Coding

Part:BBa_K1159205

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

Secretory SpyTag (SERK-SigP_SpyTag) in RFC[25] N-Part

This part encodes the SpyTag BBa_K1159201 with a N-terminal signal peptide for secretion (SERK receptor signal peptide from 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)
 AACAATGCCTGGT ... CCAACTAAGACCGGT
 ORF from nucleotide position 5 to 151 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)
MPGEVAWWRPLFLIALMPIGVLSNAEGDALNTTGAHIVMVDAYKPTKTG*
Sequence features: (with their position in the amino acid sequence, see the list of supported features)
RFC25 scar (shown in bold): 33 to 34
Spy-tag: 35 to 47
Amino acid composition:
Ala (A)6 (12.2%)
Arg (R)1 (2.0%)
Asn (N)2 (4.1%)
Asp (D)2 (4.1%)
Cys (C)0 (0.0%)
Gln (Q)0 (0.0%)
Glu (E)2 (4.1%)
Gly (G)5 (10.2%)
His (H)1 (2.0%)
Ile (I)3 (6.1%)
Leu (L)5 (10.2%)
Lys (K)2 (4.1%)
Met (M)3 (6.1%)
Phe (F)1 (2.0%)
Pro (P)4 (8.2%)
Ser (S)1 (2.0%)
Thr (T)4 (8.2%)
Trp (W)2 (4.1%)
Tyr (Y)1 (2.0%)
Val (V)4 (8.2%)
Amino acid counting
Total number:49
Positively charged (Arg+Lys):3 (6.1%)
Negatively charged (Asp+Glu):4 (8.2%)
Aromatic (Phe+His+Try+Tyr):5 (10.2%)
Biochemical parameters
Atomic composition:C240H376N60O66S3
Molecular mass [Da]:5254.2
Theoretical pI:5.46
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 (0.69)good (0.70)good (0.71)excellent (0.88)excellent (0.87)good (0.74)
Alignments (obtained from PredictProtein.org)
SwissProt: -
TrEML: -
PDB: -
Predictions (obtained from PredictProtein.org)
Subcellular Localization (reliability in brackets)
Archaea:cytosol (100%)
Bacteria:secreted (29%)
Eukarya:secreted (63%)
Gene Ontology (reliability in brackets)
Molecular Function Ontology: -
Biological Process Ontology: -
 
Predicted features:
Disulfid bridges: -
Transmembrane helices:8 to 25 going inwards
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[edit]
Categories
//chassis/eukaryote/ppatens
Parameters
None