Coding

Part:BBa_K1150005

Designed by: Freiburg 2013   Group: iGEM13_Freiburg   (2013-09-16)

PIF6


PIF6
Function Phytochrome interaction factor
Use in Mammalian cells
RFC standard RFC 25
Backbone pSB1C3
Organism Arabidopsis thaliana
Source AG Weber, Freiburg
Submitted by [http://2013.igem.org/Team:Freiburg Freiburg 2013]

The Phytochrome Interaction Factor 6 (PIF6) is derived from Arabidopsis thaliana.

Illumination with red light (660 nm wavelength) leads to binding of Phytochrome Interaction Factor 6 (PIF6) to Phytochrome B. The interaction can be abolished by illumination with far-red light (740 nm) wavelength.

A framework using the unique properties of PhyB and PIF6 was designed to enable the recruitment of different effector proteins to any desired gene locus. The system was designed to work as described: Fusion proteins linking dCas9 to PIF6 were designed, as well as fusion proteins linking the interaction partner of PIF6 – PhyB – to several effector molecules such as VP16 or KRAB.


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_K1150005 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)
 ATGGCCGGCATGATGTTC ... CATGTTGACACCGGT
 ORF from nucleotide position -8 to 306 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)
1 
101 		MAGMMFLPTDYCCRLSDQEYMELVFENGQILAKGQRSNVSLHNQRTKSIMDLYEAEYNEDFMKSIIHGGGGAITNLGDTQVVPQSHVAAAHETNMLESNK
HVDTG*
Sequence features: (with their position in the amino acid sequence, see the list of supported features)
None of the supported features appeared in the sequence
Amino acid composition:
Ala (A)7 (6.7%)
Arg (R)3 (2.9%)
Asn (N)7 (6.7%)
Asp (D)6 (5.7%)
Cys (C)2 (1.9%)
Gln (Q)6 (5.7%)
Glu (E)8 (7.6%)
Gly (G)9 (8.6%)
His (H)5 (4.8%)
Ile (I)5 (4.8%)
Leu (L)8 (7.6%)
Lys (K)4 (3.8%)
Met (M)7 (6.7%)
Phe (F)3 (2.9%)
Pro (P)2 (1.9%)
Ser (S)7 (6.7%)
Thr (T)6 (5.7%)
Trp (W)0 (0.0%)
Tyr (Y)4 (3.8%)
Val (V)6 (5.7%)
Amino acid counting
Total number:105
Positively charged (Arg+Lys):7 (6.7%)
Negatively charged (Asp+Glu):14 (13.3%)
Aromatic (Phe+His+Try+Tyr):12 (11.4%)
Biochemical parameters
Atomic composition:C500H783N141O164S9
Molecular mass [Da]:11682.1
Theoretical pI:4.98
Extinction coefficient at 280 nm [M-1 cm-1]:5960 / 6085 (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 (0.76)good (0.76)good (0.71)good (0.78)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.
If you have any questions, comments or suggestions, please leave us a comment.


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Categories
Parameters
None