IGEM:Cambridge/2008/Turing Pattern Formation/Vectors: Difference between revisions
No edit summary |
|||
Line 24: | Line 24: | ||
! Vector Map | ! Vector Map | ||
! Vector Sequence | ! Vector Sequence | ||
! Possible Uses | |||
! Necessity (1-5 stars) | |||
! Checked on Gel | |||
|- | |- | ||
| ECE112 | | ECE112 | ||
Line 36: | Line 39: | ||
| map | | map | ||
| [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=1185558 U46196] | | [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=1185558 U46196] | ||
| | |||
* Quantitative testing promoters using Beta-galactosidase assay | |||
* Biobricking: Has multiple pstI sites, Xba is not in cassette | |||
| <nowiki>****</nowiki> | |||
| bgcolor="green" | Yes | |||
|- | |- | ||
| ECE171 | | ECE171 | ||
Line 46: | Line 54: | ||
| map | | map | ||
| [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=39726213 AY464559] | | [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=39726213 AY464559] | ||
| | |||
* Inserting genes at neutral site | |||
* Can be used for biobrick insertion, all sites within cassette, PstI site is not at MCS | |||
| <nowiki>****</nowiki> | |||
| bgcolor="green" | Yes | |||
|- | |- | ||
| ECE172 | | ECE172 | ||
Line 56: | Line 69: | ||
| map | | map | ||
| [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=39726216 AY464560] | | [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=39726216 AY464560] | ||
| | |||
* Inserting genes at neutral site | |||
* 4 pstI sites, EcoRI site, no Xba site | |||
| <nowiki>**</nowiki>: can use PyrD & AmyE instead | |||
| bgcolor="green" | No: did not appear on gel | |||
|- | |- | ||
| ECE176 | | ECE176 |
Revision as of 06:49, 6 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 | map | U46196 |
|
**** | Yes |
ECE171 | pPyr-Kan |
|
pyrD | map | AY464559 |
|
**** | Yes |
ECE172 | pGlt-Cm |
|
gltA | map | AY464560 |
|
**: can use PyrD & AmyE instead | No: did not appear on gel |
ECE176 | pBCJ164 |
|
rspD | map | BGSC sequence | |||
ECE153 | pSG1154 |
|
amyE | map | not available | |||
ECE162 | pSG1193 |
|
amyE | map | not available |
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 |
---|---|---|---|---|
ECE149 | pMUTIN-GFP+ |
|
map | From Bayreuth |
ECE150 | pMUTIN-CFP |
|
map | From Bayreuth (sequence incomplete) |
ECE151 | pMUTIN-YFP |
|
map | From Bayreuth |
ECE147 | pMUTIN-cMyc |
|
map | From Bayreuth |
Shuttle Vectors
BGSC Accession | Vector Name | Features | Vector Map | Vector Sequence |
---|---|---|---|---|
ECE165 | pAD123 |
|
map | BGSC sequence |
ECE166 | pAD43-25 |
|
map | BGSC sequence |
ECE188 | pHCMC02 |
|
map | From Bayreuth |
ECE189 | pHCMC04 |
|
map | From Bayreuth |
ECE190 | pHCMC05 |
|
map | From Bayreuth |
Plasmid Gels
- We ran these plasmids on gels, using double and single digests, to check them