Difference between revisions of "Part:BBa K5228000"

(added title and overview)
(attempting to code clickable table of contents)
Line 1: Line 1:
 
<h1 id="thermostable-terminal-deoxynucleotidyl-transferase-thtdt-">Thermostable Terminal deoxynucleotidyl Transferase (ThTdT)</h1>
 
<h1 id="thermostable-terminal-deoxynucleotidyl-transferase-thtdt-">Thermostable Terminal deoxynucleotidyl Transferase (ThTdT)</h1>
 
<p>Thermostable Terminal Deoxynucleotidyl Transferase (ThTdT) is a mutated template-independent DNA polymerase originating from <em>Bos taurus</em>. This protein coding basic part performs template-free nucleotide (dNTP) addition at the 3’ end, requiring a starting primer. Recombinantly isolated from DH5α <em>E. coli</em>, the enzyme is a thermostable variant of its wild type counterpart enabling an increased working temperature range of 37°C to 55°C (as tested) and requires a divalent cation such as Co<sup>2+</sup> to increase its 3’-extension efficiency (used in characterization experiments).</p>
 
<p>Thermostable Terminal Deoxynucleotidyl Transferase (ThTdT) is a mutated template-independent DNA polymerase originating from <em>Bos taurus</em>. This protein coding basic part performs template-free nucleotide (dNTP) addition at the 3’ end, requiring a starting primer. Recombinantly isolated from DH5α <em>E. coli</em>, the enzyme is a thermostable variant of its wild type counterpart enabling an increased working temperature range of 37°C to 55°C (as tested) and requires a divalent cation such as Co<sup>2+</sup> to increase its 3’-extension efficiency (used in characterization experiments).</p>
 +
 +
__NOTOC__
 +
<h2 id="Sequence-and-Features-">Sequence and Features</h2>
 +
<partinfo>BBa_K5228000 SequenceAndFeatures</partinfo>
 +
 +
<!-- Table of Contents -->
 +
<div id="toc" class="toc">
 +
<div id="toctitle">
 +
<h2>Table of Contents</h2>
 +
</div>
 +
 +
<ul>
 +
<li class="toclevel-1 tocsection-1">
 +
<a href="#usage">
 +
<span class="tocnumber">1</span>
 +
<span class="toctext">Usage and Biology</span>
 +
</a>
 +
 +
<ul>
 +
<li class="toclevel-2 tocsection-2">
 +
<a href="#wtTdT">
 +
<span class="tocnumber">1.1</span>
 +
<span class="toctext">Terminal deoxynucleotidyl Transferase (TdT)</span>
 +
</a>
 +
 +
<ul>
 +
<li class="toclevel-3 tocsection-3">
 +
<a href="#wtTdT_overview">
 +
<span class="tocnumber">1.1.1</span>
 +
<span class="toctext">Wild type TdT (WT TdT) Overview</span>
 +
</a>
 +
 +
</li>
 +
 +
<li class="toclevel-3 tocsection-4">
 +
<a href="#ThTdT_overview">
 +
<span class="tocnumber">1.1.2</span>
 +
<span class="toctext">Thermostable TdT (ThTdT) Overview</span>
 +
</a>
 +
</li>
 +
</ul>
 +
</li>
 +
</ul>
 +
</li>
 +
<li class="toclevel-1 tocsection-5">
 +
<a href="#characterization">
 +
<span class="tocnumber">2</span>
 +
<span class="toctext">Characterization of ThTdT</span>
 +
</a>
 +
 +
<ul>
 +
<li class="toclevel-2 tocsection-6">
 +
<a href="#design">
 +
<span class="tocnumber">2.1</span>
 +
<span class="toctext">Designing the ThTdT Gene Fragment</span>
 +
</a>
 +
 +
</li>
 +
<li class="toclevel-2 tocsection-7">
 +
<a href="#cloning">
 +
<span class="tocnumber">2.2</span>
 +
<span class="toctext">Cloning of ThTdT</span>
 +
</a>
 +
 +
</li>
 +
<li class="toclevel-2 tocsection-8">
 +
<a href="#purification">
 +
<span class="tocnumber">2.3</span>
 +
<span class="toctext">Purification of ThTdT</span>
 +
</a>
 +
 +
</li>
 +
<li class="toclevel-2 tocsection-9">
 +
<a href="#pof">
 +
<span class="tocnumber">2.4</span>
 +
<span class="toctext">Proof of Function</span>
 +
</a>
 +
 +
 +
<ul>
 +
<li class="toclevel-3 tocsection-10">
 +
<a href="#lps">
 +
<span class="tocnumber">2.4.1</span>
 +
<span class="toctext">Liquid Phase Synthesis (LPS)</span>
 +
</a>
 +
</li>
 +
</ul>
 +
</li>
 +
</ul>
 +
</li>
 +
<li class="toclevel-1 tocsection-11">
 +
<a href="#potential">
 +
<span class="tocnumber">3</span>
 +
<span class="toctext">Potential Applications</span>
 +
</a>
 +
</li>
 +
</ul>
 +
<li class="toclevel-1 tocsection-12">
 +
<a href="#ref">
 +
<span class="tocnumber">4</span>
 +
<span class="toctext">References</span>
 +
</a>
 +
</li>
 +
</ul>
 +
</div>
 +
 +
 +
<!-- Content Sections -->
 +
<h2 id="usage">Usage and Biology</h2>
 +
<p>This part is designed to...</p>
 +
 +
<h2 id="characterization">Characterization of ThTdT</h2>
 +
<p>This part is designed to...</p>
 +
 +
<h2 id="applications">Potential Applications</h2>
 +
<p>This part has the following annotations...</p>
 +
 +
<h2 id="references">References</h2>
 +
<ul>
 +
    <li>Reference 1</li>
 +
    <li>Reference 2</li>
 +
</ul>
 +
 +
<!-- Additional Content Sections -->
 +
<h2 id="wtTdT_overview">Wild type TdT (WT TdT) Overview</h2>
 +
<p>Details about WT TdT...</p>
 +
 +
<h2 id="ThTdT_overview">Thermostable TdT (ThTdT) Overview</h2>
 +
<p>Details about ThTdT...</p>
 +
 +
<h2 id="design">Designing the ThTdT Gene Fragment</h2>
 +
<p>Information on the design...</p>
 +
 +
<h2 id="cloning">Cloning of ThTdT</h2>
 +
<p>Information on cloning...</p>
 +
 +
<h2 id="purification">Purification of ThTdT</h2>
 +
<p>Information on purification...</p>
 +
 +
<h2 id="pof">Proof of Function</h2>
 +
<p>Details on proof of function...</p>
 +
 +
<h2 id="lps">Liquid Phase Synthesis (LPS)</h2>
 +
<p>Details about LPS...</p>
 +
 +
</body>
 +
</html>

Revision as of 12:40, 30 September 2024

Thermostable Terminal deoxynucleotidyl Transferase (ThTdT)

Thermostable Terminal Deoxynucleotidyl Transferase (ThTdT) is a mutated template-independent DNA polymerase originating from Bos taurus. This protein coding basic part performs template-free nucleotide (dNTP) addition at the 3’ end, requiring a starting primer. Recombinantly isolated from DH5α E. coli, the enzyme is a thermostable variant of its wild type counterpart enabling an increased working temperature range of 37°C to 55°C (as tested) and requires a divalent cation such as Co2+ to increase its 3’-extension efficiency (used in characterization experiments).


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 275
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 275
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 275
    Illegal BglII site found at 164
    Illegal BglII site found at 431
    Illegal BglII site found at 799
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 275
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 275
  • 1000
    COMPATIBLE WITH RFC[1000]

Table of Contents

  • <a href="#usage"> 1 Usage and Biology </a>
    • <a href="#wtTdT"> 1.1 Terminal deoxynucleotidyl Transferase (TdT) </a>
      • <a href="#wtTdT_overview"> 1.1.1 Wild type TdT (WT TdT) Overview </a>
      • <a href="#ThTdT_overview"> 1.1.2 Thermostable TdT (ThTdT) Overview </a>
  • <a href="#characterization"> 2 Characterization of ThTdT </a>
    • <a href="#design"> 2.1 Designing the ThTdT Gene Fragment </a>
    • <a href="#cloning"> 2.2 Cloning of ThTdT </a>
    • <a href="#purification"> 2.3 Purification of ThTdT </a>
    • <a href="#pof"> 2.4 Proof of Function </a>
      • <a href="#lps"> 2.4.1 Liquid Phase Synthesis (LPS) </a>
  • <a href="#potential"> 3 Potential Applications </a>
  • <a href="#ref"> 4 References </a>

    • </ul>


    Usage and Biology

    This part is designed to...

    Characterization of ThTdT

    This part is designed to...

    Potential Applications

    This part has the following annotations...

    References

    • Reference 1
    • Reference 2

    Wild type TdT (WT TdT) Overview

    Details about WT TdT...

    Thermostable TdT (ThTdT) Overview

    Details about ThTdT...

    Designing the ThTdT Gene Fragment

    Information on the design...

    Cloning of ThTdT

    Information on cloning...

    Purification of ThTdT

    Information on purification...

    Proof of Function

    Details on proof of function...

    Liquid Phase Synthesis (LPS)

    Details about LPS...

    </body> </html>