Part:BBa_K1159202

Secretory SpyCatcher (IgKappa-SigP_SpyCatcher) in RFC[25] N-Part

This part encodes the SpyCatcher BBa_K1159200 with a N-terminal signal peptide for secretion (Ig Kappa signal peptide from Mus musculus) which 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
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)
AACAATGGAAACC ... GCTCATATTACCGGT
ORF from nucleotide position 5 to 418 (excluding stop-codon)

Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)

1
101

METDTLLLWVLLLWVPGSTGDTGVDTLSGLSSEQGQSGDMTIEEDSATHIKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYPGKYTFVE
TAAPDGYEVATAITFTVNEQGQVTVNGKATKGDAHITG*

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)	9 (6.5%)
Arg (R)	2 (1.4%)
Asn (N)	2 (1.4%)
Asp (D)	12 (8.7%)

Cys (C)	0 (0.0%)
Gln (Q)	5 (3.6%)
Glu (E)	10 (7.2%)
Gly (G)	16 (11.6%)

His (H)	2 (1.4%)
Ile (I)	6 (4.3%)
Leu (L)	11 (8.0%)
Lys (K)	8 (5.8%)

Met (M)	3 (2.2%)
Phe (F)	4 (2.9%)
Pro (P)	3 (2.2%)
Ser (S)	11 (8.0%)

Thr (T)	18 (13.0%)
Trp (W)	3 (2.2%)
Tyr (Y)	4 (2.9%)
Val (V)	9 (6.5%)

Amino acid counting

	Total number:	138
	Positively charged (Arg+Lys):	10 (7.2%)
	Negatively charged (Asp+Glu):	22 (15.9%)
	Aromatic (Phe+His+Try+Tyr):	13 (9.4%)

Biochemical parameters

	Atomic composition:	C₆₄₉H₁₀₁₀N₁₆₆O₂₂₃S₃
	Molecular mass [Da]:	14802.3
	Theoretical pI:	4.32
	Extinction coefficient at 280 nm [M^-1 cm^-1]:	22460 / 22460 (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 (0.80)	good (0.75)	good (0.75)	excellent (0.80)	good (0.78)	good (0.67)

Alignments (obtained from PredictProtein.org)

	SwissProt:	-
	TrEML:	Q879P6 (97% identity on 114 AAs), Q8G9G1 (97% identity on 114 AAs)
	PDB:	2x5p (92% identity on 100 AAs)

Predictions (obtained from PredictProtein.org)

Subcellular Localization (reliability in brackets)

	Archaea:	secreted (100%)
	Bacteria:	secreted (29%)
	Eukarya:	cytosol (41%)

Gene Ontology (reliability in brackets)

	Molecular Function Ontology:	-
	Biological Process Ontology:	-

Predicted features:

	Disulfid bridges:	-
	Transmembrane helices:	-

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