LOCUS       Exported                3147 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTn7-M, complete sequence.
ACCESSION   KR920750
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3147)
  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 3147)
  AUTHORS   Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank 
            LM.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAY-2015) System Biology, Centro Nacional de 
            Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 3147)
  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 3147)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On Nov 13, 2015 this sequence version replaced KR920750.1.
            ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3147
                     /organism="Cloning vector pTn7-M"
                     /mol_type="other DNA"
                     /db_xref="taxon:1679949"
     misc_feature    56..112
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     regulatory      151..253
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      155..249
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    254..452
                     /label=Tn7R
                     /note="Tn7R"
     CDS             562..1377
                     /codon_start=1
                     /gene="aphA"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aphA
                     /note="kanamycin resistance marker"
                     /protein_id="AKP56254.2"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            562..1377
                     /gene="aphA"
                     /label=aphA
     oriT            1398..1643
     oriT            1533..1641
                     /note="incP origin of transfer"
     rep_origin      1660..2051
                     /label=R6K
                     /note="R6K"
     rep_origin      1660..2048
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    2060..2225
                     /label=Tn7L
                     /note="Tn7L"
     mobile_element  2060..2225
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     regulatory      2226..2330
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      2239..2325
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2391..2924)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamicin 3-N-acetyltransferase"
                     /label=aacC1
                     /note="gentamicin resistance marker"
                     /protein_id="AKP56255.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(2391..2924)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(3113..3141)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"