Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V003385 | pSET2 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pSET1, pSET2, and pSET3 carry cat, spc, and both of these genes, respectively, as selectable markers. These vectors could be introduced into S. suis, E. coli, Salmonella typhimurium, S. pneumoniae, and S. equi ssp. equi by electrotransformation.
- Vector Name:
- pSET2
- Antibiotic Resistance:
- Spectinomycin
- Length:
- 5016 bp
- Type:
- Shuttle vector
- Source/Author:
- Takamatsu D, Osaki M, Sekizaki T.
- Promoter:
- lac
- Growth Temperature:
- 37℃
pSET2 vector Vector Map
Plasmid Protocol
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5. Store the plasmid at -20 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
References
- Zheng C, Wei M, Qiu J, Li J. A Markerless Gene Deletion System in Streptococcus suis by Using the Copper-Inducible Vibrio parahaemolyticus YoeB Toxin as a Counterselectable Marker. Microorganisms. 2021 May 19;9(5):1095.
pSET2 vector Sequence
LOCUS Exported 5016 bp DNA circular SYN 12-MAR-2024
DEFINITION Shuttle vector pSET2 DNA, complete sequence.
ACCESSION AB042430
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5016)
AUTHORS Takamatsu D, Osaki M, Sekizaki T.
TITLE Construction and characterization of Streptococcus suis-Escherichia
coli shuttle cloning vectors
JOURNAL Plasmid 45 (2), 101-113 (2001)
PUBMED 11322824
REFERENCE 2 (bases 1 to 5016)
AUTHORS Takamatsu D, Osaki M, Sekizaki T.
TITLE Direct Submission
JOURNAL Submitted (08-MAY-2000) Daisuke Takamatsu, National Institute of
Animal Health, Laboratory of Molecular Bacteriology; Kannondai
3-1-1, Tukuba, Ibaraki 305-0856, Japan
(E-mail:p1013dt@niah.affrc.go.jp, Tel:81-298-38-7743,
Fax:81-298-38-7743)
REFERENCE 3 (bases 1 to 5016)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid";
date: "2001"; volume: "45"; issue: "2"; pages: "101-113"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(08-MAY-2000) Daisuke Takamatsu, National Institute of Animal
Health, Laboratory of Molecular Bacteriology"; pages:
"81-298-38-7743"
FEATURES Location/Qualifiers
source 1..5016
/lab_host="Streptococcus suis, Escherichia coli"
/mol_type="other DNA"
/label=synonym:Shuttle Cloning Vector pSET2
/note="synonym:Shuttle Cloning Vector pSET2"
/db_xref="taxon:125568"
/organism="Shuttle vector pSET2"
gene 156..176
/gene="dso"
/label=dso
misc_feature 156..176
/gene="dso"
/label=double-stranded replication origin
/note="double-stranded replication origin"
gene 367..504
/gene="copG"
/label=copG
CDS 367..504
/codon_start=1
/gene="copG"
/product="CopG, transcriptional repressor protein"
/function="transcriptional repressor protein"
/label=copG
/protein_id="BAB03239.1"
/translation="MKKRLTITLSESVLENLEKMAKEMGLSKSALISVALENYKKGQVK
"
gene 566..1246
/gene="repB"
/label=repB
CDS 566..1246
/codon_start=1
/gene="repB"
/product="RepB, replication initiation¨Ctermination
protein."
/function="The plasmid-encoded initiator of replication.The
protein introduces a strand- and site-specific nick in the
double-stranded replication origin, on supercoiled DNA."
/label=repB
/protein_id="BAB03240.1"
/translation="MAKEKARYFTFLLYPESIPSDWELKLELLGVPIAVSPLHDRDKSD
VEGQQYKKPHYHVIYVSKNPVTADSVRMKIKRSLGDNSVALVQIIRTSIENTYLYLTHE
SKDAIEKKKHVYDKADITLLSNFDIDRYITLDVEEKDDMLNEVCDLIDEYDIANMRELR
RFIKLHGAEHGLPSIKIINSVLRSHTGLIRLYFDAVYQERKYGSNIPNVNQETGEILSE
DSDD"
gene complement(1703..2470)
/gene="add9"
/label=add9
CDS complement(1703..2470)
/codon_start=1
/gene="add9"
/product="spectinomycin adenyltransferase, aso known as
aad9."
/function="spectinomycin resistance"
/label=spc
/protein_id="BAB03241.1"
/translation="MRRIYLNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNS
DLDFLVVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQ
EFIYGEWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVR
RAIMDSSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERI
LLAVRSYLGENIEWTNENVNLTINYLNNRLKKL"
gene complement(2872..3195)
/gene="lacZ'"
/label=lacZ'
CDS complement(2872..3195)
/codon_start=1
/gene="lacZ'"
/product="beta-galactosidase alpha fragment"
/label=lacZ'
/protein_id="BAB03242.1"
/translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
DAA"
primer_bind 3105..3121
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 3122..3178
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(3191..3207)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3215..3231
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(3239..3269)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3284..3305
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
gene 4111..4252
/gene="ssoA"
/label=ssoA
misc_feature 4111..4252
/gene="ssoA"
/label=single-stranded replication origin
/note="single-stranded replication origin"