LOCUS       Exported                5017 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRMR6K-Km DNA, complete sequence.
ACCESSION   AB777648
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5017)
  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 5017)
  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 5017)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5017
                     /organism="Cloning vector pRMR6K-Km"
                     /mol_type="other DNA"
                     /db_xref="taxon:1284602"
     CDS             complement(175..969)
                     /codon_start=1
                     /gene="npt"
                     /product="kanamycin phosphotransferase"
                     /label=npt
                     /protein_id="BAN28915.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(175..969)
                     /gene="npt"
                     /label=npt
     misc_feature    1238..1491
                     /label=oriT
                     /note="oriT"
     oriT            1248..1356
                     /note="incP origin of transfer"
     misc_feature    1496..1543
                     /label=FRT
                     /note="FRT"
     protein_bind    1496..1543
                     /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)."
     rep_origin      1550..1941
                     /label=oriR6K
                     /note="oriR6K"
     rep_origin      1550..1938
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    1940..1975
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      1976..2264
                     /regulatory_class="terminator"
                     /note="bacteriophage lambda T0, Escherichia coli rrnB T1"
     terminator      1976..2070
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      2173..2259
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      2321..2349
                     /regulatory_class="promoter"
                     /note="Pcirc"
     misc_feature    2418..2435
                     /label=attP
                     /note="attP"
     misc_feature    2527..2545
                     /label=lacO
                     /note="lacO"
     protein_bind    2527..2543
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     CDS             2639..4612
                     /codon_start=1
                     /gene="intB13"
                     /product="P4-type integrase"
                     /label=intB13
                     /protein_id="BAN28916.1"
                     /translation="MTSIGPKSLLRSLEMALSDLIVRQAKTTGKRYTLYDNDCLSLMVS
                     AAGGKSWMFRYSWLGKQKRMALGGYPALSLREARAERDKAQALIARGIDPQIERDQRRH
                     AAKLAGEYTFKNVFDAWVEHRRKELKEGRQSTLSQILRIFNKDVLPTLGKMSIYDIRRP
                     QLLGVLAAIEKRKAFTTAEKVRTWFNQMFRYALVIAEGLEVNPAADLDVVAEPKPPVAH
                     NPYLHLPELPEFLQKLRRYNPRGWQTQLGVRLLFLTGVRTGELRLAEPEQFDLDRGFWI
                     IPPEVVKQLQDEMRKAGKRPQDVPPYIVPLSLQAIEIVRYLLGVMRPAQKYLLSHRSEL
                     KKRISENTLNKAVQLMGYEGRLTGHGIRGTISTALNEIGYPKIWVDAQLSHSDPNKVSS
                     AYNHAKYVEPRRRMMQDWADRLDLLEQGEVQAASAHLTIRIDGVPAMAEVEEAVDVAPT
                     VAEPGVSGSPPVAATPIVVTPNSGGITFQRLSQVPPPPAHAPESEVSAIQREREEMLAM
                     YESPNNLPVALFGKLAGKSKDQINRELKAGKLLSISLGNRGQRVPDWQLVPLKRRLAQA
                     LMNQCPHVDSWALYRLLTKPHSNLGNRAAIDVVTPTNVGKVLQAVTPYKEFERSSTDET
                     PQFSEFVRQLQRHVNALEEAPC"
     gene            2639..4612
                     /gene="intB13"
                     /label=intB13
     misc_feature    4970..5017
                     /label=FRT
                     /note="FRT"
     protein_bind    4970..5017
                     /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)."