LOCUS       40924_37433        7306 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRMTn-Gm DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7306)
  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 7306)
  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 7306)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7306)
  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..7306
                     /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"
     promoter        347..375
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             564..1094
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     protein_bind    complement(1172..1219)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1577..3550)
                     /codon_start=1
                     /gene="intB13"
                     /product="P4-type integrase"
                     /label=intB13
                     /protein_id="BAN28928.1"
                     /translation="MTSIGPKSLLRSLEMALSDLIVRQAKTTGKRYTLYDNDCLSLMVS
                     AAGGKSWMFRYSWLGKQKRMALGGYPALSLREARAERDKAQALIARGIDPQIERDQRRH
                     AAKLAGEYTFKNVFDAWVEHRRKELKEGRQSTLSQILRIFNKDVLPTLGKMSIYDIRRP
                     QLLGVLAAIEKRKAFTTAEKVRTWFNQMFRYALVIAEGLEVNPAADLDVVAEPKPPVAH
                     NPYLHLPELPEFLQKLRRYNPRGWQTQLGVRLLFLTGVRTGELRLAEPEQFDLDRGFWI
                     IPPEVVKQLQDEMRKAGKRPQDVPPYIVPLSLQAIEIVRYLLGVMRPAQKYLLSHRSEL
                     KKRISENTLNKAVQLMGYEGRLTGHGIRGTISTALNEIGYPKIWVDAQLSHSDPNKVSS
                     AYNHAKYVEPRRRMMQDWADRLDLLEQGEVQAASAHLTIRIDGVPAMAEVEEAVDVAPT
                     VAEPGVSGSPPVAATPIVVTPNSGGITFQRLSQVPPPPAHAPESEVSAIQREREEMLAM
                     YESPNNLPVALFGKLAGKSKDQINRELKAGKLLSISLGNRGQRVPDWQLVPLKRRLAQA
                     LMNQCPHVDSWALYRLLTKPHSNLGNRAAIDVVTPTNVGKVLQAVTPYKEFERSSTDET
                     PQFSEFVRQLQRHVNALEEAPC"
     gene            complement(1577..3550)
                     /gene="intB13"
                     /label=intB13
     protein_bind    complement(3646..3662)
                     /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    3754..3771
                     /label=attP
                     /note="attP"
     regulatory      3840..3868
                     /label=Pcirc
                     /note="Pcirc"
                     /regulatory_class="promoter"
     terminator      complement(3930..4016)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(4119..4213)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    4214..4258
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    complement(4296..4314)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     rep_origin      complement(4329..4717)
                     /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(4734..4773)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(4780..5637)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5638..5729)
                     /label=AmpR promoter
     CDS             complement(5786..7213)
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
ORIGIN
        1 ctgtctctta tacacatctg agcttgcatg cctgcatgaa gttcctatac tttctagaga
       61 ataggaactt cggaatagga acttcttttc ttttccgctg cataaccctg cttcggggtc
      121 attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa
      181 agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa
      241 gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg
      301 ctcaacggga atcctgctct gcgaggctgg ccgtaggccg gccgaattga cataagcctg
      361 ttcggttcgt aaactgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt
      421 gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt
      481 ttttgtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg
      541 tttgatgtta tggagcagca acgatgttac gcagcagcaa cgatgttacg cagcagggca
      601 gtcgccctaa aacaaagtta ggtggctcaa gtatgggcat cattcgcaca tgtaggctcg
      661 gccctgacca agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt gagttcggag
      721 acgtagccac ctactcccaa catcagccgg actccgatta cctcgggaac ttgctccgta
      781 gtaagacatt catcgcgctt gctgccttcg accaagaagc ggttgttggc gctctcgcgg
      841 cttacgttct gcccaagttt gagcagccgc gtagtgagat ctatatctat gatctcgcag
      901 tctccggcga gcaccggagg cagggcattg ccaccgcgct catcaatctc ctcaagcatg
      961 aggccaacgc gcttggtgct tatgtgatct acgtgcaagc agattacggt gacgatcccg
     1021 cagtggctct ctatacaaag ttgggcatac gggaagaagt gatgcacttt gatatcgacc
     1081 caagtaccgc cacctaacaa ttcgttcaag ccgagatcgg cttcccggcc gcggagttgt
     1141 tcggtaaatt gtcacaacgc cgcggagtac tgaagttcct atactttcta gagaatagga
     1201 acttcggaat aggtacttcc catggatggc acgcttctcg ttgtgcaggc ggcgcatatc
     1261 gacgacgtgc acgccttcgt ggcccacgac ccctacgtac aggcaggtct gtttgccgag
     1321 ctgtcggttc gcacatggga atggaccttg gggcggggaa cctcataaat cctgggtaca
     1381 ggcctcctac ccgccgctca aggaacggac atccggacgc ggaggcgaaa aaccttaccg
     1441 ggaggaattg cagtcggtct tggccgtctg gttgatcagt tcgttgcaat agccgcccac
     1501 gacgggattt aacgcactcc tagcgtctag gtaaacacgc gttgcggcac gccgttggcg
     1561 tcgaagtcct ggctcgtcag cacggcgctt cttcgagcgc attcacgtgg cgttgcaact
     1621 ggcgaacgaa ctcggagaac tggggtgtct cgtctgtact ggagcgctcg aactccttat
     1681 agggcgtgac ggcttgcaat actttgccca cgttggtcgg tgtgacgaca tcgatggctg
     1741 cacggtttcc gaggttggaa tggggcttgg tcaacagacg atacagcgcc caagaatcta
     1801 catgcgggca ttggttcatg agcgcttggg ccagccggcg cttgagcggc accagttgcc
     1861 agtcgggaac gcgctgcccc cggttgccca agctgatgga cagtagcttg cctgccttca
     1921 gctcgcggtt gatctggtct ttggacttcc cagccagctt cccgaacagc gcgaccggca
     1981 gattgttcgg cgactcgtac atggccagca tttcctcacg ctcgcgctgg atggcagaga
     2041 cctccgactc aggagcatgc gccggaggcg gcggcacctg cgacaggcgc tggaacgtga
     2101 ttccgccgct gttgggggtc acgacgatgg gcgtggccgc cacgggcggt gagccggaga
     2161 caccgggctc ggcgaccgtc ggggccacgt ccaccgcttc ctccacttcc gccatcgctg
     2221 gcaccccgtc gatacggatg gtcaggtgcg cgctcgcagc ttgcacttcg ccctgttcga
     2281 gcaggtccag gcgatcggcc cagtcctgca tcatgcggcg acgcggctcc acgtacttgg
     2341 cgtggttgta ggccgaactc accttgttgg ggtcggagtg cgaaagctgc gcgtccaccc
     2401 aaatcttcgg gtagccgatc tcgttgagtg ccgtcgagat ggtgccgcgg atgccgtgcc
     2461 cggtcaggcg cccctcgtag cccatgagct gcacggcctt gttgagggtg ttctcgctga
     2521 tgcgtttctt gagttcgctg cggtgcgaca gcaggtactt ctgcgccggc cgcatcacgc
     2581 ccaggagata gcgcacgatc tcgatggcct gcagggacag gggcacgatg tagggcggca
     2641 cgtcctgcgg ccgcttgccg gccttgcgca tttcgtcctg gagttgcttg acgacctccg
     2701 gcgggatgat ccaaaagccc cggtcgaggt cgaactgctc gggttcggcc aaccgcagct
     2761 cgcccgtgcg cacccccgtc aggaacagta gccgcacgcc aagctgggtc tgccagcccc
     2821 gggggttgta gcgtcgcagc ttctgaagga actcgggcag ttcgggcagg tgcagatagg
     2881 ggttgtgggc caccgggggc ttgggctcgg ccaccacgtc caggtccgca gccgggttga
     2941 cttccagccc ctcggcgatg accaaggcat agcggaacat ctggttgaac caggtgcgga
     3001 ccttctcggc cgtggtgaac gccttgcgct tctcgatcgc cgccaggacg cccaggagct
     3061 ggggccggcg aatgtcatag atggacatct tccctagggt gggcagcacg tccttgttga
     3121 agatgcgcag gatctgcgaa agcgtactct ggcggccttc cttgagttcc ttgcggcgat
     3181 gctcgaccca ggcatcgaag acgttcttga aggtgtattc gccggccagc ttggccgcgt
     3241 gccgacgctg atcgcgttcg atttggggat cgatccccct ggcgatcagg gcctgggcct
     3301 tgtcccgctc ggcgcgggcc tcgcgcaggc tgagggcagg atagccgccc agggccatgc
     3361 gcttttgctt gcccagccag gagtagcgga acatccaaga cttgccgcct gcggcagaga
     3421 ccatcaggct caggcagtcg ttgtcgtaga gggtgtagcg tttgccggtg gtcttggcct
     3481 gccggacgat cagatcagag agtgccattt cgaggctcct aagtaatgac ttaggcccta
     3541 tgctcgtcat atgtcccgct cagccccagc aactatccgg aagccgcctc acccgtattc
     3601 ctgtgcctag attggcgcct aaaaaaggct ggctgtgggt ggaaattgtt atccgctcac
     3661 aatttccgtg gcgctcgctg gaatgaaaaa aggggccgaa gcccctcttt tcaacgactt
     3721 acagacgtca gtagaagtct gtagatcata acttggagcg ggaaacgaga cgtttactag
     3781 ggcagtaagt cgttgattcc caagctcttt cttcgccgag gtggccagcg aagacctaga
     3841 ttgtagccta ttttttcgtc cccgccaaca aagatcgtcc aaagagttcc actggcatcc
     3901 atagacccac acccagcaga attcggcgga tttgtcctac tcaggagagc gttcaccgac
     3961 aaacaacaga taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc
     4021 ctcaagctag agagtcatta ccccaggcgt ttaagggcac caataactgc cttaaaaaaa
     4081 ttacgccccg ccctgccact catcgcagtc tagcttggat tctcaccaat aaaaaacgcc
     4141 cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct
     4201 atcaacagga gtcactagta gatctctcga ctctagagga tccccgggta ccgagctcga
     4261 attcgcgcgg ccgcggccta ggcggcctta attaaagatg tgtataagag acagctgggc
     4321 gcgccccatg tcagccgtta agtgttcctg tgtcactcaa aattgctttg agaggctcta
     4381 agggcttctc agtgcgttac atccctggct tgttgtccac aaccgttaaa ccttaaaggc
     4441 tttaaaagcc ttatatattc ttttttttct tataaaactt aaaaccttag aggctattta
     4501 agttgctgat ttatattaat tttattgttc aaacatgaga gcttagtacg tgaaacatga
     4561 gagcttagta cgttagccat gagagcttag tacgttagcc atgagggttt agttcgttaa
     4621 acatgagagc ttagtacgtt aaacatgaga gcttagtacg tgaaacatga gagcttagta
     4681 cgtactatca acaggttgaa ctgctgatct tcagatcggc gcgccggggg tcgtccaaaa
     4741 aaaaaggctc caaaaggagc ctttaattgt atcggtttac caatgtttaa tcaggctcgc
     4801 gccaatttcc gcaatctgac ggttacgttc atccatcgtc gcctggctgc cggtggtata
     4861 aatcaccaca atacggctcg gtttaccatc cgggcccagc gccgcaataa tgccacggct
     4921 gccacgttcg cccgcaccgc ttttatccgc aataaaccag ccagccggca gcgcgctacg
     4981 cagcagcgga cccgccactt tatccgcttc catccaatca atcagttgct ggcggcttgc
     5041 cagggtcagc agctcacccg tcagcagttt acgcagggtg gtcgccattg ctgccggcat
     5101 ggtggtatca cgttcatcgt tcggaatcgc ttcgttcagt tccggttccc aacgatccag
     5161 acgggtcaca tgatcgccca tgttatgcag aaacgcggtc agttctttcg gaccgccaat
     5221 ggtcgtcagc agcaggttcg ccgcggtgtt atcgctcatg gtaatcgccg cgctgcacag
     5281 ttcacgcacg gtcatgccat cggtcagatg tttttcggtc accgggctat attccaccag
     5341 atcgttctgg ctataatgaa tacgacggcc cagttgttcc tggcccgcat ccacacggct
     5401 cagaaccgca ccgcacagca gcactttaaa ggtgctcatc atcggaaaac gttcttccgg
     5461 acgaaaagat tccagaattt tgccgctgtt cagatccagt tcaatatagc ccacgcgcgc
     5521 acccagttga tcttccgcat ctttcacttt caccagggtt tccggatgcg caaacaccgg
     5581 caggcaaaac gccgcaaaaa acggaatcag cgccacacga aaatgctgaa tgctcatact
     5641 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5701 atttgaatgt atttagaaaa ataaacaaat aggggaatta aaaaaaagcc cgctcattag
     5761 gcgggctact aatttaaatc gattagattt taatgccctg cgccatcagg tctttcgcgg
     5821 ccagaaagcc atccagtttg ctttgcagcg cttcccaacc ttcccacagc gcaccccagc
     5881 tcgcaatgcc ggtacgtttg ctatccataa agccgcccag acgcgcaatc gccatatacg
     5941 cccattgcag gctgcccgct ttttctttgc gtttgcgttt gcctttatcc agatagccca
     6001 gcagttggca ttcatccggg gtcagcacgg tttccgcgct ctggctttca acgtgttccg
     6061 cttctttcag caggccctgc gcacgcagtg cttgcggcgg agtaaaagat tcacgcagtt
     6121 gcagcagacg caccgccaca aagctcagaa tgctcaccat acgttccagg ttatccggtt
     6181 cttccatacg ctgacgttcc gcacccgcac ccgttttcca cgctttgtga aattcttcaa
     6241 tgcgccaacg atgggtataa atatcaatca cacgcagcgc ttgggccaga ctttccaccg
     6301 gctcgctggt cagcagcagc catttcagcg gggtttcgcc tttcggcgga ttaatttctt
     6361 cggccagcac cgcgttcagg gtaatgttgc cctgtttcag ggtaatacgg ccgctacgca
     6421 ggctcaggct cgctttacgc gccggacggt ttttacgttt gccacgttta tccaccacgc
     6481 ctttctgcgg aatgctaatc tgatagccgc ccagttccgg ctggtttttc aggtgatcat
     6541 acagatacag gccgctttcc acatctttac gcggatgttt gctacgcacc acaaaacgtt
     6601 cgttatgggc cagtttatct tgcagatacg catgaatatc cgcttcacga tcgcacaccg
     6661 caatcacgtt gctcatcatg ctgcccattc tcagacgcga agttgcagca gcggccagcc
     6721 atttgccgct ttctttttca tccgcatccg ccggatcatc cggacgcatc caccattctt
     6781 gatgcagcag gcccacggta cgaaaggtgg tcgcttccag cagcagcacg ctatgcaccc
     6841 accaaccacg gcttttatcc tgaatgctac ccagtttgcc cagttcttcc gccacctgat
     6901 gacgatagct cagagaggtg gtatcttcaa ttgccagcag ttccggaaat tcctgggcca
     6961 gtttcacggt ctgcatggca cccgctttac gaatcgcttc cgcgctcacg ttcggattac
     7021 gaataaaacg atacgcgcct tcctgcatgg ctttgctgcc ttcgctgcta atggtaatgc
     7081 ttttgccgct atatttggcc agttgcgccg caacattcac cagacgcgcg gtacgacgcg
     7141 gatcacccag cgcagcacta gaaaacacgc ttttcgccca atccgccgca cgatgcagtg
     7201 cactggtaat catgaacgtt accatgttag gaggtcacat ggaacatcag attctggaaa
     7261 acgggaaagg ttccgttcag gacgctactt gtgtagttta aaccag
//
