LOCUS       Exported                5229 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pYL1, complete sequence.
ACCESSION   MH594278
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5229)
  AUTHORS   Liu Y, Charpin-El Hamri G, Ye H, Fussenegger M.
  TITLE     A synthetic free fatty acid-regulated transgene switch in mammalian 
            cells and mice
  JOURNAL   Nucleic Acids Res. (2018) In press
  PUBMED    30219861
REFERENCE   2  (bases 1 to 5229)
  AUTHORS   Liu Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JUL-2018) Department of Biosystems Science and 
            Engineering, ETH Zurich, Mattenstrasse 26, basel 4058, Switzerland
REFERENCE   3  (bases 1 to 5229)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5229
                     /organism="Expression vector pYL1"
                     /mol_type="other DNA"
                     /db_xref="taxon:2340779"
     misc_feature    14..21
                     /label=cAMP-response element (CRE)
                     /note="cAMP-response element (CRE)"
     misc_feature    40..44
                     /label=CRE
                     /note="CRE"
     misc_feature    50..54
                     /label=CRE
                     /note="CRE"
     misc_feature    72..79
                     /label=CRE
                     /note="CRE"
     misc_feature    109..118
                     /label=serum response factor binding element (SRE)
                     /note="serum response factor binding element (SRE)"
     misc_feature    132..141
                     /label=SRE
                     /note="SRE"
     misc_feature    155..164
                     /label=SRE
                     /note="SRE"
     protein_bind    189..218
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     misc_feature    191..199
                     /note="nuclear factor of activated T cell binding sites 
                     (NFAT)"
     protein_bind    219..248
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     misc_feature    221..229
                     /label=NFAT
                     /note="NFAT"
     protein_bind    249..278
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     misc_feature    251..259
                     /label=NFAT
                     /note="NFAT"
     promoter        291..322
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    292..322
                     /label=minimal promoter
                     /note="minimal promoter"
     CDS             350..1909
                     /codon_start=1
                     /gene="SEAP"
                     /product="secreted embryonic alkaline phosphatase"
                     /label=SEAP
                     /protein_id="AYA60341.1"
                     /translation="MLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQP
                     AQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDK
                     HVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVV
                     TTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRK
                     YMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTH
                     LMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESR
                     AYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKA
                     RDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFA
                     RGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTDAAHPGYSRVGAAGRFE
                     QT"
     gene            350..1909
                     /gene="SEAP"
                     /label=SEAP
     CDS             350..1852
                     /codon_start=1
                     /product="secreted alkaline phosphatase from human 
                     placenta"
                     /label=SEAP
                     /note="The natural enzyme has a C-terminal transmembrane 
                     sequence that is replaced by a GPI anchor."
                     /translation="MLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQP
                     AQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDK
                     HVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVV
                     TTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRK
                     YMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTH
                     LMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESR
                     AYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKA
                     RDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFA
                     RGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     polyA_signal    1910..2031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2020..2025
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late mRNA polyadenylation signal"
     rep_origin      2226..2869
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      complement(2696..3284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3455..4315)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /protein_id="AYA60342.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3455..4315)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(4316..4420)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4447..4902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     misc_feature    5033..5186
                     /label=transcription blocker
                     /note="transcription blocker"
     polyA_signal    5033..5081
                     /note="synthetic polyadenylation signal"
     misc_feature    5095..5186
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"