LOCUS       Exported                3947 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBG, complete sequence.
ACCESSION   KT192136
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On Nov 13, 2015 this sequence version replaced KT192136.1.
            ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3947
                     /organism="Cloning vector pBG"
                     /mol_type="other DNA"
                     /note="derived from cloning vector pSEVA211 as backbone to 
                     clone mini-Tn7 fragments bearing Tn7 extremes, 
                     multi-cloning site, and in some cases reporter (gfp) and 
                     synthetic promoters"
                     /db_xref="taxon:1673862"
     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             127..855
                     /codon_start=1
                     /gene="msfgfp"
                     /product="msfGFP"
                     /label=msfgfp
                     /note="monomeric superfolder green fluorescent protein"
                     /protein_id="AKQ70999.1"
                     /translation="MIMGIHKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGK
                     LTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISF
                     KDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKN
                     GIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHM
                     VLLEFVTAAGITHGMDELYK"
     gene            127..855
                     /gene="msfgfp"
                     /label=msfgfp
                     /note="monomeric superfolder green fluorescent protein"
     misc_feature    856..912
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     regulatory      951..1053
                     /regulatory_class="terminator"
                     /note="T0 transcriptional terminator"
     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..2177
                     /codon_start=1
                     /gene="aphA"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aphA
                     /note="kanamycin resistance marker"
                     /protein_id="AKQ71000.2"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            1362..2177
                     /gene="aphA"
                     /label=aphA
                     /note="aminoglycoside 3-phosphotransferase"
     misc_feature    2198..2443
                     /label=oriT origin of transfer
                     /note="oriT origin of transfer"
     oriT            2333..2441
                     /note="incP origin of transfer"
     rep_origin      2460..2851
                     /label=ori R6K
                     /note="ori R6K"
     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"
     misc_feature    2860..3025
                     /label=Tn7L transposase recognition site
                     /note="Tn7L transposase recognition site"
     mobile_element  2860..3025
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     regulatory      3026..3130
                     /regulatory_class="terminator"
                     /note="T1 transcriptional terminator"
     terminator      3039..3125
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3191..3724)
                     /codon_start=1
                     /gene="aacC1"
                     /product="3-N-acetyltransferase"
                     /label=aacC1
                     /note="gentamicin resistance marker"
                     /protein_id="AKQ71001.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(3191..3724)
                     /gene="aacC1"
                     /label=aacC1
                     /note="3-N-acetyltransferase"
     promoter        complement(3913..3941)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"