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)"
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 gccacctggg atgaatgtca
      361 gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca
      421 gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat
      481 tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt
      541 tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggatgag
      601 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
      661 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
      721 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
      781 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
      841 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
      901 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
      961 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     1021 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     1081 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     1141 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     1201 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     1261 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     1321 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     1381 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     1441 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     1501 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     1561 ggagttcttc gcccagtact gaagttccta tactttctag agaataggaa cttcggaata
     1621 ggtacttccc atggatggca cgcttctcgt tgtgcaggcg gcgcatatcg acgacgtgca
     1681 cgccttcgtg gcccacgacc cctacgtaca ggcaggtctg tttgccgagc tgtcggttcg
     1741 cacatgggaa tggaccttgg ggcggggaac ctcataaatc ctgggtacag gcctcctacc
     1801 cgccgctcaa ggaacggaca tccggacgcg gaggcgaaaa accttaccgg gaggaattgc
     1861 agtcggtctt ggccgtctgg ttgatcagtt cgttgcaata gccgcccacg acgggattta
     1921 acgcactcct agcgtctagg taaacacgcg ttgcggcacg ccgttggcgt cgaagtcctg
     1981 gctcgtcagc acggcgcttc ttcgagcgca ttcacgtggc gttgcaactg gcgaacgaac
     2041 tcggagaact ggggtgtctc gtctgtactg gagcgctcga actccttata gggcgtgacg
     2101 gcttgcaata ctttgcccac gttggtcggt gtgacgacat cgatggctgc acggtttccg
     2161 aggttggaat ggggcttggt caacagacga tacagcgccc aagaatctac atgcgggcat
     2221 tggttcatga gcgcttgggc cagccggcgc ttgagcggca ccagttgcca gtcgggaacg
     2281 cgctgccccc ggttgcccaa gctgatggac agtagcttgc ctgccttcag ctcgcggttg
     2341 atctggtctt tggacttccc agccagcttc ccgaacagcg cgaccggcag attgttcggc
     2401 gactcgtaca tggccagcat ttcctcacgc tcgcgctgga tggcagagac ctccgactca
     2461 ggagcatgcg ccggaggcgg cggcacctgc gacaggcgct ggaacgtgat tccgccgctg
     2521 ttgggggtca cgacgatggg cgtggccgcc acgggcggtg agccggagac accgggctcg
     2581 gcgaccgtcg gggccacgtc caccgcttcc tccacttccg ccatcgctgg caccccgtcg
     2641 atacggatgg tcaggtgcgc gctcgcagct tgcacttcgc cctgttcgag caggtccagg
     2701 cgatcggccc agtcctgcat catgcggcga cgcggctcca cgtacttggc gtggttgtag
     2761 gccgaactca ccttgttggg gtcggagtgc gaaagctgcg cgtccaccca aatcttcggg
     2821 tagccgatct cgttgagtgc cgtcgagatg gtgccgcgga tgccgtgccc ggtcaggcgc
     2881 ccctcgtagc ccatgagctg cacggccttg ttgagggtgt tctcgctgat gcgtttcttg
     2941 agttcgctgc ggtgcgacag caggtacttc tgcgccggcc gcatcacgcc caggagatag
     3001 cgcacgatct cgatggcctg cagggacagg ggcacgatgt agggcggcac gtcctgcggc
     3061 cgcttgccgg ccttgcgcat ttcgtcctgg agttgcttga cgacctccgg cgggatgatc
     3121 caaaagcccc ggtcgaggtc gaactgctcg ggttcggcca accgcagctc gcccgtgcgc
     3181 acccccgtca ggaacagtag ccgcacgcca agctgggtct gccagccccg ggggttgtag
     3241 cgtcgcagct tctgaaggaa ctcgggcagt tcgggcaggt gcagataggg gttgtgggcc
     3301 accgggggct tgggctcggc caccacgtcc aggtccgcag ccgggttgac ttccagcccc
     3361 tcggcgatga ccaaggcata gcggaacatc tggttgaacc aggtgcggac cttctcggcc
     3421 gtggtgaacg ccttgcgctt ctcgatcgcc gccaggacgc ccaggagctg gggccggcga
     3481 atgtcataga tggacatctt ccctagggtg ggcagcacgt ccttgttgaa gatgcgcagg
     3541 atctgcgaaa gcgtactctg gcggccttcc ttgagttcct tgcggcgatg ctcgacccag
     3601 gcatcgaaga cgttcttgaa ggtgtattcg ccggccagct tggccgcgtg ccgacgctga
     3661 tcgcgttcga tttggggatc gatccccctg gcgatcaggg cctgggcctt gtcccgctcg
     3721 gcgcgggcct cgcgcaggct gagggcagga tagccgccca gggccatgcg cttttgcttg
     3781 cccagccagg agtagcggaa catccaagac ttgccgcctg cggcagagac catcaggctc
     3841 aggcagtcgt tgtcgtagag ggtgtagcgt ttgccggtgg tcttggcctg ccggacgatc
     3901 agatcagaga gtgccatttc gaggctccta agtaatgact taggccctat gctcgtcata
     3961 tgtcccgctc agccccagca actatccgga agccgcctca cccgtattcc tgtgcctaga
     4021 ttggcgccta aaaaaggctg gctgtgggtg gaaattgtta tccgctcaca atttccgtgg
     4081 cgctcgctgg aatgaaaaaa ggggccgaag cccctctttt caacgactta cagacgtcag
     4141 tagaagtctg tagatcataa cttggagcgg gaaacgagac gtttactagg gcagtaagtc
     4201 gttgattccc aagctctttc ttcgccgagg tggccagcga agacctagat tgtagcctat
     4261 tttttcgtcc ccgccaacaa agatcgtcca aagagttcca ctggcatcca tagacccaca
     4321 cccagcagaa ttcggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     4381 aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tcaagctaga
     4441 gagtcattac cccaggcgtt taagggcacc aataactgcc ttaaaaaaat tacgccccgc
     4501 cctgccactc atcgcagtct agcttggatt ctcaccaata aaaaacgccc ggcggcaacc
     4561 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     4621 tcactagtag atctctcgac tctagaggat ccccgggtac cgagctcgaa ttcgcgcggc
     4681 cgcggcctag gcggccttaa ttaaagatgt gtataagaga cagctgggcg cgccccatgt
     4741 cagccgttaa gtgttcctgt gtcactcaaa attgctttga gaggctctaa gggcttctca
     4801 gtgcgttaca tccctggctt gttgtccaca accgttaaac cttaaaggct ttaaaagcct
     4861 tatatattct tttttttctt ataaaactta aaaccttaga ggctatttaa gttgctgatt
     4921 tatattaatt ttattgttca aacatgagag cttagtacgt gaaacatgag agcttagtac
     4981 gttagccatg agagcttagt acgttagcca tgagggttta gttcgttaaa catgagagct
     5041 tagtacgtta aacatgagag cttagtacgt gaaacatgag agcttagtac gtactatcaa
     5101 caggttgaac tgctgatctt cagatcggcg cgccgggggt cgtccaaaaa aaaaggctcc
     5161 aaaaggagcc tttaattgta tcggtttacc aatgtttaat caggctcgcg ccaatttccg
     5221 caatctgacg gttacgttca tccatcgtcg cctggctgcc ggtggtataa atcaccacaa
     5281 tacggctcgg tttaccatcc gggcccagcg ccgcaataat gccacggctg ccacgttcgc
     5341 ccgcaccgct tttatccgca ataaaccagc cagccggcag cgcgctacgc agcagcggac
     5401 ccgccacttt atccgcttcc atccaatcaa tcagttgctg gcggcttgcc agggtcagca
     5461 gctcacccgt cagcagttta cgcagggtgg tcgccattgc tgccggcatg gtggtatcac
     5521 gttcatcgtt cggaatcgct tcgttcagtt ccggttccca acgatccaga cgggtcacat
     5581 gatcgcccat gttatgcaga aacgcggtca gttctttcgg accgccaatg gtcgtcagca
     5641 gcaggttcgc cgcggtgtta tcgctcatgg taatcgccgc gctgcacagt tcacgcacgg
     5701 tcatgccatc ggtcagatgt ttttcggtca ccgggctata ttccaccaga tcgttctggc
     5761 tataatgaat acgacggccc agttgttcct ggcccgcatc cacacggctc agaaccgcac
     5821 cgcacagcag cactttaaag gtgctcatca tcggaaaacg ttcttccgga cgaaaagatt
     5881 ccagaatttt gccgctgttc agatccagtt caatatagcc cacgcgcgca cccagttgat
     5941 cttccgcatc tttcactttc accagggttt ccggatgcgc aaacaccggc aggcaaaacg
     6001 ccgcaaaaaa cggaatcagc gccacacgaa aatgctgaat gctcatactc ttcctttttc
     6061 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6121 tttagaaaaa taaacaaata ggggaattaa aaaaaagccc gctcattagg cgggctacta
     6181 atttaaatcg attagatttt aatgccctgc gccatcaggt ctttcgcggc cagaaagcca
     6241 tccagtttgc tttgcagcgc ttcccaacct tcccacagcg caccccagct cgcaatgccg
     6301 gtacgtttgc tatccataaa gccgcccaga cgcgcaatcg ccatatacgc ccattgcagg
     6361 ctgcccgctt tttctttgcg tttgcgtttg cctttatcca gatagcccag cagttggcat
     6421 tcatccgggg tcagcacggt ttccgcgctc tggctttcaa cgtgttccgc ttctttcagc
     6481 aggccctgcg cacgcagtgc ttgcggcgga gtaaaagatt cacgcagttg cagcagacgc
     6541 accgccacaa agctcagaat gctcaccata cgttccaggt tatccggttc ttccatacgc
     6601 tgacgttccg cacccgcacc cgttttccac gctttgtgaa attcttcaat gcgccaacga
     6661 tgggtataaa tatcaatcac acgcagcgct tgggccagac tttccaccgg ctcgctggtc
     6721 agcagcagcc atttcagcgg ggtttcgcct ttcggcggat taatttcttc ggccagcacc
     6781 gcgttcaggg taatgttgcc ctgtttcagg gtaatacggc cgctacgcag gctcaggctc
     6841 gctttacgcg ccggacggtt tttacgtttg ccacgtttat ccaccacgcc tttctgcgga
     6901 atgctaatct gatagccgcc cagttccggc tggtttttca ggtgatcata cagatacagg
     6961 ccgctttcca catctttacg cggatgtttg ctacgcacca caaaacgttc gttatgggcc
     7021 agtttatctt gcagatacgc atgaatatcc gcttcacgat cgcacaccgc aatcacgttg
     7081 ctcatcatgc tgcccattct cagacgcgaa gttgcagcag cggccagcca tttgccgctt
     7141 tctttttcat ccgcatccgc cggatcatcc ggacgcatcc accattcttg atgcagcagg
     7201 cccacggtac gaaaggtggt cgcttccagc agcagcacgc tatgcaccca ccaaccacgg
     7261 cttttatcct gaatgctacc cagtttgccc agttcttccg ccacctgatg acgatagctc
     7321 agagaggtgg tatcttcaat tgccagcagt tccggaaatt cctgggccag tttcacggtc
     7381 tgcatggcac ccgctttacg aatcgcttcc gcgctcacgt tcggattacg aataaaacga
     7441 tacgcgcctt cctgcatggc tttgctgcct tcgctgctaa tggtaatgct tttgccgcta
     7501 tatttggcca gttgcgccgc aacattcacc agacgcgcgg tacgacgcgg atcacccagc
     7561 gcagcactag aaaacacgct tttcgcccaa tccgccgcac gatgcagtgc actggtaatc
     7621 atgaacgtta ccatgttagg aggtcacatg gaacatcaga ttctggaaaa cgggaaaggt
     7681 tccgttcagg acgctacttg tgtagtttaa accag
//
