LOCUS       Exported                4828 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCK65, complete sequence.
ACCESSION   HQ644135
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4828)
  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 4828)
  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 4828)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4828
                     /organism="Mammalian expression vector pCK65"
                     /mol_type="other DNA"
                     /note="designed for urate-inducible Purex8-driven smUox 
                     expression; codon optimized for expression in Homo sapiens"
                     /db_xref="taxon:943931"
     regulatory      34..245
                     /regulatory_class="promoter"
                     /note="simian virus 40 promoter"
     promoter        48..244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      95..230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     repeat_region   258..519
                     /label=synthetic octameric hucO repeat (hucO8)
                     /note="synthetic octameric hucO repeat (hucO8)"
     CDS             537..1505
                     /codon_start=1
                     /gene="smUox"
                     /product="secreted mammalian urate oxidase precursor"
                     /label=smUox
                     /protein_id="ADV16000.1"
                     /translation="METDTLLLWVLLLWVPGSTGDSAVKAARYGKDNVRVYKVHKDEKT
                     GVQTVYEMTVCVLLEGEIETSYTKADNSVIVATDSIKNTIYITAKQNPVTPPELFGSIL
                     GTHFIEKYNHIHAAHVNIVCHRWTRMDIDGKPHPHSFIRDSEEKRNVQVDVVEGKGIDI
                     KSSLSGLTVLKSTNSQFWGFLRDEYTTLKETWDRILSTDVDATWQWKNFSGLQEVRSHV
                     PKFDATWATAREVTLKTFAEDNSASVQATMYKMAEQILARQQLIETVEYSLPNKHYFEI
                     DLSWHKGLQNTGKNAEVFAPQSDPNGLIKCTVGRSSLKSKL"
     gene            537..1505
                     /gene="smUox"
                     /label=smUox
     sig_peptide     537..599
                     /gene="smUox"
     polyA_signal    1545..1666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      1655..1660
                     /regulatory_class="polyA_signal_sequence"
     regulatory      1769..2014
                     /regulatory_class="enhancer"
                     /note="simian virus 40 enhancer"
     rep_origin      complement(2331..2919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     gene            complement(3090..3950)
                     /gene="AmpR"
                     /label=AmpR
                     /note="ampicillin resistance protein"
     CDS             complement(3090..3950)
                     /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(3951..4055)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4082..4537
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    4668..4716
                     /note="synthetic polyadenylation signal"
     misc_feature    4730..4821
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"