LOCUS       Exported                4867 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCK53, complete sequence.
ACCESSION   KF528989
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4867)
  AUTHORS   Kemmer C, Fluri DA, Witschi U, Passeraub A, Gutzwiller A, 
            Fussenegger M.
  TITLE     A designer network coordinating bovine artificial insemination by 
            ovulation-triggered release of implanted sperms
  JOURNAL   J Control Release 150 (1), 23-29 (2011)
  PUBMED    21108977
REFERENCE   2  (bases 1 to 4867)
  AUTHORS   Kemmer C, Fluri DA, Witschi U, Passeraub A, Gutzwiller A, 
            Fussenegger M.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-AUG-2013) BSSE, ETH Zurich, Mattenstrasse 26, Basel, 
            BS 4058, Switzerland
REFERENCE   3  (bases 1 to 4867)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4867
                     /organism="Mammalian expression vector pCK53"
                     /mol_type="other DNA"
                     /db_xref="taxon:1419745"
     CDS             3..1562
                     /codon_start=1
                     /gene="SEAP"
                     /product="SEAP"
                     /label=SEAP
                     /note="secreted alkaline phosphatase"
                     /protein_id="AHA35652.1"
                     /translation="MLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQP
                     AQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDK
                     HVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVV
                     TTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRK
                     YMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTH
                     LMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESR
                     AYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKA
                     RDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFA
                     RGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTDAAHPGYSRVGAAGRFE
                     QT"
     gene            3..1562
                     /gene="SEAP"
                     /label=SEAP
     CDS             3..1505
                     /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    1563..1684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2088..2707
     rep_origin      complement(2103..2691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2862..3722)
                     /codon_start=1
                     /gene="ampR"
                     /product="ampicillin resistance marker"
                     /label=ampR
                     /protein_id="AHA35653.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2862..3722)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(3723..3827)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3764..3792
                     /regulatory_class="promoter"
     rep_origin      3854..4309
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      3871..4177
     polyA_signal    4440..4488
                     /note="synthetic polyadenylation signal"
     misc_feature    4502..4593
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     regulatory      4627..4866
                     /regulatory_class="promoter"
                     /note="CRE promoter"