LOCUS       40924_37428        6188 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRMR6K-Tc DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6188)
  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 6188)
  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 6188)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6188)
  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..6188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1069..2256)
                     /label=TcR
                     /note="tetracycline efflux protein"
     promoter        complement(2304..2332)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     oriT            complement(2419..2527)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     protein_bind    2667..2714
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     rep_origin      2721..3109
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     misc_feature    3111..3146
                     /label=multiple cloning site
                     /note="multiple cloning site"
     terminator      3147..3241
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      3344..3430
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     regulatory      3492..3520
                     /label=Pcirc
                     /note="Pcirc"
                     /regulatory_class="promoter"
     misc_feature    3589..3606
                     /label=attP
                     /note="attP"
     protein_bind    3698..3714
                     /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             3810..5783
                     /codon_start=1
                     /gene="intB13"
                     /product="P4-type integrase"
                     /label=intB13
                     /protein_id="BAN28914.1"
                     /translation="MTSIGPKSLLRSLEMALSDLIVRQAKTTGKRYTLYDNDCLSLMVS
                     AAGGKSWMFRYSWLGKQKRMALGGYPALSLREARAERDKAQALIARGIDPQIERDQRRH
                     AAKLAGEYTFKNVFDAWVEHRRKELKEGRQSTLSQILRIFNKDVLPTLGKMSIYDIRRP
                     QLLGVLAAIEKRKAFTTAEKVRTWFNQMFRYALVIAEGLEVNPAADLDVVAEPKPPVAH
                     NPYLHLPELPEFLQKLRRYNPRGWQTQLGVRLLFLTGVRTGELRLAEPEQFDLDRGFWI
                     IPPEVVKQLQDEMRKAGKRPQDVPPYIVPLSLQAIEIVRYLLGVMRPAQKYLLSHRSEL
                     KKRISENTLNKAVQLMGYEGRLTGHGIRGTISTALNEIGYPKIWVDAQLSHSDPNKVSS
                     AYNHAKYVEPRRRMMQDWADRLDLLEQGEVQAASAHLTIRIDGVPAMAEVEEAVDVAPT
                     VAEPGVSGSPPVAATPIVVTPNSGGITFQRLSQVPPPPAHAPESEVSAIQREREEMLAM
                     YESPNNLPVALFGKLAGKSKDQINRELKAGKLLSISLGNRGQRVPDWQLVPLKRRLAQA
                     LMNQCPHVDSWALYRLLTKPHSNLGNRAAIDVVTPTNVGKVLQAVTPYKEFERSSTDET
                     PQFSEFVRQLQRHVNALEEAPC"
     gene            3810..5783
                     /gene="intB13"
                     /label=intB13
     misc_feature    6141..6188
                     /label=FRT
                     /note="FRT"
     protein_bind    6141..6188
                     /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 agtactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt
       61 aaacgcctgg tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa
      121 agcaaattcg acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat
      181 tgctggttta ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc
      241 tgaggccagt ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcatttat
      301 tctgcctccc agagcctgat aaaaacggtt agcgcttcgt taatacagat gtaggtgttc
      361 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctt
      421 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gctgccggca
      481 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
      541 cccgcgccca ccggaaggag ctaccggaca gcggtgcgga ctgttgtaac tcagaataag
      601 aaatgaggcc gctcatggcg ttgactctca gtcatagtat cgtggtatca ccggttggtt
      661 ccactctctg ttgcgggcaa cttcagcagc acgtagggga cttccgcgtt tccagacttt
      721 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
      781 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
      841 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg
      901 acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata
      961 tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa
     1021 ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc
     1081 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac
     1141 aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag
     1201 cggtccagtg atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg
     1261 atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc
     1321 ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa
     1381 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     1441 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     1501 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     1561 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     1621 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     1681 catcggtcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     1741 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     1801 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     1861 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     1921 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatccaca ggacgggtgt
     1981 ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca ggactgggcg
     2041 gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt gcatcaacgc
     2101 atatagcgct agcagcacgc catagtgact ggcgatgctg tcggaatgga cgatatcccg
     2161 caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca gggtgacggt
     2221 gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac tgcgttagca
     2281 atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc aaacatgaga
     2341 attacaactt atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca
     2401 ctgaaatcgg ccggcctacg gccagcctcg cagagcagga ttcccgttga gcaccgccag
     2461 gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta cttcacctat
     2521 cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc tttggcaaaa
     2581 tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa tgaccccgaa
     2641 gcagggttat gcagcggaaa agaaaagaag ttcctattcc gaagttccta ttctctagaa
     2701 agtataggaa cttcatgcat gatctgaaga tcagcagttc aacctgttga tagtacgtac
     2761 taagctctca tgtttcacgt actaagctct catgtttaac gtactaagct ctcatgttta
     2821 acgaactaaa ccctcatggc taacgtacta agctctcatg gctaacgtac taagctctca
     2881 tgtttcacgt actaagctct catgtttgaa caataaaatt aatataaatc agcaacttaa
     2941 atagcctcta aggttttaag ttttataaga aaaaaaagaa tatataaggc ttttaaagcc
     3001 tttaaggttt aacggttgtg gacaacaagc cagggatgta acgcactgag aagcccttag
     3061 agcctctcaa agcaattttg agtgacacag gaacacttaa cggctgacat gggcccgagc
     3121 tcggtaccct cgagagatct actagtgact cctgttgata gatccagtaa tgacctcaga
     3181 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa
     3241 tccaagctag actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg
     3301 tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa aacgaaaggc
     3361 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     3421 taggacaaat ccgccgaatt ctgctgggtg tgggtctatg gatgccagtg gaactctttg
     3481 gacgatcttt gttggcgggg acgaaaaaat aggctacaat ctaggtcttc gctggccacc
     3541 tcggcgaaga aagagcttgg gaatcaacga cttactgccc tagtaaacgt ctcgtttccc
     3601 gctccaagtt atgatctaca gacttctact gacgtctgta agtcgttgaa aagaggggct
     3661 tcggcccctt ttttcattcc agcgagcgcc acggaaattg tgagcggata acaatttcca
     3721 cccacagcca gcctttttta ggcgccaatc taggcacagg aatacgggtg aggcggcttc
     3781 cggatagttg ctggggctga gcgggacata tgacgagcat agggcctaag tcattactta
     3841 ggagcctcga aatggcactc tctgatctga tcgtccggca ggccaagacc accggcaaac
     3901 gctacaccct ctacgacaac gactgcctga gcctgatggt ctctgccgca ggcggcaagt
     3961 cttggatgtt ccgctactcc tggctgggca agcaaaagcg catggccctg ggcggctatc
     4021 ctgccctcag cctgcgcgag gcccgcgccg agcgggacaa ggcccaggcc ctgatcgcca
     4081 gggggatcga tccccaaatc gaacgcgatc agcgtcggca cgcggccaag ctggccggcg
     4141 aatacacctt caagaacgtc ttcgatgcct gggtcgagca tcgccgcaag gaactcaagg
     4201 aaggccgcca gagtacgctt tcgcagatcc tgcgcatctt caacaaggac gtgctgccca
     4261 ccctagggaa gatgtccatc tatgacattc gccggcccca gctcctgggc gtcctggcgg
     4321 cgatcgagaa gcgcaaggcg ttcaccacgg ccgagaaggt ccgcacctgg ttcaaccaga
     4381 tgttccgcta tgccttggtc atcgccgagg ggctggaagt caacccggct gcggacctgg
     4441 acgtggtggc cgagcccaag cccccggtgg cccacaaccc ctatctgcac ctgcccgaac
     4501 tgcccgagtt ccttcagaag ctgcgacgct acaacccccg gggctggcag acccagcttg
     4561 gcgtgcggct actgttcctg acgggggtgc gcacgggcga gctgcggttg gccgaacccg
     4621 agcagttcga cctcgaccgg ggcttttgga tcatcccgcc ggaggtcgtc aagcaactcc
     4681 aggacgaaat gcgcaaggcc ggcaagcggc cgcaggacgt gccgccctac atcgtgcccc
     4741 tgtccctgca ggccatcgag atcgtgcgct atctcctggg cgtgatgcgg ccggcgcaga
     4801 agtacctgct gtcgcaccgc agcgaactca agaaacgcat cagcgagaac accctcaaca
     4861 aggccgtgca gctcatgggc tacgaggggc gcctgaccgg gcacggcatc cgcggcacca
     4921 tctcgacggc actcaacgag atcggctacc cgaagatttg ggtggacgcg cagctttcgc
     4981 actccgaccc caacaaggtg agttcggcct acaaccacgc caagtacgtg gagccgcgtc
     5041 gccgcatgat gcaggactgg gccgatcgcc tggacctgct cgaacagggc gaagtgcaag
     5101 ctgcgagcgc gcacctgacc atccgtatcg acggggtgcc agcgatggcg gaagtggagg
     5161 aagcggtgga cgtggccccg acggtcgccg agcccggtgt ctccggctca ccgcccgtgg
     5221 cggccacgcc catcgtcgtg acccccaaca gcggcggaat cacgttccag cgcctgtcgc
     5281 aggtgccgcc gcctccggcg catgctcctg agtcggaggt ctctgccatc cagcgcgagc
     5341 gtgaggaaat gctggccatg tacgagtcgc cgaacaatct gccggtcgcg ctgttcggga
     5401 agctggctgg gaagtccaaa gaccagatca accgcgagct gaaggcaggc aagctactgt
     5461 ccatcagctt gggcaaccgg gggcagcgcg ttcccgactg gcaactggtg ccgctcaagc
     5521 gccggctggc ccaagcgctc atgaaccaat gcccgcatgt agattcttgg gcgctgtatc
     5581 gtctgttgac caagccccat tccaacctcg gaaaccgtgc agccatcgat gtcgtcacac
     5641 cgaccaacgt gggcaaagta ttgcaagccg tcacgcccta taaggagttc gagcgctcca
     5701 gtacagacga gacaccccag ttctccgagt tcgttcgcca gttgcaacgc cacgtgaatg
     5761 cgctcgaaga agcgccgtgc tgacgagcca ggacttcgac gccaacggcg tgccgcaacg
     5821 cgtgtttacc tagacgctag gagtgcgtta aatcccgtcg tgggcggcta ttgcaacgaa
     5881 ctgatcaacc agacggccaa gaccgactgc aattcctccc ggtaaggttt ttcgcctccg
     5941 cgtccggatg tccgttcctt gagcggcggg taggaggcct gtacccagga tttatgaggt
     6001 tccccgcccc aaggtccatt cccatgtgcg aaccgacagc tcggcaaaca gacctgcctg
     6061 tacgtagggg tcgtgggcca cgaaggcgtg cacgtcgtcg atatgcgccg cctgcacaac
     6121 gagaagcgtg ccatccatgg gaagtaccta ttccgaagtt cctattctct agaaagtata
     6181 ggaacttc
//
