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pTn7xK vector (V002583#)

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

FRT site nptII FRT site phage lambda Escherichia coli rrnB operon KS primer Tn7R R6K Tn7L attB1 Pseudomonas aeruginosa PA4974 gene

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

Copy Sequence

Download GeneBank File(.gb)

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  JOURNAL   Unpublished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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