Basic Vector Information
FH-TET1-pEF 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 ℃.
FH-TET1-pEF vector Sequence
LOCUS FH-TET1-pEF. 12598 bp DNA circular SYN 13-MAY-2021
DEFINITION This plasmid expresses the full-length human TET1 gene with a Flag
and HA tag..
ACCESSION .
VERSION .
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"
COMMENT 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=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"
This page is informational only.