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Team:Harvard/fences/parts
From 2010.igem.org
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<tr><td>Barnase</td><td> Fatal gene for the genetic fence </td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f1">link</a></td></tr> | <tr><td>Barnase</td><td> Fatal gene for the genetic fence </td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f1">link</a></td></tr> | ||
<tr><td>Barstar</td><td>Protein inhibitor of Barnase</td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f2">link</a></td></tr> | <tr><td>Barstar</td><td>Protein inhibitor of Barnase</td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f2">link</a></td></tr> | ||
| - | <tr><td>LacIN</td><td>Lac Inhibitor with Nuclear Localization sequence - transcriptional inhibitor of the Act2LacO promoter</td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f3"></td></tr> | + | <tr><td>LacIN</td><td>Lac Inhibitor with Nuclear Localization sequence - transcriptional inhibitor of the Act2LacO promoter</td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f3">link</a></td></tr> |
<tr><td>RXRLc-VP16</td><td> Fusion protein of Locust Retinoic Acid Receptor and VP16 activation domain - binds EcR-Gal4 in presence of Methoxyfenozide</td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f4">link</a></td></tr> | <tr><td>RXRLc-VP16</td><td> Fusion protein of Locust Retinoic Acid Receptor and VP16 activation domain - binds EcR-Gal4 in presence of Methoxyfenozide</td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f4">link</a></td></tr> | ||
<tr><td>EcR-Gal4</td><td> Fusion protein of Ecdysone receptor and Gal4 DNA binding domain - binds RXRLc-VP16 and initiates transcription downstream of Gal Upstream Activating Sequence in presence of Methoxyfenozide </td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f5">link</a></td></tr> | <tr><td>EcR-Gal4</td><td> Fusion protein of Ecdysone receptor and Gal4 DNA binding domain - binds RXRLc-VP16 and initiates transcription downstream of Gal Upstream Activating Sequence in presence of Methoxyfenozide </td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f5">link</a></td></tr> | ||
Revision as of 14:59, 25 October 2010
parts and primers
Parts
| Name | Description | Resistance | Sequence |
| Barnase | Fatal gene for the genetic fence | chloramphenicol | link |
| Barstar | Protein inhibitor of Barnase | chloramphenicol | link |
| LacIN | Lac Inhibitor with Nuclear Localization sequence - transcriptional inhibitor of the Act2LacO promoter | chloramphenicol | link |
| RXRLc-VP16 | Fusion protein of Locust Retinoic Acid Receptor and VP16 activation domain - binds EcR-Gal4 in presence of Methoxyfenozide | chloramphenicol | link |
| EcR-Gal4 | Fusion protein of Ecdysone receptor and Gal4 DNA binding domain - binds RXRLc-VP16 and initiates transcription downstream of Gal Upstream Activating Sequence in presence of Methoxyfenozide | chloramphenicol | link |
Promoters
| Name | Description | Active conditions | Resistance | Sequence |
| pENT CUP | Constitutive Plant Promoter | Constitutive | chloramphenicol | |
| 5xGal UAS | 5 time repeat of Gal4 upstream activating sequence | On in presense of dimerized EcR-Gal and RXRLc-VP16 | chloramphenicol | |
| Act2lacO | Actin2 promoter with Lac Inhibitor responsive sites (Lac O sites | Constitutive, inhibited by LacIN | chloramphenicol |
Primers
| Name | Description | Sequence |
| Arp2mutForward | Point mutagenesis to remove restriction enzyme site | 5'-CAA GAA GCT TGA CTT GTC TAC GAA CTC TTC CAT TTC-3' |
| Arp2mutReverse | Point mutagenesis to remove restriction enzyme site | 5'-GAA ATG GAA GAA CTC GTA GAC AAG TCA AGC TCC TTG-3' |
| Exp2MutForward | Point mutagenesis to remove restriction enzyme site | 5'-CCT ACA CAC GGA ACT CCT GGA AAT TAA GAC CG-3' |
| Exp2mutReverse | Point mutagenesis to remove restriction enzyme site | 5'-CGG TCT TAA TTT CCA GGA ACT CCG TGT GTA GG-3' |
| Bam.Gal4.Fwd | Attaching BamH1 sites to Gal4 in preparation for yeast vector ligation | 5'-AAA AGG ATC CAA AAA AAT GAA GCT ACT GTC TTC TAT C-3' |
| HmRXR.Hind.Rev | Attaching HindIII sites to HmRXR in preparation for yeast vector ligation | 5'-AAT TAA GCT TGG ATC CCT CGA GCT GCA GC-3' |
| LcRXR.Hind.Rev | Attaching HindIII sites to LcRXR in preparation for yeast vector ligation | 5'-GAT GAA GCT TGG ATC CCT CGA GCT GC-3' |
| VP16.Hind.Fwd | Attaching HindIII sites to VP16 in preparation for yeast vector ligation | 5'-AAA AAA GCT TAA AAA AAT GGC TAG CGC CGC CAC CAT G-3' |
| VP16.Rev | Reverse primer for VP16.Hind.Fwd PCR - no modification | 5'-CTG CAG CGG CCG CTA CTA GTG CCC TTG GAA TTG ACG AGT AC-3' |
| NLS.serine.BB.Fwd | Oligo to anneal NLS, with one bp difference to avoid self-complementarity | 5'-/5Phos/CTA GAA GTT CTG TTG TTC ATC CTA AGA AGA AGA GAA AGG TTT GAA CTA GTA GCG GCC GCT GCA-3' |
| NLS.serine.BB.Rev | Complement to NLS.serine.BB.Fwd. for NLS annealing | 5'-/5Phos/GCG GCC GCT ACT AGT TCA AAC CTT TCT CTT CTT CTT AGG ATG AAC AAC AGA ACT T-3' |
| Fwd.BB.AtEXP2 | for amplification/extraction of an arabidopsis germination-specific promoter | 5'-CCT TTC TAG ACC TGG TGT GGT TTC TTT G-3' |
| Rev.BB.AtEXP2 | for amplification/extraction of an arabidopsis germination-specific promoter | 5'-AAG GCT GCA GCG GCC GCT ACT GAT ATT TGC TGA TGG GCT AAA G-3' |
| Fwd.BB.AtARP2 | for amplification/extraction of the arabidopsis ARP2 promoter | 5'-CCT TTC TAG ACC ATC ACA TAT TTG TAG AAT G-3' |
| Rev.BB.AtARP2 | for amplification/extraction of the arabidopsis ARP2 promoter | 5'-AAG GCT GCA GCG GCC GCT ACT AGT CTT CTC CGA TTT CTA TAG AG-3' |
| barnase2.BB.fwd | addition of biobrick sites/ends to barnase gene | 5'-CCT TTC TAG AAT GGT ACC GGT TAT CAA CAC G-3' |
| BB.Barnase.Rev | addition of biobrick sites/ends to barnase gene | 5'-AAG GCT GCA GCG GCC GCT ACT AGT TTA TCT GAT TTT TGT AAA GGT CTG-3' |
| NLS.BB.Fwd | addition of biobrick sites/ends to the arabidopsis NLS | 5'-/5Phos/CTA GAT CTT CTG TTG TTC ATC CAT AGA AGA AGA GAA AGG TTT GAA CTA GTA GCG GCC GCT GCA-3' |
| NLS.BB.Rev | addition of biobrick sites/ends to the arabidopsis NLS | 5'/5Phos/GCG GCC GCT ACT AGT TCA AAC CTT TCT CTT CTT CTT AGG ATG AAC AAC AGA AGA-3' |
| LacIn.BB.Fwd | Addition of NLS to the Lac Inhibitor gene to produce LacIN | 5'-CCT TGA ATT CGC GGC CGC ATC TAG AAT GAA AC AGT AAC GTT ATA CGA TGT C-3' |
| LacIN.BB.Rev | Addition of NLS to the Lac Inhibitor gene to produce LacIN | 5' AAG GTC GCA GCA GCG GCC GCT ACT AGT TCA AAC CTT TCT CTT CTT CTT AGG ATG AAC AAC AGA AGA CTG CCC GCT TTC CAG TCG GGA AA-3' |
| BB.Gal4DBD.Fwd | Adding biobrick sites to Gal4 DNA Binding Domain | 5'-CTT TTC TAG AAT GAA GCT ACT GTC TTC TAT C-3' |
| BB.Gal4DBD.Rev | Adding biobrick sites to Gal4 DNA Binding Domain | 5'-AAG GCT GCA GCG GCC GCT ACT AGT CGA TAC AGT CAA CTG TCT TTG-3' |
| BB.Barstar.Fwd | addition of biobrick sites/ends to barstar gene | 5'CCT TTC TAG AAT GAA AAA AGC AGT CAT TAA C-3' |
| BB.Barstar.Rev | addition of biobrick sites/ends to barstar gene | 5'-AAG GCT GCA GCG GCC GCT ACT AGT TTA AGA AAG TATA GAT GGT GAT GTC-3' |
