Coding

Part:BBa_K1159017

Designed by: TU Munich 2013   Group: iGEM13_TU-Munich   (2013-05-17)
Revision as of 15:01, 8 October 2013 by JohannaB (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

Membrane-anchored FluA triple mutant in RFC[25] N-Part

This part codes for a membrane-anchored version of the Fluorescein binding anticalin FluA (triple mutant version for higher affinity) for the chassis Physcomitrella patens. Please see subparts for for further description. 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 BglII site found at 354
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 642
    Illegal BsaI.rc site found at 1560

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)
 AACAATGCCTGGT ... CTCTACAAAACCGGT
 ORF from nucleotide position 5 to 1636 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)
1 
101 
201 
301 
401 
501 		MPGEVAWWRPLFLIALMPIGVLSNAEGDALNTTGDVYHDGACPEVKPVDNFDWSQYHGKWWEVAKYPSPNGKYGKCGWIEYTPEGKSVKVSRYDVIHGKE
YFMEGTAYPVGDSKIGKIYHSRTVGGYTKKTVFNVLSTDNKNYIIGYTCRYDEDKKGHWDHVWVLSRSMVLTGEAKTAVENYLIGSPVVDSQKLVYSDFS
EAACKVNNTGSAWSHPQFEKGENLYFQSGTGPGQPPFPPPPPFTPPPPQTPNGASGENSTGAIAGGVAAGAALLFAAPAIGFAWWRRRRPIEATGGGSGG
GSGTGSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQER
TIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLSP
DNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKTG*
Sequence features: (with their position in the amino acid sequence, see the list of supported features)
RFC25 scar (shown in bold): 33 to 34, 209 to 210, 230 to 231, 294 to 295, 304 to 305
Strep-tag II: 213 to 220
TEV cleavage site: 222 to 228
Amino acid composition:
Ala (A)32 (5.9%)
Arg (R)15 (2.8%)
Asn (N)26 (4.8%)
Asp (D)31 (5.7%)
Cys (C)6 (1.1%)
Gln (Q)14 (2.6%)
Glu (E)32 (5.9%)
Gly (G)63 (11.6%)
His (H)17 (3.1%)
Ile (I)24 (4.4%)
Leu (L)32 (5.9%)
Lys (K)39 (7.2%)
Met (M)9 (1.7%)
Phe (F)23 (4.2%)
Pro (P)36 (6.6%)
Ser (S)32 (5.9%)
Thr (T)35 (6.4%)
Trp (W)12 (2.2%)
Tyr (Y)27 (5.0%)
Val (V)39 (7.2%)
Amino acid counting
Total number:544
Positively charged (Arg+Lys):54 (9.9%)
Negatively charged (Asp+Glu):63 (11.6%)
Aromatic (Phe+His+Try+Tyr):79 (14.5%)
Biochemical parameters
Atomic composition:C2698H4070N714O805S15
Molecular mass [Da]:59869.2
Theoretical pI:6.03
Extinction coefficient at 280 nm [M-1 cm-1]:106230 / 106605 (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 (0.71)good (0.72)good (0.71)good (0.80)excellent (0.81)good (0.73)
Alignments (obtained from PredictProtein.org)
SwissProt:P42212 (98% identity on 238 AAs), Q9U6Y3 (25% identity on 217 AAs)
TrEML:Q71RY9 (98% identity on 238 AAs), B6F2F5 (97% identity on 241 AAs)
PDB:2y0g (98% identity on 226 AAs), 1c4f (97% identity on 225 AAs)
Predictions (obtained from PredictProtein.org)
Subcellular Localization (reliability in brackets)
Archaea:secreted (99%)
Bacteria:secreted (73%)
Eukarya:secreted (36%)
Gene Ontology (reliability in brackets)
Molecular Function Ontology:GO:0019841 (29%), GO:0005515 (23%)
Biological Process Ontology:GO:0018298 (46%), GO:0008218 (30%)
 
Predicted features:
Disulfid bridges:42 to 204
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
Parameters
None