Basic Vector Information
- Vector Name:
- pUC18R6KT-mini-Tn7T-Km
- Antibiotic Resistance:
- Ampicillin, Kanamycin
- Length:
- 4911 bp
- Type:
- Bacterial Expression
- Replication origin:
- R6K γ ori
- Cloning Method:
- Restriction Enzyme
pUC18R6KT-mini-Tn7T-Km 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 ℃.
pUC18R6KT-mini-Tn7T-Km vector Sequence
LOCUS 40924_45043 4911 bp DNA circular SYN 13-MAY-2021
DEFINITION mini-Tn7 base vector with transcriptional terminator and oriT.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4911)
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. 2005 Jun;2(6):443-8.
PUBMED 15908923
REFERENCE 2 (bases 1 to 4911)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 4911)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods."; date: "2005-06"; volume: "2(6)"; pages: "443-8"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4911
/mol_type="other DNA"
/organism="synthetic DNA construct"
mobile_element 369..534
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
CDS 1083..1874
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
protein_bind complement(1985..2032)
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
terminator complement(2080..2166)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator complement(2269..2363)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
primer_bind complement(2391..2407)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter 3087..3191
/label=AmpR promoter
CDS 3192..4049
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(4461..4845)
/direction=LEFT
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
oriT complement(join(4868..4911,1..65))
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
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