Difference between revisions of "Part:BBa K2332315"
 
(8 intermediate revisions by the same user not shown)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K2332315 short</partinfo>
 
<partinfo>BBa_K2332315 short</partinfo>
Line 15: Line 14:
 
|-
 
|-
 
|'''RFC standard'''
 
|'''RFC standard'''
−
|[https://parts.igem.org/Help:Assembly_standard_10 RFC10], [https://parts.igem.org/Help:Assembly_standard_23 RFC23] & [https://parts.igem.org/Help:Assembly_standard_25 RFC25] compatible  
+
|[https://parts.igem.org/Help:Assembly_standard_10 RFC10], [https://parts.igem.org/Help:Standards/Assembly/RFC12 RFC12], [https://parts.igem.org/Help:Assembly_standard_23 RFC23] & [https://parts.igem.org/Help:Assembly_standard_25 RFC25] compatible  
 
|-
 
|-
 
|'''Backbone'''
 
|'''Backbone'''
Line 22: Line 21:
 
|'''Submitted by'''
 
|'''Submitted by'''
 
|[http://2017.igem.org/Team:UCL UCL iGEM 2017]
 
|[http://2017.igem.org/Team:UCL UCL iGEM 2017]
−
This coding region translates a transmembrane domain which is destined to go to the plasma membrane via the secretory pathway. While doing so it delivers any 'cytosolic domain insert' that has been cloned into this BioBrick in a plug-and-play via the KpnI-BamHI restriction sites.
+
|}
 +
 
 +
 
 +
This coding region translates a transmembrane domain which is destined to go to the plasma membrane via the secretory pathway. While doing so it delivers any 'cytosolic domain insert' that has been cloned into this BioBrick in a plug-and-play fashion (via the KpnI-BamHI restriction sites) to the plasma membrane. The delivered protein is localised to the plasma membrane but activation of the photocleavable linker PhoCl with 400 nm, violet light allows irreversible breakage of the connection with the PM and releases the 'cytosolic domain insert' into the cytoplasm.
 +
For characterisation purposes mCherry has been chosen 'cytosolic domain insert' in this BioBrick.
  
  
Line 35: Line 38:
  
  
−
<!-- -->
+
===Sequence and Features===
−
<span class='h3bb'>Sequence and Features</span>
+
 
<partinfo>BBa_K2332315 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2332315 SequenceAndFeatures</partinfo>
  
  
−
<!-- Uncomment this to enable Functional Parameter display
+
 
 
===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K2332315 parameters</partinfo>
 
<partinfo>BBa_K2332315 parameters</partinfo>
−
<!-- -->
+
 
 +
<html><!--- Please copy this table containing parameters for BBa_ at the end of the parametrs section ahead of the references. ---><style type="text/css">table#AutoAnnotator {border:1px solid black; width:100%; border-collapse:collapse;} th#AutoAnnotatorHeader { border:1px solid black; width:100%; background-color: rgb(221, 221, 221);} td.AutoAnnotator1col { width:100%; border:1px solid black; } span.AutoAnnotatorSequence { font-family:'Courier New', Arial; } td.AutoAnnotatorSeqNum { text-align:right; width:2%; } td.AutoAnnotatorSeqSeq { width:98% } td.AutoAnnotatorSeqFeat1 { width:3% } td.AutoAnnotatorSeqFeat2a { width:27% } td.AutoAnnotatorSeqFeat2b { width:97% } td.AutoAnnotatorSeqFeat3 { width:70% } table.AutoAnnotatorNoBorder { border:0px; width:100%; border-collapse:collapse; } table.AutoAnnotatorWithBorder { border:1px solid black; width:100%; border-collapse:collapse; } td.AutoAnnotatorOuterAmino { border:0px solid black; width:20% } td.AutoAnnotatorInnerAmino { border:1px solid black; width:50% } td.AutoAnnotatorAminoCountingOuter { border:1px solid black; width:40%;  } td.AutoAnnotatorBiochemParOuter { border:1px solid black; width:60%; } td.AutoAnnotatorAminoCountingInner1 { width: 7.5% } td.AutoAnnotatorAminoCountingInner2 { width:62.5% } td.AutoAnnotatorAminoCountingInner3 { width:30% } td.AutoAnnotatorBiochemParInner1 { width: 5% } td.AutoAnnotatorBiochemParInner2 { width:55% } td.AutoAnnotatorBiochemParInner3 { width:40% } td.AutoAnnotatorCodonUsage1 { width: 3% } td.AutoAnnotatorCodonUsage2 { width:14.2% } td.AutoAnnotatorCodonUsage3 { width:13.8% } td.AutoAnnotatorAlignment1 { width: 3% } td.AutoAnnotatorAlignment2 { width: 10% } td.AutoAnnotatorAlignment3 { width: 87% } td.AutoAnnotatorLocalizationOuter {border:1px solid black; width:40%} td.AutoAnnotatorGOOuter {border:1px solid black; width:60%} td.AutoAnnotatorLocalization1 { width: 7.5% } td.AutoAnnotatorLocalization2 { width: 22.5% } td.AutoAnnotatorLocalization3 { width: 70% } td.AutoAnnotatorGO1 { width: 5% } td.AutoAnnotatorGO2 { width: 35% } td.AutoAnnotatorGO3 { width: 60% } td.AutoAnnotatorPredFeat1 { width:3% } td.AutoAnnotatorPredFeat2a { width:27% } td.AutoAnnotatorPredFeat3 { width:70% } div.AutoAnnotator_trans { position:absolute; background:rgb(11,140,143); background-color:rgba(11,140,143, 0.8); height:5px; top:100px; } div.AutoAnnotator_sec_helix { position:absolute; background:rgb(102,0,102); background-color:rgba(102,0,102, 0.8); height:5px; top:110px; } div.AutoAnnotator_sec_strand { position:absolute; background:rgb(245,170,26); background-color:rgba(245,170,26, 1); height:5px; top:110px; } div.AutoAnnotator_acc_buried { position:absolute; background:rgb(89,168,15); background-color:rgba(89,168,15, 0.8); height:5px; top:120px; } div.AutoAnnotator_acc_exposed { position:absolute; background:rgb(0, 0, 255); background-color:rgba(0, 0, 255, 0.8); height:5px; top:120px; } div.AutoAnnotator_dis { position:absolute; text-align:center; font-family:Arial,Helvetica,sans-serif; background:rgb(255, 200, 0); background-color:rgba(255, 200, 0, 1); height:16px; width:16px; top:80px; border-radius:50%; } </style><div id='AutoAnnotator_container_1508714402355'><table id="AutoAnnotator"><tr><!-- Time stamp in ms since 1/1/1970 1508714402355 --><th id="AutoAnnotatorHeader" colspan="2">Protein data table for BioBrick <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_<!------------------------Enter BioBrick number here------------------------>">BBa_<!------------------------Enter BioBrick number here------------------------></a> automatically created by the <a href="http://2013.igem.org/Team:TU-Munich/Results/AutoAnnotator">BioBrick-AutoAnnotator</a> version 1.0</th></tr><tr><td class="AutoAnnotator1col" colspan="2"><strong>Nucleotide sequence</strong> in <strong>RFC 10</strong>: (underlined part encodes the protein)<br><span class="AutoAnnotatorSequence">&nbsp;ACC<u>ATGCGGGCC&nbsp;...&nbsp;GTCGGATCC</u>TAG</span><br>&nbsp;<strong>ORF</strong> from nucleotide position 4 to 2490 (excluding stop-codon)</td></tr><tr><td class="AutoAnnotator1col" colspan="2"><strong>Amino acid sequence:</strong> (RFC 25 scars in shown in bold, other sequence features underlined; both given below)<br><span class="AutoAnnotatorSequence"><table class="AutoAnnotatorNoBorder"><tr><td class="AutoAnnotatorSeqNum">1&nbsp;<br>101&nbsp;<br>201&nbsp;<br>301&nbsp;<br>401&nbsp;<br>501&nbsp;<br>601&nbsp;<br>701&nbsp;<br>801&nbsp;</td><td class="AutoAnnotatorSeqSeq">MRAWIFFLLCLAGRALAA<b>TG</b>RKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQH<br>DFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADH<br>YQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK<u>DYK<u>DDDDK</u></u><b>TG</b>GGGGSGGGGSPSIATGMVGALLLLLVVALGIGL<br>FMRRRHIVRKRGGGGSGGGGSGSVIPDYFKQSFPEGYSWERSMTYEDGGICIATNDITMEGDSFINKIHFKGTNFPPNGPVMQKRTVGWEASTEKMYERD<br>GVLKGDVKMKLLLKGGGHYRCDYRTTYKVKQKPVKLPDYHFVDHRIEILSHDKDYNKVKLYEHAVARNSTDSMDELYKGGSGGMVSKGEETITSVIKPDM<br>KNKLRMEGNVNGHAFVIEGEGSGKPFEGIQTIDLEVKEGAPLPFAYDILTTAFHYGNRVFTKYPRGGGGTGGGGSGGGGSVSKGEEDNMAIIKEFMRFKV<br>HMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDG<br>EFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYER<br>AEGRHS<b>TG</b>GMDELYKGGGGS<u>PKKKRKV</u>GS*</td></tr></table></span></td></tr><tr><td class="AutoAnnotator1col" colspan="2"><strong>Sequence features:</strong> (with their position in the amino acid sequence, see the <a href="http://2013.igem.org/Team:TU-Munich/Results/Software/FeatureList">list of supported features</a>)<table class="AutoAnnotatorNoBorder"><tr><td class="AutoAnnotatorSeqFeat1"></td><td class="AutoAnnotatorSeqFeat2a">RFC25 scar (shown in bold):&nbsp;</td><td class="AutoAnnotatorSeqFeat3">19 to 20, 266 to 267, 807 to 808</td></tr><tr><td class="AutoAnnotatorSeqFeat1"></td><td class="AutoAnnotatorSeqFeat2a">Flag-tag:&nbsp;</td><td class="AutoAnnotatorSeqFeat3">258 to 265</td></tr><tr><td class="AutoAnnotatorSeqFeat1"></td><td class="AutoAnnotatorSeqFeat2a">Enterokinase cleavage site:&nbsp;</td><td class="AutoAnnotatorSeqFeat3">261 to 265</td></tr><tr><td class="AutoAnnotatorSeqFeat1"></td><td class="AutoAnnotatorSeqFeat2a">SV40 nuclear localization sequence:&nbsp;</td><td class="AutoAnnotatorSeqFeat3">821 to 827</td></tr></table></td></tr><tr><td class="AutoAnnotator1col" colspan="2"><strong>Amino acid composition:</strong><table class="AutoAnnotatorNoBorder"><tr><td class="AutoAnnotatorOuterAmino"><table class="AutoAnnotatorWithBorder"><tr><td class="AutoAnnotatorInnerAmino">Ala (A)</td><td class="AutoAnnotatorInnerAmino">36 (4.3%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Arg (R)</td><td class="AutoAnnotatorInnerAmino">33 (4.0%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Asn (N)</td><td class="AutoAnnotatorInnerAmino">30 (3.6%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Asp (D)</td><td class="AutoAnnotatorInnerAmino">53 (6.4%)</td></tr></table></td><td class="AutoAnnotatorOuterAmino"><table class="AutoAnnotatorWithBorder"><tr><td class="AutoAnnotatorInnerAmino">Cys (C)</td><td class="AutoAnnotatorInnerAmino">5 (0.6%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Gln (Q)</td><td class="AutoAnnotatorInnerAmino">20 (2.4%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Glu (E)</td><td class="AutoAnnotatorInnerAmino">58 (7.0%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Gly (G)</td><td class="AutoAnnotatorInnerAmino">114 (13.8%)</td></tr></table></td><td class="AutoAnnotatorOuterAmino"><table class="AutoAnnotatorWithBorder"><tr><td class="AutoAnnotatorInnerAmino">His (H)</td><td class="AutoAnnotatorInnerAmino">25 (3.0%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Ile (I)</td><td class="AutoAnnotatorInnerAmino">40 (4.8%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Leu (L)</td><td class="AutoAnnotatorInnerAmino">55 (6.6%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Lys (K)</td><td class="AutoAnnotatorInnerAmino">75 (9.0%)</td></tr></table></td><td class="AutoAnnotatorOuterAmino"><table class="AutoAnnotatorWithBorder"><tr><td class="AutoAnnotatorInnerAmino">Met (M)</td><td class="AutoAnnotatorInnerAmino">26 (3.1%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Phe (F)</td><td class="AutoAnnotatorInnerAmino">37 (4.5%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Pro (P)</td><td class="AutoAnnotatorInnerAmino">36 (4.3%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Ser (S)</td><td class="AutoAnnotatorInnerAmino">42 (5.1%)</td></tr></table></td><td class="AutoAnnotatorOuterAmino"><table class="AutoAnnotatorWithBorder"><tr><td class="AutoAnnotatorInnerAmino">Thr (T)</td><td class="AutoAnnotatorInnerAmino">46 (5.5%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Trp (W)</td><td class="AutoAnnotatorInnerAmino">7 (0.8%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Tyr (Y)</td><td class="AutoAnnotatorInnerAmino">38 (4.6%)</td></tr><tr><td class="AutoAnnotatorInnerAmino">Val (V)</td><td class="AutoAnnotatorInnerAmino">53 (6.4%)</td></tr></table></td></tr></table></td></tr><tr><td class="AutoAnnotatorAminoCountingOuter"><strong>Amino acid counting</strong><table class="AutoAnnotatorNoBorder"><tr><td class="AutoAnnotatorAminoCountingInner1"></td><td class="AutoAnnotatorAminoCountingInner2">Total number:</td><td class="AutoAnnotatorAminoCountingInner3">829</td></tr><tr><td class="AutoAnnotatorAminoCountingInner1"></td><td class="AutoAnnotatorAminoCountingInner2">Positively charged (Arg+Lys):</td><td class="AutoAnnotatorAminoCountingInner3">108 (13.0%)</td></tr><tr><td class="AutoAnnotatorAminoCountingInner1"></td><td class="AutoAnnotatorAminoCountingInner2">Negatively charged (Asp+Glu):</td><td class="AutoAnnotatorAminoCountingInner3">111 (13.4%)</td></tr><tr><td class="AutoAnnotatorAminoCountingInner1"></td><td class="AutoAnnotatorAminoCountingInner2">Aromatic (Phe+His+Try+Tyr):</td><td class="AutoAnnotatorAminoCountingInner3">107 (12.9%)</td></tr></table></td><td class="AutoAnnotatorBiochemParOuter"><strong>Biochemical parameters</strong><table class="AutoAnnotatorNoBorder"><tr><td class="AutoAnnotatorBiochemParInner1"></td><td class="AutoAnnotatorBiochemParInner2">Atomic composition:</td><td class="AutoAnnotatorBiochemParInner3">C<sub>4078</sub>H<sub>6316</sub>N<sub>1110</sub>O<sub>1228</sub>S<sub>31</sub></td></tr><tr><td class="AutoAnnotatorBiochemParInner1"></td><td class="AutoAnnotatorBiochemParInner2">Molecular mass [Da]:</td><td class="AutoAnnotatorBiochemParInner3">91535.6</td></tr><tr><td class="AutoAnnotatorBiochemParInner1"></td><td class="AutoAnnotatorBiochemParInner2">Theoretical pI:</td><td class="AutoAnnotatorBiochemParInner3">6.82</td></tr><tr><td class="AutoAnnotatorBiochemParInner1"></td><td class="AutoAnnotatorBiochemParInner2">Extinction coefficient at 280 nm [M<sup>-1</sup> cm<sup>-1</sup>]:</td><td class="AutoAnnotatorBiochemParInner3">95120 / 95433 (all Cys red/ox)</td></tr></table></td></tr><tr><td class="AutoAnnotator1col" colspan="2"><strong>Plot for hydrophobicity, charge, predicted secondary structure, solvent accessability, transmembrane helices and disulfid bridges</strong>&nbsp;<input type='button' id='hydrophobicity_charge_button' onclick='show_or_hide_plot_1508714402355()' value='Show'><span id="hydrophobicity_charge_explanation"></span><div id="hydrophobicity_charge_container" style='display:none'><div id="hydrophobicity_charge_placeholder0" style="width:100%;height:150px"></div><div id="hydrophobicity_charge_placeholder1" style="width:100%;height:150px"></div><div id="hydrophobicity_charge_placeholder2" style="width:100%;height:150px"></div><div id="hydrophobicity_charge_placeholder3" style="width:100%;height:150px"></div><div id="hydrophobicity_charge_placeholder4" style="width:100%;height:150px"></div></div></td></tr><tr><td class="AutoAnnotator1col" colspan="2"><strong>Codon usage</strong><table class="AutoAnnotatorNoBorder"><tr><td class="AutoAnnotatorCodonUsage1"></td><td class="AutoAnnotatorCodonUsage2">Organism:</td><td class="AutoAnnotatorCodonUsage3"><i>E. coli</i></td><td class="AutoAnnotatorCodonUsage3"><i>B. subtilis</i></td><td class="AutoAnnotatorCodonUsage3"><i>S. cerevisiae</i></td><td class="AutoAnnotatorCodonUsage3"><i>A. thaliana</i></td><td class="AutoAnnotatorCodonUsage3"><i>P. patens</i></td><td class="AutoAnnotatorCodonUsage3">Mammals</td></tr><tr><td class="AutoAnnotatorCodonUsage1"></td><td class="AutoAnnotatorCodonUsage2">Codon quality (<a href="http://en.wikipedia.org/wiki/Codon_Adaptation_Index">CAI</a>):</td><td class="AutoAnnotatorCodonUsage3">good (0.69)</td><td class="AutoAnnotatorCodonUsage3">good (0.68)</td><td class="AutoAnnotatorCodonUsage3">acceptable (0.57)</td><td class="AutoAnnotatorCodonUsage3">good (0.67)</td><td class="AutoAnnotatorCodonUsage3">excellent (0.85)</td><td class="AutoAnnotatorCodonUsage3">excellent (0.85)</td></tr></table></td></tr><tr><td class="AutoAnnotator1col" colspan="2"><strong>Alignments</strong> (obtained from <a href='http://predictprotein.org'>PredictProtein.org</a>)<br>&nbsp;&nbsp;&nbsp;There were no alignments for this protein in the data base. The BLAST search was initialized and should be ready in a few hours.</td></tr><tr><th id='AutoAnnotatorHeader' colspan="2"><strong>Predictions</strong> (obtained from <a href='http://predictprotein.org'>PredictProtein.org</a>)</th></tr><tr><td class="AutoAnnotator1col" colspan="2">&nbsp;&nbsp;&nbsp;There were no predictions for this protein in the data base. The prediction was initialized and should be ready in a few hours.</td><tr><td class="AutoAnnotator1col" colspan="2"> The BioBrick-AutoAnnotator was created by <a href="http://2013.igem.org/Team:TU-Munich">TU-Munich 2013</a> iGEM team. For more information please see the <a href="http://2013.igem.org/Team:TU-Munich/Results/Software">documentation</a>.<br>If you have any questions, comments or suggestions, please leave us a <a href="http://2013.igem.org/Team:TU-Munich/Results/AutoAnnotator">comment</a>.</td></tr></table></div><br><!-- IMPORTANT: DON'T REMOVE THIS LINE, OTHERWISE NOT SUPPORTED FOR IE BEFORE 9 --><!--[if lte IE 8]><script language="javascript" type="text/javascript" src="http://2013.igem.org/Team:TU-Munich/excanvas.js"></script><![endif]--><script type='text/javascript' src='http://code.jquery.com/jquery-1.10.0.min.js'></script><script type='text/javascript' src='http://2013.igem.org/Team:TU-Munich/Flot.js?action=raw&ctype=text/js'></script><script>var jqAutoAnnotator = jQuery.noConflict(true);function show_or_hide_plot_1508714402355(){hydrophobicity_datapoints = [[2.5,0.56],[3.5,0.74],[4.5,2.20],[5.5,2.60],[6.5,3.54],[7.5,3.14],[8.5,3.34],[9.5,3.14],[10.5,2.30],[11.5,0.64],[12.5,0.50],[13.5,0.50],[14.5,0.50],[15.5,0.94],[16.5,1.70],[17.5,1.26],[18.5,-0.40],[19.5,-1.54],[20.5,-1.98],[21.5,-2.54],[22.5,-3.16],[23.5,-1.50],[24.5,-0.16],[25.5,-0.22],[26.5,0.40],[27.5,1.94],[28.5,2.02],[29.5,1.14],[30.5,2.18],[31.5,3.02],[32.5,3.02],[33.5,1.48],[34.5,2.56],[35.5,0.96],[36.5,0.12],[37.5,-1.42],[38.5,0.12],[39.5,-1.34],[40.5,-0.72],[41.5,-1.28],[42.5,-1.36],[43.5,-1.64],[44.5,-1.10],[45.5,-0.18],[46.5,0.30],[47.5,1.00],[48.5,-0.26],[49.5,-0.18],[50.5,-1.72],[51.5,-1.64],[52.5,-2.26],[53.5,-1.20],[54.5,-1.26],[55.5,-0.82],[56.5,-0.82],[57.5,-0.90],[58.5,-0.50],[59.5,-0.50],[60.5,0.52],[61.5,-0.18],[62.5,1.16],[63.5,1.30],[64.5,1.94],[65.5,1.04],[66.5,1.68],[67.5,1.04],[68.5,-0.64],[69.5,-0.38],[70.5,-0.56],[71.5,0.42],[72.5,0.18],[73.5,0.78],[74.5,-0.30],[75.5,-0.12],[76.5,-0.20],[77.5,0.96],[78.5,1.00],[79.5,1.18],[80.5,1.88],[81.5,1.04],[82.5,-0.06],[83.5,0.00],[84.5,0.98],[85.5,-0.28],[86.5,0.30],[87.5,1.12],[88.5,1.56],[89.5,-0.18],[90.5,0.26],[91.5,-0.56],[92.5,-1.82],[93.5,-2.82],[94.5,-1.54],[95.5,-2.06],[96.5,-2.44],[97.5,-2.38],[98.5,-2.44],[99.5,-2.26],[100.5,-0.92],[101.5,-1.00],[102.5,-0.52],[103.5,0.54],[104.5,0.36],[105.5,-0.52],[106.5,-0.44],[107.5,-0.36],[108.5,-0.98],[109.5,-0.52],[110.5,-0.90],[111.5,-0.90],[112.5,-1.72],[113.5,-1.60],[114.5,-1.54],[115.5,-0.28],[116.5,0.98],[117.5,1.10],[118.5,0.54],[119.5,-1.06],[120.5,-1.70],[121.5,-2.96],[122.5,-2.44],[123.5,-2.52],[124.5,-1.96],[125.5,-2.78],[126.5,-1.72],[127.5,-2.16],[128.5,-0.54],[129.5,-1.18],[130.5,0.28],[131.5,-0.78],[132.5,-0.16],[133.5,-1.70],[134.5,-1.06],[135.5,-0.86],[136.5,0.68],[137.5,0.06],[138.5,-0.14],[139.5,0.90],[140.5,-0.56],[141.5,-0.64],[142.5,-0.72],[143.5,0.10],[144.5,0.10],[145.5,0.10],[146.5,-0.10],[147.5,-0.10],[148.5,-0.72],[149.5,-2.32],[150.5,-1.70],[151.5,-2.96],[152.5,-1.28],[153.5,0.18],[154.5,0.80],[155.5,0.24],[156.5,0.16],[157.5,0.02],[158.5,-1.44],[159.5,-1.62],[160.5,-1.68],[161.5,-1.16],[162.5,-2.62],[163.5,-2.08],[164.5,-2.46],[165.5,-2.46],[166.5,-1.36],[167.5,-0.92],[168.5,0.14],[169.5,1.16],[170.5,2.22],[171.5,0.68],[172.5,0.16],[173.5,-1.44],[174.5,-2.60],[175.5,-3.66],[176.5,-3.04],[177.5,-1.36],[178.5,-1.44],[179.5,0.18],[180.5,0.18],[181.5,0.82],[182.5,-0.86],[183.5,0.82],[184.5,-0.92],[185.5,-0.86],[186.5,-2.12],[187.5,-0.44],[188.5,-2.04],[189.5,-1.84],[190.5,-1.28],[191.5,-0.74],[192.5,-0.80],[193.5,-0.80],[194.5,0.66],[195.5,1.10],[196.5,0.56],[197.5,-0.92],[198.5,-0.48],[199.5,-1.94],[200.5,-3.00],[201.5,-3.00],[202.5,-2.50],[203.5,-2.56],[204.5,-0.96],[205.5,-0.34],[206.5,-0.34],[207.5,-0.28],[208.5,-0.28],[209.5,-0.34],[210.5,0.50],[211.5,1.96],[212.5,1.72],[213.5,1.34],[214.5,-0.20],[215.5,-1.60],[216.5,-2.62],[217.5,-1.54],[218.5,-1.00],[219.5,-0.44],[220.5,-0.50],[221.5,-0.40],[222.5,-0.80],[223.5,0.12],[224.5,0.10],[225.5,0.02],[226.5,-0.52],[227.5,-1.20],[228.5,-2.66],[229.5,-3.20],[230.5,-3.20],[231.5,-3.40],[232.5,-3.78],[233.5,-3.72],[234.5,-2.64],[235.5,-1.02],[236.5,0.64],[237.5,2.10],[238.5,2.04],[239.5,2.22],[240.5,2.22],[241.5,1.32],[242.5,0.92],[243.5,1.98],[244.5,1.34],[245.5,1.40],[246.5,1.40],[247.5,0.40],[248.5,-0.04],[249.5,0.42],[250.5,-1.18],[251.5,-1.74],[252.5,-0.34],[253.5,-0.52],[254.5,-1.68],[255.5,-1.68],[256.5,-1.24],[257.5,-2.78],[258.5,-3.22],[259.5,-3.14],[260.5,-3.14],[261.5,-3.58],[262.5,-3.58],[263.5,-3.02],[264.5,-2.40],[265.5,-1.78],[266.5,-1.16],[267.5,-0.46],[268.5,-0.40],[269.5,-0.48],[270.5,-0.48],[271.5,-0.48],[272.5,-0.48],[273.5,-0.48],[274.5,-0.48],[275.5,-0.72],[276.5,-0.80],[277.5,0.18],[278.5,0.62],[279.5,0.64],[280.5,0.88],[281.5,1.42],[282.5,1.36],[283.5,0.92],[284.5,1.42],[285.5,2.26],[286.5,2.64],[287.5,2.56],[288.5,3.40],[289.5,3.80],[290.5,3.88],[291.5,3.96],[292.5,3.56],[293.5,3.56],[294.5,2.72],[295.5,2.78],[296.5,1.86],[297.5,2.26],[298.5,2.06],[299.5,2.52],[300.5,0.72],[301.5,-0.10],[302.5,-1.76],[303.5,-2.96],[304.5,-2.44],[305.5,-0.70],[306.5,-0.70],[307.5,-0.58],[308.5,-0.84],[309.5,-1.82],[310.5,-2.74],[311.5,-1.92],[312.5,-1.22],[313.5,-0.48],[314.5,-0.48],[315.5,-0.48],[316.5,-0.48],[317.5,-0.48],[318.5,-0.48],[319.5,-0.48],[320.5,-0.56],[321.5,0.36],[322.5,1.34],[323.5,1.18],[324.5,0.56],[325.5,0.46],[326.5,0.18],[327.5,-1.50],[328.5,-1.88],[329.5,-1.34],[330.5,-0.52],[331.5,-1.40],[332.5,-1.32],[333.5,-0.70],[334.5,-0.80],[335.5,-1.52],[336.5,-1.38],[337.5,-1.38],[338.5,-2.20],[339.5,-2.10],[340.5,-1.56],[341.5,-1.52],[342.5,-1.08],[343.5,-0.88],[344.5,-1.42],[345.5,-1.88],[346.5,-1.82],[347.5,-0.66],[348.5,0.54],[349.5,2.14],[350.5,2.58],[351.5,2.52],[352.5,0.92],[353.5,-0.28],[354.5,-0.28],[355.5,-0.78],[356.5,-0.26],[357.5,-0.26],[358.5,0.36],[359.5,-1.24],[360.5,-1.26],[361.5,-1.08],[362.5,0.52],[363.5,-0.10],[364.5,-0.18],[365.5,0.88],[366.5,-0.32],[367.5,-0.66],[368.5,-0.74],[369.5,-0.04],[370.5,-1.08],[371.5,-1.14],[372.5,-1.14],[373.5,-0.68],[374.5,-0.92],[375.5,-1.48],[376.5,-0.86],[377.5,-1.74],[378.5,-0.58],[379.5,0.12],[380.5,0.12],[381.5,-0.58],[382.5,-1.16],[383.5,-2.14],[384.5,-1.68],[385.5,-1.06],[386.5,-0.46],[387.5,-0.26],[388.5,0.24],[389.5,-0.76],[390.5,-0.82],[391.5,-1.34],[392.5,-1.42],[393.5,-1.40],[394.5,-1.50],[395.5,-2.06],[396.5,-2.26],[397.5,-2.18],[398.5,-2.64],[399.5,-1.54],[400.5,-0.08],[401.5,0.04],[402.5,0.66],[403.5,0.04],[404.5,0.04],[405.5,-1.50],[406.5,-0.34],[407.5,-1.04],[408.5,0.42],[409.5,0.34],[410.5,1.88],[411.5,0.72],[412.5,1.42],[413.5,0.58],[414.5,-0.26],[415.5,-1.66],[416.5,-1.14],[417.5,-1.96],[418.5,-1.38],[419.5,-2.00],[420.5,-1.62],[421.5,-2.26],[422.5,-1.50],[423.5,-2.14],[424.5,-1.70],[425.5,-2.22],[426.5,-0.48],[427.5,-1.12],[428.5,-1.68],[429.5,-2.20],[430.5,-1.74],[431.5,-1.74],[432.5,-1.74],[433.5,-0.28],[434.5,0.18],[435.5,-0.20],[436.5,-1.30],[437.5,-1.16],[438.5,-1.36],[439.5,-0.20],[440.5,-0.20],[441.5,-0.58],[442.5,-0.84],[443.5,-0.50],[444.5,-2.04],[445.5,-0.44],[446.5,0.96],[447.5,1.70],[448.5,0.16],[449.5,0.16],[450.5,-1.52],[451.5,-2.98],[452.5,-3.08],[453.5,-3.14],[454.5,-3.22],[455.5,-1.60],[456.5,-1.68],[457.5,-0.66],[458.5,-0.22],[459.5,-0.14],[460.5,-1.62],[461.5,-0.48],[462.5,-0.40],[463.5,0.22],[464.5,0.02],[465.5,-0.04],[466.5,-0.56],[467.5,-1.54],[468.5,-2.60],[469.5,-1.86],[470.5,-0.78],[471.5,-1.32],[472.5,-1.88],[473.5,-0.42],[474.5,-0.52],[475.5,-1.68],[476.5,-1.06],[477.5,-0.44],[478.5,-1.36],[479.5,-1.18],[480.5,-0.48],[481.5,-0.02],[482.5,0.90],[483.5,0.90],[484.5,0.20],[485.5,0.20],[486.5,-0.88],[487.5,-2.42],[488.5,-2.40],[489.5,-0.72],[490.5,-0.78],[491.5,-0.24],[492.5,1.30],[493.5,2.34],[494.5,0.66],[495.5,0.48],[496.5,-0.06],[497.5,-0.52],[498.5,-2.20],[499.5,-2.12],[500.5,-2.58],[501.5,-1.12],[502.5,-2.40],[503.5,-1.24],[504.5,-1.24],[505.5,-0.54],[506.5,-2.00],[507.5,-0.26],[508.5,-1.34],[509.5,-0.72],[510.5,-1.28],[511.5,-0.22],[512.5,-0.50],[513.5,1.04],[514.5,2.02],[515.5,1.96],[516.5,1.52],[517.5,0.26],[518.5,-0.66],[519.5,-1.72],[520.5,-1.10],[521.5,-1.80],[522.5,-1.42],[523.5,-0.78],[524.5,-1.32],[525.5,-1.32],[526.5,0.36],[527.5,-0.02],[528.5,-0.72],[529.5,0.88],[530.5,0.26],[531.5,0.12],[532.5,0.12],[533.5,1.10],[534.5,-0.58],[535.5,-0.58],[536.5,-1.42],[537.5,-0.36],[538.5,-1.52],[539.5,0.02],[540.5,0.40],[541.5,1.04],[542.5,1.04],[543.5,1.10],[544.5,-0.36],[545.5,0.86],[546.5,1.06],[547.5,0.56],[548.5,0.68],[549.5,1.74],[550.5,1.40],[551.5,-0.00],[552.5,-0.12],[553.5,-0.06],[554.5,-1.12],[555.5,-2.58],[556.5,-1.10],[557.5,-0.28],[558.5,-0.34],[559.5,-0.42],[560.5,0.22],[561.5,-0.94],[562.5,-2.40],[563.5,-2.34],[564.5,-1.64],[565.5,-1.46],[566.5,-1.22],[567.5,-0.46],[568.5,-0.46],[569.5,-0.46],[570.5,-0.46],[571.5,-0.46],[572.5,-0.48],[573.5,-0.48],[574.5,-0.48],[575.5,-0.48],[576.5,-0.48],[577.5,-0.48],[578.5,0.44],[579.5,0.36],[580.5,-0.34],[581.5,-0.34],[582.5,-0.88],[583.5,-2.42],[584.5,-2.96],[585.5,-2.88],[586.5,-2.42],[587.5,-1.36],[588.5,0.24],[589.5,1.84],[590.5,1.76],[591.5,0.68],[592.5,0.88],[593.5,0.36],[594.5,-1.44],[595.5,-0.10],[596.5,-0.18],[597.5,0.10],[598.5,-0.92],[599.5,0.36],[600.5,-0.90],[601.5,-0.20],[602.5,-1.20],[603.5,0.28],[604.5,-0.80],[605.5,-0.18],[606.5,-0.74],[607.5,-1.28],[608.5,-1.56],[609.5,-1.56],[610.5,-0.58],[611.5,-0.64],[612.5,-0.02],[613.5,-1.28],[614.5,-0.66],[615.5,-2.26],[616.5,-1.64],[617.5,-2.46],[618.5,-2.08],[619.5,-2.26],[620.5,-2.26],[621.5,-2.26],[622.5,-1.50],[623.5,-1.88],[624.5,-1.76],[625.5,-0.70],[626.5,-1.40],[627.5,-0.50],[628.5,-0.58],[629.5,0.40],[630.5,-0.10],[631.5,-0.10],[632.5,-0.94],[633.5,-0.24],[634.5,-1.40],[635.5,-0.50],[636.5,-0.04],[637.5,0.60],[638.5,1.04],[639.5,1.18],[640.5,-0.28],[641.5,0.94],[642.5,1.14],[643.5,0.62],[644.5,0.48],[645.5,0.48],[646.5,0.14],[647.5,-0.24],[648.5,-0.34],[649.5,-0.10],[650.5,0.44],[651.5,-0.90],[652.5,-0.92],[653.5,-0.92],[654.5,-0.00],[655.5,-0.62],[656.5,-0.48],[657.5,-1.16],[658.5,-0.54],[659.5,-2.08],[660.5,-0.40],[661.5,-0.08],[662.5,-0.46],[663.5,-1.08],[664.5,0.38],[665.5,-1.30],[666.5,-0.22],[667.5,0.32],[668.5,1.14],[669.5,0.06],[670.5,0.14],[671.5,-0.70],[672.5,0.02],[673.5,-1.32],[674.5,-1.18],[675.5,-1.18],[676.5,-2.00],[677.5,-1.72],[678.5,-0.56],[679.5,-1.08],[680.5,0.18],[681.5,0.38],[682.5,-1.16],[683.5,-1.62],[684.5,-1.00],[685.5,-0.72],[686.5,0.82],[687.5,1.38],[688.5,2.30],[689.5,2.24],[690.5,0.70],[691.5,-0.84],[692.5,-0.86],[693.5,-1.86],[694.5,-0.96],[695.5,-0.96],[696.5,-0.96],[697.5,-0.88],[698.5,-1.42],[699.5,-1.62],[700.5,-0.02],[701.5,0.42],[702.5,-0.28],[703.5,1.26],[704.5,-0.08],[705.5,-0.22],[706.5,-0.86],[707.5,-0.16],[708.5,-1.14],[709.5,-1.06],[710.5,-1.26],[711.5,-0.68],[712.5,-0.76],[713.5,-1.32],[714.5,-0.70],[715.5,-1.58],[716.5,-0.42],[717.5,0.12],[718.5,0.12],[719.5,-0.58],[720.5,-1.04],[721.5,-2.02],[722.5,-2.02],[723.5,-1.40],[724.5,-0.80],[725.5,-0.72],[726.5,-0.22],[727.5,-0.76],[728.5,-0.84],[729.5,-1.36],[730.5,-1.56],[731.5,-1.54],[732.5,-1.64],[733.5,-1.80],[734.5,-1.80],[735.5,-1.60],[736.5,-2.06],[737.5,-1.44],[738.5,-0.36],[739.5,-0.44],[740.5,0.18],[741.5,-0.44],[742.5,0.10],[743.5,-1.44],[744.5,-1.36],[745.5,-2.18],[746.5,-0.72],[747.5,-2.40],[748.5,-0.86],[749.5,-0.94],[750.5,-0.74],[751.5,-1.58],[752.5,-0.88],[753.5,-2.28],[754.5,-1.76],[755.5,-1.76],[756.5,-1.32],[757.5,-1.94],[758.5,-0.46],[759.5,-0.98],[760.5,-0.42],[761.5,-0.92],[762.5,-0.48],[763.5,-2.10],[764.5,-0.96],[765.5,-1.60],[766.5,-2.24],[767.5,-2.30],[768.5,-0.68],[769.5,-1.74],[770.5,-0.20],[771.5,0.26],[772.5,0.50],[773.5,0.02],[774.5,0.46],[775.5,-1.00],[776.5,0.16],[777.5,-0.46],[778.5,0.08],[779.5,-0.44],[780.5,1.02],[781.5,-0.52],[782.5,1.08],[783.5,0.04],[784.5,0.66],[785.5,-0.74],[786.5,-0.74],[787.5,-2.34],[788.5,-2.90],[789.5,-3.00],[790.5,-2.50],[791.5,-0.90],[792.5,0.64],[793.5,0.64],[794.5,0.20],[795.5,0.08],[796.5,-1.52],[797.5,-3.26],[798.5,-2.20],[799.5,-2.20],[800.5,-2.02],[801.5,-2.22],[802.5,-1.96],[803.5,-2.48],[804.5,-1.92],[805.5,-1.92],[806.5,-1.10],[807.5,-0.08],[808.5,-0.62],[809.5,-1.18],[810.5,-0.34],[811.5,-0.52],[812.5,-1.68],[813.5,-1.06],[814.5,-0.44],[815.5,-1.28],[816.5,-1.10],[817.5,-0.48],[818.5,-0.72],[819.5,-1.42],[820.5,-2.12],[821.5,-2.82],[822.5,-3.56],[823.5,-4.02],[824.5,-2.40],[825.5,-1.70],[826.5,-1.08]];charge_datapoints = [[2.5,0.20],[3.5,0.20],[4.5,0.00],[5.5,0.00],[6.5,0.00],[7.5,0.00],[8.5,0.00],[9.5,0.00],[10.5,0.00],[11.5,0.20],[12.5,0.20],[13.5,0.20],[14.5,0.20],[15.5,0.20],[16.5,0.00],[17.5,0.00],[18.5,0.20],[19.5,0.40],[20.5,0.40],[21.5,0.20],[22.5,0.00],[23.5,-0.20],[24.5,-0.40],[25.5,-0.40],[26.5,-0.20],[27.5,0.00],[28.5,0.00],[29.5,0.00],[30.5,0.00],[31.5,0.00],[32.5,0.00],[33.5,-0.20],[34.5,-0.20],[35.5,-0.40],[36.5,-0.40],[37.5,-0.60],[38.5,-0.40],[39.5,-0.40],[40.5,-0.20],[41.5,-0.10],[42.5,0.30],[43.5,0.30],[44.5,0.30],[45.5,0.30],[46.5,0.20],[47.5,-0.00],[48.5,-0.20],[49.5,-0.20],[50.5,-0.40],[51.5,-0.40],[52.5,-0.60],[53.5,-0.40],[54.5,-0.40],[55.5,-0.20],[56.5,-0.20],[57.5,0.20],[58.5,0.20],[59.5,0.20],[60.5,0.20],[61.5,0.40],[62.5,0.20],[63.5,0.20],[64.5,0.20],[65.5,0.20],[66.5,-0.00],[67.5,-0.00],[68.5,0.20],[69.5,0.20],[70.5,0.20],[71.5,0.20],[72.5,0.20],[73.5,-0.00],[74.5,-0.00],[75.5,-0.00],[76.5,-0.00],[77.5,-0.00],[78.5,-0.00],[79.5,-0.00],[80.5,-0.00],[81.5,-0.00],[82.5,-0.00],[83.5,-0.00],[84.5,-0.00],[85.5,-0.00],[86.5,-0.00],[87.5,-0.00],[88.5,-0.00],[89.5,0.20],[90.5,0.20],[91.5,0.20],[92.5,-0.00],[93.5,0.10],[94.5,-0.10],[95.5,0.10],[96.5,0.10],[97.5,0.40],[98.5,0.10],[99.5,0.10],[100.5,-0.10],[101.5,0.10],[102.5,-0.00],[103.5,0.20],[104.5,0.20],[105.5,0.20],[106.5,-0.20],[107.5,-0.20],[108.5,-0.20],[109.5,-0.20],[110.5,-0.20],[111.5,-0.20],[112.5,-0.00],[113.5,-0.00],[114.5,-0.00],[115.5,-0.00],[116.5,0.20],[117.5,0.20],[118.5,-0.00],[119.5,-0.20],[120.5,-0.20],[121.5,-0.20],[122.5,-0.40],[123.5,-0.00],[124.5,0.20],[125.5,0.40],[126.5,0.40],[127.5,0.20],[128.5,-0.00],[129.5,0.20],[130.5,-0.00],[131.5,-0.20],[132.5,-0.00],[133.5,-0.20],[134.5,-0.40],[135.5,-0.40],[136.5,-0.20],[137.5,-0.20],[138.5,0.20],[139.5,0.20],[140.5,-0.00],[141.5,-0.00],[142.5,0.20],[143.5,-0.00],[144.5,-0.00],[145.5,-0.00],[146.5,-0.00],[147.5,-0.00],[148.5,-0.20],[149.5,-0.40],[150.5,-0.20],[151.5,-0.20],[152.5,-0.40],[153.5,-0.20],[154.5,-0.00],[155.5,0.10],[156.5,0.30],[157.5,0.30],[158.5,0.10],[159.5,0.10],[160.5,-0.00],[161.5,-0.20],[162.5,-0.20],[163.5,-0.00],[164.5,0.10],[165.5,0.10],[166.5,0.10],[167.5,0.10],[168.5,0.10],[169.5,-0.00],[170.5,-0.00],[171.5,-0.20],[172.5,-0.00],[173.5,-0.00],[174.5,0.20],[175.5,0.20],[176.5,0.40],[177.5,0.20],[178.5,0.40],[179.5,0.20],[180.5,0.20],[181.5,0.20],[182.5,0.40],[183.5,0.20],[184.5,0.40],[185.5,0.50],[186.5,0.50],[187.5,0.30],[188.5,0.10],[189.5,-0.30],[190.5,-0.40],[191.5,-0.40],[192.5,-0.40],[193.5,-0.20],[194.5,-0.00],[195.5,-0.00],[196.5,-0.20],[197.5,-0.10],[198.5,-0.10],[199.5,-0.10],[200.5,-0.10],[201.5,0.10],[202.5,-0.00],[203.5,-0.00],[204.5,-0.00],[205.5,-0.00],[206.5,-0.20],[207.5,-0.20],[208.5,-0.20],[209.5,-0.20],[210.5,-0.20],[211.5,-0.00],[212.5,-0.00],[213.5,-0.20],[214.5,-0.20],[215.5,-0.10],[216.5,-0.10],[217.5,-0.10],[218.5,0.10],[219.5,0.10],[220.5,-0.00],[221.5,-0.00],[222.5,-0.00],[223.5,-0.00],[224.5,-0.00],[225.5,0.20],[226.5,-0.00],[227.5,-0.00],[228.5,-0.00],[229.5,-0.20],[230.5,-0.20],[231.5,0.20],[232.5,-0.00],[233.5,0.10],[234.5,0.30],[235.5,0.10],[236.5,-0.10],[237.5,0.10],[238.5,-0.20],[239.5,-0.20],[240.5,-0.20],[241.5,-0.20],[242.5,-0.20],[243.5,-0.00],[244.5,-0.00],[245.5,-0.00],[246.5,-0.00],[247.5,0.10],[248.5,0.10],[249.5,0.10],[250.5,-0.10],[251.5,-0.30],[252.5,-0.40],[253.5,-0.40],[254.5,-0.20],[255.5,-0.20],[256.5,-0.00],[257.5,0.20],[258.5,-0.00],[259.5,-0.40],[260.5,-0.40],[261.5,-0.60],[262.5,-0.60],[263.5,-0.40],[264.5,-0.20],[265.5,-0.00],[266.5,0.20],[267.5,-0.00],[268.5,-0.00],[269.5,-0.00],[270.5,-0.00],[271.5,-0.00],[272.5,-0.00],[273.5,-0.00],[274.5,-0.00],[275.5,-0.00],[276.5,-0.00],[277.5,-0.00],[278.5,-0.00],[279.5,-0.00],[280.5,-0.00],[281.5,-0.00],[282.5,-0.00],[283.5,-0.00],[284.5,-0.00],[285.5,-0.00],[286.5,-0.00],[287.5,-0.00],[288.5,-0.00],[289.5,-0.00],[290.5,-0.00],[291.5,-0.00],[292.5,-0.00],[293.5,-0.00],[294.5,-0.00],[295.5,-0.00],[296.5,-0.00],[297.5,-0.00],[298.5,-0.00],[299.5,-0.00],[300.5,0.20],[301.5,0.40],[302.5,0.60],[303.5,0.70],[304.5,0.70],[305.5,0.50],[306.5,0.50],[307.5,0.50],[308.5,0.60],[309.5,0.60],[310.5,0.60],[311.5,0.40],[312.5,0.20],[313.5,-0.00],[314.5,-0.00],[315.5,-0.00],[316.5,-0.00],[317.5,-0.00],[318.5,-0.00],[319.5,-0.00],[320.5,-0.00],[321.5,-0.00],[322.5,-0.00],[323.5,-0.00],[324.5,-0.20],[325.5,-0.20],[326.5,-0.20],[327.5,-0.00],[328.5,-0.00],[329.5,0.20],[330.5,0.20],[331.5,0.20],[332.5,-0.20],[333.5,-0.20],[334.5,-0.20],[335.5,-0.20],[336.5,-0.20],[337.5,-0.20],[338.5,-0.00],[339.5,-0.00],[340.5,-0.00],[341.5,-0.00],[342.5,0.20],[343.5,-0.20],[344.5,-0.40],[345.5,-0.40],[346.5,-0.40],[347.5,-0.40],[348.5,-0.20],[349.5,-0.00],[350.5,-0.00],[351.5,-0.00],[352.5,-0.00],[353.5,-0.20],[354.5,-0.20],[355.5,-0.20],[356.5,-0.20],[357.5,-0.40],[358.5,-0.20],[359.5,-0.40],[360.5,-0.40],[361.5,-0.40],[362.5,-0.20],[363.5,-0.20],[364.5,0.20],[365.5,0.20],[366.5,0.30],[367.5,0.30],[368.5,0.50],[369.5,0.30],[370.5,0.30],[371.5,0.20],[372.5,0.20],[373.5,-0.00],[374.5,-0.00],[375.5,-0.00],[376.5,-0.00],[377.5,-0.00],[378.5,-0.00],[379.5,-0.00],[380.5,-0.00],[381.5,0.20],[382.5,0.40],[383.5,0.40],[384.5,0.40],[385.5,0.40],[386.5,0.20],[387.5,-0.20],[388.5,-0.20],[389.5,-0.20],[390.5,-0.20],[391.5,-0.40],[392.5,-0.00],[393.5,-0.00],[394.5,-0.00],[395.5,-0.20],[396.5,0.20],[397.5,-0.20],[398.5,-0.20],[399.5,-0.20],[400.5,-0.00],[401.5,-0.00],[402.5,0.20],[403.5,-0.00],[404.5,-0.00],[405.5,0.20],[406.5,-0.00],[407.5,0.20],[408.5,0.40],[409.5,0.40],[410.5,0.20],[411.5,0.40],[412.5,0.20],[413.5,0.20],[414.5,0.20],[415.5,0.30],[416.5,0.10],[417.5,0.30],[418.5,0.30],[419.5,0.10],[420.5,-0.00],[421.5,0.20],[422.5,-0.00],[423.5,-0.00],[424.5,0.20],[425.5,0.40],[426.5,0.20],[427.5,0.40],[428.5,0.40],[429.5,0.60],[430.5,0.40],[431.5,0.40],[432.5,0.40],[433.5,0.40],[434.5,0.20],[435.5,-0.00],[436.5,-0.00],[437.5,-0.10],[438.5,-0.10],[439.5,-0.10],[440.5,-0.10],[441.5,-0.00],[442.5,0.10],[443.5,0.10],[444.5,-0.10],[445.5,0.10],[446.5,-0.00],[447.5,-0.20],[448.5,-0.10],[449.5,-0.10],[450.5,0.10],[451.5,-0.10],[452.5,-0.10],[453.5,-0.20],[454.5,0.20],[455.5,-0.00],[456.5,0.40],[457.5,0.40],[458.5,0.40],[459.5,-0.00],[460.5,0.10],[461.5,-0.10],[462.5,-0.10],[463.5,-0.10],[464.5,0.30],[465.5,0.20],[466.5,0.20],[467.5,0.20],[468.5,-0.00],[469.5,-0.20],[470.5,-0.20],[471.5,-0.40],[472.5,-0.60],[473.5,-0.40],[474.5,-0.40],[475.5,-0.20],[476.5,-0.00],[477.5,0.20],[478.5,0.20],[479.5,0.20],[480.5,-0.00],[481.5,-0.00],[482.5,-0.00],[483.5,-0.00],[484.5,0.20],[485.5,0.20],[486.5,-0.00],[487.5,-0.20],[488.5,-0.20],[489.5,-0.40],[490.5,-0.40],[491.5,-0.20],[492.5,-0.00],[493.5,-0.00],[494.5,0.20],[495.5,0.20],[496.5,-0.00],[497.5,-0.00],[498.5,0.20],[499.5,-0.00],[500.5,0.20],[501.5,0.40],[502.5,0.60],[503.5,0.40],[504.5,0.20],[505.5,-0.00],[506.5,-0.00],[507.5,-0.20],[508.5,-0.20],[509.5,-0.00],[510.5,0.10],[511.5,0.10],[512.5,0.10],[513.5,0.10],[514.5,0.10],[515.5,-0.20],[516.5,-0.20],[517.5,-0.40],[518.5,-0.40],[519.5,-0.40],[520.5,-0.20],[521.5,-0.00],[522.5,0.20],[523.5,0.20],[524.5,-0.00],[525.5,-0.00],[526.5,-0.20],[527.5,-0.20],[528.5,-0.20],[529.5,-0.00],[530.5,-0.20],[531.5,-0.20],[532.5,-0.40],[533.5,-0.40],[534.5,-0.20],[535.5,-0.20],[536.5,-0.20],[537.5,-0.00],[538.5,-0.00],[539.5,-0.20],[540.5,-0.00],[541.5,-0.00],[542.5,-0.00],[543.5,-0.00],[544.5,-0.20],[545.5,-0.20],[546.5,-0.20],[547.5,-0.20],[548.5,-0.20],[549.5,-0.00],[550.5,-0.00],[551.5,0.10],[552.5,0.10],[553.5,0.10],[554.5,0.10],[555.5,0.30],[556.5,0.20],[557.5,0.20],[558.5,0.20],[559.5,0.40],[560.5,0.20],[561.5,0.20],[562.5,0.40],[563.5,0.40],[564.5,0.20],[565.5,0.20],[566.5,0.20],[567.5,-0.00],[568.5,-0.00],[569.5,-0.00],[570.5,-0.00],[571.5,-0.00],[572.5,-0.00],[573.5,-0.00],[574.5,-0.00],[575.5,-0.00],[576.5,-0.00],[577.5,-0.00],[578.5,-0.00],[579.5,-0.00],[580.5,0.20],[581.5,0.20],[582.5,-0.00],[583.5,-0.20],[584.5,-0.40],[585.5,-0.60],[586.5,-0.60],[587.5,-0.40],[588.5,-0.20],[589.5,-0.00],[590.5,0.20],[591.5,-0.00],[592.5,-0.00],[593.5,-0.00],[594.5,0.20],[595.5,-0.00],[596.5,0.40],[597.5,0.40],[598.5,0.50],[599.5,0.30],[600.5,0.10],[601.5,-0.10],[602.5,-0.10],[603.5,-0.20],[604.5,-0.20],[605.5,-0.00],[606.5,0.10],[607.5,-0.10],[608.5,-0.10],[609.5,-0.30],[610.5,-0.30],[611.5,-0.60],[612.5,-0.40],[613.5,-0.60],[614.5,-0.40],[615.5,-0.60],[616.5,-0.40],[617.5,-0.20],[618.5,-0.00],[619.5,-0.00],[620.5,-0.00],[621.5,-0.00],[622.5,-0.20],[623.5,-0.20],[624.5,-0.20],[625.5,-0.00],[626.5,0.20],[627.5,0.20],[628.5,0.40],[629.5,0.40],[630.5,0.40],[631.5,0.40],[632.5,0.40],[633.5,0.20],[634.5,0.20],[635.5,0.20],[636.5,-0.00],[637.5,-0.00],[638.5,-0.00],[639.5,-0.00],[640.5,-0.20],[641.5,-0.20],[642.5,-0.20],[643.5,-0.20],[644.5,-0.20],[645.5,-0.00],[646.5,-0.00],[647.5,-0.00],[648.5,-0.00],[649.5,-0.00],[650.5,-0.00],[651.5,0.20],[652.5,0.20],[653.5,0.20],[654.5,0.20],[655.5,0.40],[656.5,0.30],[657.5,0.30],[658.5,0.30],[659.5,0.10],[660.5,-0.10],[661.5,-0.20],[662.5,-0.40],[663.5,-0.40],[664.5,-0.20],[665.5,-0.00],[666.5,-0.00],[667.5,0.20],[668.5,0.20],[669.5,0.20],[670.5,-0.20],[671.5,-0.20],[672.5,-0.20],[673.5,-0.00],[674.5,-0.00],[675.5,-0.00],[676.5,0.20],[677.5,0.20],[678.5,-0.00],[679.5,-0.00],[680.5,0.20],[681.5,-0.20],[682.5,-0.40],[683.5,-0.40],[684.5,-0.40],[685.5,-0.40],[686.5,-0.20],[687.5,-0.00],[688.5,-0.00],[689.5,-0.00],[690.5,-0.00],[691.5,-0.20],[692.5,-0.20],[693.5,-0.20],[694.5,-0.20],[695.5,-0.20],[696.5,-0.20],[697.5,-0.20],[698.5,-0.40],[699.5,-0.40],[700.5,-0.40],[701.5,-0.20],[702.5,-0.00],[703.5,0.20],[704.5,0.40],[705.5,0.40],[706.5,0.60],[707.5,0.40],[708.5,0.40],[709.5,0.20],[710.5,0.20],[711.5,-0.00],[712.5,-0.00],[713.5,-0.20],[714.5,-0.20],[715.5,-0.20],[716.5,-0.20],[717.5,-0.20],[718.5,-0.00],[719.5,0.20],[720.5,0.40],[721.5,0.40],[722.5,0.40],[723.5,0.40],[724.5,0.20],[725.5,-0.20],[726.5,-0.20],[727.5,-0.20],[728.5,-0.20],[729.5,-0.40],[730.5,-0.00],[731.5,-0.00],[732.5,-0.00],[733.5,-0.00],[734.5,-0.00],[735.5,-0.40],[736.5,-0.40],[737.5,-0.40],[738.5,-0.40],[739.5,-0.00],[740.5,0.20],[741.5,-0.00],[742.5,-0.00],[743.5,0.20],[744.5,-0.00],[745.5,0.20],[746.5,0.40],[747.5,0.60],[748.5,0.40],[749.5,0.60],[750.5,0.20],[751.5,0.20],[752.5,-0.00],[753.5,0.10],[754.5,-0.10],[755.5,-0.10],[756.5,-0.10],[757.5,-0.30],[758.5,-0.40],[759.5,-0.20],[760.5,-0.00],[761.5,-0.00],[762.5,0.20],[763.5,0.40],[764.5,0.20],[765.5,0.40],[766.5,0.60],[767.5,0.60],[768.5,0.40],[769.5,0.40],[770.5,0.20],[771.5,-0.00],[772.5,-0.00],[773.5,-0.00],[774.5,-0.00],[775.5,-0.00],[776.5,-0.00],[777.5,-0.00],[778.5,-0.00],[779.5,0.20],[780.5,0.20],[781.5,-0.00],[782.5,-0.00],[783.5,-0.00],[784.5,-0.20],[785.5,-0.10],[786.5,0.10],[787.5,-0.10],[788.5,-0.30],[789.5,-0.30],[790.5,-0.40],[791.5,-0.40],[792.5,-0.20],[793.5,-0.20],[794.5,-0.20],[795.5,-0.20],[796.5,-0.40],[797.5,-0.20],[798.5,-0.00],[799.5,-0.20],[800.5,-0.20],[801.5,0.20],[802.5,0.10],[803.5,0.10],[804.5,0.30],[805.5,0.30],[806.5,0.10],[807.5,-0.00],[808.5,-0.20],[809.5,-0.40],[810.5,-0.40],[811.5,-0.40],[812.5,-0.20],[813.5,-0.00],[814.5,0.20],[815.5,0.20],[816.5,0.20],[817.5,-0.00],[818.5,-0.00],[819.5,0.20],[820.5,0.40],[821.5,0.60],[822.5,0.80],[823.5,1.00],[824.5,0.80],[825.5,0.60],[826.5,0.40]];dis_datapoints = undefined;trans_datapoints = undefined;sec_helix_datapoints = undefined;sec_strand_datapoints = undefined;acc_exposed_datapoints = undefined;acc_buried_datapoints = undefined;flot_plot_options = []; flot_plot_options[0] = {grid: {borderWidth: {top: 0,right: 0,bottom: 0,left: 0}},legend: {show: false},xaxes: [{show: true,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: [{show: true,ticks: [[0, '0'], [4.5,'hydro-<br>phobic&nbsp;&nbsp;'], [-4.5,'hydro-<br>philic&nbsp;&nbsp;']],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)'}},{show: true,ticks: [[0, ''], [1,'positive<br>&nbsp;charge'], [-1,'negative<br>&nbsp;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)'}}]};number_of_plots = 5;for ( plot_num = 1 ; plot_num < number_of_plots ; plot_num ++){flot_plot_options[plot_num] = jqAutoAnnotator.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}];};try {if( jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_button').val() =='Show' ){jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_container').css('display','block');jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_button').val('Hide');var description_html = '<div id=\'AutoAnnotator_plot_selectors\'>';description_html = description_html + '<br>&nbsp;<input type=\'checkbox\' id=\'hydrophobicity_checkbox\' checked=\'checked\'>&nbsp;Moving average over 5 amino acids for hydrophobicity (<img src=\'https://static.igem.org/mediawiki/2013/e/e9/TUM13_hydrophobicity_icon.png\' alt=\'blue graph\' height=\'10\'></img>)';description_html = description_html + '<br>&nbsp;<input type=\'checkbox\' id=\'charge_checkbox\' checked=\'checked\'>&nbsp;Moving average over 5 amino acids for charge (<img src=\'https://static.igem.org/mediawiki/2013/3/3e/TUM13_charge_icon.png\' alt=\'red graph\' height=\'10\'></img>)';description_html = description_html + '<br>&nbsp;<input type=\'checkbox\' id=\'dis_checkbox\' checked=\'checked\'>&nbsp;Predicted disulfid bridges (<img src=\'https://static.igem.org/mediawiki/2013/2/28/TUM13_dis_icon.png\' alt=\'yellow circle\' height=\'10\'></img>) with the number of the bridge in the center';description_html = description_html + '<br>&nbsp;<input type=\'checkbox\' id=\'trans_checkbox\' checked=\'checked\'>&nbsp;Predicted transmembrane helices (<img src=\'https://static.igem.org/mediawiki/2013/7/78/TUM13_trans_icon.png\' alt=\'turquois bars\' height=\'10\'></img>)';description_html = description_html + '<br>&nbsp;<input type=\'checkbox\' id=\'sec_checkbox\' checked=\'checked\'>&nbsp;Predicted secondary structure: Helices (<img src=\'https://static.igem.org/mediawiki/2013/b/bf/TUM13_helix_icon.png\' alt=\'violet bars\' height=\'10\'></img>) and beta-strands (<img src=\'https://static.igem.org/mediawiki/2013/b/bf/TUM13_strand_icon.png\' alt=\'yellow bars\' height=\'10\'></img>)';description_html = description_html + '<br>&nbsp;<input type=\'checkbox\' id=\'acc_checkbox\' checked=\'checked\'>&nbsp;Predicted solvent accessability: Exposed (<img src=\'https://static.igem.org/mediawiki/2013/1/16/TUM13_exposed_icon.png\' alt=\'blue bars\' height=\'10\'></img>) and buried (<img src=\'https://static.igem.org/mediawiki/2013/0/0b/TUM13_buried_icon.png\' alt=\'green bars\' height=\'10\'></img>) residues';description_html = description_html + '<br></div>';jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_explanation').html(description_html);plot_according_to_selectors_1508714402355();jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #AutoAnnotator_plot_selectors').find('input').click(plot_according_to_selectors_1508714402355);}else{jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_container').css('display','none');jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_button').val('Show');jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_explanation').html('');}}catch(err){txt='There was an error with the button controlling the visibility of the plot.\n';txt=txt+'The originating error is:\n' + err + '\n\n';alert(txt);}};function plot_according_to_selectors_1508714402355(){try{var plot_datasets = [[],[]];if(jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_checkbox').prop('checked') == true){plot_datasets[0] = { 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};}if(jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #charge_checkbox').prop('checked') == true){plot_datasets[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};}for (plot_num = 0 ; plot_num < number_of_plots ; plot_num ++){jqAutoAnnotator.plot('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_placeholder'+ plot_num.toString(), plot_datasets, flot_plot_options[plot_num] );}var screen_width = jqAutoAnnotator('canvas.flot-base').width(); var pos_of_first_tick = 46;var pos_of_last_tick = screen_width - 51;var tick_diff = (screen_width - 97)/199;if(jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #dis_checkbox').prop('checked') == true){for ( j = 0 ; j < dis_datapoints.length ; j++ ){jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_placeholder' + Math.floor((dis_datapoints[j][0] - 1)/200) ).append('<div class=\'AutoAnnotator_dis\' style=\'left:' + ((pos_of_first_tick - 8 + (dis_datapoints[j][0] - 1)*tick_diff - Math.floor((dis_datapoints[j][0] - 1)/200)*200*tick_diff).toFixed(0)).toString() + 'px;\'><b>' + (j+1) + '</b></div>');jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_placeholder' + Math.floor((dis_datapoints[j][1] - 1)/200) ).append('<div class=\'AutoAnnotator_dis\' style=\'left:' + ((pos_of_first_tick - 8 + (dis_datapoints[j][1] - 1)*tick_diff - Math.floor((dis_datapoints[j][1] - 1)/200)*200*tick_diff).toFixed(0)).toString() + 'px;\'><b>' + (j+1) + '</b></div>');}}if(jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #trans_checkbox').prop('checked') == true){for ( j = 0 ; j < trans_datapoints.length ; j++ ){jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_placeholder' + Math.floor((trans_datapoints[j][0] - 1)/200) ).append('<div class=\'AutoAnnotator_trans\' style=\'width:' + (((trans_datapoints[j][1] - trans_datapoints[j][0] + 1)*tick_diff).toFixed(0)).toString() + 'px ;left:' + ((pos_of_first_tick + (trans_datapoints[j][0] - 1.5)*tick_diff - Math.floor((trans_datapoints[j][0] - 1)/200)*200*tick_diff).toFixed(0)).toString() + 'px\'></div>');}}if(jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #sec_checkbox').prop('checked') == true){for ( j = 0 ; j < sec_helix_datapoints.length ; j++ ){jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_placeholder' + Math.floor((sec_helix_datapoints[j][0] - 1)/200) ).append('<div class=\'AutoAnnotator_sec_helix\' style=\'width:' + (((sec_helix_datapoints[j][1] - sec_helix_datapoints[j][0] + 1)*tick_diff).toFixed(0)).toString() + 'px; left:' + ((pos_of_first_tick + (sec_helix_datapoints[j][0] - 1.5)*tick_diff - Math.floor((sec_helix_datapoints[j][0] - 1)/200)*200*tick_diff).toFixed(0)).toString() + 'px\'></div>');}for ( j = 0 ; j < sec_strand_datapoints.length ; j++ ){jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_placeholder' + Math.floor((sec_strand_datapoints[j][0] - 1)/200) ).append('<div class=\'AutoAnnotator_sec_strand\' style=\'width:' + (((sec_strand_datapoints[j][1] - sec_strand_datapoints[j][0] + 1)*tick_diff).toFixed(0)).toString() + 'px; left:' + ((pos_of_first_tick + (sec_strand_datapoints[j][0] - 1.5)*tick_diff - Math.floor((sec_strand_datapoints[j][0] - 1)/200)*200*tick_diff).toFixed(0)).toString() + 'px\'></div>');}}if(jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #acc_checkbox').prop('checked') == true){for ( j = 0 ; j < acc_buried_datapoints.length ; j++ ){jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_placeholder' + Math.floor((acc_buried_datapoints[j][0] - 1)/200) ).append('<div class=\'AutoAnnotator_acc_buried\' style=\'width:' + (((acc_buried_datapoints[j][1] - acc_buried_datapoints[j][0] + 1)*tick_diff).toFixed(0)).toString() + 'px; left:' + ((pos_of_first_tick + (acc_buried_datapoints[j][0] - 1.5)*tick_diff - Math.floor((acc_buried_datapoints[j][0] - 1)/200)*200*tick_diff).toFixed(0)).toString() + 'px\'></div>');}for ( j = 0 ; j < acc_exposed_datapoints.length ; j++ ){jqAutoAnnotator('#AutoAnnotator_container_1508714402355 #hydrophobicity_charge_placeholder' + Math.floor((acc_exposed_datapoints[j][0] - 1)/200) ).append('<div class=\'AutoAnnotator_acc_exposed\' style=\'width:' + (((acc_exposed_datapoints[j][1] - acc_exposed_datapoints[j][0] + 1)*tick_diff).toFixed(0)).toString() + 'px; left:' + ((pos_of_first_tick + (acc_exposed_datapoints[j][0] - 1.5)*tick_diff - Math.floor((acc_exposed_datapoints[j][0] - 1)/200)*200*tick_diff).toFixed(0)).toString() + 'px\'></div>');}}}catch(err){txt='There was an error while drawing the selected elements for the plot.\n';txt=txt+'The originating error is:\n' + err + '\n\n';throw(txt);}}</script></html>
  
  

Latest revision as of 23:34, 22 October 2017

Plasma membrane delivery system (PMDS, mammalian)

Plasma Membrane Delivery System (PMDS)
Function Delivery of protein (e.g. mCherry) to the PM
Use in Mammalian cells
Abstraction Hierarchy Part
RFC standard RFC10, RFC12, RFC23 & RFC25 compatible
Backbone pSB1C3
Submitted by [http://2017.igem.org/Team:UCL UCL iGEM 2017]


This coding region translates a transmembrane domain which is destined to go to the plasma membrane via the secretory pathway. While doing so it delivers any 'cytosolic domain insert' that has been cloned into this BioBrick in a plug-and-play fashion (via the KpnI-BamHI restriction sites) to the plasma membrane. The delivered protein is localised to the plasma membrane but activation of the photocleavable linker PhoCl with 400 nm, violet light allows irreversible breakage of the connection with the PM and releases the 'cytosolic domain insert' into the cytoplasm. For characterisation purposes mCherry has been chosen 'cytosolic domain insert' in this BioBrick.



Usage and Biology

As stated in the original paper:


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2485
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 704
    Illegal BsaI.rc site found at 1471


Functional Parameters

Protein data table for BioBrick BBa_ automatically created by the BioBrick-AutoAnnotator version 1.0
Nucleotide sequence in RFC 10: (underlined part encodes the protein)
 ACCATGCGGGCC ... GTCGGATCCTAG
 ORF from nucleotide position 4 to 2490 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)
1 
101 
201 
301 
401 
501 
601 
701 
801 		MRAWIFFLLCLAGRALAATGRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQH
DFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADH
YQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKDYKDDDDKTGGGGGSGGGGSPSIATGMVGALLLLLVVALGIGL
FMRRRHIVRKRGGGGSGGGGSGSVIPDYFKQSFPEGYSWERSMTYEDGGICIATNDITMEGDSFINKIHFKGTNFPPNGPVMQKRTVGWEASTEKMYERD
GVLKGDVKMKLLLKGGGHYRCDYRTTYKVKQKPVKLPDYHFVDHRIEILSHDKDYNKVKLYEHAVARNSTDSMDELYKGGSGGMVSKGEETITSVIKPDM
KNKLRMEGNVNGHAFVIEGEGSGKPFEGIQTIDLEVKEGAPLPFAYDILTTAFHYGNRVFTKYPRGGGGTGGGGSGGGGSVSKGEEDNMAIIKEFMRFKV
HMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDG
EFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYER
AEGRHSTGGMDELYKGGGGSPKKKRKVGS*
Sequence features: (with their position in the amino acid sequence, see the list of supported features)
RFC25 scar (shown in bold): 19 to 20, 266 to 267, 807 to 808
Flag-tag: 258 to 265
Enterokinase cleavage site: 261 to 265
SV40 nuclear localization sequence: 821 to 827
Amino acid composition:
Ala (A)36 (4.3%)
Arg (R)33 (4.0%)
Asn (N)30 (3.6%)
Asp (D)53 (6.4%)
Cys (C)5 (0.6%)
Gln (Q)20 (2.4%)
Glu (E)58 (7.0%)
Gly (G)114 (13.8%)
His (H)25 (3.0%)
Ile (I)40 (4.8%)
Leu (L)55 (6.6%)
Lys (K)75 (9.0%)
Met (M)26 (3.1%)
Phe (F)37 (4.5%)
Pro (P)36 (4.3%)
Ser (S)42 (5.1%)
Thr (T)46 (5.5%)
Trp (W)7 (0.8%)
Tyr (Y)38 (4.6%)
Val (V)53 (6.4%)
Amino acid counting
Total number:829
Positively charged (Arg+Lys):108 (13.0%)
Negatively charged (Asp+Glu):111 (13.4%)
Aromatic (Phe+His+Try+Tyr):107 (12.9%)
Biochemical parameters
Atomic composition:C4078H6316N1110O1228S31
Molecular mass [Da]:91535.6
Theoretical pI:6.82
Extinction coefficient at 280 nm [M-1 cm-1]:95120 / 95433 (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 (0.69)good (0.68)acceptable (0.57)good (0.67)excellent (0.85)excellent (0.85)
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.


Reference

Zhang W, Lohman AW, Zhuravlova Y, Lu X, Wiens MD, Hoi H, Yaganoglu S, Mohr MA, Kitova EN, Klassen JS, Pantazis P, Thompson RJ, Campbell RE. Optogenetic control with a photocleavable protein, PhoCl. Nat Methods. 14(4):391-394 (2017) NCBI