Difference between revisions of "Team:Sydney Australia/Hexagon"

 
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<html>
 
<html>
−
 
 
<head>
 
<head>
 +
<script src="https://2017.igem.org/Team:Sydney_Australia/Resources/angularplasmid.complete.min.js"></script>
 
<script src="http://code.jquery.com/jquery-2.1.0.min.js"></script>
 
<script src="http://code.jquery.com/jquery-2.1.0.min.js"></script>
 +
<link href="https://fonts.googleapis.com/css?family=Quicksand:300,400|Roboto:400,400i" rel="stylesheet">
 +
 +
<script type="text/javascript" src="https://use.fontawesome.com/2967839f93.js"></script>
 +
<meta name="viewport" content="width=device-width, initial-scale=1">
 
   <link href="https://fonts.googleapis.com/css?family=Quicksand" rel="stylesheet">
 
   <link href="https://fonts.googleapis.com/css?family=Quicksand" rel="stylesheet">
−
    <link rel="stylesheet" href="https://maxcdn.bootstrapcdn.com/bootstrap/3.3.7/css/bootstrap.min.css">
+
<script src="https://ajax.googleapis.com/ajax/libs/jquery/3.2.1/jquery.min.js"></script>
−
  <script src="https://ajax.googleapis.com/ajax/libs/jquery/3.2.1/jquery.min.js"></script>
+
 
−
  <script src="https://maxcdn.bootstrapcdn.com/bootstrap/3.3.7/js/bootstrap.min.js"></script>
+
 
<script>
 
<script>
−
$(".they-said").click(function(){
+
$(document).ready(function() {
−
    $(".they-said-content").fadeIn("slow");
+
  $(function() {
−
    $(".we-said").fadeIn("slow");
+
      $('#pip').click(function() {
−
});
+
          $('.pip_info').fadeToggle('slow');
 +
          $('.lacr_info').hide();
 +
          $('.eoriv_info').hide();
 +
          $('.boriv_info').hide();
 +
          $('.mcs_info').hide();
 +
          $('.anti_info').hide();
 +
          $('.laco_info').hide();
 +
          $('.super_info').hide();             
 +
      });       
 +
  });
 +
  $(function() {
 +
      $('#boriv').click(function() {
 +
          $('.boriv_info').fadeToggle('slow');
 +
          $('.lacr_info').hide();
 +
          $('.pip_info').hide();
 +
          $('.eoriv_info').hide();
 +
          $('.mcs_info').hide();
 +
          $('.anti_info').hide();
 +
          $('.laco_info').hide();
 +
          $('.super_info').hide();             
 +
      });       
 +
  });
 +
  $(function() {
 +
      $('#eoriv').click(function() {
 +
          $('.eoriv_info').fadeToggle('slow');
 +
          $('.lacr_info').hide();
 +
          $('.pip_info').hide();
 +
          $('.boriv_info').hide();
 +
          $('.mcs_info').hide();
 +
          $('.anti_info').hide();
 +
          $('.laco_info').hide();
 +
          $('.super_info').hide();             
 +
      });       
 +
  });
 +
  $(function() {
 +
      $('#mcs1').click(function() {
 +
          $('.mcs_info').fadeToggle('slow');
 +
          $('.lacr_info').hide();
 +
          $('.pip_info').hide();
 +
          $('.eoriv_info').hide();
 +
          $('.boriv_info').hide();
 +
          $('.anti_info').hide();
 +
          $('.laco_info').hide();
 +
          $('.super_info').hide();             
 +
      });       
 +
  });
 +
  $(function() {
 +
      $('#mcs2').click(function() {
 +
          $('.mcs_info').fadeToggle('slow');
 +
          $('.lacr_info').hide();
 +
          $('.pip_info').hide();
 +
          $('.eoriv_info').hide();
 +
          $('.boriv_info').hide();
 +
          $('.anti_info').hide();
 +
          $('.laco_info').hide();
 +
          $('.super_info').hide();             
 +
      });       
 +
  });
 +
  $(function() {
 +
      $('#anti').click(function() {
 +
          $('.anti_info').fadeToggle('slow');
 +
          $('.lacr_info').hide();
 +
          $('.pip_info').hide();
 +
          $('.eoriv_info').hide();
 +
          $('.boriv_info').hide();
 +
          $('.mcs_info').hide();
 +
          $('.laco_info').hide();
 +
          $('.super_info').hide();             
 +
      });       
 +
  });
 +
  $(function() {
 +
      $('#super').click(function() {
 +
          $('.super_info').fadeToggle('slow');
 +
          $('.lacr_info').hide();
 +
          $('.pip_info').hide();
 +
          $('.eoriv_info').hide();
 +
          $('.boriv_info').hide();
 +
          $('.mcs_info').hide();
 +
          $('.anti_info').hide();
 +
          $('.laco_info').hide();             
 +
      });       
 +
  });
 +
  $(function() {
 +
      $('#lacr').click(function() {
 +
          $('.lacr_info').fadeToggle('slow');
 +
          $('.pip_info').hide();
 +
          $('.eoriv_info').hide();
 +
          $('.boriv_info').hide();
 +
          $('.mcs_info').hide();
 +
          $('.anti_info').hide();
 +
          $('.laco_info').hide();
 +
          $('.super_info').hide();             
 +
      });       
 +
  });
 +
  $(function() {
 +
      $('#laco').click(function() {
 +
          $('.laco_info').fadeToggle('slow');
 +
          $('.lacr_info').hide();
 +
          $('.pip_info').hide();
 +
          $('.eoriv_info').hide();
 +
          $('.boriv_info').hide();
 +
          $('.mcs_info').hide();
 +
          $('.anti_info').hide();
 +
          $('.super_info').hide();             
 +
      });      
 +
  });
  
−
$(".we-said").click(function(){
+
  });
−
    $(".we-said-content").fadeIn("slow");
+
−
});
+
 
</script>
 
</script>
−
<style>  
+
<style>
−
 
+
  .plasmid .markerhover:hover {
−
/* our special font */
+
  stroke: #e2e2e2;
−
@font-face {
+
  stroke-width: 2px;
−
    font-family: 'banaue';
+
−
    src: url('https://static.igem.org/mediawiki/2017/b/b0/T--Sydney_Australia--Wiki_Font_Banaue-Regular.Media-File.ogg');
+
−
    font-weight: normal;
+
−
    font-style: large;
+
 
}
 
}
−
 
+
  .plasmid .track {
−
#please h1 {
+
  fill: #cfd9db;
−
font-family: "banaue", sans-serif;  
+
  stroke: #c0cdd1;
−
color: #3e3f3f
+
 
}
 
}
−
 
+
  .plasmid .tracklabelbody {
−
#please h4 {
+
   font: 400 20px "Quicksand";
−
   font-family: 'Quicksand', sans-serif;
+
   fill: #5e7980;
−
   color:#3e3f3f;
+
 
}
 
}
−
 
+
  .plasmid .tracklabeltitle {
−
.crispy {
+
  font: 700 40px "Quicksand";
−
image-rendering: -moz-crisp-edges;
+
  fill: #334145;
−
image-rendering: -o-crisp-edges;
+
−
image-rendering: -webkit-optimize-contrast;
+
−
-ms-interpolation-mode: nearest-neighbor;
+
 
}
 
}
−
 
+
  .plasmid .tracklabelsmall {
−
.img-center  {
+
  font: 400 12px "Quicksand";
−
image-rendering: -moz-crisp-edges;
+
  fill: #5e7980;
−
image-rendering: -o-crisp-edges;
+
−
image-rendering: -webkit-optimize-contrast;
+
−
-ms-interpolation-mode: nearest-neighbor;
+
−
width:100%;
+
−
    }
+
−
 
+
−
.img-circle {
+
−
    border-radius: 50%;
+
 
}
 
}
−
 
+
  .plasmid .scale-major {
−
.insert {
+
  stroke: #334145;
−
  height: 68px;
+
  stroke-width: 2px;
−
  width: 100%
+
−
  background-color:#e2e2e2;
+
 
}
 
}
−
.insert2 {
+
  .plasmid .scale-minor {
−
  height: 100px;
+
  stroke: #94aab0;
−
  width: 100%
+
−
  background-color:#fff;
+
 
}
 
}
−
 
+
  .plasmid .scale-majortext {
−
.box {
+
   font: 300 9px "Quicksand";
−
   background:#fff;
+
   fill: #94aab0;
−
   border:0px solid;
+
−
  border-color: #e2e2e2;
+
−
  margin-top: 0px;
+
−
  box-sizing: border-box;
+
−
  background-clip: padding-box;
+
−
  padding:0 20px 20px 20px;
+
−
  min-height:500px;
+
−
  min-width:400px
+
 
}
 
}
−
.they-said, .we-said {
+
  .plasmid .markerlabel {
−
display:inline-block;
+
  font: 400 9px "Quicksand";
 +
  fill: #334145;
 
}
 
}
−
.they-said:after, .we-said:after {
+
  .plasmid .featurelabel {
−
content: '';
+
  font: 700 14px "Quicksand";
−
display: block;
+
  fill: #c00;
−
height: 3px;
+
−
width: 0;
+
−
background: transparent;
+
−
transition: width .5s ease, background-color .5s ease;
+
 
}
 
}
−
.they-said:hover:after, .we-said:hover:after {
+
  .plasmid .regionlabel {
−
width: 100%;
+
  font: 400 11px "Lato";
−
background: #3e3f3f;
+
}
 +
  .plasmid .noncodingregion {
 +
  fill: none;
 +
  stroke: #a3b6bb;
 +
  stroke-dasharray: 4,2;
 +
}
 +
  .plasmid .noncodinglabel {
 +
  font: 400 9px "Quicksand";
 +
  fill: #5e7980;
 +
}
 +
  .plasmid .atracklabel {
 +
  font: 400 20px "Quicksand";
 +
  color: #5e7980;
 +
  text-anchor: middle;
 +
}
 +
  .plasmid .atracklabel h1 {
 +
  font: 700 40px "Quicksand";
 +
}
 +
  .plasmid .atracklabel .tiny {
 +
  font: 400 12px "Quicksand";
 +
  margin-top: 10px;
 
}
 
}
  
−
.they-said-content, .we-said, .we-said-content {display:none;}
+
.element_info_box_container {
 +
width:95%;
 +
height:460px;
 +
overflow:scroll;
 +
background-color:#e2e2e2;
 +
border-radius:20px;
 +
}
 +
.pus270_container {
 +
display:flex;
 +
flex-direction:row;
 +
flex-wrap: nowrap ;
 +
justify-content:  space-around;
 +
align-items:center;
 +
align-content:space-around;
 +
}
 +
.pus_270 {flex-basis:60%;}
 +
.pus_270_info {flex-grow:2;}
 +
 
 +
.lacr_info, .pip_info, .eoriv_info, .mcs_info, .anti_info, .boriv_info, .laco_info, .super_info {display:none;}
 
</style>
 
</style>
 +
 
</head>
 
</head>
−
 
 
<body>
 
<body>
−
<div class = "insert">
 
−
</div>
 
  
−
<!-- Banner-->
+
<div class="pus270_container">
−
<div class="container-fluid page-heading">
+
<div class="pus_270">
−
<div class = "row">
+
<plasmid id="pdemo" class="plasmid" plasmidheight="450" plasmidwidth="600" sequencelength="1240">
−
<div style="background-color:#e2e2e2">
+
    <plasmidtrack style="fill:#e2e2e2;stroke:none;" trackclass="track" radius="200" width="5">
−
<div class = "crispy2">
+
    <!-- plasmid title -->
−
      <center><img src="https://static.igem.org/mediawiki/2017/8/84/T--Sydney_Australia--HumanPracticesBanner.gif" class="img-center" width="70%"></center>
+
        <tracklabel text="pUS270" labelstyle="font-size:30px;font-weight:700;font-family:'Quicksand';"></tracklabel>
−
</div>
+
        <tracklabel text="5707 bp" labelstyle="font-size:18px;font-weight:400;font-family:'Quicksand';" vadjust="25"></tracklabel>
−
</div>
+
−
</div>
+
−
</div>
+
  
−
<div class ="row" style = "background-color:#e2e2e2">
 
−
<div class = "crispy">
 
−
<div class = "col-xs-2" style = "background-color:#e2e2e2">
 
−
    <a href="#nav1"><center><img src="https://static.igem.org/mediawiki/2017/3/30/T--Sydney_Australia--LogoInsulinforLife.gif" width="100%"></center></a>
 
−
</div>
 
−
<div class = "col-xs-2" style = "background-color:#e2e2e2">
 
−
    <a href="#nav2"><center><img src="https://static.igem.org/mediawiki/2017/8/86/T--Sydney_Australia--LogoIncubate.gif" width="100%"></center></a>
 
−
</div>
 
−
<div class = "col-xs-2" style = "background-color:#e2e2e2">
 
−
    <a href="#nav3"><center><img src="https://static.igem.org/mediawiki/2017/4/4b/T--Sydney_Australia--LogoMaxwells.gif" width="100%"></center></a>
 
−
</div>
 
−
<div class = "col-xs-2" style = "background-color:#e2e2e2">
 
−
    <a href="#nav4"><center><img src="https://static.igem.org/mediawiki/2017/6/6e/T--Sydney_Australia--LogoBiofoundry.gif" width="100%"></center></a>
 
−
</div>
 
−
<div class = "col-xs-2" style = "background-color:#e2e2e2">
 
−
    <a href="#nav5"><center><img src="https://static.igem.org/mediawiki/2017/5/5e/T--Sydney_Australia--LogoACCISS.gif" width="100%"></center></a>
 
−
</div>
 
−
<div class = "col-xs-2" style = "background-color:#e2e2e2">
 
−
    <a href="#nav6"><center><img src="https://static.igem.org/mediawiki/2017/8/8c/T--Sydney_Australia--LogoT1International.gif" width="100%"></center></a>
 
−
</div>
 
−
</div>
 
−
</div>
 
  
−
<div class = "image-fixed-top navbar-default">
+
        <!-- NeoR/KanR -->  
−
<div class = "insert" style="background-color:#e2e2e2">
+
  <trackmarker markerclass="markerhover" start="0" end="200" style="fill:#f2cbf1"  arrowendlength="15" arrowendwidth="10" vadjust="-10" wadjust="20">
−
</div>  
+
                <svgelement type="a" xlink:href="" id="anti" target="_blank">
−
</div>  
+
            <markerlabel labelclass="markerlabel" text="NeoR/KanR" style="fill:#3e3f3f;font-weight:700;font-family:'Quicksand';font-size:16px;" ></markerlabel>
 +
            </svgelement>
  
−
<div class = "row" id="nav1">
+
  </trackmarker>
−
<div class = "insert2">
+
        <!-- pIP501 Rep -->   
−
</div>  
+
  <trackmarker markerclass="markerhover" start="220" end="420" style="fill:#b1d7dd" arrowendlength="15" arrowendwidth="10" vadjust="-10" wadjust="20">
−
</div>
+
                <svgelement type="a" xlink:href="" id="pip" target="_blank">
 +
          <markerlabel labelclass="markerlabel" text="pIP501 Rep" style="fill:#3e3f3f;font-weight:700;font-family:'Quicksand';font-size:16px;" ></markerlabel>
 +
            </svgelement>
 +
            </trackmarker>
 +
      <!-- B. Subtilis OriV -->   
 +
  <trackmarker markerclass="markerhover" start="440" end="540" style="fill:#eca0a7" vadjust="-10" wadjust="20">
 +
                    <svgelement type="a" xlink:href="" id="boriv" target="_blank">
 +
          <markerlabel labelclass="markerlabel" text="B. Subtilis OriV" style="fill:#3e3f3f;font-weight:700;font-family:'Quicksand';font-size:16px;" ></markerlabel>
 +
            </svgelement>
 +
            </trackmarker>
 +
 
 +
        <!-- E. Coli OriV -->   
 +
  <trackmarker markerclass="markerhover" start="560" end="660" style="fill:#fbde9e" vadjust="-10" wadjust="20">
 +
                    <svgelement type="a" xlink:href="" id="eoriv" target="_blank">
 +
          <markerlabel labelclass="markerlabel" text="E. Coli OriV" style="fill:#3e3f3f;font-weight:700;font-family:'Quicksand';font-size:16px;" ></markerlabel>
 +
            </svgelement>
 +
            </trackmarker>
 +
 +
        <!-- Lac Operator -->   
 +
  <trackmarker markerclass="markerhover" start="680" end="780" style="fill:#cdf8ff" vadjust="-10" wadjust="20">
 +
                    <svgelement type="a" xlink:href="" id="laco" target="_blank">
 +
          <markerlabel labelclass="markerlabel" text="Lac Operator" style="fill:#3e3f3f;font-weight:700;font-family:'Quicksand';font-size:16px;" ></markerlabel>
 +
            </svgelement>
 +
            </trackmarker>
  
−
<div class = "row">
+
        <!-- MCS -->  
−
<div class = "crispy">
+
  <trackmarker markerclass="markerhover" start="800" end="840" style="fill:#d0cece" vadjust="-10" wadjust="20">
−
<div class="col-xs-1" align = "right">
+
                  <svgelement type="a" xlink:href="" id="mcs1" target="_blank">
−
<img src="https://static.igem.org/mediawiki/2017/0/09/T--Sydney_Australia--Handshake1.gif">
+
          <markerlabel labelclass="markerlabel" text="MCS" style="fill:#3e3f3f;font-weight:700;font-family:'Quicksand';font-size:16px;" ></markerlabel>
−
</div>
+
            </svgelement>
−
<div class = "col-xs-5" class="box" id="please">
+
            </trackmarker>
−
<center><h1 class="tag-title">Insulin for Life</h1></center>
+
      <!-- Sf-AmilCP-6 -->  
−
<center><h4>Blurb about Insulin for Life</h4></center>
+
  <trackmarker markerclass="markerhover" start="860" end="1000" style="fill:#a395c8" arrowendlength="15" arrowendwidth="10" vadjust="-10" wadjust="20">
−
</div>
+
                  <svgelement type="a" xlink:href="" id="super" target="_blank">
−
<div class = "col-xs-6" class="box" id = "please">
+
          <markerlabel labelclass="markerlabel" text="Sf-AmilCP-6" style="fill:#3e3f3f;font-weight:700;font-family:'Quicksand';font-size:16px;" ></markerlabel>
−
      <center><h1 class = "tag-title they-said">They said ></h1></center>
+
            </svgelement>
−
<center><h4 class="they-said-content">What they said</h4></center>
+
            </trackmarker>
−
        <center><h1 class="tag-title we-said">We said ></h1></center>
+
−
<center><h4 class="we-said-content">What we said</h4></center>
+
 
−
</div>
+
      <!-- MCS -->  
−
</div>
+
  <trackmarker markerclass="markerhover" start="1020" end="1060" style="fill:#d0cece" vadjust="-10" wadjust="20">
−
</div>
+
                  <svgelement type="a" xlink:href="" id="mcs2" target="_blank">
 +
          <markerlabel labelclass="markerlabel" text="MCS" style="fill:#3e3f3f;font-weight:700;font-family:'Quicksand';font-size:16px;" ></markerlabel>
 +
            </svgelement>
 +
            </trackmarker>
  
−
<div class = "row" id="nav2">
+
      <!-- LacI-->   
−
<div class = "insert2">
+
  <trackmarker markerclass="markerhover" start="1080" end="1220" style="fill:#afe0b2" arrowendlength="15" arrowendwidth="10" vadjust="-10" wadjust="20">
−
</div>  
+
                    <svgelement type="a" xlink:href="" id="lacr" target="_blank">
 +
          <markerlabel labelclass="markerlabel" text="LacI" style="fill:#3e3f3f;font-weight:700;font-family:'Quicksand';font-size:16px;" ></markerlabel>
 +
            </svgelement>
 +
            </trackmarker>
 +
 +
    </plasmidtrack>
 +
</plasmid>
 
</div>
 
</div>
 +
<div class="pus_270_info element_info_box_container">
 +
<div class="lacr_info">
 +
<h6>Lac Operon Repressor</h6>
  
−
<div class = "row">
+
<h4>Codes for the production of the repressor protein that binds to the Lac Operator to prevent expression until induction of the media with IPTG. We have designed pUS270 to have the LacI under the pVeg promoter for strong constitutive expression in Bacillus.</h4>
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<div class = "crispy">
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<img src="https://static.igem.org/mediawiki/2017/4/4f/T--Sydney_Australia--Handshake3.gif">
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<center><h1 class="tag-title">Incubate</h1></center>
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<center><h4>Blurb about Incubate</h4></center>
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      <center><h1 class = "tag-title they-said">They said ></h1></center>
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</div>
+
 
</div>
 
</div>
 +
 +
<div class="pip_info">
 +
<h6>pIP501 Rep</h6>
 +
<h4>This gene codes for the Replicator protein that pairs with the Bacillus OriV and is essential to promote the independent replication of pUS270 in Bacillus. </h4>
 
</div>
 
</div>
  
−
<div class = "row" id="nav3">
+
<div class="eoriv_info">
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<div class = "insert2">
+
<h6>E. coli OriV</h6>
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</div>  
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<h4>Taken from pSB1C3, having this origin of replication will make it easier to construct the plasmid using E. coli as well as improve cloning efficiency downstream. Chosen because of the high copy number of pSB1C3.</h4>
 
</div>
 
</div>
  
−
<div class = "row">
+
<div class="mcs_info">
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<div class = "crispy">
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<h6>Multiple Cloning Site</h6>
−
<div class="col-xs-1" align = "right">
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<h4>The multiple cloning site of pUS258 and other expression plasmids that we considered using either had too few restriction sites or sites that were incompatible for the efficient cloning with the BB Prefix and Suffix. So we have designed two highly diverse MCS’ that have 27 unique sites amongst them on either side of SF AmylCP-6 to promote simple cloning of parts from the Bio Brick Registry.</h4>
−
<img src="https://static.igem.org/mediawiki/2017/7/76/T--Sydney_Australia--Handshake4.gif">
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−
</div>
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<div class = "col-xs-5" class="box" id="please">
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<center><h1 class="tag-title">Maxwells</h1></center>
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<center><h4>Blurb about Maxwells</h4></center>
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<div class = "col-xs-6" class="box" id = "please">
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      <center><h1 class = "tag-title they-said">They said ></h1></center>
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        <center><h1 class="tag-title we-said">We said ></h1></center>
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<center><h4 class="we-said-content">What we said</h4></center>
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</div>
+
−
</div>
+
 
</div>
 
</div>
  
−
<div class = "row" id="nav4">
+
<div class="anti_info">
−
<div class = "insert2">
+
<h6>Antibiotic Resistance</h6>
−
</div>
+
−
</div>
+
  
−
<div class = "row">
+
<h4>Codes for Neomycin Phosphotransferase II that gives transformants resistance to both Neomycin and Kanamycin. Allows selective screening of transformants</h4>
−
<div class = "crispy">
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−
<div class="col-xs-1" align = "right">
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<img src="https://static.igem.org/mediawiki/2017/b/bb/T--Sydney_Australia--Handshake5.gif">
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<center><h1 class="tag-title">Biofoundry & Open Insulin</h1></center>
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<center><h4 class="we-said-content">What we said</h4></center>
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</div>
+
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</div>
+
 
</div>
 
</div>
  
−
<div class = "row" id="nav5">
+
<div class="boriv_info">
−
<div class = "insert2">
+
<h6> Antibiotic Resistance </h6>
−
</div>  
+
<h4>This region allows independent replication of the vector within Bacillus at a high copy number to provide more copies of the recombinant gene to maximise expression.</h4>
 
</div>
 
</div>
  
−
<div class = "row">
+
<div class="laco_info">
−
<div class = "crispy">
+
<h6>Lac Operator </h6>
−
<div class="col-xs-1" align = "right">
+
<h4>Consists of the target sequence for the lac repressor protein to contribute to the IPTG induction system.</h4>
−
<img src="https://static.igem.org/mediawiki/2017/9/93/T--Sydney_Australia--Handshake6.gif">
+
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<center><h1 class="tag-title">ACCISS</h1></center>
+
−
<center><h4>Blurb about ACCISS</h4></center>
+
−
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+
−
</div>
+
 
</div>
 
</div>
  
−
<div class = "row" id="nav6">
+
<div class="super_info">
−
<div class = "insert2">
+
<h6>SuperFold AmilCP-6</h6>
−
</div>  
+
<h4>SuperFold AmilCP-6 is a variant of amilCP which was used by the 2016 Sydney IGEM team and originally submitted to the registry by Team Uppsala Sweden in 2011. This variant has undergone site directed mutagenesis to increase the folding rate of the chromoprotein and enhance the blue colour. We are taking advantage of this protein to enable blue-white screening without the need for adding X-gal to the plates. Cloning with the two multiple cloning sites that flank SF AmilCP-6 essentially swaps it with the insert to leave the recombinant bacteria white rather than blue. </h4>
 
</div>
 
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Latest revision as of 10:43, 1 November 2017

Lac Operon Repressor

Codes for the production of the repressor protein that binds to the Lac Operator to prevent expression until induction of the media with IPTG. We have designed pUS270 to have the LacI under the pVeg promoter for strong constitutive expression in Bacillus.

pIP501 Rep

This gene codes for the Replicator protein that pairs with the Bacillus OriV and is essential to promote the independent replication of pUS270 in Bacillus.

E. coli OriV

Taken from pSB1C3, having this origin of replication will make it easier to construct the plasmid using E. coli as well as improve cloning efficiency downstream. Chosen because of the high copy number of pSB1C3.

Multiple Cloning Site

The multiple cloning site of pUS258 and other expression plasmids that we considered using either had too few restriction sites or sites that were incompatible for the efficient cloning with the BB Prefix and Suffix. So we have designed two highly diverse MCS’ that have 27 unique sites amongst them on either side of SF AmylCP-6 to promote simple cloning of parts from the Bio Brick Registry.

Antibiotic Resistance

Codes for Neomycin Phosphotransferase II that gives transformants resistance to both Neomycin and Kanamycin. Allows selective screening of transformants

Antibiotic Resistance

This region allows independent replication of the vector within Bacillus at a high copy number to provide more copies of the recombinant gene to maximise expression.

Lac Operator

Consists of the target sequence for the lac repressor protein to contribute to the IPTG induction system.

SuperFold AmilCP-6

SuperFold AmilCP-6 is a variant of amilCP which was used by the 2016 Sydney IGEM team and originally submitted to the registry by Team Uppsala Sweden in 2011. This variant has undergone site directed mutagenesis to increase the folding rate of the chromoprotein and enhance the blue colour. We are taking advantage of this protein to enable blue-white screening without the need for adding X-gal to the plates. Cloning with the two multiple cloning sites that flank SF AmilCP-6 essentially swaps it with the insert to leave the recombinant bacteria white rather than blue. 


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