKL2 (100% identity on 306 AAs), G5DRD2 (100% identity on 215 AAs), J9ZVS6 (100% identity on 270 AAs), J9ZVW6 (100% identity on 270 AAs), J9ZX53 (100% identity on 270 AAs), Q38Q96 (100% identity on 293 AAs), Q7AXV1 (100% identity on 306 AAs), Q7BH44 (100% identity on 306 AAs), J9ZVF5 (99% identity on 270 AAs), Q5EGN5 (99% identity on 237 AAs) PDB: 1mpy (100% identity on 306 AAs), 3hpv (48% identity on 296 AAs)
Predictions (obtained from PredictProtein.org)
Subcellular Localization (reliability in brackets) Archaea: cytosol (100%) Bacteria: cytosol (84%) Eukarya: secreted (43%)	Gene Ontology (reliability in brackets) Molecular Function Ontology: - Biological Process Ontology: GO:0019439 (49%), GO:0055114 (35%)
Predicted features: Disulfid bridges:  - Transmembrane helices:  -
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