LOCUS       Exported                5213 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCK146, complete sequence.
ACCESSION   HQ644137
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5213)
  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 5213)
  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 5213)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5213
                     /organism="Mammalian expression vector pCK146"
                     /mol_type="other DNA"
                     /note="designed for urate-inducible Purex8-2-driven smUox 
                     expression; codon optimized for expression in Homo sapiens"
                     /db_xref="taxon:943932"
     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)"
     CDS             922..1890
                     /codon_start=1
                     /gene="smUox"
                     /product="secreted mammalian urate oxidase precursor"
                     /label=smUox
                     /protein_id="ADV16001.1"
                     /translation="METDTLLLWVLLLWVPGSTGDSAVKAARYGKDNVRVYKVHKDEKT
                     GVQTVYEMTVCVLLEGEIETSYTKADNSVIVATDSIKNTIYITAKQNPVTPPELFGSIL
                     GTHFIEKYNHIHAAHVNIVCHRWTRMDIDGKPHPHSFIRDSEEKRNVQVDVVEGKGIDI
                     KSSLSGLTVLKSTNSQFWGFLRDEYTTLKETWDRILSTDVDATWQWKNFSGLQEVRSHV
                     PKFDATWATAREVTLKTFAEDNSASVQATMYKMAEQILARQQLIETVEYSLPNKHYFEI
                     DLSWHKGLQNTGKNAEVFAPQSDPNGLIKCTVGRSSLKSKL"
     gene            922..1890
                     /gene="smUox"
                     /label=smUox
     sig_peptide     922..984
                     /gene="smUox"
     polyA_signal    1930..2051
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2040..2045
                     /regulatory_class="polyA_signal_sequence"
     regulatory      2154..2399
                     /regulatory_class="enhancer"
                     /note="simian virus 40 enhancer"
     rep_origin      complement(2716..3304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     gene            complement(3475..4335)
                     /gene="AmpR"
                     /label=AmpR
                     /note="ampicillin resistance protein"
     CDS             complement(3475..4335)
                     /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(4336..4440)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4467..4922
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    5053..5101
                     /note="synthetic polyadenylation signal"
     misc_feature    5115..5206
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"