Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V016794 | pYD1-mSA | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pYD1-mSA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6490 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Yeast
- Selection Marker:
- TRP1
- Promoter:
- GAL1,10
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pYD1-mSA 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.
pYD1-mSA vector Sequence
LOCUS V016794 6490 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V016794
VERSION V016794
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6490)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6490
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 138..149
/label="Factor Xa site"
/note="Factor Xa recognition and cleavage site"
CDS 150..176
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 240..581
/label="mSA"
/note="monomeric streptavidin/rhizavidin hybrid (Lim et
al., 2013)"
CDS 603..626
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
promoter complement(957..975)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(982..998)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 1139..1594
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(1700..2371)
/label="TRP1"
/note="phosphoribosylanthranilate isomerase, required for
tryptophan biosynthesis"
promoter complement(2372..2653)
/label="TRP1 promoter"
misc_feature complement(2911..3414)
/label="CEN/ARS"
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 3451..3555
/label="AmpR promoter"
CDS 3556..4413
/label="AmpR"
/note="beta-lactamase"
rep_origin 4587..5175
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 5463..5484
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5499..5529
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 5537..5553
/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 5561..5577
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 5598..5616
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 5646..6308
/label="GAL1,10 promoter"
/note="divergent inducible promoter, regulated by Gal4"
CDS join(6349..6490,1..119)
/gene="AGA2"
/label="A-agglutinin-binding subunit"
/note="A-agglutinin-binding subunit from Saccharomyces
cerevisiae (strain ATCC 204508 / S288c). Accession#:
P32781"