Coding

Part:BBa_K1159206

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

Membrane-anchored SpyCatcher in RFC[25] N-Part

This part codes for a membrane-anchored version of SpyCatcher BBa_K1159200 for the chassis Physcomitrella patens. Please see subparts for for further description. 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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 459
    Illegal BsaI.rc site found at 1377

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 ... CTCTACAAAACCGGT
 ORF from nucleotide position 5 to 1453 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)

101 
201 
301 
401 
MPGEVAWWRPLFLIALMPIGVLSNAEGDALNTTGVDTLSGLSSEQGQSGDMTIEEDSATHIKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDF
YLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNGKATKGDAHITGSAWSHPQFEKGENLYFQSGTGPGQPPFPPPPPFTPPPPQTPNGASGENSTG
AIAGGVAAGAALLFAAPAIGFAWWRRRRPIEATGGGSGGGSGTGSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPW
PTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQ
KNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLSPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKTG*
Sequence features: (with their position in the amino acid sequence, see the list of supported features)
RFC25 scar (shown in bold): 33 to 34, 148 to 149, 169 to 170, 233 to 234, 243 to 244
Strep-tag II: 152 to 159
TEV cleavage site: 161 to 167
Amino acid composition:
Ala (A)34 (7.0%)
Arg (R)13 (2.7%)
Asn (N)20 (4.1%)
Asp (D)29 (6.0%)
Cys (C)2 (0.4%)
Gln (Q)17 (3.5%)
Glu (E)31 (6.4%)
Gly (G)59 (12.2%)
His (H)13 (2.7%)
Ile (I)23 (4.8%)
Leu (L)32 (6.6%)
Lys (K)29 (6.0%)
Met (M)9 (1.9%)
Phe (F)23 (4.8%)
Pro (P)31 (6.4%)
Ser (S)29 (6.0%)
Thr (T)39 (8.1%)
Trp (W)7 (1.4%)
Tyr (Y)16 (3.3%)
Val (V)27 (5.6%)
Amino acid counting
Total number:483
Positively charged (Arg+Lys):42 (8.7%)
Negatively charged (Asp+Glu):60 (12.4%)
Aromatic (Phe+His+Try+Tyr):59 (12.2%)
Biochemical parameters
Atomic composition:C2328H3557N621O725S11
Molecular mass [Da]:52197.2
Theoretical pI:5.19
Extinction coefficient at 280 nm [M-1 cm-1]:62340 / 62465 (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 (0.75)good (0.75)good (0.76)excellent (0.84)excellent (0.81)good (0.70)
Alignments (obtained from PredictProtein.org)
   There were no alignments for this protein in the data base. The BLAST search was initialized and should be ready in a few hours.
Predictions (obtained from PredictProtein.org)
   There were no predictions for this protein in the data base. The prediction was initialized and should be ready in a few hours.
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