Basic Vector Information
- Vector Name:
- pUC18T-miniTn7T-LAC-Zeo
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5973 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Ducas-Mowchun K, DeSilva PM, Kumar A.
pUC18T-miniTn7T-LAC-Zeo 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-Zeo vector Sequence
LOCUS 40924_45113 5973 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUC18T-miniTn7T-LAC-Zeo, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5973)
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 5973)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5973)
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..5973
/mol_type="other DNA"
/organism="synthetic DNA construct"
mobile_element 412..610
/mobile_element_type="transposon:Tn7R"
/label=n7R
primer_bind 615..631
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 659..753
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
terminator 856..942
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind 988..1035
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS complement(1148..1519)
/label=BleoR
/note="antibiotic-binding protein"
promoter complement(1538..1585)
/label=EM7 promoter
/note="synthetic bacterial promoter"
promoter complement(1764..1792)
/label=Pc promoter
/note="class 1 integron promoter"
misc_feature 1833..1880
/label=FRT-L
/note="FRT-L"
protein_bind 1833..1880
/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(1926..3005)
/label=lacI
/note="lac repressor"
promoter complement(3006..3083)
/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 3313..3341
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 3349..3365
/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)."
mobile_element complement(3468..3633)
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
oriT complement(3792..3900)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
rep_origin complement(4132..4720)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4894..5751)
/label=AmpR
/note="beta-lactamase"
promoter complement(5752..5856)
/label=AmpR promoter
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