Basic Vector Information
pUC18T-mini-Tn7T-nat 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 ℃.
pUC18T-mini-Tn7T-nat vector Sequence
LOCUS 40924_45083 4838 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pUC18T-mini-Tn7T-nat, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4838) AUTHORS Lehman SS, Mladinich KM, Boonyakanog A, Mima T, Karkhoff-Schweizer RR, Schweizer HP. TITLE Versatile nourseothricin and streptomycin/spectinomycin resistance gene cassettes and their use in chromosome integration vectors JOURNAL J. Microbiol. Methods 129, 8-13 (2016) PUBMED 27457407 REFERENCE 2 (bases 1 to 4838) AUTHORS Lehman SS, Mladinich K, Boonyakanog A, Mima T, Karkhoff-Schweizer RR, Schweizer HP. TITLE Direct Submission JOURNAL Submitted (17-MAR-2016) Molecular Genetics and Microbiology, University of Florida, 2055 Mowry Road, Gainesville, FL 32610, USA REFERENCE 3 (bases 1 to 4838) TITLE Direct Submission REFERENCE 4 (bases 1 to 4838) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Microbiol. Methods 129, 8-13 (2016)" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (17-MAR-2016) Molecular Genetics and Microbiology, University of Florida, 2055 Mowry Road, Gainesville, FL 32610, USA" COMMENT SGRef: number: 3; type: "Journal Article" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..4838 /mol_type="other DNA" /organism="synthetic DNA construct" mobile_element 382..580 /mobile_element_type="transposon:Tn7R" /label=n7R /note="right end of Tn7" primer_bind 585..601 /label=KS primer /note="common sequencing primer, one of multiple similar variants" terminator 629..723 /label=lambda t0 terminator /note="transcription terminator from phage lambda" terminator 826..912 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" misc_feature 965..987 /note="loxP; Cre recombinase site" promoter 1117..1460 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 1467..2030 /label=NrsR /note="nourseothricin acetyltransferase" protein_bind complement(2210..2243) /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." misc_feature 2269..2343 /note="MCS; multiple cloning site" mobile_element complement(2355..2520) /label=Tn7L /note="mini-Tn7 element (left end of the Tn7 transposon)" oriT complement(2679..2787) /direction=LEFT /label=oriT /note="incP origin of transfer" rep_origin complement(3019..3607) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3781..4638) /label=AmpR /note="beta-lactamase" promoter complement(4639..4743) /label=AmpR promoter
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