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Team:UCSF/Parts
From 2008.igem.org
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'''AarI Shuttle Vector''' | '''AarI Shuttle Vector''' | ||
| - | ''The bulk of our parts | + | ''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 AB | ||
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*3000 bp spacer construct (Cyc1P-GFP-Adh1t-3000 bp-8X LexA Ops) | *3000 bp spacer construct (Cyc1P-GFP-Adh1t-3000 bp-8X LexA Ops) | ||
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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.'' | ||
| - | Sir2 AB | + | *Sir2 AB |
| - | Esa1 AB (Histone Acetyltransferase) | + | *Esa1 AB (Histone Acetyltransferase) |
| - | Sas2 AB (Histone Acetylatransferase) | + | *Sas2 AB (Histone Acetylatransferase) |
| - | Sas2 BD | + | *Sas2 BD |
| - | mCherry AB | + | *mCherry AB |
| - | mCherry BD | + | *mCherry BD |
| - | GFP AB | + | *GFP AB |
| - | GFP BD | + | *GFP BD |
| - | LexA (1-87) BD | + | *LexA (1-87) BD |
| - | TetR AB (Tet Repressor DNA Binding Domain) | + | *TetR AB (Tet Repressor DNA Binding Domain) |
| - | TetR BD | + | *TetR BD |
| - | LacI AB (Lac Repressor DNA Binding Domain) | + | *LacI AB (Lac Repressor DNA Binding Domain) |
| - | Pif3 AB (Light-inducible dimerization partner) | + | *Pif3 AB (Light-inducible dimerization partner) |
| - | PhyB 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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