Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V004395 | pLOX-Ttag-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
CMV promoter is upstream of the bicistronic coding cassette with SV40-Large T and HSV1-TK. During reverse transcription U3 region of the 3'LTR, with LoxP, is duplicated; LoxP sites end up flanking the genome of the integrated provirus. Upon expression of Cre, provirus is excised.
- Vector Name:
- pLOX-Ttag-iresTK
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12290 bp
- Type:
- Lentiviral vectors
- Replication origin:
- ori
- Copy Number:
- High copy number
- Promoter:
- CMV
- Cloning Method:
- Enzyme digestion and ligation
- 5' Primer:
- CMV Fwd
- 3' Primer:
- SP6 rev
pLOX-Ttag-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-Ttag-iresTK vector Sequence
LOCUS V004395 12290 bp DNA circular SYN 13-JAN-2022
DEFINITION Exported.
ACCESSION V004395
VERSION V004395
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12290)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 12290)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12290
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 4..637
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 684..809
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1302..1535
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1720..1764
/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 1913..1954
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
enhancer 2043..2346
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2347..2550
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 2731..3252
/label="small t antigen"
/note="SV40 (simian virus 40) small t antigen"
intron 3257..3322
/note="small t intron"
/note="simian virus 40 (SV40) small t antigen intron"
CDS 3452..3472
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
primer_bind complement(5256..5272)
/label="KS primer"
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 5321..5782
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
protein_bind complement(6854..6887)
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
LTR 6907..7087
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(7112..7130)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 7918..8022
/label="AmpR promoter"
CDS 8023..8880
/label="AmpR"
/note="beta-lactamase"
rep_origin 9054..9642
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 10021..10217
/label="SV40 promoter"
/note="SV40 early promoter"
intron 11351..11416
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 11546..11566
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 11991..12125
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter 12290
/label="SV40 promoter"
/note="SV40 early promoter"