Coding

Part:BBa_K1159203

Designed by: Dong-Jiunn Jeffery TRUONG   Group: iGEM13_TU-Munich   (2013-09-17)

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)
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. coliB. subtilisS. cerevisiaeA. thalianaP. patensMammals
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: -
The BioBrick-AutoAnnotator was created by TU-Munich 2013 iGEM team. For more information please see the documentation.
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[edit]
Categories
//chassis/eukaryote/ppatens
Parameters
None