LOCUS       Exported                5768 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCK73, complete sequence.
ACCESSION   HQ644136
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5768)
  AUTHORS   Kemmer C, Gitzinger M, Daoud-El Baba M, Djonov V, Stelling J, 
            Fussenegger M.
  TITLE     Self-sufficient control of urate homeostasis in mice by a synthetic 
            circuit
  JOURNAL   Nat. Biotechnol. 28 (4), 355-360 (2010)
  PUBMED    20351688
REFERENCE   2  (bases 1 to 5768)
  AUTHORS   Kemmer C, Gitzinger M, Daoud-El Baba M, Djonov V, Stelling J, 
            Fussenegger M.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-NOV-2010) Department of Biosystems Science and 
            Engineering, D-BSSE, ETH Zurich, Mattenstrasse 20, Basel 4058, 
            Switzerland
REFERENCE   3  (bases 1 to 5768)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5768
                     /organism="Mammalian expression vector pCK73"
                     /mol_type="other DNA"
                     /note="designed for urate-inducible Purex8-2-driven SEAP 
                     expression; codon optimized for expression in Homo sapiens"
                     /db_xref="taxon:943933"
     regulatory      46..630
                     /regulatory_class="promoter"
                     /note="human cytomegalovirus derived promoter"
     enhancer        52..431
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   643..904
                     /label=synthetic octameric hucO repeat (hucO8)
                     /note="synthetic octameric hucO repeat (hucO8)"
     gene            925..2484
                     /gene="SEAP"
                     /label=SEAP
                     /note="secreted alkaline phosphatase"
     CDS             925..2427
                     /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    2485..2606
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2595..2600
                     /regulatory_class="polyA_signal_sequence"
     regulatory      2709..2954
                     /regulatory_class="enhancer"
                     /note="simian virus 40 enhancer"
     rep_origin      complement(3271..3859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     gene            complement(4030..4890)
                     /gene="AmpR"
                     /label=AmpR
                     /note="ampicillin resistance protein"
     CDS             complement(4030..4890)
                     /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(4891..4995)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5022..5477
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    5608..5656
                     /note="synthetic polyadenylation signal"
     misc_feature    5670..5761
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"