Coding

Part:BBa_K1159210

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

Secretory nLuc_SpyCatcher fusion (IgKappa-SigP_nLuc_SpyCatcher) in RFC[25] N-Part

Secretory Fusionprotein consisting N-terminal nLuc and C-terminal SpyCatcher (IgKappa-SigP_nLuc_SpyCatcher) in RFC[25] N-Part. 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 XhoI site found at 83
  • 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 934 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)
1 
101 
201 
301 		METDTLLLWVLLLWVPGSTGDTGVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRIVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKI
FKVVYPVDDHHFKVILHYGTLVIDGVTPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRLCERILATGVDTLS
GLSSEQGQSGDMTIEEDSATHIKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNGK
ATKGDAHITG*
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, 194 to 195
Amino acid composition:
Ala (A)12 (3.9%)
Arg (R)9 (2.9%)
Asn (N)10 (3.2%)
Asp (D)24 (7.7%)
Cys (C)1 (0.3%)
Gln (Q)12 (3.9%)
Glu (E)18 (5.8%)
Gly (G)37 (11.9%)
His (H)6 (1.9%)
Ile (I)24 (7.7%)
Leu (L)27 (8.7%)
Lys (K)15 (4.8%)
Met (M)5 (1.6%)
Phe (F)12 (3.9%)
Pro (P)9 (2.9%)
Ser (S)17 (5.5%)
Thr (T)29 (9.4%)
Trp (W)6 (1.9%)
Tyr (Y)10 (3.2%)
Val (V)27 (8.7%)
Amino acid counting
Total number:310
Positively charged (Arg+Lys):24 (7.7%)
Negatively charged (Asp+Glu):42 (13.5%)
Aromatic (Phe+His+Try+Tyr):34 (11.0%)
Biochemical parameters
Atomic composition:C1521H2368N392O473S6
Molecular mass [Da]:33906.2
Theoretical pI:4.63
Extinction coefficient at 280 nm [M-1 cm-1]:47900 / 47963 (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 (0.71)good (0.67)excellent (0.83)excellent (0.85)good (0.75)
Alignments (obtained from PredictProtein.org)
SwissProt:Q9GV45 (90% identity on 167 AAs)
TrEML:A2RC81 (50% identity on 251 AAs), A2T8Z5 (50% identity on 251 AAs)
PDB:2x5p (92% identity on 99 AAs)
Predictions (obtained from PredictProtein.org)
Subcellular Localization (reliability in brackets)
Archaea:secreted (100%)
Bacteria:secreted (49%)
Eukarya:secreted (52%)
Gene Ontology (reliability in brackets)
Molecular Function Ontology: -
Biological Process Ontology: -
 
Predicted features:
Disulfid bridges: -
Transmembrane helices: -
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Categories
Parameters
None