IGEM:Cambridge/2008/Turing Pattern Formation/Vectors: Difference between revisions
Line 149: | Line 149: | ||
* IPTG-inducible | * IPTG-inducible | ||
* full plasmid insertion | * full plasmid insertion | ||
| | | [[image:ECE149.jpg|thumb|ECE149 Map]] | ||
| [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pmutingfp.htm From Bayreuth] | | [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pmutingfp.htm From Bayreuth] | ||
| bgcolor="red" fontcolor="white"| No: no good bands on the gel | | bgcolor="red" fontcolor="white"| No: no good bands on the gel | ||
Line 161: | Line 161: | ||
* IPTG-inducible | * IPTG-inducible | ||
* full plasmid insertion | * full plasmid insertion | ||
| | | [[image:ECE153.jpg|thumb|ECE150 Map]] | ||
| [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pmutincfp.htm From Bayreuth] <br/> (sequence incomplete) | | [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pmutincfp.htm From Bayreuth] <br/> (sequence incomplete) | ||
| bgcolor="yellow" fontcolor="white"| Inconclusive | | bgcolor="yellow" fontcolor="white"| Inconclusive | ||
Line 173: | Line 173: | ||
* IPTG-inducible | * IPTG-inducible | ||
* full plasmid insertion | * full plasmid insertion | ||
| | | [[image:ECE151.jpg|thumb|ECE151 Map]] | ||
| [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pmutinyfp.htm From Bayreuth] | | [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pmutinyfp.htm From Bayreuth] | ||
| bgcolor="green" fontcolor="white"| Yes | | bgcolor="green" fontcolor="white"| Yes | ||
Line 184: | Line 184: | ||
* cMyc antibody tag fusion | * cMyc antibody tag fusion | ||
* IPTG-inducible Pspac promoter | * IPTG-inducible Pspac promoter | ||
| | | [[image:ECE147.jpg|thumb|ECE147 Map]] | ||
| [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pmutincmyc.htm From Bayreuth] | | [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pmutincmyc.htm From Bayreuth] | ||
| bgcolor="yellow" fontcolor="white"| Inconclusive | | bgcolor="yellow" fontcolor="white"| Inconclusive |
Revision as of 15:41, 10 August 2008
All these vectors were kindly provided by Dr. Dan Ziegler, Director of the Bacillus Genetic Stock Center at Ohio State University.
Daniel Goodman 17:01, 31 July 2008 (UTC):I just put this table together, but I have yet to add the plasmid maps to these. For now, to see the maps, go to BGSC homepage and find them in the catalog. They are also in the black Turing Group binder under "ordered vectors and strains".
Integration Vectors
These vectors integrate into the chromosome and do not have a replication origin in Bacillus. They either integrate cassettes that require double crossovers and two homologous regions, or whole-plasmid insertions that only require one crossover and one corresponding region of homology.
Ectopic Integration
The integration site of these is pre-determined
BGSC Accession | Vector Name | Features | Integration Locus | Vector Map | Vector Sequence | Possible Uses | Necessity (1-5 stars) | Checked on Gel |
---|---|---|---|---|---|---|---|---|
ECE112 | pDG1661 |
|
amyE | U46196 |
|
**** | Yes | |
ECE171 | pPyr-Kan |
|
pyrD | AY464559 |
|
**** | Yes | |
ECE172 | pGlt-Cm |
|
gltA | AY464560 |
|
*: can use PyrD & AmyE instead | No: did not appear on gel | |
ECE176 | pBCJ164 |
|
rspD | BGSC sequence |
|
*: use of strong rpsD promoter, but hard to insert into | No: unsure of enzymes/too faint | |
ECE153 | pSG1154 |
|
amyE | not available |
|
***: inducible promoter, strong gfp, no sequence | inconclusive | |
ECE162 | pSG1193 |
|
amyE | not available |
|
***: inducible promoter, strong gfp, no sequence | inconclusive |
Insert Integration
The user inserts a homologous piece of the Bacillus chromosome and the vector integrates there.
BGSC Accession | Vector Name | Features | Vector Map | Vector Sequence | Checked on gel |
---|---|---|---|---|---|
ECE149 | pMUTIN-GFP+ |
|
From Bayreuth | No: no good bands on the gel | |
ECE150 | pMUTIN-CFP |
|
From Bayreuth (sequence incomplete) |
Inconclusive | |
ECE151 | pMUTIN-YFP |
|
From Bayreuth | Yes | |
ECE147 | pMUTIN-cMyc |
|
From Bayreuth | Inconclusive |
Shuttle Vectors
BGSC Accession | Vector Name | Features | Vector Map | Vector Sequence | Checked on gel |
---|---|---|---|---|---|
ECE165 | pAD123 |
|
map | BGSC sequence | Yes |
ECE166 | pAD43-25 |
|
map | BGSC sequence | Yes |
ECE188 | pHCMC02 |
|
map | From Bayreuth | Not yet checked, DNA stocks |
ECE189 | pHCMC04 |
|
map | From Bayreuth | Not yet checked, DNA stocks |
ECE190 | pHCMC05 |
|
map | From Bayreuth | Not yet checked, DNA stocks |
Plasmid Gels
- We ran these plasmids on gels, using double and single digests, to check them