Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012885 | pLenti-Bi-cistronic | 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-Bi-cistronic
- Antibiotic Resistance:
- Kanamycin
- Length:
- 8292 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- mPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLenti-Bi-cistronic 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.
pLenti-Bi-cistronic vector Sequence
LOCUS V012885 8292 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V012885
VERSION V012885
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8292)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8292
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_signal 170..320
/label="Ad5 Psi"
/note="packaging signal for adenovirus serotype 5"
promoter 488..714
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 715..895
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 942..1067
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1557..1790
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1975..2019
/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 2168..2209
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2296..2413
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
polyA_signal complement(2441..2522)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter 2733..3232
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 3309..3335
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
promoter 3353..3650
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 3656..4252
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 4268..4855
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4865..5098
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5170..5291
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(5619..6207)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 7014..7805
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
rep_origin complement(7856..8235)
/direction=LEFT
/label="M13 ori"
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
repeat_region 8273..8292
/label="ITR"
/note="inverted terminal repeat of human adenovirus
serotype 5"