Basic Vector Information
pUC18T-miniTn7T-LAC-Apr 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-miniTn7T-LAC-Apr vector Sequence
LOCUS 40924_45108 6403 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pUC18T-miniTn7T-LAC-Apr, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6403) AUTHORS Ducas-Mowchun K, DeSilva PM, Kumar A. TITLE Direct Submission JOURNAL Submitted (02-MAY-2018) Department of Microbiology, University of Manitoba, 45, Chancellor Circle, Buller Building (Room 412), Winnipeg, MB R3T2N2, Canada REFERENCE 2 (bases 1 to 6403) TITLE Direct Submission REFERENCE 3 (bases 1 to 6403) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted (02-MAY-2018) Department of Microbiology, University of Manitoba, 45, Chancellor Circle, Buller Building (Room 412), Winnipeg, MB R3T2N2, Canada" COMMENT SGRef: number: 2; type: "Journal Article" COMMENT ##Assembly-Data-START## Assembly Method :: Geneious v. R11 Sequencing Technology :: Illumina ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..6403 /mol_type="other DNA" /organism="synthetic DNA construct" mobile_element 383..581 /mobile_element_type="transposon:Tn7R" /label=n7R primer_bind 586..602 /label=KS primer /note="common sequencing primer, one of multiple similar variants" terminator 630..724 /label=lambda t0 terminator /note="transcription terminator from phage lambda" terminator 827..913 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" protein_bind 959..1006 /label=FRT /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS complement(1124..1924) /label=ApmR /note="aminoglycoside 3-N-acetyltransferase type IV" promoter complement(2165..2193) /label=Pc promoter /note="class 1 integron promoter" misc_feature 2234..2281 /label=FRT /note="FRT" protein_bind 2234..2281 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS complement(2327..3406) /label=lacI /note="lac repressor" promoter complement(3407..3484) /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." promoter 3714..3742 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 3750..3766 /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)." misc_feature 3780..3860 /label=MCS /note="MCS" mobile_element complement(3869..4034) /label=Tn7L /note="mini-Tn7 element (left end of the Tn7 transposon)" oriT complement(4193..4301) /direction=LEFT /label=oriT /note="incP origin of transfer" rep_origin complement(4533..5121) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5295..6152) /label=AmpR /note="beta-lactamase" promoter complement(6153..6257) /label=AmpR promoter
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