LOCUS       40924_6282        3946 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBG28, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3946)
  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 3946)
  AUTHORS   Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank 
            LM.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-MAY-2015) System Biology, Centro Nacional de 
            Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 3946)
  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 3946)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3946)
  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 
            (26-MAY-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 KR872418.1.
FEATURES             Location/Qualifiers
     source          1..3946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      9..36
                     /label=promoter 14a
                     /note="promoter 14a"
                     /regulatory_class="promoter"
     misc_feature    43..119
                     /label=BCD2
                     /note="BCD2"
     RBS             111..119
                     /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             138..851
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     misc_feature    855..911
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      954..1048
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    1053..1251
                     /label=Tn7R
                     /note="Tn7R"
     CDS             1361..2173
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     oriT            2332..2440
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2459..2847
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     mobile_element  2859..3024
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     terminator      complement(3038..3124)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(3193..3723)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(3912..3940)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ttaattaact aggttgacat ggatataatg tatgtaccta gggcccaagt tcacttaaaa
       61 aggagatcaa caatgaaagc aattttcgta ctgaaacatc ttaatcatgc taaggaggtt
      121 ttctaatgat catgggaatt cataaaggtg aagaactgtt caccggtgtt gttccgatcc
      181 tggttgaact ggatggtgat gttaacggcc acaaattctc tgttcgtggt gaaggtgaag
      241 gtgatgcaac caacggtaaa ctgaccctga aattcatctg cactaccggt aaactgccgg
      301 ttccatggcc gactctggtg actaccctga cctatggtgt tcagtgtttt tctcgttacc
      361 cggatcacat gaagcagcat gatttcttca aatctgcaat gccggaaggt tatgtacagg
      421 agcgcaccat ttctttcaaa gacgatggca cctacaaaac ccgtgcagag gttaaatttg
      481 aaggtgatac tctggtgaac cgtattgaac tgaaaggcat tgatttcaaa gaggacggca
      541 acatcctggg ccacaaactg gaatataact tcaactccca taacgtttac atcaccgcag
      601 acaaacagaa gaacggtatc aaagctaact tcaaaattcg ccataacgtt gaagacggta
      661 gcgtacagct ggcggaccac taccagcaga acactccgat cggtgatggt ccggttctgc
      721 tgccggataa ccactacctg tccacccagt ctaaactgtc caaagacccg aacgaaaagc
      781 gcgaccacat ggtgctgctg gagttcgtta ctgcagcagg tatcacgcac ggcatggatg
      841 aactctacaa ataagaattc gagctcggta cccggggatc ctctagagtc gacctgcagg
      901 catgcaagct tgcggccgcg tcgtgactgg gaaaaccctg gcgactagtc ttggactcct
      961 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     1021 ccgccgggcg ttttttattg gtgagaatcc agccacgccc ctctttaata cgacgggcaa
     1081 tttgcacttc agaaaatgaa gagtttgctt tagccataac aaaagtccag tatgcttttt
     1141 cacagcataa ctggactgat ttcagtttac aactattctg tctagtttaa gactttattg
     1201 tcatagttta gatctatttt gttcagttta agactttatt gtccgcccac aatttaaatt
     1261 tgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     1321 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcagcgtga
     1381 aacgagctgt agccgtccgc gtctgaacag caacatggat gcggatctgt atggctataa
     1441 atgggcgcgt gataacgtgg gtcagagcgg cgcgaccatt tatcgtctgt atggcaaacc
     1501 ggatgcgccg gaactgtttc tgaaacatgg caaaggcagc gtggcgaacg atgtgaccga
     1561 tgaaatggtg cgtctgaact ggctgaccga atttatgccg ctgccgacca ttaaacattt
     1621 tattcgcacc ccggatgatg cgtggctgct gaccaccgcg attccgggca aaaccgcgtt
     1681 tcaggtgctg gaagaatatc cggatagcgg cgaaaacatt gtggatgcgc tggccgtgtt
     1741 tctgcgtcgt ctgcatagca ttccggtgtg caactgcccg tttaacagcg atcgtgtgtt
     1801 tcgtctggcc caggcgcaga gccgtatgaa caacggcctg gtggatgcga gcgattttga
     1861 tgatgaacgt aacggctggc cggtggaaca ggtgtggaaa gaaatgcata aactgctgcc
     1921 gtttagcccg gatagcgtgg tgacccacgg cgattttagc ctggataacc tgattttcga
     1981 tgaaggcaaa ctgattggct gcattgatgt gggccgtgtg ggcattgcgg atcgttatca
     2041 ggatctggcc attctgtgga actgcctggg cgaatttagc ccgagcctgc aaaaacgtct
     2101 gtttcagaaa tatggcattg ataatccgga tatgaacaaa ctgcaatttc atctgatgct
     2161 ggatgaattt ttctaataat taattggacc gcggtccttt tccgctgcat aaccctgctt
     2221 cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt
     2281 ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat
     2341 aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct
     2401 ggcggtgctc aacgggaatc ctgctctgcg aggctggccg taggccggcc ggcgcgccga
     2461 tctgaagatc agcagttcaa cctgttgata gtacgtacta agctctcatg tttcacgtac
     2521 taagctctca tgtttaacgt actaagctct catgtttaac gaactaaacc ctcatggcta
     2581 acgtactaag ctctcatggc taacgtacta agctctcatg tttcacgtac taagctctca
     2641 tgtttgaaca ataaaattaa tataaatcag caacttaaat agcctctaag gttttaagtt
     2701 ttataagaaa aaaaagaata tataaggctt ttaaagcctt taaggtttaa cggttgtgga
     2761 caacaagcca gggatgtaac gcactgagaa gcccttagag cctctcaaag caattttgag
     2821 tgacacagga acacttaacg gctgacatgg ggcgcgcctg tgggcggaca aaatagttgg
     2881 gaactgggag gggtggaaat ggagttttta aggattattt agggaagagt gacaaaatag
     2941 atgggaactg ggtgtagcgt cgtaagctaa tacgaaaatt aaaaatgaca aaatagtttg
     3001 gaactagatt tcacttatct ggtttctagg gcggcggatt tgtcctactc aggagagcgt
     3061 tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta
     3121 tttgatgccg acgtccaatt taccgaacaa ctccgcggcc gggaagccga tctcggcttg
     3181 aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat
     3241 gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat
     3301 cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat
     3361 gccctgcctc cggtgctctc cggagactgc gagatcatag atatagatct cactacgcgg
     3421 ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa
     3481 ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat cggagtccgg
     3541 ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat caagagcagc
     3601 ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc ccatacttga
     3661 gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa
     3721 catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg
     3781 atgcccgagg cataggctgt acaaaaaaac agtcataaca agccatgaaa accgccactg
     3841 cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct
     3901 acttgcatta cagtttacga accgaacagg cttatgtcaa gacgtc
//
