Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | pTn7xK | Antibiotic Resistance | nptII |
| Length | 4208 bp | Type | Cloning vector |
| Source | Bruckbauer ST, Kvitko BH, Karkhoff-Schweizer RR, Schweizer HP. | ||
pTn7xK 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 ℃.
pTn7xK vector Sequence
LOCUS Exported 4208 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pTn7xK, complete sequence.
ACCESSION KF813059
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4208)
AUTHORS Bruckbauer ST, Kvitko BH, Karkhoff-Schweizer RR, Schweizer HP.
TITLE Tn5/7-lux: A versatile tool for the identification and capture of
promoters in Gram-negative bacteria
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4208)
AUTHORS Bruckbauer ST, Kvitko BH, Karkhoff-Schweizer RR, Schweizer HP.
TITLE Direct Submission
JOURNAL Submitted (04-NOV-2013) Microbiology, Immunology and Pathology,
Colorado State University, 2025 Campus Delivery, Fort Collins, CO
80523, USA
REFERENCE 3 (bases 1 to 4208)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4208
/organism="Cloning vector pTn7xK"
/lab_host="Escherichia coli"
/mol_type="other DNA"
/db_xref="taxon:1454556"
misc_feature 58..105
/label=FRT site
/note="FRT site"
protein_bind complement(58..105)
/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 491..1285
/codon_start=1
/gene="nptII"
/product="neomycin phosphotransferase II"
/label=nptII
/protein_id="AHH35069.1"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
gene 491..1285
/gene="nptII"
/label=nptII
misc_feature 1384..1431
/label=FRT site
/note="FRT site"
protein_bind complement(1384..1431)
/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)."
regulatory 1472..1569
/regulatory_class="terminator"
/note="phage lambda"
terminator 1477..1563
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
regulatory 1666..1760
/regulatory_class="terminator"
/note="Escherichia coli rrnB operon"
terminator 1666..1760
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
primer_bind complement(1788..1804)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
repeat_region 1809..2008
/note="Tn7R; Tn7 right end"
oriT 2607..2868
oriT 2753..2861
/note="incP origin of transfer"
rep_origin 2892..3273
/label=R6K
/note="R6K"
rep_origin 2912..3268
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
repeat_region 3484..3649
/note="Tn7L; Tn7 left end"
mobile_element 3484..3649
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (left end of the Tn7 transposon)"
protein_bind 3680..3704
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 3735..4200
/regulatory_class="promoter"
/note="Pseudomonas aeruginosa PA4974 gene"
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