Team:SUSTC-Shenzhen-B/plasmid construction

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Sans',sans-serif;-webkit-border-top-left-radius:3px;-webkit-border-top-right-radius:3px;-moz-border-radius-topleft:3px;-moz-border-radius-topright:3px;border-top-left-radius:3px;border-top-right-radius:3px}.tab-content{padding:20px 0}ul.tabs li:hover{}ul.tabs li.active{}html ul.tabs li.active a{color:#b03121;background:#e1dfc9;-webkit-border-top-left-radius:3px;-webkit-border-top-right-radius:3px;-moz-border-radius-topleft:3px;-moz-border-radius-topright:3px;border-top-left-radius:3px;border-top-right-radius:3px}#tab-body{padding:0 20px;background:#e1dfc9;border:0;-webkit-border-radius:3px;-webkit-border-top-left-radius:0;-moz-border-radius:3px;-moz-border-radius-topleft:0;border-radius:3px;border-top-left-radius:0}#toggle{margin:0 0 20px 0}h2.trigger{padding:0 0;margin:0;font-size:13px;background:transparent;font-family:arial;color:#3f4432}h2.trigger span{text-decoration:none;display:block;background:url(/wiki/images/9/93/Sustc-b1-toggle.png) no-repeat 0 5px;padding-left:35px;cursor:pointer;line-height:35px}h2.active span{background:url(/wiki/images/c/cd/Sustc-b1-toggle-down.png) no-repeat 0 5px}h2.trigger a:hover{color:#efeed9}h2.active{background:transparent;color:#af3728}.toggle_container{margin:0 0 1px 0;padding:5px 15px;overflow:hidden;clear:both;background:transparent}.toggle_container .block{padding:0 0 0 20px}.toggle_container .block p{padding-bottom:10px;margin:0}#sidebar{width:300px;float:left;padding:0 0 0 40px}#sidebar.positionleft{float:left;padding:0 40px 0 0}#sidebar.positionright{float:right}#sidebar .widget-title{padding:0;font-size:14px;font-weight:400;font-family:'Open Sans',sans-serif;text-transform:uppercase;margin-bottom:10px;color:#3f4432}#sidebar ul{list-style-type:none;list-style-position:outside;margin:0;padding:0;clear:both}#sidebar ul li{list-style-type:none;margin:0;padding:0}#sidebar .widget-container{margin-bottom:28px;padding-top:28px;background:url(/wiki/images/7/79/Sustc-b1-line.gif) no-repeat}#sidebar .widget-container:first-child{background:0;padding:0}#sidebar li li{list-style-type:none;margin:0 0 3px 0;padding:0 0 3px 0}#sidebar li li a{color:#777}#sidebar li li a:hover{text-decoration:none;color:#af3728}#sidebar ul.sub-menu,#sidebar ul.children,#sidebar ul ul ul{margin:5px 0 0 10px}#sidebar ul.sub-menu li,#sidebar ul.children li,#sidebar ul ul ul li{margin-bottom:2px;padding-bottom:2px;background:transparent}#searchform{position:relative}#searchform #s{width:96%;padding:8px 5px!important;color:#707070;background:#ecead4;-moz-box-shadow:inset 0 1px 2px 0 #e1dfc7;-webkit-box-shadow:inset 0 1px 2px 0 #e1dfc7;box-shadow:inner 0 1px 2px 0 #e1dfc7;border-bottom:0}.rp-widget li{clear:left;margin-bottom:0;padding-bottom:10px}.rp-widget img{padding:4px}.rp-widget li h3{margin-bottom:0}.rp-widget li h3 a{font-size:13px;font-weight:600}.rp-widget li 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/* prettyPhoto.css */
div.pp_default .pp_top,div.pp_default .pp_top .pp_middle,div.pp_default .pp_top .pp_left,div.pp_default .pp_top .pp_right,div.pp_default .pp_bottom,div.pp_default .pp_bottom .pp_left,div.pp_default .pp_bottom .pp_middle,div.pp_default .pp_bottom .pp_right{height:13px}div.pp_default .pp_top .pp_left{background:url(../images/default/sprite.png) -78px -93px no-repeat}div.pp_default .pp_top .pp_middle{background:url(../images/default/sprite_x.png) top left repeat-x}div.pp_default .pp_top .pp_right{background:url(../images/default/sprite.png) -112px -93px no-repeat}div.pp_default .pp_content .ppt{color:#f8f8f8}div.pp_default .pp_content_container .pp_left{background:url(../images/default/sprite_y.png) -7px 0 repeat-y;padding-left:13px}div.pp_default .pp_content_container .pp_right{background:url(../images/default/sprite_y.png) top right repeat-y;padding-right:13px}div.pp_default .pp_next:hover{background:url(../images/default/sprite_next.png) center right no-repeat;cursor:pointer}div.pp_default .pp_previous:hover{background:url(../images/default/sprite_prev.png) center left no-repeat;cursor:pointer}div.pp_default .pp_expand{background:url(../images/default/sprite.png) 0 -29px no-repeat;cursor:pointer;height:28px;width:28px}div.pp_default .pp_expand:hover{background:url(../images/default/sprite.png) 0 -56px no-repeat;cursor:pointer}div.pp_default .pp_contract{background:url(../images/default/sprite.png) 0 -84px no-repeat;cursor:pointer;height:28px;width:28px}div.pp_default .pp_contract:hover{background:url(../images/default/sprite.png) 0 -113px no-repeat;cursor:pointer}div.pp_default .pp_close{background:url(../images/default/sprite.png) 2px 1px no-repeat;cursor:pointer;height:30px;width:30px}div.pp_default .pp_gallery ul li a{background:url(../images/default/default_thumb.png) center center #f8f8f8;border:1px solid #aaa}div.pp_default .pp_social{margin-top:7px}div.pp_default .pp_gallery a.pp_arrow_previous,div.pp_default .pp_gallery a.pp_arrow_next{left:auto;position:static}div.pp_default .pp_nav .pp_play,div.pp_default .pp_nav .pp_pause{background:url(../images/default/sprite.png) -51px 1px no-repeat;height:30px;width:30px}div.pp_default .pp_nav .pp_pause{background-position:-51px -29px}div.pp_default a.pp_arrow_previous,div.pp_default a.pp_arrow_next{background:url(../images/default/sprite.png) -31px -3px no-repeat;height:20px;margin:4px 0 0;width:20px}div.pp_default a.pp_arrow_next{background-position:-82px -3px;left:52px}div.pp_default .pp_content_container .pp_details{margin-top:5px}div.pp_default .pp_nav{clear:none;height:30px;position:relative;width:110px}div.pp_default .pp_nav .currentTextHolder{color:#999;font-family:Georgia;font-size:11px;font-style:italic;left:75px;line-height:25px;margin:0;padding:0 0 0 10px;position:absolute;top:2px}div.pp_default .pp_close:hover,div.pp_default .pp_nav .pp_play:hover,div.pp_default .pp_nav .pp_pause:hover,div.pp_default .pp_arrow_next:hover,div.pp_default .pp_arrow_previous:hover{opacity:.7}div.pp_default .pp_description{font-size:11px;font-weight:700;line-height:14px;margin:5px 50px 5px 0}div.pp_default .pp_bottom .pp_left{background:url(../images/default/sprite.png) -78px -127px no-repeat}div.pp_default .pp_bottom .pp_middle{background:url(../images/default/sprite_x.png) bottom left repeat-x}div.pp_default .pp_bottom .pp_right{background:url(../images/default/sprite.png) -112px -127px no-repeat}div.pp_default .pp_loaderIcon{background:url(../images/default/loader.gif) center center no-repeat}div.light_rounded .pp_top .pp_left{background:url(../images/light_rounded/sprite.png) -88px -53px no-repeat}div.light_rounded .pp_top .pp_right{background:url(../images/light_rounded/sprite.png) -110px -53px no-repeat}div.light_rounded .pp_next:hover{background:url(../images/light_rounded/btnNext.png) center right no-repeat;cursor:pointer}div.light_rounded .pp_previous:hover{background:url(../images/light_rounded/btnPrevious.png) center left no-repeat;cursor:pointer}div.light_rounded .pp_expand{background:url(../images/light_rounded/sprite.png) -31px -26px no-repeat;cursor:pointer}div.light_rounded .pp_expand:hover{background:url(../images/light_rounded/sprite.png) -31px -47px no-repeat;cursor:pointer}div.light_rounded .pp_contract{background:url(../images/light_rounded/sprite.png) 0 -26px no-repeat;cursor:pointer}div.light_rounded .pp_contract:hover{background:url(../images/light_rounded/sprite.png) 0 -47px no-repeat;cursor:pointer}div.light_rounded .pp_close{background:url(../images/light_rounded/sprite.png) -1px -1px no-repeat;cursor:pointer;height:22px;width:75px}div.light_rounded .pp_nav .pp_play{background:url(../images/light_rounded/sprite.png) -1px -100px no-repeat;height:15px;width:14px}div.light_rounded .pp_nav .pp_pause{background:url(../images/light_rounded/sprite.png) -24px -100px no-repeat;height:15px;width:14px}div.light_rounded .pp_arrow_previous{background:url(../images/light_rounded/sprite.png) 0 -71px no-repeat}div.light_rounded .pp_arrow_next{background:url(../images/light_rounded/sprite.png) -22px -71px no-repeat}div.light_rounded .pp_bottom .pp_left{background:url(../images/light_rounded/sprite.png) -88px -80px no-repeat}div.light_rounded .pp_bottom .pp_right{background:url(../images/light_rounded/sprite.png) -110px -80px no-repeat}div.dark_rounded .pp_top .pp_left{background:url(../images/dark_rounded/sprite.png) -88px -53px no-repeat}div.dark_rounded .pp_top .pp_right{background:url(../images/dark_rounded/sprite.png) -110px -53px no-repeat}div.dark_rounded .pp_content_container .pp_left{background:url(../images/dark_rounded/contentPattern.png) top left repeat-y}div.dark_rounded .pp_content_container .pp_right{background:url(../images/dark_rounded/contentPattern.png) top right repeat-y}div.dark_rounded .pp_next:hover{background:url(../images/dark_rounded/btnNext.png) center right no-repeat;cursor:pointer}div.dark_rounded .pp_previous:hover{background:url(../images/dark_rounded/btnPrevious.png) center left no-repeat;cursor:pointer}div.dark_rounded .pp_expand{background:url(../images/dark_rounded/sprite.png) -31px -26px no-repeat;cursor:pointer}div.dark_rounded .pp_expand:hover{background:url(../images/dark_rounded/sprite.png) -31px -47px no-repeat;cursor:pointer}div.dark_rounded .pp_contract{background:url(../images/dark_rounded/sprite.png) 0 -26px no-repeat;cursor:pointer}div.dark_rounded .pp_contract:hover{background:url(../images/dark_rounded/sprite.png) 0 -47px no-repeat;cursor:pointer}div.dark_rounded .pp_close{background:url(../images/dark_rounded/sprite.png) -1px -1px no-repeat;cursor:pointer;height:22px;width:75px}div.dark_rounded .pp_description{color:#fff;margin-right:85px}div.dark_rounded .pp_nav .pp_play{background:url(../images/dark_rounded/sprite.png) -1px -100px no-repeat;height:15px;width:14px}div.dark_rounded .pp_nav .pp_pause{background:url(../images/dark_rounded/sprite.png) -24px -100px no-repeat;height:15px;width:14px}div.dark_rounded .pp_arrow_previous{background:url(../images/dark_rounded/sprite.png) 0 -71px no-repeat}div.dark_rounded .pp_arrow_next{background:url(../images/dark_rounded/sprite.png) -22px -71px no-repeat}div.dark_rounded .pp_bottom .pp_left{background:url(../images/dark_rounded/sprite.png) -88px -80px no-repeat}div.dark_rounded .pp_bottom .pp_right{background:url(../images/dark_rounded/sprite.png) -110px -80px no-repeat}div.dark_rounded .pp_loaderIcon{background:url(../images/dark_rounded/loader.gif) center center no-repeat}div.dark_square .pp_left,div.dark_square .pp_middle,div.dark_square .pp_right,div.dark_square .pp_content{background:#000}div.dark_square .pp_description{color:#fff;margin:0 85px 0 0}div.dark_square .pp_loaderIcon{background:url(../images/dark_square/loader.gif) center center no-repeat}div.dark_square .pp_expand{background:url(../images/dark_square/sprite.png) -31px -26px no-repeat;cursor:pointer}div.dark_square .pp_expand:hover{background:url(../images/dark_square/sprite.png) -31px -47px no-repeat;cursor:pointer}div.dark_square .pp_contract{background:url(../images/dark_square/sprite.png) 0 -26px no-repeat;cursor:pointer}div.dark_square .pp_contract:hover{background:url(../images/dark_square/sprite.png) 0 -47px no-repeat;cursor:pointer}div.dark_square .pp_close{background:url(../images/dark_square/sprite.png) -1px -1px no-repeat;cursor:pointer;height:22px;width:75px}div.dark_square .pp_nav{clear:none}div.dark_square .pp_nav .pp_play{background:url(../images/dark_square/sprite.png) -1px -100px no-repeat;height:15px;width:14px}div.dark_square .pp_nav .pp_pause{background:url(../images/dark_square/sprite.png) -24px -100px no-repeat;height:15px;width:14px}div.dark_square .pp_arrow_previous{background:url(../images/dark_square/sprite.png) 0 -71px no-repeat}div.dark_square .pp_arrow_next{background:url(../images/dark_square/sprite.png) -22px -71px no-repeat}div.dark_square .pp_next:hover{background:url(../images/dark_square/btnNext.png) center right no-repeat;cursor:pointer}div.dark_square .pp_previous:hover{background:url(../images/dark_square/btnPrevious.png) center left no-repeat;cursor:pointer}div.light_square .pp_expand{background:url(../images/light_square/sprite.png) -31px -26px no-repeat;cursor:pointer}div.light_square .pp_expand:hover{background:url(../images/light_square/sprite.png) -31px -47px no-repeat;cursor:pointer}div.light_square .pp_contract{background:url(../images/light_square/sprite.png) 0 -26px no-repeat;cursor:pointer}div.light_square .pp_contract:hover{background:url(../images/light_square/sprite.png) 0 -47px no-repeat;cursor:pointer}div.light_square .pp_close{background:url(../images/light_square/sprite.png) -1px -1px no-repeat;cursor:pointer;height:22px;width:75px}div.light_square .pp_nav .pp_play{background:url(../images/light_square/sprite.png) -1px -100px no-repeat;height:15px;width:14px}div.light_square .pp_nav .pp_pause{background:url(../images/light_square/sprite.png) -24px -100px no-repeat;height:15px;width:14px}div.light_square .pp_arrow_previous{background:url(../images/light_square/sprite.png) 0 -71px no-repeat}div.light_square .pp_arrow_next{background:url(../images/light_square/sprite.png) -22px -71px no-repeat}div.light_square .pp_next:hover{background:url(../images/light_square/btnNext.png) center right no-repeat;cursor:pointer}div.light_square .pp_previous:hover{background:url(../images/light_square/btnPrevious.png) center left no-repeat;cursor:pointer}div.facebook .pp_top .pp_left{background:url(../images/facebook/sprite.png) -88px -53px no-repeat}div.facebook .pp_top .pp_middle{background:url(../images/facebook/contentPatternTop.png) top left repeat-x}div.facebook .pp_top .pp_right{background:url(../images/facebook/sprite.png) -110px -53px no-repeat}div.facebook .pp_content_container .pp_left{background:url(../images/facebook/contentPatternLeft.png) top left repeat-y}div.facebook .pp_content_container .pp_right{background:url(../images/facebook/contentPatternRight.png) top right repeat-y}div.facebook .pp_expand{background:url(../images/facebook/sprite.png) -31px -26px no-repeat;cursor:pointer}div.facebook .pp_expand:hover{background:url(../images/facebook/sprite.png) -31px -47px no-repeat;cursor:pointer}div.facebook .pp_contract{background:url(../images/facebook/sprite.png) 0 -26px no-repeat;cursor:pointer}div.facebook .pp_contract:hover{background:url(../images/facebook/sprite.png) 0 -47px no-repeat;cursor:pointer}div.facebook .pp_close{background:url(../images/facebook/sprite.png) -1px -1px no-repeat;cursor:pointer;height:22px;width:22px}div.facebook .pp_description{margin:0 37px 0 0}div.facebook .pp_loaderIcon{background:url(../images/facebook/loader.gif) center center no-repeat}div.facebook .pp_arrow_previous{background:url(../images/facebook/sprite.png) 0 -71px no-repeat;height:22px;margin-top:0;width:22px}div.facebook .pp_arrow_previous.disabled{background-position:0 -96px;cursor:default}div.facebook .pp_arrow_next{background:url(../images/facebook/sprite.png) -32px -71px no-repeat;height:22px;margin-top:0;width:22px}div.facebook .pp_arrow_next.disabled{background-position:-32px -96px;cursor:default}div.facebook .pp_nav{margin-top:0}div.facebook .pp_nav p{font-size:15px;padding:0 3px 0 4px}div.facebook .pp_nav .pp_play{background:url(../images/facebook/sprite.png) -1px -123px no-repeat;height:22px;width:22px}div.facebook .pp_nav .pp_pause{background:url(../images/facebook/sprite.png) -32px -123px no-repeat;height:22px;width:22px}div.facebook .pp_next:hover{background:url(../images/facebook/btnNext.png) center right no-repeat;cursor:pointer}div.facebook .pp_previous:hover{background:url(../images/facebook/btnPrevious.png) center left no-repeat;cursor:pointer}div.facebook .pp_bottom .pp_left{background:url(../images/facebook/sprite.png) -88px -80px no-repeat}div.facebook .pp_bottom .pp_middle{background:url(../images/facebook/contentPatternBottom.png) top left repeat-x}div.facebook .pp_bottom .pp_right{background:url(../images/facebook/sprite.png) -110px -80px no-repeat}div.pp_pic_holder a:focus{outline:0}div.pp_overlay{background:#000;display:none;left:0;position:absolute;top:0;width:100%;z-index:9500}div.pp_pic_holder{display:none;position:absolute;width:100px;z-index:10000}.pp_content{height:40px;min-width:40px}* html .pp_content{width:40px}.pp_content_container{position:relative;text-align:left;width:100%}.pp_content_container .pp_left{padding-left:20px}.pp_content_container .pp_right{padding-right:20px}.pp_content_container .pp_details{float:left;margin:10px 0 2px}.pp_description{display:none;margin:0}.pp_social{float:left;margin:0}.pp_social .facebook{float:left;margin-left:5px;overflow:hidden;width:55px}.pp_social .twitter{float:left}.pp_nav{clear:right;float:left;margin:3px 10px 0 0}.pp_nav p{float:left;margin:2px 4px;white-space:nowrap}.pp_nav .pp_play,.pp_nav .pp_pause{float:left;margin-right:4px;text-indent:-10000px}a.pp_arrow_previous,a.pp_arrow_next{display:block;float:left;height:15px;margin-top:3px;overflow:hidden;text-indent:-10000px;width:14px}.pp_hoverContainer{position:absolute;top:0;width:100%;z-index:2000}.pp_gallery{display:none;left:50%;margin-top:-50px;position:absolute;z-index:10000}.pp_gallery div{float:left;overflow:hidden;position:relative}.pp_gallery ul{float:left;height:35px;margin:0 0 0 5px;padding:0;position:relative;white-space:nowrap}.pp_gallery ul a{border:1px rgba(0,0,0,.5) solid;display:block;float:left;height:33px;overflow:hidden}.pp_gallery ul a img{border:0}.pp_gallery li{display:block;float:left;margin:0 5px 0 0;padding:0}.pp_gallery li.default a{background:url(../images/facebook/default_thumbnail.gif) 0 0 no-repeat;display:block;height:33px;width:50px}.pp_gallery .pp_arrow_previous,.pp_gallery 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/*
 * Superfish v1.4.8 - jQuery menu widget
 * Copyright (c) 2008 Joel Birch
 *
 * Dual licensed under the MIT and GPL licenses:
 *  http://www.opensource.org/licenses/mit-license.php
 *  http://www.gnu.org/licenses/gpl.html
 *
 * CHANGELOG: http://users.tpg.com.au/j_birch/plugins/superfish/changelog.txt
 */

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/*
 * Supersubs v0.2b - jQuery plugin
 * Copyright (c) 2008 Joel Birch
 *
 * Dual licensed under the MIT and GPL licenses:
 *  http://www.opensource.org/licenses/mit-license.php
 *  http://www.gnu.org/licenses/gpl.html
 *
 *
 * This plugin automatically adjusts submenu widths of suckerfish-style menus to that of
 * their longest list item children. If you use this, please expect bugs and report them
 * to the jQuery Google Group with the word 'Superfish' in the subject line.
 *
 */

(function($){ // $ will refer to jQuery within this closure

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                               <h1><p>Plasmid Construction</p></h1>


              
  <p>A goal of synthetic biology is to standardize the form of genetic components to prevent structural elements of components from changing with reactions. It provides a standard method of assembling genetic components using specified prefixes and suffixes. Prefixes include EcoRⅠ and XbaⅠ; suffixes include SpeⅠ and PstⅠ.
  <p>The recognition site of XbaⅠ:</p>
  <img src="https://static.igem.org/mediawiki/2012/2/2e/Xba1.JPG" class="img_fl img_border" />
  <p>(Figure 3)</p>
  <p>The recognition site of SpeⅠ:</p>
  <img src="https://static.igem.org/mediawiki/2012/7/77/Spe1.JPG" class="img_fl img_border" />    
  <p>(Figure 4)</p>
 <p>The same cohesive terminus enable them to connect with each other. However, after the combination, neither XbaⅠnor SpeⅠis able to recognize this sequence. Through this way we could achieve the goal of building BioBricks. </p>
<p>We can see from the picture(Figure 2) that there are 4 standard restriction enzyme cutting sites: 
EcoRⅠ, XbaⅠ, SpeⅠ,PstⅠ. </p>
  </p>
  <p>(1) As we will use pstⅠto be the restriction cutting site of terminators, if we want to ligase GFP to the plasmid, this restriction cutting site should be mutated.
  <p>The recognition site of PstⅠ:</p>
  <img src="https://static.igem.org/mediawiki/2012/c/cc/Pst1.JPG" class="img_fl img_border" /> 
   <p>(Figure 5)</p>
  <p>The recognition site of AflⅡ:</p>
  <img src="https://static.igem.org/mediawiki/2012/c/cc/Afl2.JPG" class="img_fl img_border" /> 
    <p>(Figure 6)</p>
 <p> Since AflⅡis similar to PstⅠ,they just have 2 different base pairs, we used these primers:</p>
<p>PtoA-F</p>
<p>5'-CCACCTGACGTCTAAGAAAC-3'</p>
<p>PtoA-R</p>
<p>5'-CTTAAGCGGCCGCTACTAGTA-3'</p>
 <p>to mutate the vector.</p>
  </p>

              <p>(2) After the mutation succeeded, the next step is to connect sequences of GFP & RFP to the vector. These sequences were built by PCR extension.</p>
                  <p>To digest GFP sequence, we used SpeⅠand AflⅡ. We also digested the vector with the same restriction enzyme cutting sites. During the process of base complementary pairing, GFP sequence could be ligated onto the vector with the help of T4 DNA ligase. At the same time, the recognition site of PstⅠ would be added to the vector.</p>
                  <p>To digest RFP sequence, we used NotⅠ and SpeⅠ. We also digested the vector with the same restriction enzyme cutting sites. During the process of base complementary pairing, RFP sequence could be ligated onto vector with the help of T4 DNA ligase. At the same time, the recognition site of XbaⅠwould be added to the vector.</p>
                  <p>We can either connect 3 fragments: GFP, RFP sequences and the plasmid at the same time, or connect 2 fragments first and then connect the other one. However, when carrying out the second scheme, we should decide which one to be connected first. According to the picture, GFP should be the one.</p>

                  <p>Besides:</p>
                  <p>We designed these primers to amplify RFP.</p>
                  <p>R-NPS-F</p>
                  <p>5'-TATAGCGGCCGCCTTAAGTAAGTAAGAGTATACG-3' </p> 
                  <p>R-NPS-R</p>
                  <p>5'-CGGAGACTAGTCTGCAGATCACATAAGTAAAGTGATAATC-3'</p>
                  <p>We designed these primers to amplify GFP.</p>
                  <p>G-SXA-F</p>
                  <p>5'-CTAGACTAGTTCTAGAGGCGGACTCACTATAGA-3'  </p>
                  <p>G-SXA-R</p>
                  <p>5'-CCGACTTAAGGGATCCTATAAACGCAG-3'  </p>
                <p>(3) Since we built up the vector through the steps before, we decided to use XbaⅠ,PstⅠto digest the vector.</p> 
                  <p>The terminators were built by PCR extension.</p>
                  <p>Then we can ligated terminator into the backbone and transformed into competent cells.</p>
                  <p>Characterization devices testing the performance of the terminators utilized fluorescent proteins to measure input and output and altered the arabinose transport system to control inputs. The fluorescence produced by the characterization devices were then measured using flow cytometry to calculate the termination efficiency of the terminators. </p>




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