Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000475 | OKSIM | 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:
- OKSIM
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12417 bp
- Type:
- Mammalian Expression, Lentiviral, Cre/Lox
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- EF-1α
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- EF-1a Forward
OKSIM 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.
OKSIM vector Sequence
LOCUS V000475 12417 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000475
VERSION V000475
KEYWORDS OKSIM
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12417)
AUTHORS Ross PJ, Suhr S, Rodriguez RM, Chang EA, Wang K, Siripattarapravat
K, Ko T, Cibelli JB
TITLE Human Induced Pluripotent Stem Cells Produced Under Xeno-Free
Conditions.
JOURNAL Stem Cells Dev. 2009 Dec 23. ():.
PUBMED 20030562
REFERENCE 2 (bases 1 to 12417)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 12417)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cells
Dev. 2009 Dec 23. ():."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12417
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 13..563
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 608..1924
/label="c-Myc"
/note="human c-Myc proto-oncogene"
primer_bind complement(1978..1994)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 2008..2596
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
protein_bind complement(2740..2773)
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
LTR 2793..2973
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 3002..3226
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
regulatory 3269..3278
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter 3322..3426
/label="AmpR promoter"
CDS 3427..4284
/label="AmpR"
/note="beta-lactamase"
rep_origin 4458..5046
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LTR 5205..5838
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 5885..6010
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6503..6736
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 6921..6965
/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 7114..7155
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 7252..7369
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 7498..8676
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
promoter 8692..8710
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
CDS 8736..9815
/label="hOct4"
/note="Homo sapiens Oct-4 gene. Encodes a transcription
factor containing a POU homeodomain that plays a key role
in embryonic development and stem cell pluripotency.
Aberrant expression in adult tissues is associated with
tumorigenesis."
CDS 9840..9905
/codon_start=1
/product="2A peptide from foot-and-mouth disease virus
polyprotein"
/label="F2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="VKQTLNFDLLKLAGDVESNPGP"
misc_feature 10000..10576
/label="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 11352..11417
/codon_start=1
/product="2A peptide from foot-and-mouth disease virus
polyprotein"
/label="F2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="VKQTLNFDLLKLAGDVESNPGP"
CDS 11430..12380
/label="hSOX2"
/note="Homo sapiens 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"