From 2008.igem.org

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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.

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	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	ECE112 Map	[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	****	Yes
ECE171	pPyr-Kan	Kanamycin selectable in EC or BS Ampicillin selectable only in EC cassette insertion	pyrD	ECE171 Map	[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	****	Yes
ECE172	pGlt-Cm	Cloramphenicol selectable in EC or BS Ampicillin selectable only in EC cassette insertion	gltA	ECE172 Map	[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	*: can use PyrD & AmyE instead	No: wrong size
ECE176	pBCJ164	Cloramphenicol selectable in EC or BS Ampicillin selectable only in EC cassette insertion contains strong rspD promoter and RBS	rspD	ECE176 Map	[http://www.bgsc.org/sequences/pBCJ164.htm BGSC sequence]	Inserting genes at neutral site Using strong rspD promoter 2 pstI sites, X and S before promoter, E after terminator	*: use of strong rpsD promoter, but hard to insert into	Yes
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	ECE153 Map	not available	GFP-fusion tracking of proteins (for agr components?) multiple Xbas, multiple pstIs	***: inducible promoter, strong gfp, no sequence	yes
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	ECE162 Map	not available	GFP-fusion tracking of proteins (for agr components?) multiple Xbas, multiple pstIs	***: inducible promoter, strong gfp, no sequence	inconclusive : only one band

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+	Erythromycin selectable in EC or BS Ampicillin selectable only in EC GFP+ full coding sequence, including RBS and Pspac IPTG-inducible full plasmid insertion	ECE149 Map	[http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pmutingfp.htm From Bayreuth]	No: 3 bands on the gel
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	ECE150 Map	[http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pmutincfp.htm From Bayreuth] (sequence incomplete)	yes
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	ECE151 Map	[http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pmutinyfp.htm From Bayreuth]	Yes
ECE147	pMUTIN-cMyc	Erythromycin selectable in EC or BS Ampicillin selectable only in EC cMyc antibody tag fusion IPTG-inducible Pspac promoter	ECE147 Map	[http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pmutincmyc.htm From Bayreuth]	yes

Shuttle Vectors

BGSC Accession	Vector Name	Features	Vector Map	Vector Sequence	Checked on gel
ECE165	pAD123	Chloramphenicol selectable in EC or BS Ampicillin selectable only in EC gfpmut3a in front of MCS, no promoter, but gram-positive RBS	ECE165 Map	[http://www.bgsc.org/sequences/pad123.htm BGSC sequence]	Yes
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	ECE166 Map	[http://www.bgsc.org/sequences/pad43-25.htm BGSC sequence ]	Yes
ECE188	pHCMC02	Chloramphenicol selectable in EC or BS Ampicillin selectable only in EC weak constitutive PlepA promoter in front of MCS	ECE188 Map	[http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pHCMC02.htm From Bayreuth]	Not yet checked, DNA stocks
ECE189	pHCMC04	Chloramphenicol selectable in EC or BS Ampicillin selectable only in EC strong xylose inducible PxylA promoter in front of MCS	ECE189 Map	[http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pHCMC04.htm From Bayreuth]	Not yet checked, DNA stocks
ECE190	pHCMC05	Chloramphenicol selectable in EC or BS Ampicillin selectable only in EC strong xylose inducible PxylA promoter in front of MCS	ECE190 Map	[http://www.genetik.uni-bayreuth.de/LSGenetik1/schumann_pHCMC05.htm From Bayreuth]	Yes

Plasmid Gels

We ran these plasmids on gels, using double and single digests, to check them

Team:Cambridge/Signaling/Vectors