Part:BBa_K1159316

SERK-Receptor TMD with C-terminal GFP and N-terminal Strep-tag II TEV-site linker in RFC[25]

This part codes for the Transmembrane Domain of SERK Receptor from Physcomitrella patens with a C-terminal (intracellular) GFPmut1 fusion and a N-terminal fusion to a linker containing Strep-tag II and a TEV cleavage site; SERK TMD and GFPmut1 are seperated by a middle long linker. This part is flanked by RFC[25] pre- and suffix for further protein fusions.

Note: A signal peptide is needed for membrane integration.

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 8
Illegal BsaI.rc site found at 926

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)
ATGGCCGGCAGCGCTTGG ... CTCTACAAAACCGGT
ORF from nucleotide position -8 to 1002 (excluding stop-codon)

Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)

1
101
201
301

MAGSAWSHPQFEKGENLYFQSGTGPGQPPFPPPPPFTPPPPQTPNGASGENSTGAIAGGVAAGAALLFAAPAIGFAWWRRRRPIEATGGGSGGGSGTGSK
GEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLSPDNHYLST
QSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKTG*

Sequence features: (with their position in the amino acid sequence, see the list of supported features)

	Strep-tag II:	6 to 13
	TEV cleavage site:	15 to 21
	RFC25 scar (shown in bold):	23 to 24, 87 to 88, 97 to 98

Amino acid composition:

Ala (A)	22 (6.5%)
Arg (R)	10 (3.0%)
Asn (N)	16 (4.7%)
Asp (D)	18 (5.3%)

Cys (C)	2 (0.6%)
Gln (Q)	12 (3.6%)
Glu (E)	20 (5.9%)
Gly (G)	43 (12.8%)

His (H)	11 (3.3%)
Ile (I)	15 (4.5%)
Leu (L)	22 (6.5%)
Lys (K)	21 (6.2%)

Met (M)	6 (1.8%)
Phe (F)	18 (5.3%)
Pro (P)	26 (7.7%)
Ser (S)	18 (5.3%)

Thr (T)	23 (6.8%)
Trp (W)	4 (1.2%)
Tyr (Y)	12 (3.6%)
Val (V)	18 (5.3%)

Amino acid counting

	Total number:	337
	Positively charged (Arg+Lys):	31 (9.2%)
	Negatively charged (Asp+Glu):	38 (11.3%)
	Aromatic (Phe+His+Try+Tyr):	45 (13.4%)

Biochemical parameters

	Atomic composition:	C₁₆₃₈H₂₄₈₈N₄₄₂O₄₉₅S₈
	Molecular mass [Da]:	36548.9
	Theoretical pI:	5.88
	Extinction coefficient at 280 nm [M^-1 cm^-1]:	39880 / 40005 (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 (0.75)	good (0.76)	excellent (0.84)	excellent (0.81)	good (0.71)

Alignments (obtained from PredictProtein.org)

	SwissProt:	P42212 (98% identity on 237 AAs), Q9U6Y3 (25% identity on 217 AAs)
	TrEML:	B6UPG7 (98% identity on 237 AAs), Q71RY9 (98% identity on 237 AAs), Q8GHE2 (98% identity on 237 AAs)
	PDB:	1emm (97% identity on 221 AAs), 1gfl (97% identity on 230 AAs)

Predictions (obtained from PredictProtein.org)

Subcellular Localization (reliability in brackets)

	Archaea:	secreted (100%)
	Bacteria:	secreted (77%)
	Eukarya:	cytosol (47%)

Gene Ontology (reliability in brackets)

	Molecular Function Ontology:	-
	Biological Process Ontology:	GO:0018298 (44%), GO:0008218 (31%)

Predicted features:

	Disulfid bridges:	-
	Transmembrane helices:	57 to 74 going inwards

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