Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V013796 | pMLAV-EF1a-MCS-IRES-EGFP-Puro | 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:
- pMLAV-EF1a-MCS-IRES-EGFP-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7819 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Adeno-associated virus
- Selection Marker:
- Puro
- Promoter:
- EF1a
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pMLAV-EF1a-MCS-IRES-EGFP-Puro 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.
pMLAV-EF1a-MCS-IRES-EGFP-Puro vector Sequence
LOCUS V013796 7819 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V013796
VERSION V013796
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7819)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..7819
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(60..435)
/label="ISS"
/note="immunostimulatory sequence from the AmpR gene;
contains unmethylated CpG dinucleotides in the context of
5'-AACGTT-3' (Sato et al., 1996)"
promoter complement(620..724)
/label="AmpR promoter"
repeat_region 838..940
/label="ITR"
/note="inverted terminal repeat of human adenovirus
serotype 5"
misc_signal 1028..1178
/label="Ad5 Psi"
/note="packaging signal for adenovirus serotype 5"
primer_bind 1212..1228
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter 1447..1658
/label="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
misc_feature 1916..2499
/label="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 2515..3231
/label="EGFP"
/note="enhanced GFP"
polyA_signal complement(3857..3991)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS 4559..4978
/gene="IX"
/label="Hexon-interlacing protein"
/note="Hexon-interlacing protein from Human adenovirus C
serotype 5. Accession#: P03281"
CDS complement(5044..6378)
/gene="IVa2"
/label="Packaging protein 1"
/note="Packaging protein 1 from Human adenovirus C serotype
5. Accession#: P03271"
rep_origin complement(7231..7819)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"