Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006619 | FH-TET1-pEF | 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:
- FH-TET1-pEF
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12598 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Promoter:
- EF-1α
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- T7 promoter
- 3' Primer:
- BGH Reverse
FH-TET1-pEF 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.
FH-TET1-pEF vector Sequence
LOCUS V006619 12598 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V006619
VERSION V006619
KEYWORDS FH-TET1-pEF
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12598)
AUTHORS Tahiliani M, Koh KP, Shen Y, Pastor WA, Bandukwala H, Brudno Y,
Agarwal S, Iyer LM, Liu DR, Aravind L, Rao A
TITLE Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in
mammalian DNA by MLL partner TET1.
JOURNAL Science. 2009 May 15;324(5929):930-5. Epub 2009 Apr 16.
PUBMED 19372391
REFERENCE 2 (bases 1 to 12598)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 12598)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science.";
date: "2009-05-15"; volume: "324(5929)"; pages: "930-5. Epub 2009
Apr 16"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12598
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 163..1341
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
promoter 1358..1376
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 1409..1418
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1418..1441
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 1454..1480
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 1493..7897
/gene="TET1"
/label="Methylcytosine dioxygenase TET1"
/note="Methylcytosine dioxygenase TET1 from Homo sapiens.
Accession#: Q8NFU7"
CDS 7919..7960
/label="V5 tag"
/note="epitope tag from simian virus 5"
CDS 7970..7987
/label="6xHis"
/note="6xHis affinity tag"
polyA_signal 8016..8240
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 8286..8714
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 8728..9057
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 9124..9915
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
polyA_signal 10094..10227
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(10264..10280)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10264..10280)
/label="M13 Reverse"
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(10277..10299)
/label="M13/pUC Reverse"
/note="In lacZ gene"
protein_bind 10288..10304
/label="lac operator"
/bound_moiety="lac repressor encoded by lacI"
/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(10312..10342)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(10357..10378)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(10495..10512)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(10655..11243)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11417..12274)
/label="AmpR"
/note="beta-lactamase"
promoter complement(12275..12379)
/label="AmpR promoter"
primer_bind complement(12454..12473)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"