Team:UCSF/Parts

From 2008.igem.org

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'''AarI TOPO clones''' (digestion of these plasmids with AarI will release a fragment for AarI cloning).
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'''UCSF iGEM 2008 Parts'''
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GFP BD
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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
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Sir4
 
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Sir2 AB
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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.''
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Sir2 BD
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*AarI--Biobrick Shuttle AB
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mCherry AB
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*AarI--Biobrick Shuttle BD
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mCherry BD
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more details: [[Everything_you_ever_wanted_to_know_about_AarI]]
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LexA (1-87) AB
 
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LexA (1-87) BD
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'''Project-relevant AarI Parts'''
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''digestion of these plasmids with AarI will release a fragment for AarI cloning.''
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Pif3 AB
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*LexA (1-87 a.a) AB
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PhyB AB
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*Sir2 BD
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Sas2 AB
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*GFP AD
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Sas2 BD
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*Ssn8 AD
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Esa1 AB
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*Sir3 A!D
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*Sir4 AD
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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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 +
EFFECTORS
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*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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*LexA-Sir2 under a medium constitutive promoter (Cyc1P-LexA(1-87)-Sir2-Adh1t)
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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)
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REPORTERS
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*GFP under medium constitutive promoter, 5' LexA Operators (8X LexA Ops-Cyc1P-GFP-Adh1t)
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*GFP under medium constitutive promoter, 3' LexA Operators (Cyc1P-GFP-Adh1t-8X LexA Ops)
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*GFP under pheromone-inducible promoter, 5' LexA Operators (8X LexA Ops-Fig1P-GFP-Adh1t)
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*Regional Silencing test construct (Cyc1P-mCherry-Adh1t-8X LexA Ops-Cyc1P-GFP-Adh1t)
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*250 bp spacer construct (Cyc1P-GFP-Adh1t-250 bp-8X LexA Ops)
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*500 bp spacer construct (Cyc1P-GFP-Adh1t-500 bp-8X LexA Ops)
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*1000 bp spacer construct (Cyc1P-GFP-Adh1t-1000 bp-8X LexA Ops)
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*2000 bp spacer construct (Cyc1P-GFP-Adh1t-2000 bp-8X LexA Ops)
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*3000 bp spacer construct (Cyc1P-GFP-Adh1t-3000 bp-8X LexA Ops)
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 +
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'''AarI acceptor vectors (empty)'''
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''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 AD Acceptor (pRS305, Gal1P, Adh1t)
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*AarI AD Acceptor (pRS315, Adh1P, Adh1t)
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*AarI AD Acceptor (pRS315, Cyc1P, Adh1t)
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*AarI AD Acceptor (pRS315, 8X LexA Ops-Cyc1P, Adh1t)
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*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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'''Bonus AarI Parts'''
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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.''
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*Sir2 AB
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*Esa1 AB (Histone Acetyltransferase)
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*Sas2 AB (Histone Acetylatransferase)
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*Sas2 BD
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*mCherry AB
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*mCherry BD
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*GFP AB
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*GFP BD
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*LexA (1-87) BD
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*TetR AB (Tet Repressor DNA Binding Domain)
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*TetR BD
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*LacI AB (Lac Repressor DNA Binding Domain)
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*Pif3 AB (Light-inducible dimerization partner)
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*PhyB AB (Light-inducible dimerization partner)
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'''AarI Acceptor Vectors'''
 
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pRS315, Adh1P, Adh1t: CEN ARS vector for yeast expression (LEU2 marker), with Adh1 promoter and terminator.
 

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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