LOCUS       40924_37423        5017 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRMR6K-Km DNA, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5017)
  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..5017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(178..969)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     oriT            complement(1248..1356)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     protein_bind    1496..1543
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     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"
     terminator      1976..2070
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      2173..2259
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     regulatory      2321..2349
                     /label=Pcirc
                     /note="Pcirc"
                     /regulatory_class="promoter"
     misc_feature    2418..2435
                     /label=attP
                     /note="attP"
     protein_bind    2527..2543
                     /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)."
     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)."
ORIGIN
        1 agtactgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt
       61 cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc
      121 gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa
      181 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
      241 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
      301 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
      361 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
      421 atcctcgccg tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc
      481 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
      541 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
      601 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
      661 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
      721 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
      781 ctcgtcctgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
      841 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
      901 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
      961 ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca
     1021 tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc
     1081 agagggcgcc ccagctggca attccggttc gcttgctgtc cataaaaccg cccagtctag
     1141 ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgttttccct
     1201 tgtccagata gcccagtagc tgacattcat cccaggtggc cggcctacgg ccagcctcgc
     1261 agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac
     1321 ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa
     1381 ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat
     1441 accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaaa gaaaagaagt
     1501 tcctattccg aagttcctat tctctagaaa gtataggaac ttcatgcatg atctgaagat
     1561 cagcagttca acctgttgat agtacgtact aagctctcat gtttcacgta ctaagctctc
     1621 atgtttaacg tactaagctc tcatgtttaa cgaactaaac cctcatggct aacgtactaa
     1681 gctctcatgg ctaacgtact aagctctcat gtttcacgta ctaagctctc atgtttgaac
     1741 aataaaatta atataaatca gcaacttaaa tagcctctaa ggttttaagt tttataagaa
     1801 aaaaaagaat atataaggct tttaaagcct ttaaggttta acggttgtgg acaacaagcc
     1861 agggatgtaa cgcactgaga agcccttaga gcctctcaaa gcaattttga gtgacacagg
     1921 aacacttaac ggctgacatg ggcccgagct cggtaccctc gagagatcta ctagtgactc
     1981 ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt
     2041 tgccgccggg cgttttttat tggtgagaat ccaagctaga ctgcgatgag tggcagggcg
     2101 gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggggta atgactctct
     2161 agcttgaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     2221 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgaattc tgctgggtgt
     2281 gggtctatgg atgccagtgg aactctttgg acgatctttg ttggcgggga cgaaaaaata
     2341 ggctacaatc taggtcttcg ctggccacct cggcgaagaa agagcttggg aatcaacgac
     2401 ttactgccct agtaaacgtc tcgtttcccg ctccaagtta tgatctacag acttctactg
     2461 acgtctgtaa gtcgttgaaa agaggggctt cggccccttt tttcattcca gcgagcgcca
     2521 cggaaattgt gagcggataa caatttccac ccacagccag ccttttttag gcgccaatct
     2581 aggcacagga atacgggtga ggcggcttcc ggatagttgc tggggctgag cgggacatat
     2641 gacgagcata gggcctaagt cattacttag gagcctcgaa atggcactct ctgatctgat
     2701 cgtccggcag gccaagacca ccggcaaacg ctacaccctc tacgacaacg actgcctgag
     2761 cctgatggtc tctgccgcag gcggcaagtc ttggatgttc cgctactcct ggctgggcaa
     2821 gcaaaagcgc atggccctgg gcggctatcc tgccctcagc ctgcgcgagg cccgcgccga
     2881 gcgggacaag gcccaggccc tgatcgccag ggggatcgat ccccaaatcg aacgcgatca
     2941 gcgtcggcac gcggccaagc tggccggcga atacaccttc aagaacgtct tcgatgcctg
     3001 ggtcgagcat cgccgcaagg aactcaagga aggccgccag agtacgcttt cgcagatcct
     3061 gcgcatcttc aacaaggacg tgctgcccac cctagggaag atgtccatct atgacattcg
     3121 ccggccccag ctcctgggcg tcctggcggc gatcgagaag cgcaaggcgt tcaccacggc
     3181 cgagaaggtc cgcacctggt tcaaccagat gttccgctat gccttggtca tcgccgaggg
     3241 gctggaagtc aacccggctg cggacctgga cgtggtggcc gagcccaagc ccccggtggc
     3301 ccacaacccc tatctgcacc tgcccgaact gcccgagttc cttcagaagc tgcgacgcta
     3361 caacccccgg ggctggcaga cccagcttgg cgtgcggcta ctgttcctga cgggggtgcg
     3421 cacgggcgag ctgcggttgg ccgaacccga gcagttcgac ctcgaccggg gcttttggat
     3481 catcccgccg gaggtcgtca agcaactcca ggacgaaatg cgcaaggccg gcaagcggcc
     3541 gcaggacgtg ccgccctaca tcgtgcccct gtccctgcag gccatcgaga tcgtgcgcta
     3601 tctcctgggc gtgatgcggc cggcgcagaa gtacctgctg tcgcaccgca gcgaactcaa
     3661 gaaacgcatc agcgagaaca ccctcaacaa ggccgtgcag ctcatgggct acgaggggcg
     3721 cctgaccggg cacggcatcc gcggcaccat ctcgacggca ctcaacgaga tcggctaccc
     3781 gaagatttgg gtggacgcgc agctttcgca ctccgacccc aacaaggtga gttcggccta
     3841 caaccacgcc aagtacgtgg agccgcgtcg ccgcatgatg caggactggg ccgatcgcct
     3901 ggacctgctc gaacagggcg aagtgcaagc tgcgagcgcg cacctgacca tccgtatcga
     3961 cggggtgcca gcgatggcgg aagtggagga agcggtggac gtggccccga cggtcgccga
     4021 gcccggtgtc tccggctcac cgcccgtggc ggccacgccc atcgtcgtga cccccaacag
     4081 cggcggaatc acgttccagc gcctgtcgca ggtgccgccg cctccggcgc atgctcctga
     4141 gtcggaggtc tctgccatcc agcgcgagcg tgaggaaatg ctggccatgt acgagtcgcc
     4201 gaacaatctg ccggtcgcgc tgttcgggaa gctggctggg aagtccaaag accagatcaa
     4261 ccgcgagctg aaggcaggca agctactgtc catcagcttg ggcaaccggg ggcagcgcgt
     4321 tcccgactgg caactggtgc cgctcaagcg ccggctggcc caagcgctca tgaaccaatg
     4381 cccgcatgta gattcttggg cgctgtatcg tctgttgacc aagccccatt ccaacctcgg
     4441 aaaccgtgca gccatcgatg tcgtcacacc gaccaacgtg ggcaaagtat tgcaagccgt
     4501 cacgccctat aaggagttcg agcgctccag tacagacgag acaccccagt tctccgagtt
     4561 cgttcgccag ttgcaacgcc acgtgaatgc gctcgaagaa gcgccgtgct gacgagccag
     4621 gacttcgacg ccaacggcgt gccgcaacgc gtgtttacct agacgctagg agtgcgttaa
     4681 atcccgtcgt gggcggctat tgcaacgaac tgatcaacca gacggccaag accgactgca
     4741 attcctcccg gtaaggtttt tcgcctccgc gtccggatgt ccgttccttg agcggcgggt
     4801 aggaggcctg tacccaggat ttatgaggtt ccccgcccca aggtccattc ccatgtgcga
     4861 accgacagct cggcaaacag acctgcctgt acgtaggggt cgtgggccac gaaggcgtgc
     4921 acgtcgtcga tatgcgccgc ctgcacaacg agaagcgtgc catccatggg aagtacctat
     4981 tccgaagttc ctattctcta gaaagtatag gaacttc
//
