Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012737 | TetO-FUW-OSKM | 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
Lentiviral plasmid for tet-inducible expression of mouse Oct4, Sox2, Klf4 and Myc for iPS cell generation
- Vector Name:
- TetO-FUW-OSKM
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13457 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Source/Author:
- Rudolf Jaenisch
- Selection Marker:
- Zeocin
- Copy Number:
- High copy number
- Promoter:
- EM7
- Expression Method:
- Constiutive, Stable / Transient
TetO-FUW-OSKM 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.
TetO-FUW-OSKM vector Sequence
LOCUS V012737 13457 bp DNA circular SYN 11-SEP-2021
DEFINITION Exported.
ACCESSION V012737
VERSION V012737
KEYWORDS TetO-FUW-OSKM
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13457)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 13457)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13457
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 74..453
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 454..656
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 671..851
/label="5' LTR (truncated)"
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 898..1023
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1516..1749
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1934..1978
/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 2127..2168
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2276..2393
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 2446..2716
/label="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 2749..2786
/note="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 2889..3944
/label="mPou5f1"
/note="Mus musculus Oct-4 gene. Encodes a transcription
factor containing a POU homeodomain that plays a key role
in embryonic development and stem cell pluripotency."
CDS 3960..4016
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label="2A peptide from porcine teschovirus-1 polyprotein"
/note="P2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 4023..4979
/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 4995..5048
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label="2A peptide from Thosea asigna virus capsid protein"
/note="T2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 5055..6503
/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 6519..6578
/codon_start=1
/product="2A peptide from equine rhinitis A virus
polyprotein"
/label="2A peptide from equine rhinitis A virus polypro"
/note="E2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="QCTNYALLKLAGDVESNPGP"
CDS 6588..7949
/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 7985..8573
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(8576..8592)
/label="KS primer"
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
LTR 9098..9278
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 9310..9534
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 9580..10000
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 10022..10351
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 10399..10446
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 10465..10836
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 10969..11102
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(11139..11155)
/label="M13 rev"
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 11163..11179
/label="lac repressor encoded by lacI binding site"
/bound_moiety="lac repressor encoded by lacI"
/note="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(11187..11217)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(11232..11253)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(11541..12129)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(12303..13160)
/label="AmpR"
/note="beta-lactamase"
promoter complement(13161..13265)
/label="AmpR promoter"