LOCUS       40924_6287        3959 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBG35, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3959)
  AUTHORS   Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank 
            LM.
  TITLE     A Tn7-based device for calibrated heterologous gene expression in 
            Pseudomonas putida
  JOURNAL   ACS Synth Biol (2015) In press
  PUBMED    26133359
REFERENCE   2  (bases 1 to 3959)
  AUTHORS   Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank 
            LM.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUN-2015) System Biology, Centro Nacional de 
            Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 3959)
  AUTHORS   Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank 
            LM.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-NOV-2015) System Biology, Centro Nacional de 
            Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE   4  (bases 1 to 3959)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3959)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol (2015) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-JUN-2015) System Biology, Centro Nacional de Biotecnologia, 
            C/Darwin 3, Madrid 28049, Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (13-NOV-2015) System Biology, Centro Nacional de Biotecnologia, 
            C/Darwin 3, Madrid 28049, Spain"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
            On Nov 13, 2015 this sequence version replaced KT192141.1.
FEATURES             Location/Qualifiers
     source          1..3959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      9..49
                     /label=promoter 14b
                     /note="promoter 14b"
                     /regulatory_class="promoter"
     misc_feature    56..132
                     /label=BCD2 bicistronic linker
                     /note="BCD2 bicistronic linker"
     RBS             124..132
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             151..864
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     misc_feature    868..924
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      967..1061
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    1066..1264
                     /label=Tn7R transposase recognition site
                     /note="Tn7R transposase recognition site"
     CDS             1374..2186
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     oriT            2345..2453
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2472..2860
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     mobile_element  2872..3037
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     terminator      complement(3051..3137)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(3206..3736)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(3925..3953)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ttaattaatt tatttgacat gcgtgatgtt tagaattata atttggggac ctagggccca
       61 agttcactta aaaaggagat caacaatgaa agcaattttc gtactgaaac atcttaatca
      121 tgctaaggag gttttctaat gatcatggga attcataaag gtgaagaact gttcaccggt
      181 gttgttccga tcctggttga actggatggt gatgttaacg gccacaaatt ctctgttcgt
      241 ggtgaaggtg aaggtgatgc aaccaacggt aaactgaccc tgaaattcat ctgcactacc
      301 ggtaaactgc cggttccatg gccgactctg gtgactaccc tgacctatgg tgttcagtgt
      361 ttttctcgtt acccggatca catgaagcag catgatttct tcaaatctgc aatgccggaa
      421 ggttatgtac aggagcgcac catttctttc aaagacgatg gcacctacaa aacccgtgca
      481 gaggttaaat ttgaaggtga tactctggtg aaccgtattg aactgaaagg cattgatttc
      541 aaagaggacg gcaacatcct gggccacaaa ctggaatata acttcaactc ccataacgtt
      601 tacatcaccg cagacaaaca gaagaacggt atcaaagcta acttcaaaat tcgccataac
      661 gttgaagacg gtagcgtaca gctggcggac cactaccagc agaacactcc gatcggtgat
      721 ggtccggttc tgctgccgga taaccactac ctgtccaccc agtctaaact gtccaaagac
      781 ccgaacgaaa agcgcgacca catggtgctg ctggagttcg ttactgcagc aggtatcacg
      841 cacggcatgg atgaactcta caaataagaa ttcgagctcg gtacccgggg atcctctaga
      901 gtcgacctgc aggcatgcaa gcttgcggcc gcgtcgtgac tgggaaaacc ctggcgacta
      961 gtcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     1021 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccagccacg cccctcttta
     1081 atacgacggg caatttgcac ttcagaaaat gaagagtttg ctttagccat aacaaaagtc
     1141 cagtatgctt tttcacagca taactggact gatttcagtt tacaactatt ctgtctagtt
     1201 taagacttta ttgtcatagt ttagatctat tttgttcagt ttaagacttt attgtccgcc
     1261 cacaatttaa atttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     1321 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     1381 atattcagcg tgaaacgagc tgtagccgtc cgcgtctgaa cagcaacatg gatgcggatc
     1441 tgtatggcta taaatgggcg cgtgataacg tgggtcagag cggcgcgacc atttatcgtc
     1501 tgtatggcaa accggatgcg ccggaactgt ttctgaaaca tggcaaaggc agcgtggcga
     1561 acgatgtgac cgatgaaatg gtgcgtctga actggctgac cgaatttatg ccgctgccga
     1621 ccattaaaca ttttattcgc accccggatg atgcgtggct gctgaccacc gcgattccgg
     1681 gcaaaaccgc gtttcaggtg ctggaagaat atccggatag cggcgaaaac attgtggatg
     1741 cgctggccgt gtttctgcgt cgtctgcata gcattccggt gtgcaactgc ccgtttaaca
     1801 gcgatcgtgt gtttcgtctg gcccaggcgc agagccgtat gaacaacggc ctggtggatg
     1861 cgagcgattt tgatgatgaa cgtaacggct ggccggtgga acaggtgtgg aaagaaatgc
     1921 ataaactgct gccgtttagc ccggatagcg tggtgaccca cggcgatttt agcctggata
     1981 acctgatttt cgatgaaggc aaactgattg gctgcattga tgtgggccgt gtgggcattg
     2041 cggatcgtta tcaggatctg gccattctgt ggaactgcct gggcgaattt agcccgagcc
     2101 tgcaaaaacg tctgtttcag aaatatggca ttgataatcc ggatatgaac aaactgcaat
     2161 ttcatctgat gctggatgaa tttttctaat aattaattgg accgcggtcc ttttccgctg
     2221 cataaccctg cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac
     2281 gatatacagg attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc
     2341 agccgggcag gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc
     2401 cttattcgca cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccgtaggccg
     2461 gccggcgcgc cgatctgaag atcagcagtt caacctgttg atagtacgta ctaagctctc
     2521 atgtttcacg tactaagctc tcatgtttaa cgtactaagc tctcatgttt aacgaactaa
     2581 accctcatgg ctaacgtact aagctctcat ggctaacgta ctaagctctc atgtttcacg
     2641 tactaagctc tcatgtttga acaataaaat taatataaat cagcaactta aatagcctct
     2701 aaggttttaa gttttataag aaaaaaaaga atatataagg cttttaaagc ctttaaggtt
     2761 taacggttgt ggacaacaag ccagggatgt aacgcactga gaagccctta gagcctctca
     2821 aagcaatttt gagtgacaca ggaacactta acggctgaca tggggcgcgc ctgtgggcgg
     2881 acaaaatagt tgggaactgg gaggggtgga aatggagttt ttaaggatta tttagggaag
     2941 agtgacaaaa tagatgggaa ctgggtgtag cgtcgtaagc taatacgaaa attaaaaatg
     3001 acaaaatagt ttggaactag atttcactta tctggtttct agggcggcgg atttgtccta
     3061 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga
     3121 gcctttcgtt ttatttgatg ccgacgtcca atttaccgaa caactccgcg gccgggaagc
     3181 cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca
     3241 cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg
     3301 cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga
     3361 gcgcggtggc aatgccctgc ctccggtgct ctccggagac tgcgagatca tagatataga
     3421 tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg
     3481 cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt
     3541 aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa
     3601 gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga
     3661 tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt
     3721 tgctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     3781 gcttgctgct tggatgcccg aggcataggc tgtacaaaaa aacagtcata acaagccatg
     3841 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     3901 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caagacgtc
//
