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  JOURNAL   Unpublished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"