pRMTn-Km vector (V003668)

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LOCUS       40924_37438        7715 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRMTn-Km DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7715)
  AUTHORS   Miyazaki R, van der Meer JR.
  TITLE     A New Large-DNA-Fragment Delivery System Based on Integrase Activity
            from an Integrative and Conjugative Element
  JOURNAL   Appl. Environ. Microbiol. 79 (14), 4440-4447 (2013)
  PUBMED    23686268
REFERENCE   2  (bases 1 to 7715)
  AUTHORS   Miyazaki R, van der Meer JR.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JAN-2013) Contact:Ryo Miyazaki University of Lausanne,
            Department of Fundamental Microbiology; Batiment Biophore, Quartier 
            UNIL-Sorge, Lausanne 1015, Switzerland
REFERENCE   3  (bases 1 to 7715)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7715)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2013"; volume: "79"; issue: "14"; 
            pages: "4440-4447"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-JAN-2013) Contact:Ryo Miyazaki University of Lausanne, 
            Department of Fundamental Microbiology; Batiment Biophore, Quartier 
            UNIL-Sorge, Lausanne 1015, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..19
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     misc_feature    38..85
                     /label=FRT
                     /note="FRT"
     protein_bind    complement(38..85)
                     /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)."
     oriT            225..333
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             612..1403
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     protein_bind    complement(1581..1628)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1986..3959)
                     /codon_start=1
                     /gene="intB13"
                     /product="P4-type integrase"
                     /label=intB13
                     /protein_id="BAN28924.1"
                     /translation="MTSIGPKSLLRSLEMALSDLIVRQAKTTGKRYTLYDNDCLSLMVS
                     AAGGKSWMFRYSWLGKQKRMALGGYPALSLREARAERDKAQALIARGIDPQIERDQRRH
                     AAKLAGEYTFKNVFDAWVEHRRKELKEGRQSTLSQILRIFNKDVLPTLGKMSIYDIRRP
                     QLLGVLAAIEKRKAFTTAEKVRTWFNQMFRYALVIAEGLEVNPAADLDVVAEPKPPVAH
                     NPYLHLPELPEFLQKLRRYNPRGWQTQLGVRLLFLTGVRTGELRLAEPEQFDLDRGFWI
                     IPPEVVKQLQDEMRKAGKRPQDVPPYIVPLSLQAIEIVRYLLGVMRPAQKYLLSHRSEL
                     KKRISENTLNKAVQLMGYEGRLTGHGIRGTISTALNEIGYPKIWVDAQLSHSDPNKVSS
                     AYNHAKYVEPRRRMMQDWADRLDLLEQGEVQAASAHLTIRIDGVPAMAEVEEAVDVAPT
                     VAEPGVSGSPPVAATPIVVTPNSGGITFQRLSQVPPPPAHAPESEVSAIQREREEMLAM
                     YESPNNLPVALFGKLAGKSKDQINRELKAGKLLSISLGNRGQRVPDWQLVPLKRRLAQA
                     LMNQCPHVDSWALYRLLTKPHSNLGNRAAIDVVTPTNVGKVLQAVTPYKEFERSSTDET
                     PQFSEFVRQLQRHVNALEEAPC"
     gene            complement(1986..3959)
                     /gene="intB13"
                     /label=intB13
     protein_bind    complement(4055..4071)
                     /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)."
     misc_feature    4163..4180
                     /label=attP
                     /note="attP"
     regulatory      4249..4277
                     /label=Pcirc
                     /note="Pcirc"
                     /regulatory_class="promoter"
     terminator      complement(4339..4425)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(4528..4622)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    4623..4667
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    complement(4705..4723)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     rep_origin      complement(4738..5126)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     terminator      complement(5143..5182)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(5189..6046)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6047..6138)
                     /label=AmpR promoter
     CDS             complement(6195..7622)
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"