Part:BBa_K1159011

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. coli	B. subtilis	S. cerevisiae	A. thaliana	P. patens	Mammals
	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:	-

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