Basic Vector Information
pcDNA3/Flag-METTL3 vector Vector Map
Plasmid Resuspension 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 ℃.
pcDNA3/Flag-METTL3 vector Sequence
LOCUS pcDNA3/Flag-METT 7052 bp DNA circular SYN 21-DEC-2021
DEFINITION For expression of METTL3 (methyltransferase-like 3) in human cells.
ACCESSION .
VERSION .
KEYWORDS pcDNA3/Flag-METTL3.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7052)
AUTHORS Liu J, Yue Y, Han D, Wang X, Fu Y, Zhang L, Jia G, Yu M, Lu Z, Deng
X, Dai Q, Chen W, He C
TITLE A METTL3-METTL14 complex mediates mammalian nuclear RNA N6-adenosine
methylation.
JOURNAL Nat Chem Biol. 2014 Feb;10(2):93-5. doi: 10.1038/nchembio.1432. Epub
2013 Dec 6.
PUBMED 24316715
REFERENCE 2 (bases 1 to 7052)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7052)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/nchembio.1432"; journalName: "Nat Chem Biol"; date:
"2014-02"; volume: "10"; issue: "2"; pages: "93-5"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7052
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 23..44
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(163..183)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 207..417
/label=SV40 promoter
/note="SV40 early promoter"
CDS 492..1283
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
polyA_signal 1460..1593
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1630..1646)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1654..1670)
/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(1678..1708)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1723..1744)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1861..1878)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2032..2620)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2794..3651)
/label=AmpR
/note="beta-lactamase"
promoter complement(3652..3756)
/label=AmpR promoter
primer_bind complement(3831..3850)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 4022..4401
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4402..4605
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 4650..4668
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 4685..4708
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 4709..6445
/gene="METTL3"
/label=METTL3
/note="N6-adenosine-methyltransferase catalytic subunit
from Homo sapiens. Accession#: Q86U44"
promoter complement(6464..6482)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
polyA_signal 6508..6732
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 6778..7052
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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