Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000813 | pLPCX2-MT1-MMP-eGFP | 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:
- pLPCX2-MT1-MMP-eGFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8743 bp
- Type:
- Retroviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- Low Copy
pLPCX2-MT1-MMP-eGFP 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.
pLPCX2-MT1-MMP-eGFP vector Sequence
LOCUS V000813 8743 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000813
VERSION V000813
KEYWORDS pLPCX2-MT1-MMP-eGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8743)
AUTHORS Sun J, Lu F, He H, Shen J, Messina J, Mathew R, Wang D, Sarnaik AA,
Chang WC, Kim M, Cheng H, Yang S
TITLE STIM1- and Orai1-mediated Ca(2+) oscillation orchestrates
invadopodium formation and melanoma invasion.
JOURNAL J Cell Biol. 2014 Nov 24;207(4):535-48. doi: 10.1083/jcb.201407082.
Epub 2014 Nov 17.
PUBMED 25404747
REFERENCE 2 (bases 1 to 8743)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8743)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1083/jcb.201407082"; journalName: "J Cell Biol"; date:
"2014-11-24- 24"; volume: "207"; issue: "4"; pages: "535-48"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8743
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..304
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 305..508
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 583..2328
/gene="MMP14"
/label="Matrix metalloproteinase-14"
/note="Matrix metalloproteinase-14 from Homo sapiens.
Accession#: P50281"
CDS 2359..3075
/label="EGFP"
/note="enhanced GFP"
LTR 3144..3737
/label="LTR"
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(3840..3862)
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
protein_bind 3990..4023
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
protein_bind complement(4083..4116)
/label="loxP"
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind 4136..4155
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4236..4253)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(4407..4995)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5169..6026)
/label="AmpR"
/note="beta-lactamase"
promoter complement(6027..6131)
/label="AmpR promoter"
primer_bind 6199..6217
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
LTR 6407..6994
/label="5' LTR"
/note="long terminal repeat from Moloney murine sarcoma
virus"
misc_feature 7057..7256
/label="MMLV Psi"
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 7457..7873
/label="gag (truncated)"
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
CDS 7970..8566
/label="PuroR"
/note="puromycin N-acetyltransferase"
promoter 8687..8743
/label="CMV"
/note="Human cytomegalovirus immediate early
enhancer/promoter"