Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006914 | phage-ubc-nls-ha-tdMCP-gfp | 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:
- phage-ubc-nls-ha-tdMCP-gfp
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7772 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Promoter:
- UbC
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- FUGW (5'-ATTACAGGGACAGCAGAGATCC-3')
- 3' Primer:
- WPRE-R (5'CATAGCGTAAAAGGAGCAACA-3')
phage-ubc-nls-ha-tdMCP-gfp 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.
phage-ubc-nls-ha-tdMCP-gfp vector Sequence
LOCUS V006914 7772 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V006914
VERSION V006914
KEYWORDS phage-ubc-nls-ha-tdMCP-gfp
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7772)
AUTHORS Wu B, Chao JA, Singer RH
TITLE Fluorescence fluctuation spectroscopy enables quantitative imaging
of single mRNAs in living cells.
JOURNAL Biophys J. 2012 Jun 20;102(12):2936-44. Epub 2012 Jun 19.
PUBMED 22735544
REFERENCE 2 (bases 1 to 7772)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7772)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biophys
J."; date: "2012-06-20"; volume: "102(12)"; pages: "2936-44. Epub
2012 Jun 19"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7772
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/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 1914..1955
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2058..2175
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2218..2616
/label="UbC promoter"
/note="human ubiquitin C promoter"
CDS 2631..2651
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 2655..2681
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 2682..2693
/label="Factor Xa site"
/note="Factor Xa recognition and cleavage site"
CDS 3429..4145
/label="EGFP"
/note="enhanced GFP"
misc_feature 4162..4750
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4825..5058
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(5177..5195)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5263..5367
/label="AmpR promoter"
CDS 5368..6225
/label="AmpR"
/note="beta-lactamase"
rep_origin 6399..6987
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7141..7158
/label="L4440"
/note="L4440 vector, forward primer"
primer_bind 7250..7269
/label="SV40pro-F"
/note="SV40 promoter/origin, forward primer"
protein_bind 7398..7419
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7434..7464
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 7472..7488
/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 7496..7512
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7562..7581
/label="EBV-rev"
/note="SV40 polyA terminator, reverse primer"