Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V009174 | pETcon(-) | 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
The pETcon(-) is a Yeast Surface Display Plasmid. Designed by Andrew Scharenberg Lab.
- Vector Name:
- pETcon(-)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6173 bp
- Type:
- Yeast Expression
- Replication origin:
- ori
- Host:
- Yeast
- Selection Marker:
- TRP1
- Copy Number:
- High Copy
- Promoter:
- GAL1,10
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- GAL1
- 3' Primer:
- T7
- Growth Strain(s):
- Top10
pETcon(-) 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
- Özçelik CE, Beğli Ö, Hınçer A, Ahan RE, Kesici MS, Oğuz O, Kasırga TS, Özçubukçu S, Şeker UÖŞ. Synergistic Screening of Peptide-Based Biotechnological Drug Candidates for Neurodegenerative Diseases Using Yeast Display and Phage Display. ACS Chem Neurosci. 2023 Oct 4;14(19):3609-3621. doi: 10.1021/acschemneuro.3c00248. Epub 2023 Aug 28. PMID: 37638647; PMCID: PMC10557061.
pETcon(-) vector Sequence
LOCUS V009174 6173 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V009174
VERSION V009174
KEYWORDS pETcon(-)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6173)
TITLE pETCON Yeast Surface Display Plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6173)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6173)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6173
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 2..18
/label="lac operator"
/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)."
primer_bind 26..42
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 63..81
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 111..763
/label="GAL1,10 promoter"
/note="divergent inducible promoter, regulated by Gal4"
CDS 803..1063
/gene="AGA2"
/label="A-agglutinin-binding subunit"
/note="A-agglutinin-binding subunit from Saccharomyces
cerevisiae (strain ATCC 204508 / S288c). Accession#:
P32781"
CDS 1082..1093
/label="Factor Xa site"
/note="Factor Xa recognition and cleavage site"
CDS 1094..1120
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 1236..1265
/label="Myc"
/note="Myc (human c-Myc proto-oncogene) epitope tag"
promoter complement(1595..1613)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1620..1636)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 1777..2232
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(2338..3009)
/label="TRP1"
/note="phosphoribosylanthranilate isomerase, required for
tryptophan biosynthesis"
promoter complement(3010..3291)
/label="TRP1 promoter"
primer_bind complement(3311..3330)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3430..3452
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind complement(3490..3508)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_feature complement(3549..4052)
/label="CEN/ARS"
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 4089..4193
/label="AmpR promoter"
CDS 4194..5051
/label="AmpR"
/note="beta-lactamase"
rep_origin 5225..5813
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5967..5984
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 6101..6122
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6137..6167
/label="lac promoter"
/note="promoter for the E. coli lac operon"