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"