LOCUS       40924_37443        8886 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRMTn-Tc DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8886)
  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 8886)
  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 8886)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8886)
  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..8886
                     /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        420..448
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             496..1683
                     /label=TcR
                     /note="tetracycline efflux protein"
     protein_bind    complement(2752..2799)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(3157..5130)
                     /codon_start=1
                     /gene="intB13"
                     /product="P4-type integrase"
                     /label=intB13
                     /protein_id="BAN28920.1"
                     /translation="MTSIGPKSLLRSLEMALSDLIVRQAKTTGKRYTLYDNDCLSLMVS
                     AAGGKSWMFRYSWLGKQKRMALGGYPALSLREARAERDKAQALIARGIDPQIERDQRRH
                     AAKLAGEYTFKNVFDAWVEHRRKELKEGRQSTLSQILRIFNKDVLPTLGKMSIYDIRRP
                     QLLGVLAAIEKRKAFTTAEKVRTWFNQMFRYALVIAEGLEVNPAADLDVVAEPKPPVAH
                     NPYLHLPELPEFLQKLRRYNPRGWQTQLGVRLLFLTGVRTGELRLAEPEQFDLDRGFWI
                     IPPEVVKQLQDEMRKAGKRPQDVPPYIVPLSLQAIEIVRYLLGVMRPAQKYLLSHRSEL
                     KKRISENTLNKAVQLMGYEGRLTGHGIRGTISTALNEIGYPKIWVDAQLSHSDPNKVSS
                     AYNHAKYVEPRRRMMQDWADRLDLLEQGEVQAASAHLTIRIDGVPAMAEVEEAVDVAPT
                     VAEPGVSGSPPVAATPIVVTPNSGGITFQRLSQVPPPPAHAPESEVSAIQREREEMLAM
                     YESPNNLPVALFGKLAGKSKDQINRELKAGKLLSISLGNRGQRVPDWQLVPLKRRLAQA
                     LMNQCPHVDSWALYRLLTKPHSNLGNRAAIDVVTPTNVGKVLQAVTPYKEFERSSTDET
                     PQFSEFVRQLQRHVNALEEAPC"
     gene            complement(3157..5130)
                     /gene="intB13"
                     /label=intB13
     protein_bind    complement(5226..5242)
                     /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    5334..5351
                     /label=attP
                     /note="attP"
     regulatory      5420..5448
                     /label=Pcirc
                     /note="Pcirc"
                     /regulatory_class="promoter"
     terminator      complement(5510..5596)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(5699..5793)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    5794..5838
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    complement(5876..5894)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     rep_origin      complement(5909..6297)
                     /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(6314..6353)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(6360..7217)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7218..7309)
                     /label=AmpR promoter
     CDS             complement(7366..8793)
                     /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 gccgatttca gtgcaattta
      361 tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt
      421 tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa ttgctaacgc
      481 agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg caccgtcacc
      541 ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt gcgggatatc
      601 gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata tgcgttgatg
      661 caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg ccgcccagtc
      721 ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac cacacccgtc
      781 ctgtggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
      841 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
      901 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
      961 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
     1021 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat gcccttgaga
     1081 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     1141 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     1201 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     1261 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     1321 gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg
     1381 ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc
     1441 gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag
     1501 cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc gctgatcgtc
     1561 acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc
     1621 gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg
     1681 acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga attggagcca
     1741 atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg
     1801 cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg tcctggccac
     1861 gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
     1921 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
     1981 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
     2041 aaacgcggaa gtcccctacg tgctgctgaa gttgcccgca acagagagtg gaaccaaccg
     2101 gtgataccac gatactatga ctgagagtca acgccatgag cggcctcatt tcttattctg
     2161 agttacaaca gtccgcaccg ctgtccggta gctccttccg gtgggcgcgg ggcatgacta
     2221 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
     2281 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgccct
     2341 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
     2401 catctgtatt aacgaagcgc taaccgtttt tatcaggctc tgggaggcag aataaatgat
     2461 catatcgtca attattacct ccacggggag agcctgagca aactggcctc aggcatttga
     2521 gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata
     2581 agcggctatt taacgaccct gccctgaacc gacgaccggg tcgaatttgc tttcgaattt
     2641 ctgccattca tccgcttatt atcacttatt caggcgtagc accaggcgtt taagggcacc
     2701 aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgaagttcct
     2761 atactttcta gagaatagga acttcggaat aggtacttcc catggatggc acgcttctcg
     2821 ttgtgcaggc ggcgcatatc gacgacgtgc acgccttcgt ggcccacgac ccctacgtac
     2881 aggcaggtct gtttgccgag ctgtcggttc gcacatggga atggaccttg gggcggggaa
     2941 cctcataaat cctgggtaca ggcctcctac ccgccgctca aggaacggac atccggacgc
     3001 ggaggcgaaa aaccttaccg ggaggaattg cagtcggtct tggccgtctg gttgatcagt
     3061 tcgttgcaat agccgcccac gacgggattt aacgcactcc tagcgtctag gtaaacacgc
     3121 gttgcggcac gccgttggcg tcgaagtcct ggctcgtcag cacggcgctt cttcgagcgc
     3181 attcacgtgg cgttgcaact ggcgaacgaa ctcggagaac tggggtgtct cgtctgtact
     3241 ggagcgctcg aactccttat agggcgtgac ggcttgcaat actttgccca cgttggtcgg
     3301 tgtgacgaca tcgatggctg cacggtttcc gaggttggaa tggggcttgg tcaacagacg
     3361 atacagcgcc caagaatcta catgcgggca ttggttcatg agcgcttggg ccagccggcg
     3421 cttgagcggc accagttgcc agtcgggaac gcgctgcccc cggttgccca agctgatgga
     3481 cagtagcttg cctgccttca gctcgcggtt gatctggtct ttggacttcc cagccagctt
     3541 cccgaacagc gcgaccggca gattgttcgg cgactcgtac atggccagca tttcctcacg
     3601 ctcgcgctgg atggcagaga cctccgactc aggagcatgc gccggaggcg gcggcacctg
     3661 cgacaggcgc tggaacgtga ttccgccgct gttgggggtc acgacgatgg gcgtggccgc
     3721 cacgggcggt gagccggaga caccgggctc ggcgaccgtc ggggccacgt ccaccgcttc
     3781 ctccacttcc gccatcgctg gcaccccgtc gatacggatg gtcaggtgcg cgctcgcagc
     3841 ttgcacttcg ccctgttcga gcaggtccag gcgatcggcc cagtcctgca tcatgcggcg
     3901 acgcggctcc acgtacttgg cgtggttgta ggccgaactc accttgttgg ggtcggagtg
     3961 cgaaagctgc gcgtccaccc aaatcttcgg gtagccgatc tcgttgagtg ccgtcgagat
     4021 ggtgccgcgg atgccgtgcc cggtcaggcg cccctcgtag cccatgagct gcacggcctt
     4081 gttgagggtg ttctcgctga tgcgtttctt gagttcgctg cggtgcgaca gcaggtactt
     4141 ctgcgccggc cgcatcacgc ccaggagata gcgcacgatc tcgatggcct gcagggacag
     4201 gggcacgatg tagggcggca cgtcctgcgg ccgcttgccg gccttgcgca tttcgtcctg
     4261 gagttgcttg acgacctccg gcgggatgat ccaaaagccc cggtcgaggt cgaactgctc
     4321 gggttcggcc aaccgcagct cgcccgtgcg cacccccgtc aggaacagta gccgcacgcc
     4381 aagctgggtc tgccagcccc gggggttgta gcgtcgcagc ttctgaagga actcgggcag
     4441 ttcgggcagg tgcagatagg ggttgtgggc caccgggggc ttgggctcgg ccaccacgtc
     4501 caggtccgca gccgggttga cttccagccc ctcggcgatg accaaggcat agcggaacat
     4561 ctggttgaac caggtgcgga ccttctcggc cgtggtgaac gccttgcgct tctcgatcgc
     4621 cgccaggacg cccaggagct ggggccggcg aatgtcatag atggacatct tccctagggt
     4681 gggcagcacg tccttgttga agatgcgcag gatctgcgaa agcgtactct ggcggccttc
     4741 cttgagttcc ttgcggcgat gctcgaccca ggcatcgaag acgttcttga aggtgtattc
     4801 gccggccagc ttggccgcgt gccgacgctg atcgcgttcg atttggggat cgatccccct
     4861 ggcgatcagg gcctgggcct tgtcccgctc ggcgcgggcc tcgcgcaggc tgagggcagg
     4921 atagccgccc agggccatgc gcttttgctt gcccagccag gagtagcgga acatccaaga
     4981 cttgccgcct gcggcagaga ccatcaggct caggcagtcg ttgtcgtaga gggtgtagcg
     5041 tttgccggtg gtcttggcct gccggacgat cagatcagag agtgccattt cgaggctcct
     5101 aagtaatgac ttaggcccta tgctcgtcat atgtcccgct cagccccagc aactatccgg
     5161 aagccgcctc acccgtattc ctgtgcctag attggcgcct aaaaaaggct ggctgtgggt
     5221 ggaaattgtt atccgctcac aatttccgtg gcgctcgctg gaatgaaaaa aggggccgaa
     5281 gcccctcttt tcaacgactt acagacgtca gtagaagtct gtagatcata acttggagcg
     5341 ggaaacgaga cgtttactag ggcagtaagt cgttgattcc caagctcttt cttcgccgag
     5401 gtggccagcg aagacctaga ttgtagccta ttttttcgtc cccgccaaca aagatcgtcc
     5461 aaagagttcc actggcatcc atagacccac acccagcaga attcggcgga tttgtcctac
     5521 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag
     5581 cctttcgttt tatttgatgc ctcaagctag agagtcatta ccccaggcgt ttaagggcac
     5641 caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagtc tagcttggat
     5701 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     5761 ctgaggtcat tactggatct atcaacagga gtcactagta gatctctcga ctctagagga
     5821 tccccgggta ccgagctcga attcgcgcgg ccgcggccta ggcggcctta attaaagatg
     5881 tgtataagag acagctgggc gcgccccatg tcagccgtta agtgttcctg tgtcactcaa
     5941 aattgctttg agaggctcta agggcttctc agtgcgttac atccctggct tgttgtccac
     6001 aaccgttaaa ccttaaaggc tttaaaagcc ttatatattc ttttttttct tataaaactt
     6061 aaaaccttag aggctattta agttgctgat ttatattaat tttattgttc aaacatgaga
     6121 gcttagtacg tgaaacatga gagcttagta cgttagccat gagagcttag tacgttagcc
     6181 atgagggttt agttcgttaa acatgagagc ttagtacgtt aaacatgaga gcttagtacg
     6241 tgaaacatga gagcttagta cgtactatca acaggttgaa ctgctgatct tcagatcggc
     6301 gcgccggggg tcgtccaaaa aaaaaggctc caaaaggagc ctttaattgt atcggtttac
     6361 caatgtttaa tcaggctcgc gccaatttcc gcaatctgac ggttacgttc atccatcgtc
     6421 gcctggctgc cggtggtata aatcaccaca atacggctcg gtttaccatc cgggcccagc
     6481 gccgcaataa tgccacggct gccacgttcg cccgcaccgc ttttatccgc aataaaccag
     6541 ccagccggca gcgcgctacg cagcagcgga cccgccactt tatccgcttc catccaatca
     6601 atcagttgct ggcggcttgc cagggtcagc agctcacccg tcagcagttt acgcagggtg
     6661 gtcgccattg ctgccggcat ggtggtatca cgttcatcgt tcggaatcgc ttcgttcagt
     6721 tccggttccc aacgatccag acgggtcaca tgatcgccca tgttatgcag aaacgcggtc
     6781 agttctttcg gaccgccaat ggtcgtcagc agcaggttcg ccgcggtgtt atcgctcatg
     6841 gtaatcgccg cgctgcacag ttcacgcacg gtcatgccat cggtcagatg tttttcggtc
     6901 accgggctat attccaccag atcgttctgg ctataatgaa tacgacggcc cagttgttcc
     6961 tggcccgcat ccacacggct cagaaccgca ccgcacagca gcactttaaa ggtgctcatc
     7021 atcggaaaac gttcttccgg acgaaaagat tccagaattt tgccgctgtt cagatccagt
     7081 tcaatatagc ccacgcgcgc acccagttga tcttccgcat ctttcacttt caccagggtt
     7141 tccggatgcg caaacaccgg caggcaaaac gccgcaaaaa acggaatcag cgccacacga
     7201 aaatgctgaa tgctcatact cttccttttt caatattatt gaagcattta tcagggttat
     7261 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggaatta
     7321 aaaaaaagcc cgctcattag gcgggctact aatttaaatc gattagattt taatgccctg
     7381 cgccatcagg tctttcgcgg ccagaaagcc atccagtttg ctttgcagcg cttcccaacc
     7441 ttcccacagc gcaccccagc tcgcaatgcc ggtacgtttg ctatccataa agccgcccag
     7501 acgcgcaatc gccatatacg cccattgcag gctgcccgct ttttctttgc gtttgcgttt
     7561 gcctttatcc agatagccca gcagttggca ttcatccggg gtcagcacgg tttccgcgct
     7621 ctggctttca acgtgttccg cttctttcag caggccctgc gcacgcagtg cttgcggcgg
     7681 agtaaaagat tcacgcagtt gcagcagacg caccgccaca aagctcagaa tgctcaccat
     7741 acgttccagg ttatccggtt cttccatacg ctgacgttcc gcacccgcac ccgttttcca
     7801 cgctttgtga aattcttcaa tgcgccaacg atgggtataa atatcaatca cacgcagcgc
     7861 ttgggccaga ctttccaccg gctcgctggt cagcagcagc catttcagcg gggtttcgcc
     7921 tttcggcgga ttaatttctt cggccagcac cgcgttcagg gtaatgttgc cctgtttcag
     7981 ggtaatacgg ccgctacgca ggctcaggct cgctttacgc gccggacggt ttttacgttt
     8041 gccacgttta tccaccacgc ctttctgcgg aatgctaatc tgatagccgc ccagttccgg
     8101 ctggtttttc aggtgatcat acagatacag gccgctttcc acatctttac gcggatgttt
     8161 gctacgcacc acaaaacgtt cgttatgggc cagtttatct tgcagatacg catgaatatc
     8221 cgcttcacga tcgcacaccg caatcacgtt gctcatcatg ctgcccattc tcagacgcga
     8281 agttgcagca gcggccagcc atttgccgct ttctttttca tccgcatccg ccggatcatc
     8341 cggacgcatc caccattctt gatgcagcag gcccacggta cgaaaggtgg tcgcttccag
     8401 cagcagcacg ctatgcaccc accaaccacg gcttttatcc tgaatgctac ccagtttgcc
     8461 cagttcttcc gccacctgat gacgatagct cagagaggtg gtatcttcaa ttgccagcag
     8521 ttccggaaat tcctgggcca gtttcacggt ctgcatggca cccgctttac gaatcgcttc
     8581 cgcgctcacg ttcggattac gaataaaacg atacgcgcct tcctgcatgg ctttgctgcc
     8641 ttcgctgcta atggtaatgc ttttgccgct atatttggcc agttgcgccg caacattcac
     8701 cagacgcgcg gtacgacgcg gatcacccag cgcagcacta gaaaacacgc ttttcgccca
     8761 atccgccgca cgatgcagtg cactggtaat catgaacgtt accatgttag gaggtcacat
     8821 ggaacatcag attctggaaa acgggaaagg ttccgttcag gacgctactt gtgtagttta
     8881 aaccag
//
