Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V016805 | pHAGE2-TetOminiCMV-SKM | 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:
- pHAGE2-TetOminiCMV-SKM
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10186 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- Tet
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pHAGE2-TetOminiCMV-SKM 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.
pHAGE2-TetOminiCMV-SKM vector Sequence
LOCUS V016805 10186 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V016805
VERSION V016805
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10186)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..10186
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 173..1129
/label="mSox2"
/note="Mus musculus transcription factor SOX-2 gene.
Belongs to the SRY-related HMG-box (SOX) family of
transcription factors, which is involved in the regulation
of embryonic development and in the determination of cell
fate"
CDS 1205..2653
/label="mKlf4"
/note="Mouse Kruppel-like factor (Klf4) gene. Belongs to
the relatively large family of SP1-like transcription
factors and is involved in the regulation of proliferation,
differentiation, apoptosis and somatic cell reprogramming."
CDS 2738..4099
/label="mMyc"
/note="Mus musculus myc proto-oncogene. Belongs to
myelocytomatosis (Myc) family of transcription factors.
Plays a role in cell cycle procession, apotosis and
cellular transformation"
misc_feature 4116..4704
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4852..5032
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter 5165..5269
/label="AmpR promoter"
CDS 5270..6127
/label="AmpR"
/note="beta-lactamase"
rep_origin 6301..6889
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 7300..7321
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7336..7366
/label="lac promoter"
/note="promoter for the E. coli lac operon"
LTR 7675..8308
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 8355..8480
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8977..9210
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 9395..9439
/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 9588..9629
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 9732..9849
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 9925..9943
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 10009..10027
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"