IGEM:Cambridge/2008/Turing Pattern Formation/Vectors: Difference between revisions

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(New page: 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 ...)
 
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All these vectors were kindly provided by Dr. Dan Ziegler, Director of the Bacillus Genetic Stock Center at Ohio State University.
= 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.
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 ====
== Ectopic Integration ==
The integration site of these is pre-determined
The integration site of these is pre-determined


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==== Insert Integration ====
== Insert Integration ==
The user inserts a homologous piece of the Bacillus chromosome and the vector integrates there.
The user inserts a homologous piece of the Bacillus chromosome and the vector integrates there.


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=== Shuttle Vectors ===
= Shuttle Vectors =


{| class="wikitable" border="1"
{| class="wikitable" border="1"
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* gfpmut3a in front of MCS, strong Pupp promoter, gram-positive RBS
* gfpmut3a in front of MCS, strong Pupp promoter, gram-positive RBS
| map
| map
| [http://www.bgsc.org/sequences/pad43-25.htm BGSC sequence]
| [http://www.bgsc.org/sequences/pad43-25.htm BGSC sequence ]
|-
|-
| ECE188
| ECE188
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* weak constitutive PlepA promoter in front of MCS
* weak constitutive PlepA promoter in front of MCS
| map
| map
| [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pHCMC02.htm]
| [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pHCMC02.htm From Bayreuth]
|-
|-
| ECE189
| ECE189
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* strong xylose inducible PxylA promoter in front of MCS
* strong xylose inducible PxylA promoter in front of MCS
| map
| map
| [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pHCMC04.htm]
| [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pHCMC04.htm From Bayreuth]
|-
|-
| ECE190
| ECE190
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* strong xylose inducible PxylA promoter in front of MCS
* strong xylose inducible PxylA promoter in front of MCS
| map
| map
| [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pHCMC05.htm]
| [http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pHCMC05.htm From Bayreuth]
|}
|}

Revision as of 09:59, 31 July 2008

All these vectors were kindly provided by Dr. Dan Ziegler, Director of the Bacillus Genetic Stock Center at Ohio State University.

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
ECE112 pDG1661
  • Chloramphenicol selectable in EC and BS
  • Spectinomycin selectable only in EC
  • spoVG-lacZ fusion: contains spoVG RBS
  • insert must contain a promoter
  • cassette insertion
 amyE map U46196
ECE171 pPyr-Kan
  • Kanamycin selectable in EC or BS
  • Ampicillin selectable only in EC
  • cassette insertion
 pyrD map AY464559
ECE172 pGlt-Cm
  • Cloramphenicol selectable in EC or BS
  • Ampicillin selectable only in EC
  • cassette insertion
 gltA map AY464560
ECE176 pBCJ164
  • Cloramphenicol selectable in EC or BS
  • Ampicillin selectable only in EC
  • cassette insertion
  • contains strong rspD promoter and RBS
 rspD map BGSC sequence
ECE153 pSG1154
  • Spectinomycin selectable in EC or BS
  • Ampicillin selectable only in EC
  • cassette insertion
  • xylose-inducible promoter
  • high fluorescence GFP variant for protein fusions
 amyE map not available
ECE162 pSG1193
  • Spectinomycin selectable in EC or BS
  • Ampicillin selectable only in EC
  • YFP full coding sequence, including RBS and Pxyl
  • xylose-inducible
  • cassette insertion
 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+
  • Erythromycin selectable in EC or BS
  • Ampicillin selectable only in EC
  • GFP+ full coding sequence, including RBS and Pspac
  • IPTG-inducible
  • full plasmid insertion
 map From Bayreuth
ECE150 pMUTIN-CFP
  • Erythromycin selectable in EC or BS
  • Ampicillin selectable only in EC
  • CFP full coding sequence, including RBS and Pspac
  • IPTG-inducible
  • full plasmid insertion
 map From Bayreuth
ECE151 pMUTIN-YFP
  • Erythromycin selectable in EC or BS
  • Ampicillin selectable only in EC
  • YFP full coding sequence, including RBS and Pspac
  • IPTG-inducible
  • full plasmid insertion
 map From Bayreuth
ECE147 pMUTIN-cMyc
  • Erythromycin selectable in EC or BS
  • Ampicillin selectable only in EC
  • cMyc antibody tag fusion
  • IPTG-inducible Pspac promoter
 map From Bayreuth

Shuttle Vectors

BGSC Accession Vector Name Features Vector Map Vector Sequence
ECE165 pAD123
  • Chloramphenicol selectable in EC or BS
  • Ampicillin selectable only in EC
  • gfpmut3a in front of MCS, no promoter, but gram-positive RBS
 map BGSC sequence
ECE166 pAD43-25
  • Chloramphenicol selectable in EC or BS
  • Ampicillin selectable only in EC
  • gfpmut3a in front of MCS, strong Pupp promoter, gram-positive RBS
 map BGSC sequence
ECE188 pHCMC02
  • Chloramphenicol selectable in EC or BS
  • Ampicillin selectable only in EC
  • weak constitutive PlepA promoter in front of MCS
 map From Bayreuth
ECE189 pHCMC04
  • Chloramphenicol selectable in EC or BS
  • Ampicillin selectable only in EC
  • strong xylose inducible PxylA promoter in front of MCS
 map From Bayreuth
ECE190 pHCMC05
  • Chloramphenicol selectable in EC or BS
  • Ampicillin selectable only in EC
  • strong xylose inducible PxylA promoter in front of MCS
 map From Bayreuth
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