Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006706 | 7TFP CDH1 reporter | 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:
- 7TFP CDH1 reporter
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9174 bp
- Type:
- Mammalian Expression, Lentiviral, Luciferase
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- High Copy
- Promoter:
- RSV
- Cloning Method:
- Restriction Enzyme
- 3' Primer:
- LucNRev
7TFP CDH1 reporter 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.
7TFP CDH1 reporter vector Sequence
LOCUS V006706 9174 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V006706
VERSION V006706
KEYWORDS 7TFP CDH1 reporter
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9174)
AUTHORS Pattabiraman DR, Bierie B, Kober KI, Thiru P, Krall JA, Zill C,
Reinhardt F, Tam WL, Weinberg RA
TITLE Activation of PKA leads to mesenchymal-to-epithelial transition and
loss of tumor-initiating ability.
JOURNAL Science. 2016 Mar 4;351(6277):aad3680. doi: 10.1126/science.aad3680.
PUBMED 26941323
REFERENCE 2 (bases 1 to 9174)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 9174)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science.";
date: "2016-03-4"; pages: "
10.1126/science.aad3680"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9174
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 325..356
/label="minP"
/note="minimal TATA-box promoter with low basal activity"
CDS 418..2067
/label="luciferase"
/note="firefly luciferase"
promoter 2094..2423
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 2433..3029
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 3099..3687
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 3774..4007
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4079..4213
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 4240..4375
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(4396..4414)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4424..4440)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4582..5037
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5063..5167
/label="AmpR promoter"
CDS 5168..6025
/label="AmpR"
/note="beta-lactamase"
rep_origin 6199..6787
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6941..6958
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 7075..7096
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7111..7141
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 7149..7165
/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 7173..7189
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 7210..7228
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 7256..7482
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 7483..7663
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 7710..7835
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8328..8561
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 8746..8790
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 8939..8980
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 9057..9174
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"