LOCUS       40924_6262        3986 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBG13, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3986)
  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 3986)
  AUTHORS   Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank 
            LM.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUN-2015) System Biology, Centro Nacional de 
            Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 3986)
  AUTHORS   Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank 
            LM.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-AUG-2015) System Biology, Centro Nacional de 
            Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE   4  (bases 1 to 3986)
  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   5  (bases 1 to 3986)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 3986)
  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 
            (15-JUN-2015) System Biology, Centro Nacional de Biotecnologia, 
            C/Darwin 3, Madrid 28049, Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (31-AUG-2015) System Biology, Centro Nacional de Biotecnologia, 
            C/Darwin 3, Madrid 28049, Spain"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: "Submitted 
            (13-NOV-2015) System Biology, Centro Nacional de Biotecnologia, 
            C/Darwin 3, Madrid 28049, Spain"
COMMENT     SGRef: number: 5; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
            On Nov 13, 2015 this sequence version replaced KT192137.2.
FEATURES             Location/Qualifiers
     source          1..3986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..58
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     misc_feature    83..159
                     /label=BCD2 bicistronic linker
                     /note="BCD2 bicistronic linker"
     RBS             151..159
                     /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             178..891
                     /codon_start=1
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
                     /translation="IHKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    895..951
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      994..1088
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    1093..1291
                     /label=Tn7R transposase recognition site
                     /note="Tn7R transposase recognition site"
     CDS             1401..2213
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2372..2480
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2499..2887
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     mobile_element  2899..3064
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     terminator      complement(3078..3164)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(3233..3763)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(3952..3980)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ttaattaatt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
       61 ggtgaggaac taaaccccta gggcccaagt tcacttaaaa aggagatcaa caatgaaagc
      121 aattttcgta ctgaaacatc ttaatcatgc taaggaggtt ttctaatgat catgggaatt
      181 cataaaggtg aagaactgtt caccggtgtt gttccgatcc tggttgaact ggatggtgat
      241 gttaacggcc acaaattctc tgttcgtggt gaaggtgaag gtgatgcaac caacggtaaa
      301 ctgaccctga aattcatctg cactaccggt aaactgccgg ttccatggcc gactctggtg
      361 actaccctga cctatggtgt tcagtgtttt tctcgttacc cggatcacat gaagcagcat
      421 gatttcttca aatctgcaat gccggaaggt tatgtacagg agcgcaccat ttctttcaaa
      481 gacgatggca cctacaaaac ccgtgcagag gttaaatttg aaggtgatac tctggtgaac
      541 cgtattgaac tgaaaggcat tgatttcaaa gaggacggca acatcctggg ccacaaactg
      601 gaatataact tcaactccca taacgtttac atcaccgcag acaaacagaa gaacggtatc
      661 aaagctaact tcaaaattcg ccataacgtt gaagacggta gcgtacagct ggcggaccac
      721 taccagcaga acactccgat cggtgatggt ccggttctgc tgccggataa ccactacctg
      781 tccacccagt ctaaactgtc caaagacccg aacgaaaagc gcgaccacat ggtgctgctg
      841 gagttcgtta ctgcagcagg tatcacgcac ggcatggatg aactctacaa ataagaattc
      901 gagctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct tgcggccgcg
      961 tcgtgactgg gaaaaccctg gcgactagtc ttggactcct gttgatagat ccagtaatga
     1021 cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg
     1081 gtgagaatcc agccacgccc ctctttaata cgacgggcaa tttgcacttc agaaaatgaa
     1141 gagtttgctt tagccataac aaaagtccag tatgcttttt cacagcataa ctggactgat
     1201 ttcagtttac aactattctg tctagtttaa gactttattg tcatagttta gatctatttt
     1261 gttcagttta agactttatt gtccgcccac aatttaaatt tgtgtctcaa aatctctgat
     1321 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     1381 acagtaatac aaggggtgtt atgagccata ttcagcgtga aacgagctgt agccgtccgc
     1441 gtctgaacag caacatggat gcggatctgt atggctataa atgggcgcgt gataacgtgg
     1501 gtcagagcgg cgcgaccatt tatcgtctgt atggcaaacc ggatgcgccg gaactgtttc
     1561 tgaaacatgg caaaggcagc gtggcgaacg atgtgaccga tgaaatggtg cgtctgaact
     1621 ggctgaccga atttatgccg ctgccgacca ttaaacattt tattcgcacc ccggatgatg
     1681 cgtggctgct gaccaccgcg attccgggca aaaccgcgtt tcaggtgctg gaagaatatc
     1741 cggatagcgg cgaaaacatt gtggatgcgc tggccgtgtt tctgcgtcgt ctgcatagca
     1801 ttccggtgtg caactgcccg tttaacagcg atcgtgtgtt tcgtctggcc caggcgcaga
     1861 gccgtatgaa caacggcctg gtggatgcga gcgattttga tgatgaacgt aacggctggc
     1921 cggtggaaca ggtgtggaaa gaaatgcata aactgctgcc gtttagcccg gatagcgtgg
     1981 tgacccacgg cgattttagc ctggataacc tgattttcga tgaaggcaaa ctgattggct
     2041 gcattgatgt gggccgtgtg ggcattgcgg atcgttatca ggatctggcc attctgtgga
     2101 actgcctggg cgaatttagc ccgagcctgc aaaaacgtct gtttcagaaa tatggcattg
     2161 ataatccgga tatgaacaaa ctgcaatttc atctgatgct ggatgaattt ttctaataat
     2221 taattggacc gcggtccttt tccgctgcat aaccctgctt cggggtcatt atagcgattt
     2281 tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag
     2341 actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg
     2401 cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc
     2461 ctgctctgcg aggctggccg taggccggcc ggcgcgccga tctgaagatc agcagttcaa
     2521 cctgttgata gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt
     2581 actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc
     2641 taacgtacta agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa
     2701 tataaatcag caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata
     2761 tataaggctt ttaaagcctt taaggtttaa cggttgtgga caacaagcca gggatgtaac
     2821 gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg
     2881 gctgacatgg ggcgcgcctg tgggcggaca aaatagttgg gaactgggag gggtggaaat
     2941 ggagttttta aggattattt agggaagagt gacaaaatag atgggaactg ggtgtagcgt
     3001 cgtaagctaa tacgaaaatt aaaaatgaca aaatagtttg gaactagatt tcacttatct
     3061 ggtttctagg gcggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     3121 aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgccg acgtccaatt
     3181 taccgaacaa ctccgcggcc gggaagccga tctcggcttg aacgaattgt taggtggcgg
     3241 tacttgggtc gatatcaaag tgcatcactt cttcccgtat gcccaacttt gtatagagag
     3301 ccactgcggg atcgtcaccg taatctgctt gcacgtagat cacataagca ccaagcgcgt
     3361 tggcctcatg cttgaggaga ttgatgagcg cggtggcaat gccctgcctc cggtgctctc
     3421 cggagactgc gagatcatag atatagatct cactacgcgg ctgctcaaac ttgggcagaa
     3481 cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg
     3541 tcttactacg gagcaagttc ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg
     3601 ctacgtctcc gaactcacga ccgaaaagat caagagcagc ccgcatggat ttgacttggt
     3661 cagggccgag cctacatgtg cgaatgatgc ccatacttga gccacctaac tttgttttag
     3721 ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa catcgttgct gctccataac
     3781 atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg cataggctgt
     3841 acaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt
     3901 tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga
     3961 accgaacagg cttatgtcaa gacgtc
//
