Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017219 | pLEX307-DPP4-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:
- pLEX307-DPP4-puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10717 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- EF-1α
- Growth Temperature:
- 30℃
pLEX307-DPP4-puro 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.
pLEX307-DPP4-puro vector Sequence
LOCUS V017219 10717 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017219
VERSION V017219
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10717)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..10717
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 96..1274
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
protein_bind 1294..1318
/label="attB1"
/note="recombination site for the Gateway(R) BP reaction"
CDS 1324..3621
/gene="DPP4"
/label="Dipeptidyl peptidase 4"
/note="Dipeptidyl peptidase 4 from Homo sapiens.
Accession#: P27487"
protein_bind complement(3626..3650)
/label="attB2"
/note="recombination site for the Gateway(R) BP reaction"
CDS 3658..3699
/label="V5 tag"
/note="epitope tag from simian virus 5"
misc_feature 3750..4338
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4410..4643
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4721..4855
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(5038..5056)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin 5224..5679
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5705..5809
/label="AmpR promoter"
CDS 5810..6667
/label="AmpR"
/note="beta-lactamase"
rep_origin 6841..7429
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 7717..7738
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7753..7783
/label="lac promoter"
/note="promoter for the E. coli lac operon"
promoter 7852..7870
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 7898..8124
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
misc_feature 8352..8477
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8970..9203
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 9388..9432
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
promoter 9663..9873
/label="SV40 promoter"
/note="SV40 early promoter"
CDS 9913..10509
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 10558..10675
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"