LOCUS       Exported                5769 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pUT3 DNA, complete sequence.
ACCESSION   LC276367
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5769)
  AUTHORS   Kasai Y, Matsuzaki K, Ikeda F, Yoshimitsu Y, Harayama S.
  TITLE     Precise excision of a selectable marker gene in transgenic 
            Pseudococcomyxa strains by the piggyBac transposase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5769)
  AUTHORS   Kasai Y, Harayama S.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUN-2017) Contact:Yuki Kasai Chuo University, Research
            and Development Initiative; 1-13-27 Kasuga, Bunkyo-ku, Tokyo 
            112-8551, Japan
REFERENCE   3  (bases 1 to 5769)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5769
                     /organism="Expression vector pUT3"
                     /mol_type="other DNA"
                     /db_xref="taxon:2014526"
     misc_feature    1..693
                     /label=delived from plasmid pBluescript SK(+)
                     /note="delived from plasmid pBluescript SK(+)"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      694..1341
                     /regulatory_class="promoter"
                     /gene="EF1a"
                     /note="Includes promoter region of Pseudococcomyxa sp. 
                     strain KJ nuclear gene EF1a for elongation factor 1 alpha"
     gene            694..1341
                     /gene="EF1a"
                     /label=EF1a
     regulatory      1336..1345
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1342..2847
                     /codon_start=1
                     /gene="UMPS"
                     /product="uridine monophosphate synthase"
                     /label=UMPS
                     /protein_id="BAY00744.1"
                     /translation="MATSTPSVATAAELKPKEGPITSQEFEELVLRLHEIEAVKFGNFK
                     LKSGLMSPIYIDLRVIVSYPDVLRRVAEVMWHQVSGAGFDVMCGVPYTALPIATCMSLL
                     HGTPMLMRRKEVKEYGTKKAIEGAFKKGQTCLIVEDLVTSGASVMETVEPLEVEGLKVA
                     DVVVLIDREQGGAARMASNGLRLHSAFTLSFIIKTLQAHGLVSAEVADSVAAFIAANQT
                     FSPSAAAAPTPASAPPAQPKRLPFGERAALCQNAAGRKLLELMARKRTNLAVAADVATV
                     EEMLRIADAAGPHIAVFKTHVDIFDEWDDGIAAQLRHLADKHEFLIFEDRKFADIGNTV
                     VSQYGGGIYKIADWSDITNAHLVPGPGIIDGLRKVGQEKGRGLLLLAEMSSKGALATGA
                     YTEKVAEAAAANQDFVMGFICQSPAKWATSVPPGLVHMTPGVQLASGSDALGQQYNTPA
                     SVIGQGGSDVIIVGRGIIKAADPAAAAAQYREAGWAAYEATLA"
     gene            1342..2847
                     /gene="UMPS"
                     /label=UMPS
     regulatory      2853..3544
                     /regulatory_class="terminator"
                     /gene="EF1a"
                     /note="Includes terminator region of Pseudococcomyxa sp. 
                     strain KJ nuclear gene EF1a for elongation factor 1 alpha"
     gene            2853..3544
                     /gene="EF1a"
                     /label=EF1a
     misc_feature    3545..5769
                     /label=delived from plasmid pBluescript SK (+)
                     /note="delived from plasmid pBluescript SK (+)"
     promoter        complement(3581..3599)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3620..3636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3644..3660
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3668..3698)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3713..3734
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4022..4610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4781..5641)
                     /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(5642..5746)
                     /gene="bla"
                     /label=AmpR promoter