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pTn7-M vector (V002592#)

Basic Vector Information

Basic Vector Information
Vector Name pTn7-M Antibiotic Resistance Kanamycin
Length 3147 bp Type Cloning vector
Source Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank LM.

pTn7-M vector Vector Map

MCS T0 Tn7R aphA R6K Tn7L T1 aacC1 Pc promoter

Plasmid Resuspension protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pTn7-M vector Sequence

Copy Sequence

Download GeneBank File(.gb)

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"

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