Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V013241 | pHAGE-CMV-attR2 C XbaI EGFP taa XbaI | 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-CMV-attR2 C XbaI EGFP taa XbaI
- Antibiotic Resistance:
- Ampicillin, Chloramphenicol
- Length:
- 10758 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- CMV
- Growth Strain(s):
- DB3.1
- Growth Temperature:
- 37℃
pHAGE-CMV-attR2 C XbaI EGFP taa XbaI 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-CMV-attR2 C XbaI EGFP taa XbaI vector Sequence
LOCUS V013241 10758 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V013241
VERSION V013241
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10758)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..10758
/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 2062..2179
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 2231..2534
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2535..2738
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 2834..2852
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 2882..3006
/label="attR1"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(3438..3740)
/label="ccdB"
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
CDS complement(4085..4741)
/label="CmR"
/note="chloramphenicol acetyltransferase"
promoter complement(4742..4844)
/label="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
protein_bind complement(5022..5146)
/label="attR2"
/note="recombination site for the Gateway(R) LR reaction"
CDS 5154..5870
/label="EGFP"
/note="enhanced GFP"
CDS 5880..5903
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 5919..5945
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
misc_feature 5956..6531
/label="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 6532..7128
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 7148..7736
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 7811..8044
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter 8249..8353
/label="AmpR promoter"
CDS 8354..9211
/label="AmpR"
/note="beta-lactamase"
rep_origin 9385..9973
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 10384..10405
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 10420..10450
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 10458..10474
/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 10482..10498
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"