Coding

Part:BBa_K1159202

Designed by: Dong-Jiunn Jeffery TRUONG   Group: iGEM13_TU-Munich   (2013-09-16)

Secretory SpyCatcher (IgKappa-SigP_SpyCatcher) in RFC[25] N-Part

This part encodes the SpyCatcher BBa_K1159200 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 ... GCTCATATTACCGGT
 ORF from nucleotide position 5 to 418 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)

101 
METDTLLLWVLLLWVPGSTGDTGVDTLSGLSSEQGQSGDMTIEEDSATHIKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYPGKYTFVE
TAAPDGYEVATAITFTVNEQGQVTVNGKATKGDAHITG*
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)9 (6.5%)
Arg (R)2 (1.4%)
Asn (N)2 (1.4%)
Asp (D)12 (8.7%)
Cys (C)0 (0.0%)
Gln (Q)5 (3.6%)
Glu (E)10 (7.2%)
Gly (G)16 (11.6%)
His (H)2 (1.4%)
Ile (I)6 (4.3%)
Leu (L)11 (8.0%)
Lys (K)8 (5.8%)
Met (M)3 (2.2%)
Phe (F)4 (2.9%)
Pro (P)3 (2.2%)
Ser (S)11 (8.0%)
Thr (T)18 (13.0%)
Trp (W)3 (2.2%)
Tyr (Y)4 (2.9%)
Val (V)9 (6.5%)
Amino acid counting
Total number:138
Positively charged (Arg+Lys):10 (7.2%)
Negatively charged (Asp+Glu):22 (15.9%)
Aromatic (Phe+His+Try+Tyr):13 (9.4%)
Biochemical parameters
Atomic composition:C649H1010N166O223S3
Molecular mass [Da]:14802.3
Theoretical pI:4.32
Extinction coefficient at 280 nm [M-1 cm-1]:22460 / 22460 (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 (0.80)good (0.75)good (0.75)excellent (0.80)good (0.78)good (0.67)
Alignments (obtained from PredictProtein.org)
SwissProt: -
TrEML:Q879P6 (97% identity on 114 AAs), Q8G9G1 (97% identity on 114 AAs)
PDB:2x5p (92% identity on 100 AAs)
Predictions (obtained from PredictProtein.org)
Subcellular Localization (reliability in brackets)
Archaea:secreted (100%)
Bacteria:secreted (29%)
Eukarya:cytosol (41%)
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