pUT3 vector (V002268)

Basic Vector Information

      • Vector Name:
      • pUT3
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 5769 bp
      • Type:
      • Expression vector
      • Source/Author:
      • Kasai Y, Matsuzaki K, Ikeda F, Yoshimitsu Y, Harayama S.

pUT3 vector Vector Map

pUT35769 bp60012001800240030003600420048005400f1 oriM13 fwdT7 promoterKS primerIncludes promoter region of Pseudococcomyxa sp. strain KJ nuclear gene EF1a for elongation factor 1 alphaUMPSIncludes terminator region of Pseudococcomyxa sp. strain KJ nuclear gene EF1a for elongation factor 1 alphaT3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pUT3 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_45818        5769 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pUT3 DNA, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5769)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-JUN-2017) Contact:Yuki Kasai Chuo University, Research and 
            Development Initiative; 1-13-27 Kasuga, Bunkyo-ku, Tokyo 112-8551, 
            Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /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
                     /gene="EF1a"
                     /note="Includes promoter region of Pseudococcomyxa sp.
                     strain KJ nuclear gene EF1a for elongation factor 1 alpha"
                     /regulatory_class="promoter"
     gene            694..1341
                     /gene="EF1a"
                     /label=EF1a
     regulatory      1336..1345
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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
                     /gene="EF1a"
                     /note="Includes terminator region of Pseudococcomyxa sp.
                     strain KJ nuclear gene EF1a for elongation factor 1 alpha"
                     /regulatory_class="terminator"
     gene            2853..3544
                     /gene="EF1a"
                     /label=EF1a
     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    complement(3644..3660)
                     /label=lac operator
                     /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    complement(3713..3734)
                     /label=CAP binding site
                     /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(4784..5641)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5642..5746)
                     /label=AmpR promoter

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