Coding

Part:BBa_K1159209

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

Secretory nLuc_SpyTag fusion (IgKappa-SigP_nLuc_SpyTag) in RFC[25] N-Part

Secretory Fusionprotein consisting N-terminal nLuc and C-terminal SpyTag (IgKappa-SigP_nLuc_SpyTag) 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 ... CCAACTAAGACCGGT
 ORF from nucleotide position 5 to 634 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)
1 
101 
201 		METDTLLLWVLLLWVPGSTGDTGVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRIVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKI
FKVVYPVDDHHFKVILHYGTLVIDGVTPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRLCERILATGAHIVM
VDAYKPTKTG*
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
Spy-tag: 196 to 208
Amino acid composition:
Ala (A)5 (2.4%)
Arg (R)7 (3.3%)
Asn (N)8 (3.8%)
Asp (D)15 (7.1%)
Cys (C)1 (0.5%)
Gln (Q)7 (3.3%)
Glu (E)9 (4.3%)
Gly (G)25 (11.9%)
His (H)5 (2.4%)
Ile (I)19 (9.0%)
Leu (L)22 (10.5%)
Lys (K)9 (4.3%)
Met (M)4 (1.9%)
Phe (F)8 (3.8%)
Pro (P)8 (3.8%)
Ser (S)7 (3.3%)
Thr (T)17 (8.1%)
Trp (W)5 (2.4%)
Tyr (Y)7 (3.3%)
Val (V)22 (10.5%)
Amino acid counting
Total number:210
Positively charged (Arg+Lys):16 (7.6%)
Negatively charged (Asp+Glu):24 (11.4%)
Aromatic (Phe+His+Try+Tyr):25 (11.9%)
Biochemical parameters
Atomic composition:C1061H1656N270O305S5
Molecular mass [Da]:23234.7
Theoretical pI:5.01
Extinction coefficient at 280 nm [M-1 cm-1]:37930 / 37993 (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 (0.66)good (0.68)good (0.63)excellent (0.84)excellent (0.89)good (0.80)
Alignments (obtained from PredictProtein.org)
SwissProt:Q9GV45 (90% identity on 169 AAs)
TrEML: -
PDB: -
Predictions (obtained from PredictProtein.org)
Subcellular Localization (reliability in brackets)
Archaea:secreted (100%)
Bacteria:secreted (44%)
Eukarya:secreted (31%)
Gene Ontology (reliability in brackets)
Molecular Function Ontology: -
Biological Process Ontology: -
 
Predicted features:
Disulfid bridges: -
Transmembrane helices: -
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Parameters
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