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Coding

Part:BBa_K801060:Design

Designed by: Lara Kuntz   Group: iGEM12_TU_Munich   (2012-09-21)
Revision as of 22:11, 7 September 2013 by ChristopherW (Talk | contribs)

(+)-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. coliB. subtilisS. cerevisiaeA. thalianaP. patensMammals
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.
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var flot_plot_options = []; flot_plot_options[0] = { grid: { borderWidth: { top: 0, right: 0, bottom: 0, left: 0 } }, legend: {show: false}, xaxes: [{ min: 0, max: 200, ticks: [[0.5, '1'], [24.5, '25'], [49.5, '50'], [74.5, '75'], [99.5, '100'], [124.5, '125'], [149.5, '150'], [174.5, '175'], [199.5, '200']], tickLength: -5 }], yaxes: [{ ticks: [[0, '0'], [4.5,'hydro-
phobic  '], [-4.5,'hydro-
philic  ']], min: -4.5, max: +4.5, font: { size: 12, lineHeight: 14, style: 'italic', weight: 'bold', family: 'sans-serif', variant: 'small-caps', color: 'rgba(100,149,237,1)' } }, { ticks: [[0, ''], [1,'positive
 charge'], [-1,'negative
 charge']], position: 'right', min: -1, max: 1, font: { size: 12, lineHeight: 14, style: 'italic', weight: 'bold', family: 'sans-serif', variant: 'small-caps', color: 'rgba(255,99,71,1)' } }]};var number_of_plots = Math.ceil(amino_sequence.length/200);for ( plot_num = 1 ; plot_num < number_of_plots ; plot_num ++){flot_plot_options[plot_num] = $.extend(true, {} ,flot_plot_options[0]);flot_plot_options[plot_num].xaxes = [{ min: plot_num*200, max: (plot_num + 1)*200, ticks: [ [plot_num*200 + 0.5, (plot_num*200 + 1).toString()], [plot_num*200 + 24.5, (plot_num*200 + 25).toString()], [plot_num*200 + 49.5, (plot_num*200 + 50).toString()], [plot_num*200 + 74.5, (plot_num*200 + 75).toString()], [plot_num*200 + 99.5, (plot_num*200 + 100).toString()], [plot_num*200 + 124.5, (plot_num*200 + 125).toString()], [plot_num*200 + 149.5, (plot_num*200 + 150).toString()], [plot_num*200 + 174.5, (plot_num*200 + 175).toString()], [plot_num*200 + 199.5, (plot_num*200 + 200).toString()] ], tickLength: -5 }];}; alert("Here"); function show_or_hide_plot(){ try { if( $('#hydrophobicity_charge_button').val() =='Show' ){ $('#hydrophobicity_charge_container').css('display','block'); for (plot_num = 0 ; plot_num < number_of_plots ; plot_num ++){ $.plot('#hydrophobicity_charge_placeholder'+ plot_num.toString(), [{ color: 'rgba(100,149,237,1)', data: hydrophobicity_datapoints, label: 'Hydrophobicity', lines: { show: true, fill: true, fillColor: 'rgba(100,149,237,0.1)' }, yaxis: 1 }, { color: 'rgba(255,99,71,1)', data: charge_datapoints, label: 'Charge', lines: { show: true, fill: true, fillColor: 'rgba(255,99,71,0.1)' }, yaxis: 2 }], flot_plot_options[plot_num] ); } $('#hydrophobicity_charge_button').val('Hide'); $('#hydrophobicity_charge_explanation').text(': Moving average over 5 amino acids for hydrophobicity (blue) and charge (red)'); } else{ $('#hydrophobicity_charge_container').css('display','none'); $('#hydrophobicity_charge_button').val('Show'); $('#hydrophobicity_charge_explanation').text(''); } } catch(err){ txt='There was an error with the button controlling the visibility of the plot. '; txt=txt+'The originating error is: ' + err + ' '; alert(txt); }}


Keywords: limonene synthase, limonene


Abbreviations:

Design Notes

Related BioBrick:

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]