Basic Vector Information
- Vector Name:
- pUC18-mini-Tn7-LACM15
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8347 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer RR, Schweizer HP.
pUC18-mini-Tn7-LACM15 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 ℃.
pUC18-mini-Tn7-LACM15 vector Sequence
LOCUS 40924_44973 8347 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUC18-mini-Tn7-LACM15, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8347)
AUTHORS Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer
RR, Schweizer HP.
TITLE A Tn7-based broad-range bacterial cloning and expression system
JOURNAL Nat. Methods 2 (6), 443-448 (2005)
PUBMED 15908923
REFERENCE 2 (bases 1 to 8347)
AUTHORS Choi K-H., Gaynor JB, Karkhoff-Schweizer RR, Schweizer HP.
TITLE Direct Submission
JOURNAL Submitted (06-MAY-2004) Microbiology, Immunology and Pathology,
Colorado State University, Fort Collins, CO 80523, USA
REFERENCE 3 (bases 1 to 8347)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8347)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
Methods"; date: "2005"; volume: "2"; issue: "6"; pages: "443-448"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-MAY-2004) Microbiology, Immunology and Pathology, Colorado State
University, Fort Collins, CO 80523, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8347
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 382..580
/note="Tn7R; right end of Tn7"
protein_bind 600..647
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS complement(780..1310)
/label=GmR
/note="gentamycin acetyltransferase"
promoter complement(1499..1527)
/label=Pc promoter
/note="class 1 integron promoter"
misc_feature 1568..1615
/label=FRT, Flp recombinase target site
/note="FRT, Flp recombinase target site"
protein_bind 1568..1615
/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(1765..4710)
/label=lacZ
/note="beta-galactosidase"
primer_bind complement(4739..4755)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4763..4779)
/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(4787..4817)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4832..4853)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(4869..5948)
/label=lacI
/note="lac repressor"
promoter complement(5949..6026)
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
mobile_element complement(6093..6258)
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
rep_origin complement(6528..7116)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7290..8147)
/label=AmpR
/note="beta-lactamase"
promoter complement(8148..8252)
/label=AmpR promoter
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