Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V014074 | pHR-SFFV-NO5-IRES-EGFP | 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:
- pHR-SFFV-NO5-IRES-EGFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13738 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- SFFV
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pHR-SFFV-NO5-IRES-EGFP 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.
pHR-SFFV-NO5-IRES-EGFP vector Sequence
LOCUS V014074 13738 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V014074
VERSION V014074
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13738)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..13738
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(1..17)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
LTR 108..341
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(366..384)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 1172..1276
/label="AmpR promoter"
CDS 1277..2134
/label="AmpR"
/note="beta-lactamase"
rep_origin 2308..2896
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 3142..3471
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 4605..4670
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 4800..4820
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 5245..5379
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 5547..6180
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 6227..6352
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6849..7082
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7267..7311
/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 7460..7501
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 7596..7713
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 7866..8273
/label="SFFV promoter"
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
CDS 8376..8441
/label="3xFLAG"
/note="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
CDS 9888..9944
/label="P2A"
/note="2A peptide from porcine teschovirus-1 polyprotein"
CDS 10047..10703
/gene="IL12A"
/label="Interleukin-12 subunit alpha"
/note="Interleukin-12 subunit alpha from Homo sapiens.
Accession#: P29459"
CDS 10713..10766
/label="T2A"
/note="2A peptide from Thosea asigna virus capsid protein"
CDS 10767..11750
/gene="IL12B"
/label="Interleukin-12 subunit beta"
/note="Interleukin-12 subunit beta from Homo sapiens.
Accession#: P29460"
misc_feature 11776..12349
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 12368..13084
/label="EGFP"
/note="enhanced GFP"
misc_feature 13148..13736
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"