Basic Vector Information
- Vector Name:
- pUC18T-mini-Tn7T-LAC-Gm
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6047 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Choi KH, Mima T, Casart Y, Rholl D, Kumar A, Beacham IR, Schweizer HP.
pUC18T-mini-Tn7T-LAC-Gm 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-LAC-Gm vector Sequence
LOCUS 40924_45073 6047 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pUC18T-mini-Tn7T-LAC-Gm, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6047)
AUTHORS Choi KH, Mima T, Casart Y, Rholl D, Kumar A, Beacham IR, Schweizer
HP.
TITLE Genetic tools for select-agent-compliant manipulation of
Burkholderia pseudomallei
JOURNAL Appl. Environ. Microbiol. 74 (4), 1064-1075 (2008)
PUBMED 18156318
REFERENCE 2 (bases 1 to 6047)
AUTHORS Choi K-H., Schweizer HP.
TITLE Genetic tools for Pseudomonas and its related bacteria
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 6047)
AUTHORS Choi K-H., Schweizer HP.
TITLE Direct Submission
JOURNAL Submitted (23-OCT-2007) Microbiology, Immunology and Pathology,
Colorado State University, 1682 Campus delivery, Ft. Collins, CO
80523, USA
REFERENCE 4 (bases 1 to 6047)
AUTHORS Choi K-H., Schweizer HP.
TITLE Direct Submission
JOURNAL Submitted (25-MAY-2010) Microbiology, Immunology and Pathology,
Colorado State University, 1682 Campus delivery, Ft. Collins, CO
80523, USA
REFERENCE 5 (bases 1 to 6047)
TITLE Direct Submission
REFERENCE 6 (bases 1 to 6047)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2008"; volume: "74"; issue: "4"; pages:
"1064-1075"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(23-OCT-2007) Microbiology, Immunology and Pathology, Colorado State
University, 1682 Campus delivery, Ft. Collins, CO 80523, USA"
COMMENT SGRef: number: 4; type: "Journal Article"; journalName: "Submitted
(25-MAY-2010) Microbiology, Immunology and Pathology, Colorado State
University, 1682 Campus delivery, Ft. Collins, CO 80523, USA"
COMMENT SGRef: number: 5; type: "Journal Article"
COMMENT On May 25, 2010 this sequence version replaced EU240902.1.
FEATURES Location/Qualifiers
source 1..6047
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 382..580
/label=right end of Tn7
/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"
protein_bind 958..1005
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS complement(1138..1668)
/label=GmR
/note="gentamycin acetyltransferase"
promoter complement(1857..1885)
/label=Pc promoter
/note="class 1 integron promoter"
misc_feature 1926..1973
/note="FRT; Flp recombinase target site"
protein_bind 1926..1973
/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(2019..3098)
/label=lacI
/note="lac repressor"
promoter complement(3099..3176)
/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 3406..3434
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 3442..3458
/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 3475..3555
/note="MCS; multiple cloning site"
mobile_element complement(3564..3729)
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
oriT complement(3888..3996)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
rep_origin complement(4228..4816)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4990..5847)
/label=AmpR
/note="beta-lactamase"
promoter complement(5848..5952)
/label=AmpR promoter
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