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