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FH-TET1-pEF vector (V006619#)

Basic Vector Information

Basic Vector Information
Vector Name FH-TET1-pEF Antibiotic Resistance Ampicillin
Length 12598 bp Type Mammalian Expression
Selection Marker Neomycin (select with G418) Promoter EF-1alpha
Cloning Method Restriction Enzyme 5' Primer T7 promoter
3' Primer BGH Reverse

FH-TET1-pEF vector Vector Map

FH-TET1-pEF12598 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000EF-1-alpha promoterT7Kozak sequenceFLAGHAV5 tag6xHisBGH-revf1 oripBABE 3'NeoR/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       Exported               12598 bp ds-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)
  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"
FEATURES             Location/Qualifiers
     source          1..12598
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        163..1341
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          394..1332
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1289..1309
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1358..1377
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1358..1376
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1409..1418
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1418..1441
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1454..1480
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7919..7960
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7970..7987
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8010..8027)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8016..8240
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8286..8714
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8373..8392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8583..8604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8723..8743)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8728..9057
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8908..9043
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8970..8989
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             9124..9918
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(9178..9197)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9788..9807
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    10094..10215
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10131..10150)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10185..10204
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     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    10357..10378
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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"
     primer_bind     complement(10735..10754)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11414..12274)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     12037..12056
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12275..12379)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12454..12473)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"

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