Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V006787 | Mfn2-Myc | 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:
- Mfn2-Myc
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8419 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV-F
- 3' Primer:
- BGH-rev
Mfn2-Myc 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.
Mfn2-Myc vector Sequence
LOCUS V006787 8419 bp DNA circular SYN 13-MAY-2021 DEFINITION Exported. ACCESSION V006787 VERSION V006787 KEYWORDS Mfn2-Myc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 8419) AUTHORS Chen H, Detmer SA, Ewald AJ, Griffin EE, Fraser SE, Chan DC TITLE Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development. JOURNAL J Cell Biol. 2003 Jan 20. 160(2):189-200. PUBMED 12527753 REFERENCE 2 (bases 1 to 8419) TITLE Direct Submission REFERENCE 3 (bases 1 to 8419) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Cell Biol. 2003 Jan 20. 160(2):189-200." SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..8419 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(44..63) /label="pRS-marker" /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" promoter 863..881 /label="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" CDS 1002..3272 /gene="Mfn2" /label="Mitofusin-2" /note="Mitofusin-2 from Mus musculus. Accession#: Q80U63" CDS 3288..3317 /label="Myc" /note="Myc (human c-Myc proto-oncogene) epitope tag" CDS 3324..3353 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3360..3389 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3408..3437 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3444..3473 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3480..3509 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3528..3557 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3564..3593 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3600..3629 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3648..3677 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3684..3713 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3720..3749 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3768..3797 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3804..3833 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3840..3869 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3903..3932 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label="Myc" /translation="EQKLISEEDL" CDS 3948..3965 /label="6xHis" /note="6xHis affinity tag" polyA_signal 4015..4239 /label="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" rep_origin 4285..4713 /label="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 4727..5057 /label="SV40 promoter" /note="SV40 enhancer and early promoter" CDS 5124..5915 /label="NeoR/KanR" /note="aminoglycoside phosphotransferase" polyA_signal 6092..6225 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal" primer_bind complement(6262..6278) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind complement(6286..6302) /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)." promoter complement(6310..6340) /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind complement(6355..6376) /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6493..6510) /label="L4440" /note="L4440 vector, forward primer" rep_origin complement(6664..7252) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7426..8283) /label="AmpR" /note="beta-lactamase" promoter complement(8284..8388) /label="AmpR promoter"