Coding

Part:BBa_K1159302

Designed by: Dong-Jiunn Jeffery TRUONG   Group: iGEM13_TU-Munich   (2013-09-14)

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)

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. coliB. subtilisS. cerevisiaeA. thalianaP. patensMammals
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: -
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.


[edit]
Categories
//cds/reporter/cfp
//function/reporter/fluorescence
Parameters
colorCyan