Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010277 | pGADT7-Rec | 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:
- pGADT7-Rec
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8058 bp
- Type:
- Yeast Plasmids
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Clontech
- Copy Number:
- High copy number
- Promoter:
- ADH1(long)
pGADT7-Rec 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.
pGADT7-Rec vector Sequence
LOCUS pGADT7-Rec. 8058 bp DNA circular SYN 01-JAN-1980
DEFINITION High-copy yeast vector for fusing a cDNA to the GAL4 activation
domain by in vivo recombination.
ACCESSION .
VERSION .
KEYWORDS pGADT7-Rec
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8058)
AUTHORS Clontech
TITLE Direct Submission
REFERENCE 2 (bases 1 to 8058)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT Linearize with SmaI.
FEATURES Location/Qualifiers
source 1..8058
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 771..1475
/label=ADH1 promoter
/note="promoter for alcohol dehydrogenase 1"
CDS 1491..1493
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 1521..1541
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 1557..1898
/label=GAL4 activation domain
/note="activation domain of the GAL4 transcriptional
activator"
promoter 1904..1922
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1935..1937
/codon_start=1
/product="in vitro start codon"
/label=in vitro start codon
/note="ATG"
/translation="M"
CDS 1941..1967
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
misc_feature 2000..2035
/label=SMART III Oligonucleotide Sequence
/note="SMART III"
/note="matches the SMART III Oligonucleotide for cDNA
synthesis"
misc_feature 2046..2070
/label=CDS III Sequence
/note="CDS III"
/note="matches the CDS III Primer for cDNA synthesis"
terminator 2486..2673
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
CDS complement(2793..3884)
/label=LEU2
/note="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
promoter complement(3885..4290)
/label=LEU2 promoter
primer_bind complement(4332..4348)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4356..4372)
/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)."
promoter complement(4380..4410)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4425..4446)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
protein_bind 4501..4534
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
rep_origin complement(4885..5473)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5647..6504)
/label=AmpR
/note="beta-lactamase"
promoter complement(6505..6609)
/label=AmpR promoter
rep_origin 6891..8055
/label=2u ori
/note="yeast 2u plasmid origin of replication"