Difference between revisions of "Part:BBa K079045"
 
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Parts K079045 through K079048 are libraries intended to introduce standardization and modularity for operator sequences.  
 
Parts K079045 through K079048 are libraries intended to introduce standardization and modularity for operator sequences.  
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Each of the 4 collections has a different repressor specificity (Lac, Tet, Lambda and LexA) and consist of 3 different members with distinct binding affinities.
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Each of the 4 collections has a different repressor specificity (Lac, Tet, Lambda and LexA) and consists of 3 different members with distinct binding affinities.
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Since operators flanked by Biobrick ends are unpractical and costly to obtain both by dna sinthesys and phosphorylated oligos we designed a "Standard Collection Vector".
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Since operators flanked by Biobrick ends are unpractical and costly to obtain, both by dna sinthesys and phosphorylated oligos, we designed a "Standard Collection Vector".
 
[[Image:Layout.jpg | Left |320x240 ]]
 
[[Image:Layout.jpg | Left |320x240 ]]
  
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This layout allows to get multiple small sequences with a single Synthesis process and specifically enable the extraction of one or a combination of operators using exclusively Biobrick prefix and suffix restriction enzymes.
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This layout allows to get multiple small sequences with a single synthesis process and specifically enable the extraction of one or a combination of operators using exclusively Biobrick prefix and suffix restriction enzymes.
−
Digestion and religation of the Collection Vector with '''Xba''' and '''Pst''' yields a single '''Operators 1''' and '''Operator 3''' vectors respectively (Es: [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079019 Lac2] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079017 Lac Symmetric]) avoiding the troubles and optimization related to the run and purification of very small bands on agarose gel.
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Digestion and religation of the collection vector with '''Xba''' and '''Pst''' yields a single '''Operators 1''' and '''Operator 3''' vectors respectively (Es: [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079019 Lac2] and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079017 Lac Symmetric]) avoiding the troubles and optimization related to the run and purification of very small bands on agarose gel.
  
 
A similar approach using '''EcoRI''' give rise to a multiple Operator Vector comprising '''Operator 1 and Operator 2''' sequences, usable per se as a composite regulatory site or further restrictable with '''PstI''' to extract the single''' Operator 2''' (Es: [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079018 Lac1])
 
A similar approach using '''EcoRI''' give rise to a multiple Operator Vector comprising '''Operator 1 and Operator 2''' sequences, usable per se as a composite regulatory site or further restrictable with '''PstI''' to extract the single''' Operator 2''' (Es: [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079018 Lac1])
 
    
 
    
  
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The Libraries have been syntetized by Geneart in their standard  pGA18 and pMA synthesis vector, they both have ColE1 high copy number origin and Ampicillin resistance.<br>
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The Libraries have been syntetized by Geneart in their standard  pGA18 and pMA synthesis vector, with both ColE1 high copy number origin of replication and Ampicillin resistance.
  
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[[Image:LacPlasmid.jpg |left|500 px ]]
 
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[[Image:LacTitle.jpg |right|300 px ]]           
 
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[[Image:Tet.jpg |right|500 px ]]
 
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[[Image:TetTitle.jpg|left|300 px ]]
 
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<br>
 
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[[Image:CI.jpg |left|500px ]]
 
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<br>
 
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[[Image:Lex.jpg |right|500px ]]
 
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{| cellpadding="15" border="0" align="center"
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|rowspan=5|[[Image:LacPlasmid.jpg |left|500 px ]]
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|'''<font size="5">LAC LIBRARY</font>'''
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|-
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| [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079018 Lac1]
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|-
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| [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079019 Lac2]
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|-
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| [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079017 Lac SymL]
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|-   
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|
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|-
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|-
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|rowspan=5|[[Image:Tet.jpg |right|500 px ]]
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|'''<font size="5">TET LIBRARY</font>'''
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|-
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| [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079036 TeT O]
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|-
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| [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079037 TetO-4C]
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|-
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| [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079037 TetO-wt/4C5G]
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|-
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|-
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|-
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|rowspan=5|[[Image:CI.jpg |right|500 px ]]
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|'''<font size="5">LAMBDA LIBRARY</font>'''
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|-
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| [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079041 Lambda OR1]
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|-
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| [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079042 Lambda OR2]
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|-
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| [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079043 Lambda OR3]
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|rowspan=5|[[Image:LexLib.jpg |right|500 px ]]
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|'''<font size="5">LEXA LIBRARY</font>'''
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|-
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| [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079039 LexA 1]
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|-
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| [https://parts.igem.org/wiki/index.php?title=Part:BBa_K079040 LexA 2]
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|-
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|
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|-
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|}
  
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[[Image:LexTitle.jpg|left|300 px ]]
 
  
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<br><br><br><br><br><br><br><br><br><br><br><br><br><br><br>
 
  
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<br><br><br><br><br><br><br><br><br><br><br><br><br><br><br>
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<br><br><br><br>
  
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Right now we're ultimating the design and testing of an equally simple protocol to transfer operator sequences in Biobrick Standard Assembly Vectors using standard cloning reagents and avoiding as much as possible experimentally fragile or optimization dependent steps
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Right now we're ultimating the design and testing of an equally [http://2008.igem.org/Team:Bologna/Wetlab#Operator_site_cloning_in_standard_plasmids simple protocol] to transfer operator sequences in Biobrick Standard Assembly Vectors using standard cloning reagents and avoiding as much as possible experimentally fragile or optimization dependent steps.
  
  
  
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<br><br><br><br><br><br><br><br><br><br><br><br><br><br><br>
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<br><br><br>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 00:19, 30 October 2008

Lac operator library

Parts K079045 through K079048 are libraries intended to introduce standardization and modularity for operator sequences. Each of the 4 collections has a different repressor specificity (Lac, Tet, Lambda and LexA) and consists of 3 different members with distinct binding affinities. Since operators flanked by Biobrick ends are unpractical and costly to obtain, both by dna sinthesys and phosphorylated oligos, we designed a "Standard Collection Vector". 320x240

This layout allows to get multiple small sequences with a single synthesis process and specifically enable the extraction of one or a combination of operators using exclusively Biobrick prefix and suffix restriction enzymes. Digestion and religation of the collection vector with Xba and Pst yields a single Operators 1 and Operator 3 vectors respectively (Es: Lac2 and Lac Symmetric) avoiding the troubles and optimization related to the run and purification of very small bands on agarose gel.

A similar approach using EcoRI give rise to a multiple Operator Vector comprising Operator 1 and Operator 2 sequences, usable per se as a composite regulatory site or further restrictable with PstI to extract the single Operator 2 (Es: Lac1)


The Libraries have been syntetized by Geneart in their standard pGA18 and pMA synthesis vector, with both ColE1 high copy number origin of replication and Ampicillin resistance.


LacPlasmid.jpg
 LAC LIBRARY
Lac1
Lac2
Lac SymL
Tet.jpg
 TET LIBRARY
TeT O
TetO-4C
TetO-wt/4C5G
CI.jpg
 LAMBDA LIBRARY
Lambda OR1
Lambda OR2
Lambda OR3
LexLib.jpg
 LEXA LIBRARY
LexA 1
LexA 2






Right now we're ultimating the design and testing of an equally [http://2008.igem.org/Team:Bologna/Wetlab#Operator_site_cloning_in_standard_plasmids simple protocol] to transfer operator sequences in Biobrick Standard Assembly Vectors using standard cloning reagents and avoiding as much as possible experimentally fragile or optimization dependent steps.





Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]