Part:BBa_K1159203

Secretory SpyTag (IgKappa-SigP_SpyTag) in RFC[25] N-Part

This part encodes the SpyTag BBa_K1159201 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 ... CCAACTAAGACCGGT
ORF from nucleotide position 5 to 118 (excluding stop-codon)

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

1	METDTLLLWVLLLWVPGSTGDTGAHIVMVDAYKPTKTG*

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
	Spy-tag:	24 to 36

Amino acid composition:

Ala (A)	2 (5.3%)
Arg (R)	0 (0.0%)
Asn (N)	0 (0.0%)
Asp (D)	3 (7.9%)

Cys (C)	0 (0.0%)
Gln (Q)	0 (0.0%)
Glu (E)	1 (2.6%)
Gly (G)	4 (10.5%)

His (H)	1 (2.6%)
Ile (I)	1 (2.6%)
Leu (L)	6 (15.8%)
Lys (K)	2 (5.3%)

Met (M)	2 (5.3%)
Phe (F)	0 (0.0%)
Pro (P)	2 (5.3%)
Ser (S)	1 (2.6%)

Thr (T)	6 (15.8%)
Trp (W)	2 (5.3%)
Tyr (Y)	1 (2.6%)
Val (V)	4 (10.5%)

Amino acid counting

	Total number:	38
	Positively charged (Arg+Lys):	2 (5.3%)
	Negatively charged (Asp+Glu):	4 (10.5%)
	Aromatic (Phe+His+Try+Tyr):	4 (10.5%)

Biochemical parameters

	Atomic composition:	C₁₈₉H₂₉₈N₄₄O₅₅S₂
	Molecular mass [Da]:	4130.8
	Theoretical pI:	4.66
	Extinction coefficient at 280 nm [M^-1 cm^-1]:	12490 / 12490 (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 (0.68)	good (0.66)	good (0.68)	excellent (0.81)	good (0.80)	good (0.69)

Alignments (obtained from PredictProtein.org)

	SwissProt:	-
	TrEML:	-
	PDB:	-

Predictions (obtained from PredictProtein.org)

Subcellular Localization (reliability in brackets)

	Archaea:	secreted (73%)
	Bacteria:	innermembrane (37%)
	Eukarya:	secreted (85%)

Gene Ontology (reliability in brackets)

	Molecular Function Ontology:	-
	Biological Process Ontology:	-

Predicted features:

	Disulfid bridges:	-
	Transmembrane helices:	-

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