Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V005501 | pLOX-TERT-iresTK | 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:
- pLOX-TERT-iresTK
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13332 bp
- Type:
- Mammalian Expression, Lentiviral, Cre/Lox
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- SP6
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV Fwd for hTert
- 3' Primer:
- SP6 primer rev for LoxP
pLOX-TERT-iresTK 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.
pLOX-TERT-iresTK vector Sequence
LOCUS V005501 13332 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V005501
VERSION V005501
KEYWORDS pLOX-TERT-iresTK
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13332)
AUTHORS Salmon P, Oberholzer J, Occhiodoro T, Morel P, Lou J, Trono D
TITLE Reversible immortalization of human primary cells by
lentivector-mediated transfer of specific genes.
JOURNAL Mol Ther. 2000 Oct . 2(4):404-14.
PUBMED 11020357
REFERENCE 2 (bases 1 to 13332)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 13332)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther.
2000 Oct . 2(4):404-14."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13332
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 92..725
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 772..897
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1390..1623
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1808..1852
/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 2001..2042
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
enhancer 2131..2434
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2435..2638
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2635..2659
/label="LNCX"
/note="Human CMV promoter, forward primer"
CDS 2782..6177
/label="hTERT"
/note="human telomerase reverse transcriptase, the
catalytic subunit of telomerase"
misc_feature 6233..6694
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 6699..7826
/label="HSV TK"
/note="herpes simplex virus thymidine kinase"
protein_bind complement(7985..8018)
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
LTR 8038..8218
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(8243..8261)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(8551..8570)
/label="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(8784..8803)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 8903..8925
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind complement(8963..8981)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 9049..9153
/label="AmpR promoter"
CDS 9154..10011
/label="AmpR"
/note="beta-lactamase"
rep_origin 10185..10773
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10927..10944
/label="L4440"
/note="L4440 vector, forward primer"
promoter 11019..11348
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 12482..12547
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 12677..12697
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 13122..13256
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"