LOCUS       40924_6257        3947 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBG, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3947)
  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 3947)
  AUTHORS   Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank 
            LM.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-JUN-2015) System Biology, Centro Nacional de 
            Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 3947)
  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 3947)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3947)
  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 
            (12-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 KT192136.1.
FEATURES             Location/Qualifiers
     source          1..3947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    44..120
                     /label=BCD2 bicistronic linker
                     /note="BCD2 bicistronic linker"
     RBS             112..120
                     /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             139..852
                     /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    856..912
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      955..1049
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    1054..1252
                     /label=Tn7R transposase recognition site
                     /note="Tn7R transposase recognition site"
     CDS             1362..2174
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2333..2441
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2460..2848
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     mobile_element  2860..3025
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     terminator      complement(3039..3125)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(3194..3724)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(3913..3941)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct agggcccaag ttcacttaaa
       61 aaggagatca acaatgaaag caattttcgt actgaaacat cttaatcatg ctaaggaggt
      121 tttctaatga tcatgggaat tcataaaggt gaagaactgt tcaccggtgt tgttccgatc
      181 ctggttgaac tggatggtga tgttaacggc cacaaattct ctgttcgtgg tgaaggtgaa
      241 ggtgatgcaa ccaacggtaa actgaccctg aaattcatct gcactaccgg taaactgccg
      301 gttccatggc cgactctggt gactaccctg acctatggtg ttcagtgttt ttctcgttac
      361 ccggatcaca tgaagcagca tgatttcttc aaatctgcaa tgccggaagg ttatgtacag
      421 gagcgcacca tttctttcaa agacgatggc acctacaaaa cccgtgcaga ggttaaattt
      481 gaaggtgata ctctggtgaa ccgtattgaa ctgaaaggca ttgatttcaa agaggacggc
      541 aacatcctgg gccacaaact ggaatataac ttcaactccc ataacgttta catcaccgca
      601 gacaaacaga agaacggtat caaagctaac ttcaaaattc gccataacgt tgaagacggt
      661 agcgtacagc tggcggacca ctaccagcag aacactccga tcggtgatgg tccggttctg
      721 ctgccggata accactacct gtccacccag tctaaactgt ccaaagaccc gaacgaaaag
      781 cgcgaccaca tggtgctgct ggagttcgtt actgcagcag gtatcacgca cggcatggat
      841 gaactctaca aataagaatt cgagctcggt acccggggat cctctagagt cgacctgcag
      901 gcatgcaagc ttgcggccgc gtcgtgactg ggaaaaccct ggcgactagt cttggactcc
      961 tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt
     1021 gccgccgggc gttttttatt ggtgagaatc cagccacgcc cctctttaat acgacgggca
     1081 atttgcactt cagaaaatga agagtttgct ttagccataa caaaagtcca gtatgctttt
     1141 tcacagcata actggactga tttcagttta caactattct gtctagttta agactttatt
     1201 gtcatagttt agatctattt tgttcagttt aagactttat tgtccgccca caatttaaat
     1261 ttgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     1321 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcagcgtg
     1381 aaacgagctg tagccgtccg cgtctgaaca gcaacatgga tgcggatctg tatggctata
     1441 aatgggcgcg tgataacgtg ggtcagagcg gcgcgaccat ttatcgtctg tatggcaaac
     1501 cggatgcgcc ggaactgttt ctgaaacatg gcaaaggcag cgtggcgaac gatgtgaccg
     1561 atgaaatggt gcgtctgaac tggctgaccg aatttatgcc gctgccgacc attaaacatt
     1621 ttattcgcac cccggatgat gcgtggctgc tgaccaccgc gattccgggc aaaaccgcgt
     1681 ttcaggtgct ggaagaatat ccggatagcg gcgaaaacat tgtggatgcg ctggccgtgt
     1741 ttctgcgtcg tctgcatagc attccggtgt gcaactgccc gtttaacagc gatcgtgtgt
     1801 ttcgtctggc ccaggcgcag agccgtatga acaacggcct ggtggatgcg agcgattttg
     1861 atgatgaacg taacggctgg ccggtggaac aggtgtggaa agaaatgcat aaactgctgc
     1921 cgtttagccc ggatagcgtg gtgacccacg gcgattttag cctggataac ctgattttcg
     1981 atgaaggcaa actgattggc tgcattgatg tgggccgtgt gggcattgcg gatcgttatc
     2041 aggatctggc cattctgtgg aactgcctgg gcgaatttag cccgagcctg caaaaacgtc
     2101 tgtttcagaa atatggcatt gataatccgg atatgaacaa actgcaattt catctgatgc
     2161 tggatgaatt tttctaataa ttaattggac cgcggtcctt ttccgctgca taaccctgct
     2221 tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat
     2281 tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga
     2341 taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc
     2401 tggcggtgct caacgggaat cctgctctgc gaggctggcc gtaggccggc cggcgcgccg
     2461 atctgaagat cagcagttca acctgttgat agtacgtact aagctctcat gtttcacgta
     2521 ctaagctctc atgtttaacg tactaagctc tcatgtttaa cgaactaaac cctcatggct
     2581 aacgtactaa gctctcatgg ctaacgtact aagctctcat gtttcacgta ctaagctctc
     2641 atgtttgaac aataaaatta atataaatca gcaacttaaa tagcctctaa ggttttaagt
     2701 tttataagaa aaaaaagaat atataaggct tttaaagcct ttaaggttta acggttgtgg
     2761 acaacaagcc agggatgtaa cgcactgaga agcccttaga gcctctcaaa gcaattttga
     2821 gtgacacagg aacacttaac ggctgacatg gggcgcgcct gtgggcggac aaaatagttg
     2881 ggaactggga ggggtggaaa tggagttttt aaggattatt tagggaagag tgacaaaata
     2941 gatgggaact gggtgtagcg tcgtaagcta atacgaaaat taaaaatgac aaaatagttt
     3001 ggaactagat ttcacttatc tggtttctag ggcggcggat ttgtcctact caggagagcg
     3061 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt
     3121 atttgatgcc gacgtccaat ttaccgaaca actccgcggc cgggaagccg atctcggctt
     3181 gaacgaattg ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact tcttcccgta
     3241 tgcccaactt tgtatagaga gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga
     3301 tcacataagc accaagcgcg ttggcctcat gcttgaggag attgatgagc gcggtggcaa
     3361 tgccctgcct ccggtgctct ccggagactg cgagatcata gatatagatc tcactacgcg
     3421 gctgctcaaa cttgggcaga acgtaagccg cgagagcgcc aacaaccgct tcttggtcga
     3481 aggcagcaag cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa tcggagtccg
     3541 gctgatgttg ggagtaggtg gctacgtctc cgaactcacg accgaaaaga tcaagagcag
     3601 cccgcatgga tttgacttgg tcagggccga gcctacatgt gcgaatgatg cccatacttg
     3661 agccacctaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta
     3721 acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg
     3781 gatgcccgag gcataggctg tacaaaaaaa cagtcataac aagccatgaa aaccgccact
     3841 gcgccgttac caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc
     3901 tacttgcatt acagtttacg aaccgaacag gcttatgtca agacgtc
//
