Part:BBa_K1159108

N-TEV-Protease_36AALinker_PIF6 (Second part for a light-switchable TEV-Protease) in RFC[25]

This parts encodes a fusion protein consisting the N-terminal half of the TEV Protease as its terminus and first 100 amino acids of PIF6 as its C-terminus. PIF6 interacts with PhyB in the presence of red light. This reaction is reversible by far-red light. Both parts are seperated by a 36 amino acids long flexible linker. This part is flanked by RFC[25] pre- and suffix to allow further protein fusions.

By expressing this part together with its interaction partner BBa_K1159106 that contains the part, which interacts with PIF6, and the C-terminal half of the splitted TEV Protease, they form together a light-switchable TEV Protease system which can be activated by red light and inactivated by far-red light.

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 SapI.rc site found at 316
  

Protein data table for BioBrick BBa_ automatically created by the BioBrick-AutoAnnotator version 1.0
Nucleotide sequence in RFC 25, so ATGGCCGGC and ACCGGT were added (in italics) to the 5' and 3' ends: (underlined part encodes the protein)  ATGGCCGGCGGAGAAAGC ... CATGTTGACACCGGT  ORF from nucleotide position -8 to 777 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below) 1  101  201  MAGGESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKHLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQK LKFREPQREERICLVTTNFQTTGGGSAGGSGSGSSGGSSGASGTGTAGGTGSGSGTGSGTGMFLPTDYCCRLSDQEYMELVFENGQILAKGQRSNVSLHN QRTKSIMDLYEAEYNEDFMKSIIHGGGGAITNLGDTQVVPQSHVAAAHETNMLESNKHVDTG*
Sequence features: (with their position in the amino acid sequence, see the list of supported features) Factor Xa cleavage site:  80 to 83 RFC25 scar (shown in bold):  122 to 123, 160 to 161
Amino acid composition: Ala (A) 10 (3.8%) Arg (R) 11 (4.2%) Asn (N) 14 (5.3%) Asp (D) 11 (4.2%) Cys (C) 4 (1.5%) Gln (Q) 12 (4.6%) Glu (E) 13 (5.0%) Gly (G) 38 (14.5%) His (H) 10 (3.8%) Ile (I) 15 (5.7%) Leu (L) 19 (7.3%) Lys (K) 11 (4.2%) Met (M) 8 (3.1%) Phe (F) 12 (4.6%) Pro (P) 10 (3.8%) Ser (S) 24 (9.2%) Thr (T) 24 (9.2%) Trp (W) 0 (0.0%) Tyr (Y) 6 (2.3%) Val (V) 10 (3.8%)
Amino acid counting Total number: 262 Positively charged (Arg+Lys): 22 (8.4%) Negatively charged (Asp+Glu): 24 (9.2%) Aromatic (Phe+His+Try+Tyr): 28 (10.7%)	Biochemical parameters Atomic composition: C1209H1902N352O391S12 Molecular mass [Da]: 28009.2 Theoretical pI: 6.55 Extinction coefficient at 280 nm [M-1 cm-1]: 8940 / 9190 (all Cys red/ox)
Plot for hydrophobicity, charge, predicted secondary structure, solvent accessability, transmembrane helices and disulfid bridges
Codon usage Organism: E. coli B. subtilis S. cerevisiae A. thaliana P. patens Mammals Codon quality (CAI): good (0.73) good (0.72) good (0.69) good (0.74) excellent (0.82) good (0.69)
Alignments (obtained from PredictProtein.org) SwissProt: P04517 (49% identity on 239 AAs), P31999 (36% identity on 222 AAs) TrEML: Q000U0 (49% identity on 239 AAs), A4UQV2 (48% identity on 239 AAs) PDB: 1lvm (65% identity on 178 AAs), 1q31 (65% identity on 175 AAs)
Predictions (obtained from PredictProtein.org)
Subcellular Localization (reliability in brackets) Archaea: secreted (100%) Bacteria: secreted (56%) Eukarya: cytosol (20%)	Gene Ontology (reliability in brackets) Molecular Function Ontology: GO:0005515 (70%), GO:0043621 (13%) Biological Process Ontology: GO:0006508 (32%), GO:0006410 (26%)
Predicted features: Disulfid bridges:  - Transmembrane helices:  -
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