FH-TET1-pEF vector (V006619)

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 Vector Map

FH-TET1-pEF12598 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000EF-1-alpha promoterT7 promoterKozak sequenceFLAGHATET1V5 tag6xHisbGH poly(A) signalf1 oriSV40 promoterNeoR/KanRSV40 poly(A) signalM13 revlac operatorlac promoterCAP binding siteL4440oriAmpRAmpR promoterpRS-marker

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

Copy Sequence

Download GeneBank File(.gb)

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..
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)
  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)"
     CDS             1418..1441
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             1454..1480
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             1493..7897
                     /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
                     /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
                     /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 
     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)
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10655..11243)
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
     CDS             complement(11417..12274)
     promoter        complement(12275..12379)
                     /label=AmpR promoter
     primer_bind     complement(12454..12473)
                     /note="pRS vectors, use to sequence yeast selectable

This page is informational only.