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| - | {{iitm/main_new}}
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| - | {{iitm/clickhide}}
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| - | <html>
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| - | <head>
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| - | <title>People</title>
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| - | <style type="text/css">
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| - | #secondarylinks {
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| - | width: 25%;
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| - | margin-left: 4%;
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| - | padding-left: 3%;
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| - | float: left;
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| - | background: white;
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| - | margin-right: 5%;
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| - | border: 1px solid black;
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| - | webkit-border-radius: 1.5ex;
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| - | -moz-border-radius: 1.5ex;
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| - | }
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| - | #secondarytext {
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| - | width: 60%;
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| - | float: left;
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| - | text-align: justify;
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| - | margin: 5px;
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| - | }
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| - | .clear {
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| - | clear: both;
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| - | }
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| - | </style>
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| - | </head>
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| - |
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| - | <body width=80%>
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| - | <div id="secondarylinks">
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| - | <ul>
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| - | <li> <a href="2010.igem.org/Team:IIT_Madras/Project/Motivation">Motivation</a>
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| - | <li> <a href="2010.igem.org/Team:IIT_Madras/Project/Ideation">Ideation</a>
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| - | </div>
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| - | <div id="secondarytext">
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| - | This is where the people stuff comes.
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| - | </div>
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| - | <div class="clear"></div>
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| - | <div height="25px"></div>
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| - |
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| - | == Project Details==
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| - |
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| - | === Part Design ===
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| - |
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| - | ==== PCR Products ====
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| - | {|
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| - | <small>
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| - | :'''Legend'''
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| - |
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| - | :* <font style="color:green"> This color represents the portion to be added by Extension PCR / Inverse PCR / Assembly PCR.</font>
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| - |
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| - | :* <font style="color:red"> This color represents sites that lie in the middle of the parts to be submitted. These are placed so that the promoters and the inserts can be switched out.</font>
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| - |
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| - | :* <font style="color:blue">This represents an apparent restriction site, one that cannot be used due to the sequences' presence in an ORF</font>
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| - | </small>
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| - |
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| - | :'''SCM'''
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| - |
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| - | ::Gene of 295 bp to be synthesised by Assembly PCR / LCR.
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| - |
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| - | :'''Block 1'''
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| - |
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| - | :# <font style="color:green">Kpn1</font> | P170 | <font style="color:green">RBS</font> | <font style="color:green">HindIII</font>
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| - | :# <font style="color:green">Kpn1</font> | NICE | <font style="color:green">RBS</font> | <font style="color:green">HindIII</font>
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| - | :# <font style="color:green">HindIII</font>| GFP | Terminator | <font style="color:green">BamH1</font>
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| - | :# <font style="color:green">Kpn1</font>| Constitutive | RBS | GFP | Terminator | <font style="color:green">BamH1</font>
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| - |
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| - |
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| - | :'''Block 2'''
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| - |
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| - | :# Kpn1 | Constitutive | RBS | <font style="color:green">HindIII</font> | <font style="color:green">SP310mut2</font> | <font style="color:green">Nhe1</font> | GFP | Terminator | BamH1 (by Inverse PCR) // Can't design a Primer this big.
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| - | :# <font style="color:green">Kpn1</font> | <font style="color:green">Constitutive</font> | <font style="color:green">RBS</font> | <font style="color:green">HindIII</font> | SP310mut2 | <font style="color:green">Nhe1</font>
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| - | :# <font style="color:green">Nhe1</font> | GFP | Terminator | BamH1 (Extension PCR from 1.4)
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| - |
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| - |
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| - | :'''Block 3'''
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| - |
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| - | :# <font style="color:green">Kpn1</font> | Constitutive | RBS | <font style="color:green">HindIII</font>
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| - | :# <font style="color:green">HindIII</font> | CRE-NLS | <font style="color:green">Terminator</font> | <font style="color:green">Sac1</font>
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| - | :# <font style="color:green">Sac1</font> | Constitutive | RBS | <font style="color:green">loxP</font> | <font style="color:green">Nhe1</font> | GFP | Terminator | <font style="color:green">loxP</font> | <font style="color:green">Xho1</font> (by Inverse PCR + Extension PCR)
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| - | :# <font style="color:green">Xho1</font> | RFP | Terminator | <font style="color:green">BamH1</font>
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| - |
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| - | :'''Final Design'''
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| - |
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| - | :# Kpn1 | NICE | RBS | HindIII (as made in 1.1)
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| - | :# HindIII | CRE-NLS | Terminator | Sac1 (as made in 3.2)
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| - | :# <font style="color:green">Sac1</font> | <font style="color:green">loxP</font> | P170 | <font style="color:green">RBS</font> | SP310mut2 | <font style="color:green">Xho1</font> (by Inverse PCR + Extension PCR)
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| - | :# <font style="color:green">Xho1</font> | SCM | <font style="color:green">Nhe1</font> | <font style="color:green">loxP</font> | <font style="color:green">Terminator</font> | <font style="color:green">BamH1</font>
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| - |
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| - |
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| - | :'''Part Submission'''
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| - | :# <font style="color:green">EcoR1 | Not1 | Xba1</font> | NICE | RBS | <font style="color:green">Spe1 | Not1 | Pst1</font>
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| - | :# <font style="color:green">EcoR1 | Not1 | Xba1</font> | P170 | RBS | <font style="color:green">Spe1 | Not1 | Pst1</font>
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| - | :# <font style="color:green">EcoR1 | Not1 | Xba1</font> | <font style="color:red">Kpn1</font> | Constitutive | RBS | <font style="color:red">HindIII</font> | SP310mut2 | <font style="color:red">Nhe1</font> | GFP | Terminator | <font style="color:red">BamH1</font> | <font style="color:green">Spe1 | Not1 | Pst1</font>
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| - | :# <font style="color:green">EcoR1 | Not1 | Xba1</font> | <font style="color:red">Kpn1</font> | Constitutive | RBS | <font style="color:red">HindIII</font> | CRE | Terminator | <font style="color:red">Sac1</font> | Constitutive | RBS | loxP | <font style="color:red">Sma1</font> | GFP | Terminator | loxP | <font style="color:red">Xho1</font> | RFP | Terminator | <font style="color:red">BamH1</font> | <font style="color:green">Spe1 | Not1 | Xba1</font>
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| - | :# <font style="color:green">EcoR1</font> | <font style="color:blue">Not1</font> | <font style="color:green">Xba1</font> | <font style="color:red">Kpn1</font> | NICE | RBS | <font style="color:red">HindIII</font> | CRE-NLS | Terminator | <font style="color:red">Sac1</font> | loxP | P170 | RBS | SP310mut2 | <font style="color:red">Xho1</font> | SCM | <font style="color:red">BamH1</font> | loxP | Terminator | <font style="color:green">Spe1</font> | <font style="color:blue">Not1</font> | <font style="color:green">Pst1</font>
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| - | |}
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| - | ====Proof of Concept ====
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| - | {|
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| - | :'''Block 1 - To characterize promoters against a standard'''
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| - |
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| - | :*P170 | GFP | Terminator
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| - |
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| - | :*NICE | GFP | Terminator
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| - |
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| - | :*Constitutive | GFP | Terminator
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| - |
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| - | :'''Block 2 - Test of export efficiency'''
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| - |
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| - | :*Constitutive | GFP | Terminator
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| - |
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| - | :*Constitutive | SP310mut2 | GFP | Terminator
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| - |
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| - | :'''Block 3''' -'''To test CRE-loxP system'''
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| - |
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| - | :*Constitutive | RBS | CRE | Terminator | Constitutive | RBS | loxP | GFP | Terminator | loxP | RFP | Terminator
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| - | :Acid Tolerance Promoter (P170) | CRE | Terminator | loxP | Consitutive | GFP | Terminator | loxP | RFP | Terminator
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| - | |}
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| - |
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| - | === Final design ===
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| - | {|$
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| - | <big><font style="color:red">NICE | CRE | Terminator | loxP | Acid Tolerance Inducer (P170) | Export tag | SCM | loxP | Terminator</font></big>
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| - |
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| - |
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| - | |}
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| - |
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| - | === The Experiments ===
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| - |
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| - |
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| - |
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| - | === Part 3 ===
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| - |
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| - |
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| - | == Results ==
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| - |
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| - | {{iitm/footbar}}
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