LOCUS       Exported               14224 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pUMLIEP DNA, complete sequence.
ACCESSION   LC175306
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14224)
  AUTHORS   Okochi-Takada E, Hattori N, Ito A, Niwa T, Wakabayashi M, Yoshida M,
            Ushijima T.
  TITLE     Establishment of a high-throughput detection system for DNA 
            demethylating agents
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14224)
  AUTHORS   Hattori N, Ushijima T.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-AUG-2016) Contact:Naoko Hattori National Cancer Center
            Research Institute, Division of Epigenomics; 5-1-1 Tsukiji, Chuo-ku,
            Tokyo, Tokyo 104-0045, Japan
REFERENCE   3  (bases 1 to 14224)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14224
                     /organism="Expression vector pUMLIEP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1886769"
     misc_feature    1..697
                     /label=pBlueScript II SK(+)
                     /note="pBlueScript II SK(+)"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    698..6262
                     /gene="UCHL1"
                     /label=UCHL1 5region
                     /note="UCHL1 5region"
     gene            698..6262
                     /gene="UCHL1"
                     /label=UCHL1
     misc_feature    6225
                     /gene="UCHL1"
                     /label=UCHL1 TSS
                     /note="UCHL1 TSS"
     regulatory      6310..6319
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6316..6975
                     /codon_start=1
                     /gene="MetLuc"
                     /product="secreted luciferase"
                     /label=MetLuc
                     /note="Metridia longa secreted luciferase"
                     /protein_id="BAV53308.1"
                     /translation="MDIKVVFTLVFSALVQAKSTEFDPNIDIVGLEGKFGITNLETDLF
                     TIWETMEVMIKADIADTDRASNFVATETDANRGKMPGKKLPLAVIMEMEANAFKAGCTR
                     GCLICLSKIKCTAKMKVYIPGRCHDYGGDKKTGQAGIVGAIVDIPEISGFKEMAPMEQF
                     IAQVDRCASCTTGCLKGLANVKCSELLKKWLPDRCASFADKIQKEVHNIKGMAGDR"
     gene            6316..6975
                     /gene="MetLuc"
                     /label=MetLuc
     misc_feature    6987..7571
                     /label=IRES
                     /note="IRES"
     misc_feature    6988..7574
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             7575..8294
                     /codon_start=1
                     /gene="EGFP"
                     /product="enhanced green fluorescent protein"
                     /label=EGFP
                     /protein_id="BAV53309.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            7575..8294
                     /gene="EGFP"
                     /label=EGFP
     polyA_signal    8417..8538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8562..8578)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8603..9410
                     /gene="UCHL1"
                     /label=contains UCHL1 intron 4
                     /note="contains UCHL1 intron 4"
     gene            8603..9410
                     /gene="UCHL1"
                     /label=UCHL1
     promoter        9420..9749
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     regulatory      9433..9701
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     rep_origin      9600..9735
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             9842..10441
                     /codon_start=1
                     /gene="Puro(r)"
                     /product="puromycin N-acetyltransferase"
                     /label=Puro(r)
                     /note="puromycin resistance cassette"
                     /protein_id="BAV53310.1"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     gene            9842..10441
                     /gene="Puro(r)"
                     /label=Puro(r)
     polyA_signal    10671..10805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    10805..12017
                     /gene="UCHL1"
                     /note="contains UCHL1 intron 4, exon 5, intron 5, exon 6, 
                     and intron 6"
     gene            10805..12017
                     /gene="UCHL1"
                     /label=UCHL1
     misc_feature    12018..14224
                     /label=pBlueScript II SK(+)
                     /note="pBlueScript II SK(+)"
     promoter        complement(12036..12054)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12075..12091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    12099..12115
                     /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(12123..12153)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12168..12189
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(12477..13065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(13236..14096)
                     /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"
     promoter        complement(14097..14201)
                     /gene="bla"
                     /label=AmpR promoter