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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