Part:BBa_K1159006

Secretory NanoLuc Luciferase (IgKappa-SigP_nLuc) in RFC[25] N-Part

NanoLuc Luciferase is engineered ATP-independent luciferase from a deep-sea shrimp which luminescense 2 magnitues higher than these from Renilla reniformis or from Photinus pyralis (firefly). Also the molecular weight of NanoLuc luciferase is twice smaller compared to other luciferase (only 19 kDa). This part encodes for a NanoLuc Luciferase which is fused to a N-terminal signal peptide for secretion (IgKappa signal peptide from Mus musculus) with 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
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 83
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 ... TTCGTTGGAACCGGT
ORF from nucleotide position 5 to 106 (excluding stop-codon)

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

1	METDTLLLWVLLLWVPGSTGDTGVFTLEDFVGTG*

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)	0 (0.0%)
Arg (R)	0 (0.0%)
Asn (N)	0 (0.0%)
Asp (D)	3 (8.8%)

Cys (C)	0 (0.0%)
Gln (Q)	0 (0.0%)
Glu (E)	2 (5.9%)
Gly (G)	5 (14.7%)

His (H)	0 (0.0%)
Ile (I)	0 (0.0%)
Leu (L)	7 (20.6%)
Lys (K)	0 (0.0%)

Met (M)	1 (2.9%)
Phe (F)	2 (5.9%)
Pro (P)	1 (2.9%)
Ser (S)	1 (2.9%)

Thr (T)	6 (17.6%)
Trp (W)	2 (5.9%)
Tyr (Y)	0 (0.0%)
Val (V)	4 (11.8%)

Amino acid counting

	Total number:	34
	Positively charged (Arg+Lys):	0 (0.0%)
	Negatively charged (Asp+Glu):	5 (14.7%)
	Aromatic (Phe+His+Try+Tyr):	4 (11.8%)

Biochemical parameters

	Atomic composition:	C₁₇₁H₂₆₀N₃₆O₅₂S₁
	Molecular mass [Da]:	3684.2
	Theoretical pI:	3.33
	Extinction coefficient at 280 nm [M^-1 cm^-1]:	11000 / 11000 (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 (0.66)	good (0.64)	acceptable (0.58)	good (0.78)	good (0.79)	good (0.70)

Alignments (obtained from PredictProtein.org)
There were no alignments for this protein in the data base. The BLAST search was initialized and should be ready in a few hours.

Predictions (obtained from PredictProtein.org)

There were no predictions for this protein in the data base. The prediction was initialized and should be ready in a few hours.

The BioBrick-AutoAnnotator was created by TU-Munich 2013 iGEM team. For more information please see the documentation.
If you have any questions, comments or suggestions, please leave us a comment.