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Team:Harvard/fences/parts
From 2010.igem.org
(Difference between revisions)
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<h2>Parts</h2> | <h2>Parts</h2> | ||
<table cellspacing="0"> | <table cellspacing="0"> | ||
| - | <tr><td><b>Name</b></td><td><b>Description</b></td><td><b>Resistance</b></td><td><b>Sequence</b></td></tr> | + | <tr><td><b>Name</b></td><td><b>Biobrick Name</b></td><td><b>Description</b></td><td><b>Resistance</b></td><td><b>Sequence</b></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>Barnase</td><td><center>-</center></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><center>-</center></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">link</a></td></tr> | + | <tr><td>LacIN</td><td> BBa_K382035 </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</td><td> Part of the Locust Retinoic Acid Receptor and VP16 activation domain fusion protein. RXRLc binds EcR 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</td><td> Part of the Locust Retinoic Acid Receptor and VP16 activation domain fusion protein. RXRLc binds EcR in presence of Methoxyfenozide</td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f4">link</a></td></tr> | ||
<tr><td>RXRHm</td><td>Human Retinoic Acid Receptor performs a function analogous to RXRLc. RXRHm has been shown to be sensitive to lower level of methoxyfenozide, but also to have higher background activation in the absence of methoxyfenozide.</td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f5">link</a></td></tr> | <tr><td>RXRHm</td><td>Human Retinoic Acid Receptor performs a function analogous to RXRLc. RXRHm has been shown to be sensitive to lower level of methoxyfenozide, but also to have higher background activation in the absence of methoxyfenozide.</td><td>chloramphenicol</td><td><a href="https://2010.igem.org/Team:Harvard/f5">link</a></td></tr> | ||
Revision as of 01:07, 27 October 2010
parts and primers
Parts
| Name | Biobrick Name | Description | Resistance | Sequence |
| Barnase | Fatal gene for the genetic fence | chloramphenicol | link | |
| Barstar | Protein inhibitor of Barnase | chloramphenicol | link | |
| LacIN | BBa_K382035 | Lac Inhibitor with Nuclear Localization sequence - transcriptional inhibitor of the Act2LacO promoter | chloramphenicol | link |
| RXRLc | Part of the Locust Retinoic Acid Receptor and VP16 activation domain fusion protein. RXRLc binds EcR in presence of Methoxyfenozide | chloramphenicol | link | |
| RXRHm | Human Retinoic Acid Receptor performs a function analogous to RXRLc. RXRHm has been shown to be sensitive to lower level of methoxyfenozide, but also to have higher background activation in the absence of methoxyfenozide. | chloramphenicol | link | |
| VP16 | Part of the Retinoic Acid Receptor and VP16 activation domain fusion protein. VP16 recruits transcription factors, and when colocalized with the Gal4 Upstream Activating Sequence initiates transcription downstream of the 5xGal435s promoter. | chloramphenicol | link | |
| Ecdysone Receptor | Part of the fusion protein of Ecdysone receptor and Gal4 DNA binding domain. Ecdysone receptor binds RXR-VP16 in presence of Methoxyfenozide | chloramphenicol | link | |
| Gal4 DNA binding domain | Part of the Gal4-EcR fusion protein. Gal4 associates with the Gal4 upstream activating sequence, and when RXR-VP16 is bound, it localizes VP16 to the Gal4UAS35s promoter resulting in transcription downstream. | chloramphenicol | link |
Promoters
| Name | Description | Active conditions | Resistance | Sequence |
| pENT CUP | Constitutive Plant Promoter | Constitutive | chloramphenicol | link |
| 5xGal UAS | 5 time repeat of Gal4 upstream activating sequence | On in presense of dimerized EcR-Gal and RXRLc-VP16 | chloramphenicol | link |
| Act2lacO | Actin2 promoter with Lac Inhibitor responsive sites (Lac O sites | Constitutive, inhibited by LacIN | chloramphenicol | link |
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' |
