Part:BBa_K801060:Design

(+)-Limonene synthase 1 with Strep-tag and yeast consensus sequence.

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 1683
  Illegal SapI.rc site found at 107
  

Protein data table for BioBrick BBa_K801060 automatically created by the BioBrick-AutoAnnotator version 1.0
Nucleotide sequence in RFC 10: (underlined part encodes the protein)  GTACACAATGCGTCGT ... TTCGAAAAATAA  ORF from nucleotide position 8 to 1705 (excluding stop-codon)
Amino acid sequence: (RFC25 scars in shown in bold, other sequence features underlined; both given below) 1  101  201  301  401  501  MRRSANYQPSIWDHDFLQSLNSNYTDEAYKRRAEELRGKVKIAIKDVIEPLDQLDLIDNLQRLGLAHRFETEIRNILNNIYNNNKDYNWRKENLYATSLE FRLLRQHGYPVSQEVFNGFKDDQGGFICDDFKGILSLHEASYYSLEGESIMEEAWQFTSKHLKEVMISKNMEEDVFVAEQAKRALELPLHWKVPMLEARW FIHIYERREDKNHLLLELAKMEFNTLQAIYQEELKEISGWWKDTGLGEKLSFARNRLVASFLWSMGIAFEPQFAYCRRVLTISIALITVIDDIYDVYGTL DELEIFTDAVERWDINYALKHLPGYMKMCFLALYNFVNEFAYYVLKQQDFDLLLSIKNAWLGLIQAYLVEAKWYHSKYTPKLEEYLENGLVSITGPLIIT ISYLSGTNPIIKKELEFLESNPDIVHWSSKIFRLQDDLGTSSDEIQRGDVPKSIQCYMHETGASEEVARQHIKDMMRQMWKKVNAYTADKDSPLTGTTTE FLLNLVRMSHFMYLHGDGHGVQNQETIDVGFTLLFQPIPLEDKHMAFTASPGTKGTGAWSHPQFEK*
Sequence features: (with their position in the amino acid sequence, see the list of supported features) RFC25 scar (shown in bold):  556 to 557 Strep-tag II:  559 to 566
Amino acid composition: Ala (A) 33 (5.8%) Arg (R) 25 (4.4%) Asn (N) 27 (4.8%) Asp (D) 34 (6.0%) Cys (C) 4 (0.7%) Gln (Q) 24 (4.2%) Glu (E) 48 (8.5%) Gly (G) 29 (5.1%) His (H) 18 (3.2%) Ile (I) 39 (6.9%) Leu (L) 64 (11.3%) Lys (K) 36 (6.4%) Met (M) 16 (2.8%) Phe (F) 28 (4.9%) Pro (P) 17 (3.0%) Ser (S) 33 (5.8%) Thr (T) 26 (4.6%) Trp (W) 14 (2.5%) Tyr (Y) 27 (4.8%) Val (V) 24 (4.2%)
Amino acid counting Total number: 566 Positively charged (Arg+Lys): 61 (10.8%) Negatively charged (Asp+Glu): 82 (14.5%) Aromatic (Phe+His+Try+Tyr): 87 (15.4%)	Biochemical parameters Atomic composition: C3002H4586N778O868S20 Molecular mass [Da]: 66105.3 Theoretical pI: 5.38 Extinction coefficient at 280 nm [M-1 cm-1]: 117230 / 117480 (all Cys red/ox)
Hydrophobicity and charge plot
Codon usage Organism: E. coli B. subtilis S. cerevisiae A. thaliana P. patens Mammals Codon quality (CAI): good (0.71) good (0.75) good (0.69) good (0.78) excellent (0.80) good (0.68)
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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Keywords: limonene synthase, limonene

Abbreviations:

Design Notes

Related BioBrick:

• BBa_K801061: (+)-Limonene synthase 1 coding region
• BBa_K801062: (+)-limonene synthase 1 expression cassette for yeast
• BBa_K801063: (+)-limonene synthase 1 expression cassette for yeast
• BBa_K801064: (+)-limonene synthase 1 expression cassette for yeast
• BBa_K801065: (+)-limonene synthase 1 with Strep-Tag
• BBa_K801066: (+)-limonene synthase 1 with Yeast Consensus Sequence for improved expression

Quality control measures:

• Biobrick was sequenced

Backbone:

• Backbone name: pSB1C3
• Resistance: Cp
• Copynumber: high

Protein coding:

• BioBrick encodes (+)-limonene synthase
• UniProt entry: [http://www.uniprot.org/uniprot/Q8L5K3 Q8L5K3]

Enzymatic activity:

• E.C. Number: [http://enzyme.expasy.org/EC/4.2.3.20 4.2.3.20]

Cytotoxicity:

• see toxicity assay

Safety notes:

Intellectual property:

Corresponding part author/authors:

Source

Source:

• BioBrick: BBa_I742111

• Forward Primer: TUM12-LIMS1-fw-cons: 5'- GGATCCTCTAGAGTACACAATGCGTCGTTCAGCAAAC - 3'
• Reverse Primer: TUM12-LIMS1-rv: 5'- GGATCCACCGGTGCCTTTGGTGCCAGGAGA - 3'

Organism:

• Gene sequence derived from Citrus limon

References

Literature references:

• [http://www.ncbi.nlm.nih.gov/pubmed/12084056Pubmed: Lücker, J., El Tamer, M. K., Schwab, W., Verstappen, F. W. A., van der Plas, L. H. W., Bouwmeester, H. J., and Verhoeven, H. A., 2002: Monoterpene biosynthesis in lemon (Citrus limon). cDNA isolation and functional analysis of four monoterpene synthases. Eur J Biochem, 269(13):3160–71.]

Database references:

• [http://www.ncbi.nlm.nih.gov/nuccore/21435702 GenBank: source of the sequence]
• [http://www.uniprot.org/uniprot/Q8L5K3 Uniprot: (R)-limonene synthase 1]