Difference between revisions of "Template:TypeIIS/Plasmids/Level1"

(Created page with "When four Level 0 parts are assembled into a Transcriptional Unit (TU) into the following plasmid backbones (with <partinfo>BBa_J04454</partinfo>), the TU will be flanked by S...")
 
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When four Level 0 parts are assembled into a Transcriptional Unit (TU) into the following plasmid backbones (with <partinfo>BBa_J04454</partinfo>), the TU will be flanked by SapI restriction sites and these 3 bp Fusion Sites.
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====Assembling into Level 1 Plasmds====
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pSB1K01-pSB1K04 series of plasmids are based on the pOdd set of Loop Vectors and are built upon the <partinfo>pSB1K3</partinfo> plasmid backbone.
  
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'''Note:'''
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*Since <partinfo>pSB1K3</partinfo> is a high-copy plasmid backbone, these vectors should not be used for Level 3 assemblies.
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*Currently, Registry samples of these plasmid backbones have not been fully sequenced. Only their prefix and suffix along with their insert <partinfo>BBa_J04454</partinfo> have been verified.
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<center>
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[[File:TypeIIS-Level1-J04454-Simple.png]]<br>
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'''Simplified diagram of BBa_J04454 and pSB1K0# with BsaI cut site.'''
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</center>
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When four Level 0 parts are assembled into the plasmid backbone (paired with <partinfo>BBa_J04454</partinfo>) in a one-pot assembly with BsaI...
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<center>[[File:TypeIIS-Level1-L0-Part-Assembly-Simple.png|900px]]</center>
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the resulting transcriptional unit (TU) will be flanked by SapI restriction sites and the specified 3 bp Fusion Sites (NNN).
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<center>[[File:TypeIIS-Level1-TU-Simple.png]]</center>
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====Level 1 Plasmid Set====
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Level 1 transcriptional units in this set of plasmid backbones will be flanked by SapI recognition sites and specified 3bp fusion sites, enabling further assembly of up to 4 transcriptional units as a Level 2 Assembly.
 
<html>
 
<html>
 
<center>
 
<center>
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</tr>
 
</tr>
 
<tr>
 
<tr>
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<td></html><partinfo>pSB1K01</partinfo><html></td>
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<td></html>[[Part:pSB1K01|pSB1K01]]<html></td>
 
<td>pOdd1</td>
 
<td>pOdd1</td>
 
<td class="dnaSeq rightAlign">ATG</td>
 
<td class="dnaSeq rightAlign">ATG</td>
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</tr>
 
</tr>
 
<tr>
 
<tr>
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<td></html><partinfo>pSB1K02</partinfo><html></td>
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<td></html>[[Part:pSB1K02|pSB1K02]]<html></td>
 
<td>pOdd2</td>
 
<td>pOdd2</td>
 
<td class="dnaSeq rightAlign">GCA</td>
 
<td class="dnaSeq rightAlign">GCA</td>
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</tr>
 
</tr>
 
<tr>
 
<tr>
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<td></html><partinfo>pSB1K03</partinfo><html></td>
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<td></html>[[Part:pSB1K03|pSB1K03]]<html></td>
 
<td>pOdd3</td>
 
<td>pOdd3</td>
 
<td class="dnaSeq rightAlign">TAC</td>
 
<td class="dnaSeq rightAlign">TAC</td>
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</tr>
 
</tr>
 
<tr>
 
<tr>
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<td></html><partinfo>pSB1K04</partinfo><html></td>
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<td></html>[[Part:pSB1K04|pSB1K04]]<html></td>
 
<td>pOdd4</td>
 
<td>pOdd4</td>
 
<td class="dnaSeq rightAlign">CAG</td>
 
<td class="dnaSeq rightAlign">CAG</td>
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</center>
 
</center>
 
</html>
 
</html>
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'''Note:''' Currently, Registry samples of these plasmid backbones have not been fully sequenced. Only their prefix and suffix along with their insert <partinfo>BBa_J04454</partinfo> have been verified.
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====Level 1 -> Level 2 Assembly====
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Up to 4 Level 1 transcriptional units can be assembled into a Level 2 multi-transcriptional unit. By planning your assemb
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<center>
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[[File:TypeIIS-Level1-Plasmid-Set.png]]<br>
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'''Simplified diagram of pSB1K0# set with assembled transcriptional units (TUs).'''
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</center>

Latest revision as of 19:53, 23 June 2020

Assembling into Level 1 Plasmds

pSB1K01-pSB1K04 series of plasmids are based on the pOdd set of Loop Vectors and are built upon the pSB1K3 plasmid backbone.

Note:

  • Since pSB1K3 is a high-copy plasmid backbone, these vectors should not be used for Level 3 assemblies.
  • Currently, Registry samples of these plasmid backbones have not been fully sequenced. Only their prefix and suffix along with their insert BBa_J04454 have been verified.


TypeIIS-Level1-J04454-Simple.png
Simplified diagram of BBa_J04454 and pSB1K0# with BsaI cut site.


When four Level 0 parts are assembled into the plasmid backbone (paired with BBa_J04454) in a one-pot assembly with BsaI...

TypeIIS-Level1-L0-Part-Assembly-Simple.png


the resulting transcriptional unit (TU) will be flanked by SapI restriction sites and the specified 3 bp Fusion Sites (NNN).

TypeIIS-Level1-TU-Simple.png



Level 1 Plasmid Set

Level 1 transcriptional units in this set of plasmid backbones will be flanked by SapI recognition sites and specified 3bp fusion sites, enabling further assembly of up to 4 transcriptional units as a Level 2 Assembly.

Registry Name Loop Name Fusion Site 5' TU Fusion Site 3'
pSB1K01 pOdd1 ATG TU 1 GCA
pSB1K02 pOdd2 GCA TU 2 TAC
pSB1K03 pOdd3 TAC TU 3 CAG
pSB1K04 pOdd4 CAG TU 4 GGT

Level 1 -> Level 2 Assembly

Up to 4 Level 1 transcriptional units can be assembled into a Level 2 multi-transcriptional unit. By planning your assemb

TypeIIS-Level1-Plasmid-Set.png
Simplified diagram of pSB1K0# set with assembled transcriptional units (TUs).