Team:UCSF/Parts

From 2008.igem.org

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'''UCSF iGEM 2008 Parts'''
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For complete details and sequence, see the Registry: http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=UCSF&Done=1
For complete details and sequence, see the Registry: http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=UCSF&Done=1
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'''Project-relevant AarI TOPO clones'''  
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'''AarI Shuttle Vector'''
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''The bulk of our parts were generated using the AarI cloning technique. If that is not your thing, we have created a shuttle vector to facilitate conversion of our parts into biobricks.''
 +
 
 +
*AarI--Biobrick Shuttle AB
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 +
*AarI--Biobrick Shuttle BD
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 +
more details: [[Everything_you_ever_wanted_to_know_about_AarI]]
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 +
 
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'''Project-relevant AarI Parts'''  
''digestion of these plasmids with AarI will release a fragment for AarI cloning.''
''digestion of these plasmids with AarI will release a fragment for AarI cloning.''
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*Sir4 AD
*Sir4 AD
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GFP BD
 
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Sir4
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'''Project-relevant Composite Parts'''
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''AarI combinatorial cloning, combined with an occasional subclone, produced these composite parts.''
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Sir2 AB
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EFFECTORS
 +
*LexA-Sir2 under a galactose-inducible promoter (Gal1P-LexA(1-87)-Sir2-Adh1t)
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*LexA-Sir2 under a strong constitutive promoter (Adh1P-LexA(1-87)-Sir2-Adh1t)
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mCherry AB
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*LexA-Sir2 under a medium constitutive promoter (Cyc1P-LexA(1-87)-Sir2-Adh1t)
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mCherry BD
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*Ssn8, under a strong constitutive promoter (Adh1P-Ssn8-Adh1t)
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*Sir3, under a strong constitutive promoter (Adh1P!-Sir3-Adh1t)
 +
REPORTERS
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LexA (1-87) BD
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*GFP under medium constitutive promoter, 5' LexA Operators (8X LexA Ops-Cyc1P-GFP-Adh1t)
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Pif3 AB
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*GFP under medium constitutive promoter, 3' LexA Operators (Cyc1P-GFP-Adh1t-8X LexA Ops)
-
PhyB AB
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*GFP under pheromone-inducible promoter, 5' LexA Operators (8X LexA Ops-Fig1P-GFP-Adh1t)
 +
 
 +
*Regional Silencing test construct (Cyc1P-mCherry-Adh1t-8X LexA Ops-Cyc1P-GFP-Adh1t)
 +
 
 +
*250 bp spacer construct (Cyc1P-GFP-Adh1t-250 bp-8X LexA Ops)
 +
 
 +
*500 bp spacer construct (Cyc1P-GFP-Adh1t-500 bp-8X LexA Ops)
 +
 
 +
*1000 bp spacer construct (Cyc1P-GFP-Adh1t-1000 bp-8X LexA Ops)
 +
 
 +
*2000 bp spacer construct (Cyc1P-GFP-Adh1t-2000 bp-8X LexA Ops)
 +
 
 +
*3000 bp spacer construct (Cyc1P-GFP-Adh1t-3000 bp-8X LexA Ops)
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Sas2 AB
 
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Sas2 BD
 
'''AarI acceptor vectors (empty)'''
'''AarI acceptor vectors (empty)'''
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''These accept AarI digested parts, and include a promoter/terminator.''
+
''These pRS3series vectors accept AarI digested parts between a promoter/terminator. The promoter is flanked by PspOMI/XhoI sites, and the terminator by Not1/SacI sites, in case you want to change them out.''
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'''AarI Shuttle Vector'''
+
*AarI AD Acceptor (pRS305, Gal1P, Adh1t)
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''So you don't want to use AarI cloning? OK. We've provided a shuttle vector allows one-sublcone conversion of AarI parts into biobricks.
+
-
*AarI--Biobrick Shuttle AB
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*AarI AD Acceptor (pRS315, Adh1P, Adh1t)
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*AarI--Biobrick Shuttle BD
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*AarI AD Acceptor (pRS315, Cyc1P, Adh1t)
 +
 
 +
*AarI AD Acceptor (pRS315, 8X LexA Ops-Cyc1P, Adh1t)
 +
 
 +
*AarI AD acceptor (pRS315, Cyc1P, Adh1t-8XLexA Ops)
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 +
*AarI AD Acceptor (pRS315, 8X LexA Ops-Fig1P, Adh1t)
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''These are provided to the registry to facilitate getting started with AarI cloning. Some we had planned to use, but didn't get around to, and others are selected from the lab database of AarI parts.''  
''These are provided to the registry to facilitate getting started with AarI cloning. Some we had planned to use, but didn't get around to, and others are selected from the lab database of AarI parts.''  
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Esa1 AB
+
*Sir2 AB
 +
 
 +
*Esa1 AB (Histone Acetyltransferase)
 +
 
 +
*Sas2 AB (Histone Acetylatransferase)
 +
 
 +
*Sas2 BD
 +
 
 +
*mCherry AB
 +
 
 +
*mCherry BD
 +
 
 +
*GFP AB
 +
 
 +
*GFP BD
 +
 
 +
*LexA (1-87) BD
 +
 
 +
*TetR AB (Tet Repressor DNA Binding Domain)
 +
 
 +
*TetR BD
 +
 
 +
*LacI AB (Lac Repressor DNA Binding Domain)
 +
 
 +
*Pif3 AB (Light-inducible dimerization partner)
 +
 
 +
*PhyB AB (Light-inducible dimerization partner)

Latest revision as of 17:57, 29 October 2008

UCSF iGEM 2008 Parts

For complete details and sequence, see the Registry: http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=UCSF&Done=1


AarI Shuttle Vector The bulk of our parts were generated using the AarI cloning technique. If that is not your thing, we have created a shuttle vector to facilitate conversion of our parts into biobricks.

  • AarI--Biobrick Shuttle AB
  • AarI--Biobrick Shuttle BD

more details: Everything_you_ever_wanted_to_know_about_AarI


Project-relevant AarI Parts digestion of these plasmids with AarI will release a fragment for AarI cloning.

  • LexA (1-87 a.a) AB
  • Sir2 BD
  • GFP AD
  • Ssn8 AD
  • Sir3 A!D
  • Sir4 AD


Project-relevant Composite Parts AarI combinatorial cloning, combined with an occasional subclone, produced these composite parts.

EFFECTORS

  • LexA-Sir2 under a galactose-inducible promoter (Gal1P-LexA(1-87)-Sir2-Adh1t)
  • LexA-Sir2 under a strong constitutive promoter (Adh1P-LexA(1-87)-Sir2-Adh1t)
  • LexA-Sir2 under a medium constitutive promoter (Cyc1P-LexA(1-87)-Sir2-Adh1t)
  • Ssn8, under a strong constitutive promoter (Adh1P-Ssn8-Adh1t)
  • Sir3, under a strong constitutive promoter (Adh1P!-Sir3-Adh1t)

REPORTERS

  • GFP under medium constitutive promoter, 5' LexA Operators (8X LexA Ops-Cyc1P-GFP-Adh1t)
  • GFP under medium constitutive promoter, 3' LexA Operators (Cyc1P-GFP-Adh1t-8X LexA Ops)
  • GFP under pheromone-inducible promoter, 5' LexA Operators (8X LexA Ops-Fig1P-GFP-Adh1t)
  • Regional Silencing test construct (Cyc1P-mCherry-Adh1t-8X LexA Ops-Cyc1P-GFP-Adh1t)
  • 250 bp spacer construct (Cyc1P-GFP-Adh1t-250 bp-8X LexA Ops)
  • 500 bp spacer construct (Cyc1P-GFP-Adh1t-500 bp-8X LexA Ops)
  • 1000 bp spacer construct (Cyc1P-GFP-Adh1t-1000 bp-8X LexA Ops)
  • 2000 bp spacer construct (Cyc1P-GFP-Adh1t-2000 bp-8X LexA Ops)
  • 3000 bp spacer construct (Cyc1P-GFP-Adh1t-3000 bp-8X LexA Ops)


AarI acceptor vectors (empty) These pRS3series vectors accept AarI digested parts between a promoter/terminator. The promoter is flanked by PspOMI/XhoI sites, and the terminator by Not1/SacI sites, in case you want to change them out.

  • AarI AD Acceptor (pRS305, Gal1P, Adh1t)
  • AarI AD Acceptor (pRS315, Adh1P, Adh1t)
  • AarI AD Acceptor (pRS315, Cyc1P, Adh1t)
  • AarI AD Acceptor (pRS315, 8X LexA Ops-Cyc1P, Adh1t)
  • AarI AD acceptor (pRS315, Cyc1P, Adh1t-8XLexA Ops)
  • AarI AD Acceptor (pRS315, 8X LexA Ops-Fig1P, Adh1t)


Bonus AarI Parts These are provided to the registry to facilitate getting started with AarI cloning. Some we had planned to use, but didn't get around to, and others are selected from the lab database of AarI parts.

  • Sir2 AB
  • Esa1 AB (Histone Acetyltransferase)
  • Sas2 AB (Histone Acetylatransferase)
  • Sas2 BD
  • mCherry AB
  • mCherry BD
  • GFP AB
  • GFP BD
  • LexA (1-87) BD
  • TetR AB (Tet Repressor DNA Binding Domain)
  • TetR BD
  • LacI AB (Lac Repressor DNA Binding Domain)
  • Pif3 AB (Light-inducible dimerization partner)
  • PhyB AB (Light-inducible dimerization partner)




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