Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012444 | Lentiviral-TOP-dGFP-reporter | 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:
- Lentiviral-TOP-dGFP-reporter
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7151 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- RSV
- Cloning Method:
- Restriction Enzyme
Lentiviral-TOP-dGFP-reporter 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.
Lentiviral-TOP-dGFP-reporter vector Sequence
LOCUS V012444 7151 bp DNA circular SYN 17-AUG-2021
DEFINITION Exported.
ACCESSION V012444
VERSION V012444
KEYWORDS Lentiviral-TOP-dGFP-reporter
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7151)
AUTHORS Reya T, Duncan AW, Ailles L, Domen J, Scherer DC, Willert K, Hintz
L, Nusse R, Weissman IL
TITLE A role for Wnt signalling in self-renewal of haematopoietic stem
cells.
JOURNAL Nature. 2003 May 22. 423(6938):409-14.
PUBMED 12717450
REFERENCE 2 (bases 1 to 7151)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7151)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature.
2003 May 22. 423(6938):409-14."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7151
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 15..603
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 757..774
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 891..912
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 927..957
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 965..981
/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 989..1005
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 1026..1044
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 1072..1298
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 1299..1479
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1526..1651
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 2144..2377
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2562..2606
/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 2755..2796
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2873..2990
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
regulatory 3169..3178
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3200..4042
/label="d2EGFP"
/note="EGFP destabilized by fusion to residues 422-461 of
mouse ornithine decarboxylase, giving an in vivo half-life
of ~2 hours"
misc_feature 4066..4654
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4741..4974
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5046..5180
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 5207..5342
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(5363..5381)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5391..5407)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 5549..6004
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6030..6134
/label="AmpR promoter"
CDS 6135..6992
/label="AmpR"
/note="beta-lactamase"