Part:BBa_K1159302

Enhanced Cyan Fluorescent Protein (eCFP) in RFC[25]

This Biobricks is a improved version BBa_E0020 which codes for the Enhanced Cyan Fluorescent Protein. We improve the function of the biobrick, to allow protein fusions which is originally not possible due to RFC[10] pre- and suffix. This part is flanked by RFC[25] pre- and suffix for further protein fusions.

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, so ATGGCCGGC and ACCGGT were added (in italics) to the 5' and 3' ends: (underlined part encodes the protein)
ATGGCCGGCGTGAGCAAG ... CTGTACAAGACCGGT
ORF from nucleotide position -8 to 720 (excluding stop-codon)

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

1
101
201

MAGVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERT
IFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPD
NHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKTG*

Sequence features: (with their position in the amino acid sequence, see the list of supported features)

RFC25 scar (shown in bold):

53 to 54

Amino acid composition:

Ala (A)	10 (4.1%)
Arg (R)	6 (2.5%)
Asn (N)	12 (4.9%)
Asp (D)	18 (7.4%)

Cys (C)	2 (0.8%)
Gln (Q)	8 (3.3%)
Glu (E)	16 (6.6%)
Gly (G)	24 (9.9%)

His (H)	9 (3.7%)
Ile (I)	13 (5.3%)
Leu (L)	21 (8.6%)
Lys (K)	20 (8.2%)

Met (M)	5 (2.1%)
Phe (F)	12 (4.9%)
Pro (P)	10 (4.1%)
Ser (S)	10 (4.1%)

Thr (T)	18 (7.4%)
Trp (W)	2 (0.8%)
Tyr (Y)	10 (4.1%)
Val (V)	17 (7.0%)

Amino acid counting

	Total number:	243
	Positively charged (Arg+Lys):	26 (10.7%)
	Negatively charged (Asp+Glu):	34 (14.0%)
	Aromatic (Phe+His+Try+Tyr):	33 (13.6%)

Biochemical parameters

	Atomic composition:	C₁₂₂₀H₁₈₈₃N₃₂₁O₃₇₀S₇
	Molecular mass [Da]:	27191.7
	Theoretical pI:	5.58
	Extinction coefficient at 280 nm [M^-1 cm^-1]:	25900 / 26025 (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 (0.69)	good (0.63)	acceptable (0.51)	good (0.60)	excellent (0.88)	excellent (0.96)

Alignments (obtained from PredictProtein.org)

	SwissProt:	P42212 (96% identity on 238 AAs), Q9U6Y3 (25% identity on 217 AAs)
	TrEML:	B7UCZ6 (98% identity on 238 AAs), B6F2F5 (97% identity on 242 AAs)
	PDB:	1oxf (98% identity on 225 AAs), 1qxt (98% identity on 226 AAs), 1qy3 (98% identity on 227 AAs), 2awj (98% identity on 227 AAs), 2g16 (98% identity on 162 AAs), 2g2s (98% identity on 164 AAs), 2g3d (98% identity on 164 AAs), 2g5z (98% identity on 163 AAs), 2hfc (98% identity on 230 AAs), 2wsn (98% identity on 225 AAs)

Predictions (obtained from PredictProtein.org)

Subcellular Localization (reliability in brackets)

	Archaea:	secreted (100%)
	Bacteria:	cytosol (59%)
	Eukarya:	cytosol (35%)

Gene Ontology (reliability in brackets)

	Molecular Function Ontology:	-
	Biological Process Ontology:	GO:0018298 (44%), GO:0008218 (31%)

Predicted features:

	Disulfid bridges:	-
	Transmembrane helices:	-

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