LOCUS       Exported                3946 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBG28, complete sequence.
ACCESSION   KR872418
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On Nov 13, 2015 this sequence version replaced KR872418.1.
            ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3946
                     /organism="Cloning vector pBG28"
                     /mol_type="other DNA"
                     /db_xref="taxon:1679948"
     regulatory      9..36
                     /regulatory_class="promoter"
                     /note="promoter 14a"
     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             126..854
                     /codon_start=1
                     /gene="msfGFP"
                     /product="msfGFP"
                     /label=msfGFP
                     /protein_id="AKP56253.1"
                     /translation="MIMGIHKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGK
                     LTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISF
                     KDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKN
                     GIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHM
                     VLLEFVTAAGITHGMDELYK"
     gene            126..854
                     /gene="msfGFP"
                     /label=msfGFP
     misc_feature    855..911
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     regulatory      950..1052
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      954..1048
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    1053..1251
                     /label=Tn7R
                     /note="Tn7R"
     CDS             1361..2176
                     /codon_start=1
                     /gene="aphA"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aphA
                     /note="kanamycin resistance marker"
                     /protein_id="AKP56252.2"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            1361..2176
                     /gene="aphA"
                     /label=aphA
     oriT            2197..2442
     oriT            2332..2440
                     /note="incP origin of transfer"
     rep_origin      2459..2850
                     /label=R6K
                     /note="R6K"
     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"
     misc_feature    2859..3024
                     /label=Tn7L
                     /note="Tn7L"
     mobile_element  2859..3024
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     regulatory      3025..3129
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      3038..3124
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3190..3723)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamicin 3-N-acetyltransferase"
                     /label=aacC1
                     /note="gentamicin resistance marker"
                     /protein_id="AKP56251.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(3190..3723)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(3912..3940)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"