Difference between revisions of "Team:Pasteur Paris/Parts"

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{{Pasteur_Paris}}
+
<html lang="en">
−
<html>
+
  <head>
 +
    <meta charset="utf-8">
 +
    <meta name="description" content="igem">
 +
    <meta name="keywords" content="igem, biology">
 +
    <meta name="copyright" content="© Copyright 2016 - iGEM Pasteur" />
 +
    <meta name="author" content="Pasteur">
 +
    <meta name="location" content="Paris, FRANCE">
 +
    <meta http-equiv="x-ua-compatible" content="ie=edge">
 +
    <meta name="viewport" content="width=device-width, initial-scale=1">
 +
    <link rel="icon" type="image/png" href="./static/favicon.png" />
 +
<head>
 +
<link rel="stylesheet" href="https://cdnjs.cloudflare.com/ajax/libs/font-awesome/4.7.0/css/font-awesome.min.css">
 +
<link rel="stylesheet" href="https://fonts.googleapis.com/icon?family=Material+Icons">
  
  
 +
<script>
  
 +
/*Fonction apparition onglet après click sur le menu*/
 +
function apparitionTeam() {
 +
    document.getElementById("Team").style.display = "block";
 +
    document.getElementById("Project").style.display = "none";
 +
    document.getElementById("Design").style.display = "none";
 +
    document.getElementById("Labwork").style.display = "none";
 +
    document.getElementById("HumanPractice").style.display = "none";
 +
    document.getElementById("Achievements").style.display = "none";
 +
}
 +
function apparitionProject() {
 +
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    document.getElementById("Design").style.display = "none";
 +
    document.getElementById("Labwork").style.display = "none";
 +
    document.getElementById("HumanPractice").style.display = "none";
 +
    document.getElementById("Achievements").style.display = "none";
 +
}
 +
function apparitionDesign() {
 +
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 +
    document.getElementById("HumanPractice").style.display = "none";
 +
    document.getElementById("Achievements").style.display = "none";
 +
}
 +
function apparitionLabWork() {
 +
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 +
    document.getElementById("Team").style.display = "none";
 +
    document.getElementById("Project").style.display = "none";
 +
    document.getElementById("Design").style.display = "none";
 +
    document.getElementById("HumanPractice").style.display = "none";
 +
    document.getElementById("Achievements").style.display = "none";
 +
}
 +
function apparitionHumanPractice() {
 +
    document.getElementById("HumanPractice").style.display = "block";
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    document.getElementById("Team").style.display = "none";
 +
    document.getElementById("Project").style.display = "none";
 +
    document.getElementById("Design").style.display = "none";
 +
    document.getElementById("Labwork").style.display = "none";
 +
    document.getElementById("Achievements").style.display = "none";
 +
}
 +
function apparitionAchievements() {
 +
    document.getElementById("Achievements").style.display = "block";
 +
    document.getElementById("Team").style.display = "none";
 +
    document.getElementById("Project").style.display = "none";
 +
    document.getElementById("Design").style.display = "none";
 +
    document.getElementById("HumanPractice").style.display = "none";
 +
    document.getElementById("Labwork").style.display = "none";
 +
}
 +
/*////////////////////////////////////////////*/
  
 +
/* Fonction pour apparition barre rouge onglet*/
  
 +
function apparitionbaton() {
 +
    document.getElementById("batonbleu").style.display = "block";
 +
}
  
−
<div class="column full_size">
+
function deletebaton() {
 +
    document.getElementById("batonbleu").style.display = "none";
 +
}
  
−
<h1>Parts</h1>
+
function apparitionbaton1() {
 +
    document.getElementById("batonbleu1").style.display = "block";
 +
}
  
−
<p>Each team will make new parts during iGEM and will submit them to the Registry of Standard Biological Parts. The iGEM software provides an easy way to present the parts your team has created. The <code>&lt;groupparts&gt;</code> tag (see below) will generate a table with all of the parts that your team adds to your team sandbox.</p>
+
function deletebaton1() {
−
<p>Remember that the goal of proper part documentation is to describe and define a part, so that it can be used without needing to refer to the primary literature. Registry users in future years should be able to read your documentation and be able to use the part successfully. Also, you should provide proper references to acknowledge previous authors and to provide for users who wish to know more.</p>
+
    document.getElementById("batonbleu1").style.display = "none";
 +
}
  
 +
function apparitionbaton1b() {
 +
    document.getElementById("batonbleu1b").style.display = "block";
 +
}
  
−
</div>
+
function deletebaton1b() {
 +
    document.getElementById("batonbleu1b").style.display = "none";
 +
}
  
 +
function apparitionbaton2() {
 +
    document.getElementById("batonbleu2").style.display = "block";
 +
}
  
 +
function deletebaton2() {
 +
    document.getElementById("batonbleu2").style.display = "none";
 +
}
  
 +
function apparitionbaton3() {
 +
    document.getElementById("batonbleu3").style.display = "block";
 +
}
  
 +
function deletebaton3() {
 +
    document.getElementById("batonbleu3").style.display = "none";
 +
}
  
−
<div class="column half_size">
+
function apparitionbaton4() {
−
<div class="highlight">
+
    document.getElementById("batonbleu4").style.display = "block";
−
<h5>Note</h5>
+
}
−
<p>Note that parts must be documented on the <a href="http://parts.igem.org/Main_Page"> Registry</a>. This page serves to <i>showcase</i> the parts you have made. Future teams and other users and are much more likely to find parts by looking in the Registry than by looking at your team wiki.</p>
+
 
 +
function deletebaton4() {
 +
    document.getElementById("batonbleu4").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton5() {
 +
    document.getElementById("batonbleu5").style.display = "block";
 +
}
 +
 
 +
function deletebaton5() {
 +
    document.getElementById("batonbleu5").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton6() {
 +
    document.getElementById("batonbleu6").style.display = "block";
 +
}
 +
 
 +
function deletebaton6() {
 +
    document.getElementById("batonbleu6").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton7() {
 +
    document.getElementById("batonbleu7").style.display = "block";
 +
}
 +
 
 +
function deletebaton7() {
 +
    document.getElementById("batonbleu7").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton8() {
 +
    document.getElementById("batonbleu8").style.display = "block";
 +
}
 +
 
 +
function deletebaton8() {
 +
    document.getElementById("batonbleu8").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton8b() {
 +
    document.getElementById("batonbleu8b").style.display = "block";
 +
}
 +
 
 +
function deletebaton8b() {
 +
    document.getElementById("batonbleu8b").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton9() {
 +
    document.getElementById("batonbleu9").style.display = "block";
 +
}
 +
 
 +
function deletebaton9() {
 +
    document.getElementById("batonbleu9").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton10() {
 +
    document.getElementById("batonbleu10").style.display = "block";
 +
}
 +
 
 +
function deletebaton10() {
 +
    document.getElementById("batonbleu10").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton11() {
 +
    document.getElementById("batonbleu11").style.display = "block";
 +
}
 +
 
 +
function deletebaton11() {
 +
    document.getElementById("batonbleu11").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton12() {
 +
    document.getElementById("batonbleu12").style.display = "block";
 +
}
 +
 
 +
function deletebaton12() {
 +
    document.getElementById("batonbleu12").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton13() {
 +
    document.getElementById("batonbleu13").style.display = "block";
 +
}
 +
 
 +
function deletebaton13() {
 +
    document.getElementById("batonbleu13").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton14() {
 +
    document.getElementById("batonbleu14").style.display = "block";
 +
}
 +
 
 +
function deletebaton14() {
 +
    document.getElementById("batonbleu14").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton15() {
 +
    document.getElementById("batonbleu15").style.display = "block";
 +
}
 +
 
 +
function deletebaton15() {
 +
    document.getElementById("batonbleu15").style.display = "none";
 +
}
 +
 
 +
function apparitionbaton16() {
 +
    document.getElementById("batonbleu16").style.display = "block";
 +
}
 +
 
 +
function deletebaton16() {
 +
    document.getElementById("batonbleu16").style.display = "none";
 +
}
 +
 
 +
/*////////////////////////////////////////////*/
 +
 
 +
function CarnotNoiretBlanc() {
 +
  document.getElementById("CarnotBW").style.display = "block";
 +
  document.getElementById("Carnot").style.display = "none";
 +
}
 +
 
 +
function CarnotCouleur() {
 +
  document.getElementById("Carnot").style.display = "block";
 +
  document.getElementById("CarnotBW").style.display = "none";
 +
}
 +
 
 +
 
 +
function GlobalCareNoiretBlanc() {
 +
  document.getElementById("GlobalCareBW").style.display = "block";
 +
  document.getElementById("GlobalCare").style.display = "none";
 +
}
 +
 
 +
function GlobalCareCouleur() {
 +
  document.getElementById("GlobalCare").style.display = "block";
 +
  document.getElementById("GlobalCareBW").style.display = "none";
 +
}
 +
 
 +
function MettlerToledoNoiretBlanc() {
 +
  document.getElementById("MettlerToledoBW").style.display = "block";
 +
  document.getElementById("MettlerToledo").style.display = "none";
 +
}
 +
 
 +
function MettlerToledoCouleur() {
 +
  document.getElementById("MettlerToledo").style.display = "block";
 +
  document.getElementById("MettlerToledoBW").style.display = "none";
 +
}
 +
 
 +
function geniousNoiretBlanc() {
 +
  document.getElementById("geniousBW").style.display = "block";
 +
  document.getElementById("genious").style.display = "none";
 +
}
 +
 
 +
function geniousCouleur() {
 +
  document.getElementById("genious").style.display = "block";
 +
  document.getElementById("geniousBW").style.display = "none";
 +
}
 +
 
 +
function centraleNoiretBlanc() {
 +
  document.getElementById("CentraleBW").style.display = "block";
 +
  document.getElementById("Centrale").style.display = "none";
 +
}
 +
 
 +
function centraleCouleur() {
 +
  document.getElementById("Centrale").style.display = "block";
 +
  document.getElementById("CentraleBW").style.display = "none";
 +
}
 +
 
 +
function ambassadeNoiretBlanc() {
 +
  document.getElementById("ambassadeBW").style.display = "block";
 +
  document.getElementById("ambassade").style.display = "none";
 +
}
 +
 
 +
function ambassadeCouleur() {
 +
  document.getElementById("ambassade").style.display = "block";
 +
  document.getElementById("ambassadeBW").style.display = "none";
 +
}
 +
 
 +
function QuiagenNoiretBlanc() {
 +
  document.getElementById("QuiagenBW").style.display = "block";
 +
  document.getElementById("Quiagen").style.display = "none";
 +
}
 +
 
 +
function QuiagenCouleur() {
 +
  document.getElementById("Quiagen").style.display = "block";
 +
  document.getElementById("QuiagenBW").style.display = "none";
 +
}
 +
 
 +
function eurofinsNoiretBlanc() {
 +
  document.getElementById("eurofinsBW").style.display = "block";
 +
  document.getElementById("eurofins").style.display = "none";
 +
}
 +
 
 +
function eurofinsCouleur() {
 +
  document.getElementById("eurofins").style.display = "block";
 +
  document.getElementById("eurofinsBW").style.display = "none";
 +
}
 +
 
 +
function ThermofisherNoiretBlanc() {
 +
  document.getElementById("ThermofisherBW").style.display = "block";
 +
  document.getElementById("Thermofisher").style.display = "none";
 +
}
 +
 
 +
function ThermofisherCouleur() {
 +
  document.getElementById("Thermofisher").style.display = "block";
 +
  document.getElementById("ThermofisherBW").style.display = "none";
 +
}
 +
 
 +
function BarthenteimNoiretBlanc() {
 +
  document.getElementById("BarthenteimBW").style.display = "block";
 +
  document.getElementById("Barthenteim").style.display = "none";
 +
}
 +
 
 +
function BarthenteimCouleur() {
 +
  document.getElementById("Barthenteim").style.display = "block";
 +
  document.getElementById("BarthenteimBW").style.display = "none";
 +
}
 +
 
 +
</script>
 +
<style>
 +
 
 +
@font-face {
 +
  font-family: "CrimsonTextSemi";
 +
  src: url("https://static.igem.org/mediawiki/2017/7/75/T--Pasteur_Paris--CrimsonText-Semibold.ttf");
 +
}
 +
 
 +
@font-face {
 +
  font-family: "CrimsonTextRoman";
 +
  src: url("https://static.igem.org/mediawiki/2017/0/0e/T--Pasteur_Paris--CrimsonText-Roman.ttf");
 +
}
 +
 
 +
body{
 +
  margin:30px;
 +
  padding:30px;
 +
  background-color:white;
 +
  height:100%;
 +
}
 +
 
 +
a{
 +
  text-decoration: none;
 +
  color:black;
 +
}
 +
 
 +
li{
 +
  list-style:none;
 +
}
 +
 
 +
.left{
 +
  left: 30px;
 +
  margin-bottom: 7.5px;
 +
  padding-top: 37.5px;
 +
  position:fixed;
 +
  background-color: none;
 +
  top:0;
 +
}
 +
 
 +
.right{
 +
  right: 30px;
 +
  margin-bottom: 7.5px; 
 +
  padding-top: 37.5px;
 +
  position:fixed;
 +
  background-color: none;
 +
  top:0;
 +
}
 +
 
 +
.center{
 +
    margin-left: auto;
 +
    margin-right: auto;
 +
}
 +
 
 +
/*Position menu*/
 +
.menu { 
 +
    text-align: left;
 +
    width: 450px;
 +
    position: fixed;
 +
    left:30px;
 +
    line-height: 200%;
 +
    font-family: Helvetica, sans-serif;
 +
    font-weight: bold;
 +
    font-size: 7.5pt;
 +
    display: flex;
 +
    flex-direction: column-reverse;
 +
    text-align: bottom left;
 +
    bottom: 4%;
 +
}
 +
 
 +
.menu span{
 +
  cursor: pointer;
 +
}
 +
 
 +
.com{
 +
    display: flex;
 +
    flex-direction: column;
 +
    position:fixed;
 +
    text-align: left;
 +
    right:30px;
 +
    bottom: 4%;
 +
}
 +
 
 +
/*Ensemble pour trait après le menu*/
 +
 
 +
.lastword {
 +
    width: 150px;
 +
}
 +
 
 +
.lastword img {
 +
    position:absolute;
 +
    opacity:0;
 +
}
 +
 
 +
.lastword:hover:after{
 +
    display:inline-block;
 +
    vertical-align: middle;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
/*////////////////////////////////////////////*/
 +
 
 +
.com li{
 +
  line-height:200%;
 +
  vertical-align: middle;
 +
}
 +
 
 +
.underline{
 +
  display:block;
 +
  position:relative;
 +
  top:-10px;
 +
  left:0px;
 +
  border-bottom:4px dotted black;
 +
  width: 75px;
 +
}
 +
 
 +
.underline2{
 +
  display:block;
 +
  position:relative;
 +
  top:-10px;
 +
  left:0px;
 +
  border-bottom:4px dotted black;
 +
  width: 520px;
 +
}
 +
 
 +
.underline3{
 +
  display:block;
 +
  position:relative;
 +
  top:-10px;
 +
  left:0px;
 +
  border-bottom:4px dotted black;
 +
  width: 350px;
 +
}
 +
 
 +
.underline4{
 +
  display:block;
 +
  position:relative;
 +
  top:-10px;
 +
  left:0px;
 +
  border-bottom:4px dotted black;
 +
  width: 360px;
 +
}
 +
 
 +
.underline5{
 +
  display:block;
 +
  position:relative;
 +
  top:-10px;
 +
  left:0px;
 +
  border-bottom:4px dotted black;
 +
  width: 546px;
 +
}
 +
 
 +
.underline6{
 +
  display:block;
 +
  position:relative;
 +
  top:-10px;
 +
  left:0px;
 +
  border-bottom:4px dotted black;
 +
  width: 582px;
 +
}
 +
 
 +
#top_title{
 +
display:none;
 +
}
 +
 
 +
#sideMenu{
 +
display:none;
 +
position:fixed;
 +
right:120px;
 +
}
 +
 
 +
#content{
 +
  width:100%;
 +
  margin: 0px 0px 0px 0px;
 +
  padding: 0px 0px 0px 0px;
 +
}
 +
 
 +
.title{
 +
  font-size: 48.75pt;
 +
  margin-left:300px;
 +
  margin-top:112.5px;
 +
  cursor: default;
 +
}
 +
 
 +
.subtitle{
 +
  margin-left:12.25px;
 +
  margin-top:55px;
 +
  cursor: default;
 +
  font-size: 18pt;
 +
}
 +
 
 +
 
 +
.maintext{
 +
    font-size: 9pt;
 +
    font-family:Helvetica;
 +
    margin-top:10px;
 +
    text-align: justify;
 +
    cursor: default;
 +
    width:75%;
 +
    line-height:16pt;
 +
}
 +
 
 +
.maintextsuite{
 +
    font-size: 9pt;
 +
    margin-top:10px;
 +
    text-align: justify;
 +
    cursor: default;
 +
    font-family:Helvetica;
 +
    width:75%;
 +
    line-height:16pt;
 +
}
 +
 
 +
.maintextsuiteimage{
 +
    font-size: 9pt;
 +
    margin-top:10px;
 +
    text-align: justify;
 +
    cursor: default;
 +
    font-family:Helvetica;
 +
    width:40%;
 +
}
 +
 
 +
.text {
 +
  position:relative;
 +
  margin-left: 287.75px;
 +
  line-height:16pt;
 +
}
 +
 
 +
#logo{
 +
  font-size:18px;
 +
text-decoration:none;
 +
color:black;
 +
}
 +
 
 +
#logo:hover{
 +
  color: #3298CB;
 +
}
 +
 
 +
/*Flèche de remontée*/
 +
 
 +
#toparrow{
 +
    justify-content: center;
 +
    position:fixed;
 +
    vertical-align:middle;
 +
    z-index:10;
 +
    width:2%;
 +
    top:42%;
 +
    right:30px;
 +
}
 +
 
 +
#toparrow a:hover{
 +
  color: #3298CB;
 +
}
 +
 
 +
.Chili{
 +
  z-index:-1;
 +
  margin-left: 287.75px;
 +
  top:90px;
 +
  width:56vw;
 +
  height: 55vh;
 +
}
 +
 
 +
table{
 +
  border-collapse: collapse;
 +
  }
 +
 
 +
/*////////////////////////////////////////////*/
 +
/*Ensemble de class pour les onglets*/
 +
 
 +
.outer {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 28%;
 +
}
 +
 
 +
.outer1 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 38%;
 +
}
 +
 
 +
.outer1b {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 47.5%;
 +
}
 +
 
 +
.outer2 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 25%;
 +
}
 +
 
 +
.outer3 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 34%;
 +
}
 +
.outer3a {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 48%;
 +
}
 +
.outer3b {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 59%;
 +
}
 +
 
 +
.outer4 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 26%;
 +
}
 +
 
 +
.outer5 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 28%;
 +
}
 +
 
 +
.outer6 {
 +
  position: fixed;
 +
  left: 33px;
 +
  top: 42.5%;
 +
  text-align: center;
 +
}
 +
 
 +
.outer7 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 50.7%;
 +
}
 +
 
 +
.outer8 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 22%;
 +
}
 +
 
 +
.outer8b {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 31%;
 +
}
 +
 
 +
.outer9 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 42%;
 +
}
 +
 
 +
.outer10 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 49%;
 +
}
 +
 
 +
.outer11 {
 +
  position: fixed;
 +
  left: 33px;
 +
  top: 57%;
 +
  text-align: center;
 +
}
 +
 
 +
.outer12 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 22%;
 +
}
 +
 
 +
.outer13 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 28%;
 +
}
 +
 
 +
.outer13b {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 40%;
 +
}
 +
 
 +
.outer14 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 54%;
 +
}
 +
 
 +
.outer15 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 50%;
 +
}
 +
 
 +
.outer16 {
 +
  position: fixed;
 +
  left: 40px;
 +
  top: 36%;
 +
}
 +
 
 +
.inner {
 +
  position: absolute;
 +
  bottom: 100%;
 +
  -webkit-transform: rotateZ(90deg);
 +
  transform-origin: 0 100%;
 +
  white-space: nowrap;
 +
}
 +
/*////////////////////////////////////////////*/
 +
 
 +
 
 +
.rotate {
 +
  -webkit-transform: rotate(180deg);
 +
}
 +
 
 +
#Team, #Design, #Project, #HumanPractice, #Achievements, #Labwork{
 +
  font-family: Helvetica, sans-serif;
 +
    font-weight: bold;
 +
    font-size: 8pt;
 +
}
 +
 
 +
/*Trait rouge des onglets*/
 +
 
 +
#batonbleu{
 +
    position:relative;
 +
    top:5px;
 +
    left:33%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu1{
 +
    position:relative;
 +
    top:5px;
 +
    left:33%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu1b{
 +
    position:relative;
 +
    top:5px;
 +
    left:33%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu2{
 +
    position:relative;
 +
    top:5px;
 +
    left:33%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu3{
 +
    position:relative;
 +
    top:5px;
 +
    left:33%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
#batonbleu3a{
 +
    position:relative;
 +
    top:5px;
 +
    left:33%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
#batonbleu3b{
 +
    position:relative;
 +
    top:5px;
 +
    left:15%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu4{
 +
    position:relative;
 +
    top:5px;
 +
    left:33%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu5{
 +
    position:relative;
 +
    top:5px;
 +
    left:33%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu6{
 +
    position:relative;
 +
    top:5px;
 +
    left:25%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu7{
 +
    position:relative;
 +
    top:5px;
 +
    left:22%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu8{
 +
    position:relative;
 +
    top:5px;
 +
    left:30%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu8b{
 +
    position:relative;
 +
    top:5px;
 +
    left:30%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu9{
 +
    position:relative;
 +
    top:5px;
 +
    left:29%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu10{
 +
    position:relative;
 +
    top:5px;
 +
    left:33%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu11{
 +
    position:relative;
 +
    top:5px;
 +
    left:25%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu12{
 +
    position:relative;
 +
    top:5px;
 +
    left:10%;
 +
    display:none;
 +
    height:3px;
 +
    width:17.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu13{
 +
    position:relative;
 +
    top:5px;
 +
    left:33%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
#batonbleu13b{
 +
    position:relative;
 +
    top:5px;
 +
    left:33%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu14{
 +
    position:relative;
 +
    top:5px;
 +
    left:37%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu15{
 +
    position:relative;
 +
    top:5px;
 +
    left:10%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
#batonbleu16{
 +
    position:relative;
 +
    top:5px;
 +
    left:35%;
 +
    display:none;
 +
    height:3px;
 +
    width:22.5px;
 +
    background: #3298CB;
 +
    content:""
 +
}
 +
 
 +
li{margin:5px}
 +
 
 +
a:visited {
 +
    color: black;
 +
    text-decoration:none;
 +
}
 +
 
 +
 
 +
/*////////////////////////////////*/
 +
 
 +
 
 +
#title{
 +
  font-size: 60pt;
 +
  border-bottom:4px dotted black;
 +
  width: 190px;
 +
  z-index:-1;
 +
  margin-left: 287.75px;
 +
  margin-top: 90px;
 +
  line-height:59pt;
 +
}
 +
 
 +
#subtitles{
 +
  position:relative;
 +
  margin-left: 300px;
 +
  margin-top:-5vw;
 +
  font-size: 18pt;
 +
  font-weight: normal;
 +
  /*border-bottom:4px dotted black;*/
 +
  z-index:-1;
 +
  width: 530px;
 +
  border-bottom:4px dotted black;
 +
}
 +
 
 +
li{margin:5px}
 +
 
 +
/*////////////////////////////////////////////*/
 +
 
 +
</style>
 +
  </head>
 +
 
 +
 
 +
  <body>
 +
  <a class="left" href="https://2017.igem.org/Team:Pasteur_Paris"><img src="https://static.igem.org/mediawiki/2017/d/d8/T--Pasteur_Paris--aether.png"; width="220px"; height="70px"></img></a>
 +
  <a class="right" href="https://2017.igem.org/Team:Pasteur_Paris"><img src="https://static.igem.org/mediawiki/2017/1/10/T--Pasteur_Paris--iGEM.png"; width="90px"; height="70px"></img></a>
 +
<nav class="menu">
 +
  <ul>
 +
    <li><FONT size ="2.25pt" color=#00000><span class="lastword"; onclick="apparitionTeam()">Team </span></FONT></li>
 +
    <li><FONT size ="2.25pt" color=#00000><span class="lastword"; onclick="apparitionProject()">Project </span></FONT></li>
 +
    <li><FONT size ="2.25pt" color=#00000><span class="lastword"; onclick="apparitionDesign()">Design </span></FONT></li>
 +
    <li><FONT size ="2.25pt" color=#00000><span class="lastword"; onclick="apparitionLabWork()">Lab work </span></FONT></li>
 +
    <li><FONT size ="2.25pt" color=#00000><span class="lastword"; onclick="apparitionHumanPractice()">Human Practice </span></FONT></li>
 +
    <li><FONT size ="2.25pt" color=#00000><span class="lastword"; onclick="apparitionAchievements()">Achievements </span></FONT></li>
 +
  </ul>
 +
</nav>
 +
 
 +
<div id="Team" style="display:none">
 +
  <div class="outer">
 +
    <div class="inner"; onmouseover="apparitionbaton()"; onmouseout="deletebaton()">
 +
      <div class="list rotate">  
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Team">Our Team </a>
 +
        <div id="batonbleu">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
  <div class="outer1">
 +
    <div class="inner"; onmouseover="apparitionbaton1()"; onmouseout="deletebaton1()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Team#pagesponsors">Sponsors </a>
 +
        <div id="batonbleu1">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
  <div class="outer1b">
 +
    <div class="inner"; onmouseover="apparitionbaton1b()"; onmouseout="deletebaton1b()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Attributions">Attributions </a>
 +
        <div id="batonbleu1b">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 
</div>
 
</div>
 +
 +
<div id="Project" style="display:none">
 +
  <div class="outer2">
 +
    <div class="inner"; onmouseover="apparitionbaton2()"; onmouseout="deletebaton2()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Project">Context</a>
 +
        <div id="batonbleu2">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
  <div class="outer3">
 +
    <div class="inner"; onmouseover="apparitionbaton3()"; onmouseout="deletebaton3()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Entrepreneurship">Entrepreneurship</a>
 +
        <div id="batonbleu3">
 +
        </div>
 +
      </div>
 +
  </div>
 +
  </div>
 +
    <div class="outer3a">
 +
    <div class="inner"; onmouseover="apparitionbaton3a()"; onmouseout="deletebaton3a()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Engagement">Engagement</a>
 +
        <div id="batonbleu3a">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
  <div class="outer3b">
 +
    <div class="inner"; onmouseover="apparitionbaton3b()"; onmouseout="deletebaton3b()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Model">Model</a>
 +
        <div id="batonbleu3b">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 
</div>
 
</div>
  
 +
<div id="Design" style="display:none">
 +
  <div class="outer4">
 +
    <div class="inner"; onmouseover="apparitionbaton4()"; onmouseout="deletebaton4()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Design">Science</a>
 +
        <div id="batonbleu4">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
  <div class="outer5">
 +
    <div class="inner"; onmouseover="apparitionbaton5()"; onmouseout="deletebaton5()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Design">Product</a>
 +
        <div id="batonbleu5">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
  <div class="outer6">
 +
    <div class="inner"; onmouseover="apparitionbaton6()"; onmouseout="deletebaton6()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Design">Scenario</a>
 +
        <div id="batonbleu6">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
  <div class="outer7">
 +
    <div class="inner"; onmouseover="apparitionbaton7()"; onmouseout="deletebaton7()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Applied_Design">Applied<br>Design</a>
 +
        <div id="batonbleu7">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
</div>
  
 +
<div id="Labwork" style="display:none">
 +
  <div class="outer8">
 +
    <div class="inner"; onmouseover="apparitionbaton8()"; onmouseout="deletebaton8()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Notebook">Notebook</a>
 +
        <div id="batonbleu8">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
  <div class="outer8b">
 +
    <div class="inner"; onmouseover="apparitionbaton8b()"; onmouseout="deletebaton8b()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Demonstrate">Demonstrate</a>
 +
        <div id="batonbleu8b">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
  <div class="outer9">
 +
    <div class="inner"; onmouseover="apparitionbaton9()"; onmouseout="deletebaton9()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/InterLab">InterLab</a>
 +
        <div id="batonbleu9">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
  <div class="outer10">
 +
    <div class="inner"; onmouseover="apparitionbaton10()"; onmouseout="deletebaton10()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Protocol">Protocol</a>
 +
        <div id="batonbleu10">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
  <div class="outer11">
 +
    <div class="inner"; onmouseover="apparitionbaton11()"; onmouseout="deletebaton11()">
 +
      <div class="list rotate">
 +
        <a href="https://2017.igem.org/Team:Pasteur_Paris/Safety">Safety<br>Security</a>
 +
        <div id="batonbleu11">
 +
        </div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
</div>
  
 +
<div id="HumanPractice" style="display:none">
 +
  <div class="outer12">
 +
    <div class="inner"; onmouseover="apparitionbaton12()"; onmouseout="deletebaton12()">
 +
      <div class="list rotate">
 +
      <a href="https://2017.igem.org/Team:Pasteur_Paris/HP/Law">Law</a>
 +
      <div id="batonbleu12"></div>
 +
      </div>
 +
    </div>
 +
  </div>
 +
  <div class="outer13">
 +
    <div class="inner"; onmouseover="apparitionbaton13()"; onmouseout="deletebaton13()">
 +
      <div class="list rotate">
 +
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<h5>Adding parts to the registry</h5>
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<p>You can add parts to the Registry at our <a href="http://parts.igem.org/Add_a_Part_to_the_Registry">Add a Part to the Registry</a> link.</p>
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  <img src="https://static.igem.org/mediawiki/2017/b/ba/T--Pasteur_Paris--BioB_3.png"; style="position:absolute; left:390px; height: 66.5vh;width:auto;max-width: 60vw; z-index:-2;top:-70px"/>
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<p>We encourage teams to start completing documentation for their parts on the Registry as soon as you have it available. The sooner you put up your parts, the better you will remember all the details about your parts. Remember, you don't need to send us the DNA sample before you create an entry for a part on the Registry. (However, you <b>do</b> need to send us the DNA sample before the Jamboree. If you don't send us a DNA sample of a part, that part will not be eligible for awards and medal criteria.)</p>
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  <p id="title" style="font-size:50pt; font-family: MinionPro, serif;"> Parts</p>
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  <p id="subtitles" style=" font-family: MinionPro, serif; font-size:18pt">Our Biobricks</p>
 
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<div class="suitemain" style="top:-25px; font-size: 10pt;">
  
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<p class="maintext"><b style="font-size: 13pt"> BBa_K2198000:E1_1</b> The naphthalene 1,2-dioxygenase part 1 (E1_1) from <i>Pseudomonas putida</i> Ref. [1.A]</p>
  
 +
<p class="maintext">Forms a complex with naphthalene 1,2-dioxygenase part 2 (E1_2) to form an active enzyme to oxidize PAH that can be used as substrate. This oxidation with dioxygen form a vicinal diol and add two hydrogens on the ring. This step need energy to break the aromaticity. This complex enzyme need NADH,H+.</p>
  
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<p class="maintext"><b style="font-size: 13pt"> BBa_K2198001:E1_2</b> The naphthalene 1,2-dioxygenase part 1 (E1_2) from <i>Pseudomonas putida</i> Ref. [1.B]</p>
  
−
<h5>What information do I need to start putting my parts on the Registry?</h5>
+
<p class="maintext">Gets complex with naphthalene 1,2-dioxygenase part 1 (E1_1) to form an active enzyme to oxidize PAH that can be used as substrate. This oxidation with dioxygen form a vicinal diol and add two hydrogens on the ring. This step need energy to break the aromaticity. This complex enzyme need NADH,H+.</p>
−
<p>The information needed to initially create a part on the Registry is:</p>
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<ul>
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<li>Part Name</li>
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<li>Part type</li>
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<li>Creator</li>
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<li>Sequence</li>
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<li>Short Description (60 characters on what the DNA does)</li>
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<li>Long Description (Longer description of what the DNA does)</li>
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<li>Design considerations</li>
+
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</ul>
+
  
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<p>
+
<br><br>
−
We encourage you to put up <em>much more</em> information as you gather it over the summer. If you have images, plots, characterization data and other information, please also put it up on the part page. </p>
+
  
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<p style="font-size: 13pt" class="maintext"><b style="font-size: 13pt"> BBa_K2198002:E2</b> The dihydrodiol dehydrogenase from <i>Ralstonia sp. </i> Ref. [2]</p>
  
 +
<p class="maintext">This step allows the restoration of the aromatic with a dehydrogenation of the vicinal diol. Thanks to this, E3 will be able to open the ring with an oxidative cleavage.</p>
  
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<br><br>
  
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<h5>Inspiration</h5>
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<p class="maintext"><b style="font-size: 13pt"> BBa_K2198003:E3</b> The catechol 2,3-dioxygenase from <i>Pseudomonas putida</i> Ref. [3]</p>
−
<p>We have a created  a <a href="http://parts.igem.org/Well_Documented_Parts">collection of well documented parts</a> that can help you get started.</p>
+
  
−
<p> You can also take a look at how other teams have documented their parts in their wiki:</p>
+
<p class="maintext">In our degradation pathway the key step is to remove ring from PAH to reduce their toxicity. The E3 is able to do an extradiol ring cleavage. Thanks to this step, the PAH are now less toxic and available for an alkyl removal by E4. If this step is the last one of the process all the products can be handle by chemistry thanks to this alkyl chain. Furthermore E3 can begin an iterative degradation with E4 if the E4 main product enter in the range of substrate specificity.</p>
−
<ul>
+
<br><br>
−
<li><a href="https://2014.igem.org/Team:MIT/Parts"> 2014 MIT </a></li>
+
 
−
<li><a href="https://2014.igem.org/Team:Heidelberg/Parts"> 2014 Heidelberg</a></li>
+
<p class="maintext"><b style="font-size: 13pt"> BBa_K2198004:E4</b> The trans-2-carboxybenzalpyruvate hydratase-aldolase from <i>Nocardioides sp</i> Ref. [4]</p>
−
<li><a href="https://2014.igem.org/Team:Tokyo_Tech/Parts">2014 Tokyo Tech</a></li>
+
 
−
</ul>
+
<p>The last step of our degradation pathway allow us to remove a ring from the PAH by producing a py-ruvic acid. This product is a common harmless cell intermediate molecule. Moreover, this step pro-duce a smaller PAH if there is more than 2 ring at the beginning. This second generation PAH can reintegrate the pathway at the previous step thanks to a low substrate specificity E3</p>
 +
 
 +
<br><br>
 +
 
 +
<p class="maintext"><b style="font-size: 13pt"> BBa_K2198005:S1</b> The Major ampullate Spidroin 1 (MaSp1) from <i>Euprosthenops australis </i> Ref. [5]</p>
 +
 
 +
<p class="maintext">In order to carry out the reaction on a solid support, we designed a scaffold made of spider silk that can polymerize.</p>
 +
 
 +
 
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<div style="width:100% ; margin-top:20px;margin:auto; top: 70px">
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<div style="margin:auto ;width:100%">
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  <img src="https://static.igem.org/mediawiki/2017/e/e4/T--Pasteur_Paris--Parts.png"; style="height:auto; max-height:150%">
 +
  <p class="maintext"><b> Figure 1: Biochemical degradation pathway for Anthracene</b></p>
 +
  <p class="maintext" style="top: 30px; width:50vw; text-align: center"> The catalytic degradation of PAH is ensured by four enzymes. (E1_1, E1_2) homodimer catalyzes the di-hydroxylation, E2 the dehydrogenation of the vicinal diol, E3 oxidative cleavage of the aromatic cycle and E4 cleavage of a linear unsaturated &alpha;-&beta; enone to release a pyruvate molecule.</p>
 +
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<h5>Part Table </h5>
 
  
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<p>Please include a table of all the parts your team has made during your project on this page. Remember part characterization and measurement data must go on your team part pages on the Registry. </p>
+
<p class="maintext">Open reading frames for the four enzymes were designed using Geneious 11.0.2 Ref. [5] software with <a href="http://www.uniprot.org" style="color: #3298CB;">Uniprot</a> database sequences. Each sequence was designed with Biobrick prefix and suffix <a href="https://igem.org/Registry" style="color: #3298CB;">Registry</a> in the 5’ and 3’ regions respectively. In addition, His(6) tag, spacer and TeV cleavage sites were included at the N-terminus for purification and release of the His(6) -tag. BamHI site were inserted in the construct after the prefix and before the suffix respectively for insertion into pET43.1a(+) or pET32a(+) (Merck-Millipore) expression plasmids. Genes were ordered from <a href="www.eurofinsgenomics.com" style="color: #3298CB;">Eurofins Genomics</a> after sequence optimization for Biobrick and cloning into pET vector compatibility. Sequences were further optimized for expression in Escherichia coli and gene synthesis. </p>
  
−
<div class="highlight">
+
<p class="maintext"><b style="font-size: 13pt">References</b>
  
 +
<p class="maintext">1. A.: J Bacteriol. 2000 Mar;182(6):1641-9. Substrate specificity of naphthalene dioxygenase: effect of specific amino acids at the active site of the enzyme. Parales RE1, Lee K, Resnick SM, Jiang H, Lessner DJ, Gibson DT</p>
 +
<p class="maintext">1. B.: J Bacteriol. 1983 Aug;155(2):505-11. Naphthalene dioxygenase: purification and properties of a terminal oxygenase component. Ensley BD, Gibson DT</p>
 +
<p class="maintext">2. : J Bacteriol. 1998 May; 180(9): 2522–2530. A Gene Cluster Encoding Steps in Conversion of Naphthalene to Gentisate in Pseudomonas sp. Strain U2 Sergio L. Fuenmayor,1 Mark Wild,2 Alastair L. Boyes,2 and Peter A. Williams1</p>
 +
<p class="maintext">3 : Int J Biol Macromol. 2017 Aug 25. pii: S0141-8130(17)32824-6. doi: 10.1016/j.ijbiomac.2017.08.113. [Epub ahead of print] Isolation and characterization of three novel catechol 2,3-dioxygenase from three novel haloalkaliphilic BTEX-degrading Pseudomonas strains. Hassan HA1, Aly AA2.</p>
 +
<p class="maintext">4 : J Bacteriol. 1998 Feb; 180(4): 945–949. PMCID: PMC106976 - Biochemical and Genetic Characterization of trans-2′-Carboxybenzalpyruvate Hydratase-Aldolase from a Phenanthrene-Degrading Nocardioides Strain - Tokuro Iwabuchi† and Shigeaki Harayama</p>
 +
<p class="maintext">5 : “Geneious version (the version you are using) (https://www.geneious.com, Kearse et al., 2012)” Kearse, M., Moir, R., Wilson, A., Stones-Havas, S., Cheung, M., Sturrock, S., Buxton, S., Cooper, A., Markowitz, S., Duran, C., Thierer, T., Ashton, B., Mentjies, P., Drummond, A. (2012). Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data.Bioinformatics, 28(12), 1647-1649.</p>
 +
<p class="maintext">6 : Biomacromolecules 2007, 8, 1695-1701 // Macroscopic Fibers Self-Assembled from Recombinant Miniature Spider Silk Proteins; Margareta Stark, Stefan Grip, Anna Rising, My Hedhammar, Wilhelm Engström, Goran Hj&aumllm, and Jan Johansson*, Department of Anatomy, Physiology, and Biochemistry, The Biomedical Centre, Swedish University of Agricultural Sciences, SE-751 23 Uppsala, Sweden, and Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, SE-750 07 Uppsala, Sweden Received January 16, 2007; Revised Manuscript Received February 20, 2007 </p>
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Latest revision as of 21:41, 15 December 2017

Parts

Our Biobricks

BBa_K2198000:E1_1 The naphthalene 1,2-dioxygenase part 1 (E1_1) from Pseudomonas putida Ref. [1.A]

Forms a complex with naphthalene 1,2-dioxygenase part 2 (E1_2) to form an active enzyme to oxidize PAH that can be used as substrate. This oxidation with dioxygen form a vicinal diol and add two hydrogens on the ring. This step need energy to break the aromaticity. This complex enzyme need NADH,H+.



BBa_K2198001:E1_2 The naphthalene 1,2-dioxygenase part 1 (E1_2) from Pseudomonas putida Ref. [1.B]

Gets complex with naphthalene 1,2-dioxygenase part 1 (E1_1) to form an active enzyme to oxidize PAH that can be used as substrate. This oxidation with dioxygen form a vicinal diol and add two hydrogens on the ring. This step need energy to break the aromaticity. This complex enzyme need NADH,H+.



BBa_K2198002:E2 The dihydrodiol dehydrogenase from Ralstonia sp. Ref. [2]

This step allows the restoration of the aromatic with a dehydrogenation of the vicinal diol. Thanks to this, E3 will be able to open the ring with an oxidative cleavage.



BBa_K2198003:E3 The catechol 2,3-dioxygenase from Pseudomonas putida Ref. [3]

In our degradation pathway the key step is to remove ring from PAH to reduce their toxicity. The E3 is able to do an extradiol ring cleavage. Thanks to this step, the PAH are now less toxic and available for an alkyl removal by E4. If this step is the last one of the process all the products can be handle by chemistry thanks to this alkyl chain. Furthermore E3 can begin an iterative degradation with E4 if the E4 main product enter in the range of substrate specificity.



BBa_K2198004:E4 The trans-2-carboxybenzalpyruvate hydratase-aldolase from Nocardioides sp Ref. [4]

The last step of our degradation pathway allow us to remove a ring from the PAH by producing a py-ruvic acid. This product is a common harmless cell intermediate molecule. Moreover, this step pro-duce a smaller PAH if there is more than 2 ring at the beginning. This second generation PAH can reintegrate the pathway at the previous step thanks to a low substrate specificity E3



BBa_K2198005:S1 The Major ampullate Spidroin 1 (MaSp1) from Euprosthenops australis Ref. [5]

In order to carry out the reaction on a solid support, we designed a scaffold made of spider silk that can polymerize.

Figure 1: Biochemical degradation pathway for Anthracene

The catalytic degradation of PAH is ensured by four enzymes. (E1_1, E1_2) homodimer catalyzes the di-hydroxylation, E2 the dehydrogenation of the vicinal diol, E3 oxidative cleavage of the aromatic cycle and E4 cleavage of a linear unsaturated α-β enone to release a pyruvate molecule.

Open reading frames for the four enzymes were designed using Geneious 11.0.2 Ref. [5] software with Uniprot database sequences. Each sequence was designed with Biobrick prefix and suffix Registry in the 5’ and 3’ regions respectively. In addition, His(6) tag, spacer and TeV cleavage sites were included at the N-terminus for purification and release of the His(6) -tag. BamHI site were inserted in the construct after the prefix and before the suffix respectively for insertion into pET43.1a(+) or pET32a(+) (Merck-Millipore) expression plasmids. Genes were ordered from Eurofins Genomics after sequence optimization for Biobrick and cloning into pET vector compatibility. Sequences were further optimized for expression in Escherichia coli and gene synthesis.

References

1. A.: J Bacteriol. 2000 Mar;182(6):1641-9. Substrate specificity of naphthalene dioxygenase: effect of specific amino acids at the active site of the enzyme. Parales RE1, Lee K, Resnick SM, Jiang H, Lessner DJ, Gibson DT

1. B.: J Bacteriol. 1983 Aug;155(2):505-11. Naphthalene dioxygenase: purification and properties of a terminal oxygenase component. Ensley BD, Gibson DT

2. : J Bacteriol. 1998 May; 180(9): 2522–2530. A Gene Cluster Encoding Steps in Conversion of Naphthalene to Gentisate in Pseudomonas sp. Strain U2 Sergio L. Fuenmayor,1 Mark Wild,2 Alastair L. Boyes,2 and Peter A. Williams1

3 : Int J Biol Macromol. 2017 Aug 25. pii: S0141-8130(17)32824-6. doi: 10.1016/j.ijbiomac.2017.08.113. [Epub ahead of print] Isolation and characterization of three novel catechol 2,3-dioxygenase from three novel haloalkaliphilic BTEX-degrading Pseudomonas strains. Hassan HA1, Aly AA2.

4 : J Bacteriol. 1998 Feb; 180(4): 945–949. PMCID: PMC106976 - Biochemical and Genetic Characterization of trans-2′-Carboxybenzalpyruvate Hydratase-Aldolase from a Phenanthrene-Degrading Nocardioides Strain - Tokuro Iwabuchi† and Shigeaki Harayama

5 : “Geneious version (the version you are using) (https://www.geneious.com, Kearse et al., 2012)” Kearse, M., Moir, R., Wilson, A., Stones-Havas, S., Cheung, M., Sturrock, S., Buxton, S., Cooper, A., Markowitz, S., Duran, C., Thierer, T., Ashton, B., Mentjies, P., Drummond, A. (2012). Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data.Bioinformatics, 28(12), 1647-1649.

6 : Biomacromolecules 2007, 8, 1695-1701 // Macroscopic Fibers Self-Assembled from Recombinant Miniature Spider Silk Proteins; Margareta Stark, Stefan Grip, Anna Rising, My Hedhammar, Wilhelm Engström, Goran Hj&aumllm, and Jan Johansson*, Department of Anatomy, Physiology, and Biochemistry, The Biomedical Centre, Swedish University of Agricultural Sciences, SE-751 23 Uppsala, Sweden, and Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, SE-750 07 Uppsala, Sweden Received January 16, 2007; Revised Manuscript Received February 20, 2007