LOCUS       Exported                5191 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  SEAP expression vector pPST67-H6, complete sequence.
ACCESSION   KM403566
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5191)
  AUTHORS   Auslander S, Stucheli P, Rehm C, Auslander D, Hartig JS, Fussenegger
            M.
  TITLE     A general design strategy for protein-responsive riboswitches in 
            mammalian cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5191)
  AUTHORS   Auslander S, Stucheli P, Rehm C, Auslander D, Hartig JS, Fussenegger
            M.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-AUG-2014) Department of Biosystems Science and 
            Engineering, D-BSSE, ETH Zurich, Mattenstrasse 26, Basel, BS 4058, 
            Switzerland
REFERENCE   3  (bases 1 to 5191)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5191
                     /organism="SEAP expression vector pPST67-H6"
                     /mol_type="other DNA"
                     /db_xref="taxon:1549503"
     regulatory      37..245
                     /regulatory_class="promoter"
                     /gene="SV40 early promoter"
     gene            37..245
                     /gene="SV40 early promoter"
                     /label=SV40 early promoter
     promoter        48..244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      95..230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             272..1831
                     /codon_start=1
                     /gene="SEAP"
                     /product="SEAP"
                     /label=SEAP
                     /note="secreted embryonic alkaline phosphatase"
                     /protein_id="AIR09507.1"
                     /translation="MLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQP
                     AQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDK
                     HVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVV
                     TTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRK
                     YMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTH
                     LMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESR
                     AYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKA
                     RDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFA
                     RGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTDAAHPGYSRVGAAGRFE
                     QT"
     gene            272..1831
                     /gene="SEAP"
                     /label=SEAP
                     /note="secreted embryonic alkaline phosphatase"
     CDS             272..1774
                     /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"
     misc_feature    1858..1933
                     /label=HHR Env140_nutR_H6
                     /note="HHR Env140_nutR_H6"
     polyA_signal    1948..2029
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2018..2023
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late mRNA polyadenylation signal"
     rep_origin      complement(2224..2867)
                     /direction=LEFT
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      complement(2694..3282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3453..4313)
                     /codon_start=1
                     /gene="AmpR"
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AIR09508.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3453..4313)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(4314..4418)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4445..4900
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    5031..5079
                     /note="synthetic polyadenylation signal"
     misc_feature    5093..5184
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"