LOCUS       40924_37418        4608 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRMR6K-Gm DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4608)
  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 4608)
  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 4608)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4608)
  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..4608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(78..608)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(797..825)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     oriT            complement(839..947)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     protein_bind    1087..1134
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     rep_origin      1141..1529
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     misc_feature    1531..1566
                     /label=multiple cloning site
                     /note="multiple cloning site"
     terminator      1567..1661
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      1764..1850
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     regulatory      1912..1940
                     /label=Pcirc
                     /note="Pcirc"
                     /regulatory_class="promoter"
     misc_feature    2009..2026
                     /label=attP
                     /note="attP"
     protein_bind    2118..2134
                     /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             2230..4203
                     /codon_start=1
                     /gene="intB13"
                     /product="P4-type integrase"
                     /label=intB13
                     /protein_id="BAN28918.1"
                     /translation="MTSIGPKSLLRSLEMALSDLIVRQAKTTGKRYTLYDNDCLSLMVS
                     AAGGKSWMFRYSWLGKQKRMALGGYPALSLREARAERDKAQALIARGIDPQIERDQRRH
                     AAKLAGEYTFKNVFDAWVEHRRKELKEGRQSTLSQILRIFNKDVLPTLGKMSIYDIRRP
                     QLLGVLAAIEKRKAFTTAEKVRTWFNQMFRYALVIAEGLEVNPAADLDVVAEPKPPVAH
                     NPYLHLPELPEFLQKLRRYNPRGWQTQLGVRLLFLTGVRTGELRLAEPEQFDLDRGFWI
                     IPPEVVKQLQDEMRKAGKRPQDVPPYIVPLSLQAIEIVRYLLGVMRPAQKYLLSHRSEL
                     KKRISENTLNKAVQLMGYEGRLTGHGIRGTISTALNEIGYPKIWVDAQLSHSDPNKVSS
                     AYNHAKYVEPRRRMMQDWADRLDLLEQGEVQAASAHLTIRIDGVPAMAEVEEAVDVAPT
                     VAEPGVSGSPPVAATPIVVTPNSGGITFQRLSQVPPPPAHAPESEVSAIQREREEMLAM
                     YESPNNLPVALFGKLAGKSKDQINRELKAGKLLSISLGNRGQRVPDWQLVPLKRRLAQA
                     LMNQCPHVDSWALYRLLTKPHSNLGNRAAIDVVTPTNVGKVLQAVTPYKEFERSSTDET
                     PQFSEFVRQLQRHVNALEEAPC"
     gene            2230..4203
                     /gene="intB13"
                     /label=intB13
     misc_feature    4561..4608
                     /label=FRT
                     /note="FRT"
     protein_bind    4561..4608
                     /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 agtactccgc ggcgttgtga caatttaccg aacaactccg cggccgggaa gccgatctcg
       61 gcttgaacga attgttaggt ggcggtactt gggtcgatat caaagtgcat cacttcttcc
      121 cgtatgccca actttgtata gagagccact gcgggatcgt caccgtaatc tgcttgcacg
      181 tagatcacat aagcaccaag cgcgttggcc tcatgcttga ggagattgat gagcgcggtg
      241 gcaatgccct gcctccggtg ctcgccggag actgcgagat catagatata gatctcacta
      301 cgcggctgct caaacttggg cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg
      361 tcgaaggcag caagcgcgat gaatgtctta ctacggagca agttcccgag gtaatcggag
      421 tccggctgat gttgggagta ggtggctacg tctccgaact cacgaccgaa aagatcaaga
      481 gcagcccgca tggatttgac ttggtcaggg ccgagcctac atgtgcgaat gatgcccata
      541 cttgagccac ctaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctg
      601 cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg
      661 cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca tgaaaaccgc
      721 cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgagcgcat
      781 acgctacttg cattacagtt tacgaaccga acaggcttat gtcaattcgg ccggcctacg
      841 gccagcctcg cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga
      901 agaaggaaca cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt
      961 ggatacacca aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa
     1021 aggatggata taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa
     1081 agaaaagaag ttcctattcc gaagttccta ttctctagaa agtataggaa cttcatgcat
     1141 gatctgaaga tcagcagttc aacctgttga tagtacgtac taagctctca tgtttcacgt
     1201 actaagctct catgtttaac gtactaagct ctcatgttta acgaactaaa ccctcatggc
     1261 taacgtacta agctctcatg gctaacgtac taagctctca tgtttcacgt actaagctct
     1321 catgtttgaa caataaaatt aatataaatc agcaacttaa atagcctcta aggttttaag
     1381 ttttataaga aaaaaaagaa tatataaggc ttttaaagcc tttaaggttt aacggttgtg
     1441 gacaacaagc cagggatgta acgcactgag aagcccttag agcctctcaa agcaattttg
     1501 agtgacacag gaacacttaa cggctgacat gggcccgagc tcggtaccct cgagagatct
     1561 actagtgact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     1621 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagctag actgcgatga
     1681 gtggcagggc ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggggt
     1741 aatgactctc tagcttgagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     1801 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgaatt
     1861 ctgctgggtg tgggtctatg gatgccagtg gaactctttg gacgatcttt gttggcgggg
     1921 acgaaaaaat aggctacaat ctaggtcttc gctggccacc tcggcgaaga aagagcttgg
     1981 gaatcaacga cttactgccc tagtaaacgt ctcgtttccc gctccaagtt atgatctaca
     2041 gacttctact gacgtctgta agtcgttgaa aagaggggct tcggcccctt ttttcattcc
     2101 agcgagcgcc acggaaattg tgagcggata acaatttcca cccacagcca gcctttttta
     2161 ggcgccaatc taggcacagg aatacgggtg aggcggcttc cggatagttg ctggggctga
     2221 gcgggacata tgacgagcat agggcctaag tcattactta ggagcctcga aatggcactc
     2281 tctgatctga tcgtccggca ggccaagacc accggcaaac gctacaccct ctacgacaac
     2341 gactgcctga gcctgatggt ctctgccgca ggcggcaagt cttggatgtt ccgctactcc
     2401 tggctgggca agcaaaagcg catggccctg ggcggctatc ctgccctcag cctgcgcgag
     2461 gcccgcgccg agcgggacaa ggcccaggcc ctgatcgcca gggggatcga tccccaaatc
     2521 gaacgcgatc agcgtcggca cgcggccaag ctggccggcg aatacacctt caagaacgtc
     2581 ttcgatgcct gggtcgagca tcgccgcaag gaactcaagg aaggccgcca gagtacgctt
     2641 tcgcagatcc tgcgcatctt caacaaggac gtgctgccca ccctagggaa gatgtccatc
     2701 tatgacattc gccggcccca gctcctgggc gtcctggcgg cgatcgagaa gcgcaaggcg
     2761 ttcaccacgg ccgagaaggt ccgcacctgg ttcaaccaga tgttccgcta tgccttggtc
     2821 atcgccgagg ggctggaagt caacccggct gcggacctgg acgtggtggc cgagcccaag
     2881 cccccggtgg cccacaaccc ctatctgcac ctgcccgaac tgcccgagtt ccttcagaag
     2941 ctgcgacgct acaacccccg gggctggcag acccagcttg gcgtgcggct actgttcctg
     3001 acgggggtgc gcacgggcga gctgcggttg gccgaacccg agcagttcga cctcgaccgg
     3061 ggcttttgga tcatcccgcc ggaggtcgtc aagcaactcc aggacgaaat gcgcaaggcc
     3121 ggcaagcggc cgcaggacgt gccgccctac atcgtgcccc tgtccctgca ggccatcgag
     3181 atcgtgcgct atctcctggg cgtgatgcgg ccggcgcaga agtacctgct gtcgcaccgc
     3241 agcgaactca agaaacgcat cagcgagaac accctcaaca aggccgtgca gctcatgggc
     3301 tacgaggggc gcctgaccgg gcacggcatc cgcggcacca tctcgacggc actcaacgag
     3361 atcggctacc cgaagatttg ggtggacgcg cagctttcgc actccgaccc caacaaggtg
     3421 agttcggcct acaaccacgc caagtacgtg gagccgcgtc gccgcatgat gcaggactgg
     3481 gccgatcgcc tggacctgct cgaacagggc gaagtgcaag ctgcgagcgc gcacctgacc
     3541 atccgtatcg acggggtgcc agcgatggcg gaagtggagg aagcggtgga cgtggccccg
     3601 acggtcgccg agcccggtgt ctccggctca ccgcccgtgg cggccacgcc catcgtcgtg
     3661 acccccaaca gcggcggaat cacgttccag cgcctgtcgc aggtgccgcc gcctccggcg
     3721 catgctcctg agtcggaggt ctctgccatc cagcgcgagc gtgaggaaat gctggccatg
     3781 tacgagtcgc cgaacaatct gccggtcgcg ctgttcggga agctggctgg gaagtccaaa
     3841 gaccagatca accgcgagct gaaggcaggc aagctactgt ccatcagctt gggcaaccgg
     3901 gggcagcgcg ttcccgactg gcaactggtg ccgctcaagc gccggctggc ccaagcgctc
     3961 atgaaccaat gcccgcatgt agattcttgg gcgctgtatc gtctgttgac caagccccat
     4021 tccaacctcg gaaaccgtgc agccatcgat gtcgtcacac cgaccaacgt gggcaaagta
     4081 ttgcaagccg tcacgcccta taaggagttc gagcgctcca gtacagacga gacaccccag
     4141 ttctccgagt tcgttcgcca gttgcaacgc cacgtgaatg cgctcgaaga agcgccgtgc
     4201 tgacgagcca ggacttcgac gccaacggcg tgccgcaacg cgtgtttacc tagacgctag
     4261 gagtgcgtta aatcccgtcg tgggcggcta ttgcaacgaa ctgatcaacc agacggccaa
     4321 gaccgactgc aattcctccc ggtaaggttt ttcgcctccg cgtccggatg tccgttcctt
     4381 gagcggcggg taggaggcct gtacccagga tttatgaggt tccccgcccc aaggtccatt
     4441 cccatgtgcg aaccgacagc tcggcaaaca gacctgcctg tacgtagggg tcgtgggcca
     4501 cgaaggcgtg cacgtcgtcg atatgcgccg cctgcacaac gagaagcgtg ccatccatgg
     4561 gaagtaccta ttccgaagtt cctattctct agaaagtata ggaacttc
//
