Basic Vector Information
pUT3 vector Vector Map
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
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Precise excision of a selectable marker gene in transgenic Pseudococcomyxa strains by the piggyBac transposase JOURNAL Unpublished REFERENCE 2 (bases 1 to 5769) AUTHORS Kasai Y, Harayama S. TITLE Direct Submission JOURNAL Submitted (08-JUN-2017) Contact:Yuki Kasai Chuo University, Research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 TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (08-JUN-2017) Contact:Yuki Kasai Chuo University, Research and Development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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