Team:CSU Fort Collins

From 2014.igem.org

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{{CSS/Main}}
{{CSS/Main}}
-
 
<html>
<html>
-
<!--main content -->
+
 
 +
 
 +
<title>Safety</title>
 +
 
 +
<head>
<link href='http://fonts.googleapis.com/css?family=Bitter:400,700,400italic' rel='stylesheet' type='text/css'>
<link href='http://fonts.googleapis.com/css?family=Bitter:400,700,400italic' rel='stylesheet' type='text/css'>
<link href='http://fonts.googleapis.com/css?family=Muli:300,400' rel='stylesheet' type='text/css'>
<link href='http://fonts.googleapis.com/css?family=Muli:300,400' rel='stylesheet' type='text/css'>
-
 
-
<body style="background-size:100%; background-color:#EFEFEF">
 
<style type="text/css">
<style type="text/css">
 +
/* http://meyerweb.com/eric/tools/css/reset/
 +
  v2.0 | 20110126
 +
  License: none (public domain)
 +
*/
 +
 +
html, body, div, span, applet, object, iframe, p, blockquote, pre,
 +
a, abbr, acronym, address, big, cite, code,
 +
del, dfn, em, img, ins, kbd, q, s, samp,
 +
small, strike, strong, sub, sup, tt, var,
 +
b, u, i, center,
 +
dl, dt, dd, ol, ul, li,
 +
fieldset, form, label, legend,
 +
table, caption, tbody, tfoot, thead, tr, th, td,
 +
article, aside, canvas, details, embed,
 +
figure, figcaption, footer, header, hgroup,
 +
menu, nav, output, ruby, section, summary,
 +
time, mark, audio, video {
 +
  margin: 0;
 +
  padding: 0;
 +
  border: 0;
 +
  font-size: 90%;
 +
  font: inherit;
 +
  vertical-align: baseline;
 +
}
 +
/* HTML5 display-role reset for older browsers */
 +
article, aside, details, figcaption, figure,
 +
footer, header, hgroup, menu, nav, section {
 +
  display: block;
 +
}
 +
body {
 +
  line-height: 1;
 +
}
 +
ol, ul {
 +
  list-style: none;
 +
}
 +
blockquote, q {
 +
  quotes: none;
 +
}
 +
blockquote:before, blockquote:after,
 +
q:before, q:after {
 +
  content: '';
 +
  content: none;
 +
}
 +
table {
 +
  border-collapse: collapse;
 +
  border-spacing: 0;
 +
}
 +
 +
#menubar > ul{
 +
  vertical-align:center;
 +
  font-size:14px;
 +
  font-family:'Times New Roman',serif;
 +
}
 +
.firstHeading{
.firstHeading{
-
    display:none;
+
display:none;
}
}
-
A{
+
 
-
    color:#222222;
+
#textbox span{
 +
  color:#FF8533;
}
}
-
A:visited{
+
.photo{
-
    color:#222222;
+
  width:70%;
}
}
-
A:hover{
+
table{
-
    opacity:0.3;
+
  font-family:'Muli',sans-serif;
-
    text-decoration:none;
+
}
}
-
h1,h2,h3,h4,h5,h6{
+
body{
-
    font-family:'Bitter', serif;
+
  background-color:#EFEFEF;
-
    text-decoration:none;
+
  line-height:1.3;
-
    font-weight:700;
+
  font-size:90%;
-
    color:#222222;
+
-
    border:none;
+
}
}
-
.button{
+
#cssmenu {
-
    margin-left:15px;
+
  width:100%;
-
    margin-right:15px;
+
  height: 44px;
-
    padding:10px;
+
  background: #222222;
-
    font-family:'Bitter', serif;
+
  position:relative;
-
    font-size:14px;
+
  text-align:center;
-
    width:5em;
+
  margin:auto;
}
}
-
.sbutton{
+
#cssmenu ul {
-
    padding:5px 0px;
+
  list-style: none;
-
    font-family:'Bitter', serif;
+
  padding: 0;
-
    font-size:14px;
+
  margin: 0;
 +
  line-height: 1;
}
}
-
nav ul{
+
#cssmenu > ul {
-
    position:relative;
+
  position: relative;
-
    list-style:none;
+
  display: block;
-
    white-space:nowrap;
+
  background: #222222;
 +
  width: 100%;
 +
  z-index: 500;
}
}
-
nav ul li{
+
#cssmenu:after,
-
    display:inline-block;
+
#cssmenu > ul:after {
-
    white-space:nowrap;
+
  content: '.';
 +
  display: block;
 +
  clear: both;
 +
  visibility: hidden;
 +
  line-height: 0;
 +
  height: 0;
}
}
-
nav ul ul {
+
#cssmenu.align-right > ul > li {
-
    display: none;
+
  float: right;
-
    position:absolute;
+
-
    margin-left: 0;
+
-
    margin-top: 10px;
+
}
}
-
nav ul ul li{
+
#cssmenu.align-center ul {
-
    display: block;
+
  text-align: center;
-
    text-align: left;
+
-
    float: none;
+
}
}
-
nav ul ul .button {
+
#cssmenu.align-center ul ul {
-
    margin-left: 0;
+
  text-align: left;
-
    padding-left: 0;
+
}
}
-
nav ul li a{
+
#cssmenu > ul > li {
-
    display:block !important;
+
  display: inline-block;
 +
  position: relative;
 +
  margin: 0;
 +
  padding: 0;
 +
}
 +
#cssmenu > ul > #menu-button {
 +
  display: none;
 +
}
 +
#cssmenu ul li a {
 +
  display: block;
 +
  font-family: Helvetica, sans-serif;
 +
  text-decoration: none;
 +
}
 +
#cssmenu > ul > li > a {
 +
  font-size: 14px;
 +
  font-weight: bold;
 +
  padding: 15px 20px;
 +
  color: #efefef;
 +
  text-transform: uppercase;
 +
  -webkit-transition: color 0.25s ease-out;
 +
  -moz-transition: color 0.25s ease-out;
 +
  -ms-transition: color 0.25s ease-out;
 +
  -o-transition: color 0.25s ease-out;
 +
  transition: color 0.25s ease-out;
 +
}
 +
#cssmenu > ul > li.has-sub > a {
 +
  padding-right: 32px;
 +
}
 +
#cssmenu > ul > li:hover > a {
 +
  color: #f2a76d;
 +
}
 +
#cssmenu li.has-sub::after {
 +
  display: block;
 +
  content: '';
 +
  position: absolute;
 +
  width: 0;
 +
  height: 0;
 +
}
 +
#cssmenu > ul > li.has-sub::after {
 +
  right: 10px;
 +
  top: 20px;
 +
  border: 5px solid transparent;
 +
  border-top-color: #efefef;
 +
}
 +
#cssmenu > ul > li:hover::after {
 +
  border-top-color: #f2a76d;
 +
}
 +
#indicatorContainer {
 +
  position: absolute;
 +
  height: 12px;
 +
  width: 100%;
 +
  bottom: 0px;
 +
  overflow: hidden;
 +
  z-index: -1;
 +
}
 +
#pIndicator {
 +
  position: absolute;
 +
  height: 0;
 +
  width: 100%;
 +
  border: 12px solid transparent;
 +
  border-top-color: #2b2f3a;
 +
  z-index: -2;
 +
  -webkit-transition: left .25s ease;
 +
  -moz-transition: left .25s ease;
 +
  -ms-transition: left .25s ease;
 +
  -o-transition: left .25s ease;
 +
  transition: left .25s ease;
 +
}
 +
#cIndicator {
 +
  position: absolute;
 +
  height: 0;
 +
  width: 100%;
 +
  border: 12px solid transparent;
 +
  border-top-color: #2b2f3a;
 +
  top: -12px;
 +
  right: 100%;
 +
  z-index: -2;
 +
}
 +
#cssmenu ul ul {
 +
  position: absolute;
 +
  left: -9999px;
 +
  top: 70px;
 +
  opacity: 0;
 +
  -webkit-transition: opacity .3s ease, top .25s ease;
 +
  -moz-transition: opacity .3s ease, top .25s ease;
 +
  -ms-transition: opacity .3s ease, top .25s ease;
 +
  -o-transition: opacity .3s ease, top .25s ease;
 +
  transition: opacity .3s ease, top .25s ease;
 +
  z-index: 1000;
 +
}
 +
#cssmenu ul ul ul {
 +
  top: 37px;
 +
  padding-left: 5px;
 +
}
 +
#cssmenu ul ul li {
 +
  position: relative;
 +
}
 +
#cssmenu > ul > li:hover > ul {
 +
  left: auto;
 +
  top: 44px;
 +
  opacity: 1;
 +
}
 +
#cssmenu.align-right > ul > li:hover > ul {
 +
  left: auto;
 +
  right: 0;
 +
  opacity: 1;
 +
}
 +
#cssmenu ul ul li:hover > ul {
 +
  left: 170px;
 +
  top: 0;
 +
  opacity: 1;
 +
}
 +
#cssmenu.align-right ul ul li:hover > ul {
 +
  left: auto;
 +
  right: 170px;
 +
  top: 0;
 +
  opacity: 1;
 +
  padding-right: 5px;
 +
}
 +
#cssmenu ul ul li a {
 +
  width: 130px;
 +
  border-bottom: 1px solid #eeeeee;
 +
  padding: 10px 20px;
 +
  font-size: 12px;
 +
  color: #222222;
 +
  background: white;
 +
  -webkit-transition: all .35s ease;
 +
  -moz-transition: all .35s ease;
 +
  -ms-transition: all .35s ease;
 +
  -o-transition: all .35s ease;
 +
  transition: all .35s ease;
 +
}
 +
#cssmenu.align-right ul ul li a {
 +
  text-align: right;
 +
}
 +
#cssmenu ul ul li:hover > a {
 +
  background: #e2e2e2;
 +
  color: #FF8533;
 +
}
 +
#cssmenu ul ul li:last-child > a,
 +
#cssmenu ul ul li.last > a {
 +
  border-bottom: 0;
 +
}
 +
#cssmenu > ul > li > ul::after {
 +
  content: '';
 +
  border: 6px solid transparent;
 +
  width: 0;
 +
  height: 0;
 +
  border-bottom-color: #efefef;
 +
  position: absolute;
 +
  top: -12px;
 +
  left: 30px;
 +
}
 +
#cssmenu.align-right > ul > li > ul::after {
 +
  left: auto;
 +
  right: 30px;
 +
}
 +
#cssmenu ul ul li.has-sub::after {
 +
  border: 4px solid transparent;
 +
  border-left-color: #222222;
 +
  right: 10px;
 +
  top: 12px;
 +
  -moz-transition: all .2s ease;
 +
  -ms-transition: all .2s ease;
 +
  -o-transition: all .2s ease;
 +
  transition: all .2s ease;
 +
  -webkit-transition: -webkit-transform 0.2s ease, right 0.2s ease;
 +
}
 +
#cssmenu.align-right ul ul li.has-sub::after {
 +
  border-left-color: transparent;
 +
  border-right-color: #222222;
 +
  right: auto;
 +
  left: 10px;
 +
}
 +
#cssmenu ul ul li.has-sub:hover::after {
 +
  border-left-color: #efefef;
 +
  right: -5px;
 +
  -webkit-transform: rotateY(180deg);
 +
  -ms-transform: rotateY(180deg);
 +
  -moz-transform: rotateY(180deg);
 +
  -o-transform: rotateY(180deg);
 +
  transform: rotateY(180deg);
 +
}
 +
#cssmenu.align-right ul ul li.has-sub:hover::after {
 +
  border-right-color: #efefef;
 +
  border-left-color: transparent;
 +
  left: -5px;
 +
  -webkit-transform: rotateY(180deg);
 +
  -ms-transform: rotateY(180deg);
 +
  -moz-transform: rotateY(180deg);
 +
  -o-transform: rotateY(180deg);
 +
  transform: rotateY(180deg);
 +
}
 +
@media all and (max-width: 800px), only screen and (-webkit-min-device-pixel-ratio: 2) and (max-width: 1024px), only screen and (min--moz-device-pixel-ratio: 2) and (max-width: 1024px), only screen and (-o-min-device-pixel-ratio: 2/1) and (max-width: 1024px), only screen and (min-device-pixel-ratio: 2) and (max-width: 1024px), only screen and (min-resolution: 192dpi) and (max-width: 1024px), only screen and (min-resolution: 2dppx) and (max-width: 1024px) {
 +
  #cssmenu {
 +
    width: 100%;
 +
  }
 +
  #cssmenu.align-center ul {
 +
    text-align: left;
 +
  }
 +
  #cssmenu.align-right > ul > li {
 +
    float: none;
 +
  }
 +
  #cssmenu ul {
 +
    width: auto;
 +
  }
 +
  #cssmenu .submenuArrow,
 +
  #cssmenu #indicatorContainer {
 +
    display: none;
 +
  }
 +
  #cssmenu > ul {
 +
    height: auto;
 +
    display: block;
 +
    color:#EFEFEF;
 +
  }
 +
  #cssmenu > ul > li {
 +
    float: none;
 +
  }
 +
  #cssmenu li,
 +
  #cssmenu > ul > li {
 +
    display: none;
 +
    color:white;
 +
  }
 +
  #cssmenu ul ul,
 +
  #cssmenu ul ul ul,
 +
  #cssmenu ul > li:hover > ul,
 +
  #cssmenu ul ul > li:hover > ul,
 +
  #cssmenu.align-right ul ul,
 +
  #cssmenu.align-right ul ul ul,
 +
  #cssmenu.align-right ul > li:hover > ul,
 +
  #cssmenu.align-right ul ul > li:hover > ul {
 +
    position: relative;
 +
    left: auto;
 +
    top: auto;
 +
    opacity: 1;
 +
    padding-left: 0;
 +
    padding-right: 0;
 +
    right: auto;
 +
  }
 +
  #cssmenu ul .has-sub::after {
 +
    display: none;
 +
  }
 +
  #cssmenu ul li a {
 +
    padding: 12px 20px;
 +
  }
 +
  #cssmenu ul ul li a {
 +
    border: 0;
 +
    background: none;
 +
    width: auto;
 +
    padding: 8px 35px;
 +
    color:white;
 +
  }
 +
  #cssmenu.align-right ul ul li a {
 +
    text-align: left;
 +
  }
 +
  #cssmenu ul ul li:hover > a {
 +
    background:none;
 +
    color: #ec9359;
 +
  }
 +
  #cssmenu ul ul ul a {
 +
    padding: 8px 50px;
 +
  }
 +
  #cssmenu ul ul ul ul a {
 +
    padding: 8px 65px;
 +
  }
 +
  #cssmenu ul ul ul ul ul a {
 +
    padding: 8px 80px;
 +
  }
 +
  #cssmenu ul ul ul ul ul ul a {
 +
    padding: 8px 95px;
 +
  }
 +
  #cssmenu > ul > #menu-button {
 +
    display: block;
 +
    cursor: pointer;
 +
  }
 +
  #cssmenu #menu-button > a {
 +
    padding: 14px 20px;
 +
  }
 +
  #cssmenu ul.open li,
 +
  #cssmenu > ul.open > li {
 +
    display: block;
 +
  }
 +
  #cssmenu > ul.open > li#menu-button > a {
 +
    color: #fff;
 +
    border-bottom: 1px solid rgba(150, 150, 150, 0.1);
 +
  }
 +
  #cssmenu ul ul::after {
 +
    display: none;
 +
  }
 +
  #cssmenu #menu-button::after {
 +
    display: block;
 +
    content: '';
 +
    position: absolute;
 +
    height: 3px;
 +
    width: 22px;
 +
    border-top: 2px solid #efefef;
 +
    border-bottom: 2px solid #efefef;
 +
    right: 20px;
 +
    top: 15px;
 +
  }
 +
  #cssmenu #menu-button::before {
 +
    display: block;
 +
    content: '';
 +
    position: absolute;
 +
    height: 3px;
 +
    width: 22px;
 +
    border-top: 2px solid #efefef;
 +
    right: 20px;
 +
    top: 25px;
 +
  }
 +
  #cssmenu ul.open #menu-button::after,
 +
  #cssmenu ul.open #menu-button::before {
 +
    border-color: #fff;
 +
  }
 +
}
 +
.icon{
 +
  width:30px;
 +
}
 +
.wrapper2{
 +
  width:20%;
 +
  padding:20px 0px 20px 20px;
 +
  display:inline-block;
 +
  margin:auto;
 +
  vertical-align:top;
 +
}
 +
.wrapper{
 +
  width:90%;
 +
  padding:20px 0px 20px 20px;
 +
  padding-left:0;
 +
  margin:auto;
 +
  vertical-align: top;
 +
}
 +
.page, .container{
 +
  width:50%;
 +
  min-width:600px;
 +
  max-width:800px;
 +
  padding:30px;
 +
  margin:auto;
 +
  text-align:left;
 +
  font-family:'Muli',sans-serif;
 +
  display:inline-block;
 +
}
 +
.page a, .container a{
 +
  color:#222222;
 +
}
 +
.page a:hover, .container a:hover{
 +
  color:#FF8533;
 +
  text-decoration:none;
}
}
-
nav ul li:hover > ul {
+
h1{
-
    display:block;
+
  color:#222222;
 +
  font-family:'Bitter',serif;
 +
  border-bottom:none;
}
}
-
.footer{
+
h2,h3,.toc,.show,.hide,.content{
-
    background-color:#222222;
+
  color:#222222;
-
    width:100%;
+
  font-family:'Bitter',serif;
-
    margin:auto;
+
-
    padding:15px;
+
-
    white-space:nowrap;
+
}
}
-
p{
+
h4,h5,h6, .page{
-
    font-family:'Muli', sans-serif;
+
  color:#222222;
-
    color:white;
+
  font-family:'Muli',sans-serif;
-
    font-size:12px;
+
}
}
 +
.toc{
 +
  text-decoration:none;
 +
  display:none;
 +
  width:100%;
 +
  margin:auto;
 +
}
 +
.toc .month-group { margin: 20px 0; }
 +
.toc .month-group p { padding: 0; margin: 0; }
 +
#ftoc{
 +
  color:#666666;
 +
}
 +
ul{
 +
  list-style-type:none;
 +
}
 +
.month-group, a{
 +
  text-decoration:none;
 +
}
 +
.month-group{
 +
  margin-left:20px;
 +
}
 +
.month-group:hover{
 +
  color:#FF8533;
 +
}
 +
.toc a, .toc2 a, #navi a{
 +
  text-decoration:none;
 +
  color:#222222;
 +
  font-weight: bold;
 +
}
 +
.toc a:hover, #ftoc>a:hover, #navi a:hover, .show:hover, .hide:hover, .container>a:hover{
 +
  color:#FF8533 !important;
 +
  cursor:pointer;
 +
}
 +
.toc a, #ftoc, .container a, #navi{
 +
  border-bottom:2px dotted;
 +
}
 +
#navi{
 +
  text-decoration:none;
 +
  font-family:'Bitter',serif;
 +
}
 +
#logo{
 +
  width:175px;
 +
  margin:auto;
 +
  padding:40px;
 +
  border-radius:100%;
 +
}
 +
#banner{
 +
  width:auto;
 +
  height:250px;
 +
  vertical-align:top;
 +
  background-color:#222222;
 +
}
 +
.fixed {
 +
  position:fixed !important;
 +
  top:0;
 +
}
 +
.disp{
 +
  display:none;
 +
}
 +
.col {
 +
    width: 20%;
 +
    float: left;
 +
    clear: right;
 +
    margin: 20px 1.66666666666666666666666666%;
 +
    font-family:'Muli',sans-serif;
 +
    font-weight:300;
 +
    font-size:14px;
 +
    color:#222222;
 +
    text-align:left;
 +
}
 +
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Revision as of 18:46, 15 September 2014

Safety

Welcome to CSU's 2014 iGEM Wiki


Approximately 3 billion gallons per year of used frying oil are produced in the U.S. alone. While some recycling efforts are put forth to turn used frying oil into biodiesel and some companies are working on more effective ways to recycle the frying oil, the iGEM team at Colorado State University is working to turn spent frying oil into a high value product. There are four major components to this year’s project including; the breakdown of frying oil, a biosensor for detecting the breakdown of frying oil, production of a high value product, and a kill switch to kill the bacteria if they were to be released into the environment. Our team aims to put each of these components into Escherichia coli.


In order to detect that the process is working we have developed a biosensor that will detect the breakdown of Acyl-CoA, which is a byproduct of the breakdown pathway of lipids in E. coli. Our biosensor is being constructed by inserting a novel promoter in front of DNA expressing a Green Fluorescent Protein (GFP). This promoter is activated by Acyl-CoA. This will result in E. coli expressing GFP when successful breakdown is occurring.


In order to break down the frying oil we will be taking advantage of the cell’s natural ability to break down fatty acids for use in the Kreb’s cycle. We will be up-regulating the limiting enzymes that aid in the breakdown of fatty acids which will get the cell to produce more Acetyl-CoA.


This Acetyl-CoA can then be used in the mevalonate pathway to produce terpenoids. This pathway is common to most plants as well as yeast. We will be taking the mevalonate pathway genes from yeast and constructing two operons in order to produce the terpenoid intermediates isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP). While both IPP and DMAPP are both naturally produced in E. coli through the non-mevalonate pathway, the mevalonate pathway provides an efficient route from acetyl-CoA to terpenoid production than the alternative. Our high-value product will be a terpenoid that has yet to be determined.


For our kill switch will be using a gene named Killer Red which is a mutant of hydrozoan chromoprotein anm2CP from Anthomedusae sp. DC-2005. Our team plans to insert this gene into our construct and by doing so, it will provide a fail-safe in the unlikely event of our E. coli escaping into the environment. The Killer Red gene has a repressible promoter in which tryptophan acts as the repressor. There will be tryptophan present while the cell is used to break down the frying oil, but is not highly present in the environment. Without the presence of tryptophan the Killer Red gene will be activated and white light will produce reactive oxygen species within the cell, causing cell death.

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