Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V001184 | pLenti CMV/TO SV40 small + Large T (w612-1) | 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:
- pLenti CMV/TO SV40 small + Large T (w612-1)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9388 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- RSV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMVforw
pLenti CMV/TO SV40 small + Large T (w612-1) 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.
pLenti CMV/TO SV40 small + Large T (w612-1) vector Sequence
LOCUS V001184 9388 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V001184
VERSION V001184
KEYWORDS pLenti CMV/TO SV40 small + Large T (w612-1)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9388)
TITLE shMDC1 plasmids
REFERENCE 2 (bases 1 to 9388)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 9388)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9388
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 21..324
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 325..528
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 530..548
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 551..569
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 610..634
/label="attB1"
/note="recombination site for the Gateway(R) BP reaction"
CDS 741..1262
/label="small t antigen"
/note="SV40 (simian virus 40) small t antigen"
CDS 1462..1482
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
protein_bind complement(3277..3301)
/label="attB2"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 3332..3920
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4007..4240
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4312..4446
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 4452..4587
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(4608..4626)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4636..4652)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4794..5249
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5276..5380
/label="AmpR promoter"
CDS 5381..6238
/label="AmpR"
/note="beta-lactamase"
rep_origin 6412..7000
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7154..7171
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 7288..7309
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7324..7354
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 7362..7378
/label="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)."
primer_bind 7386..7402
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 7423..7441
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 7469..7695
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 7696..7876
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 7923..8048
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8541..8774
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 8959..9003
/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 9152..9193
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 9270..9386
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"