PIR2-m1 vector (V005406)

Basic Vector Information

      • Vector Name:
      • PIR2-m1
      • Antibiotic Resistance:
      • Kanamycin
      • Length:
      • 3846 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank LM.

PIR2-m1 vector Vector Map

PIR2-m13846 bp60012001800240030003600MCSmsfGFPT0 transcriptional terminatorTn7R transposase recognition siteaphAoriT origin of transferR6KTn7L transposase recognition siteT1 transcriptional terminatoraacC1Pc 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 ℃.

PIR2-m1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                3846 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector PIR2-m1, complete sequence.
ACCESSION   KT192140
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3846)
  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 3846)
  AUTHORS   Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank 
            LM.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUN-2015) System Biology, Centro Nacional de 
            Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 3846)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3846
                     /organism="Cloning vector PIR2-m1"
                     /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:1673863"
     misc_feature    56..112
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             127..843
                     /codon_start=1
                     /gene="msfgfp"
                     /product="msfGFP"
                     /label=msfgfp
                     /note="monomeric superfolder green fluorescent protein"
                     /protein_id="AKQ71011.1"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            127..843
                     /gene="msfgfp"
                     /label=msfgfp
                     /note="monomeric superfolder green fluorescent protein"
     regulatory      850..952
                     /regulatory_class="terminator"
                     /note="T0 transcriptional terminator"
     terminator      854..948
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    953..1151
                     /label=Tn7R transposase recognition site
                     /note="Tn7R transposase recognition site"
     CDS             1280..2032
                     /codon_start=1
                     /gene="aphA"
                     /product="aminoglycoside 3'-phosphotransferase"
                     /label=aphA
                     /note="kanamycin resistance marker"
                     /protein_id="AKQ71012.1"
                     /translation="MLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVT
                     DEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALA
                     VFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHK
                     LLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSL
                     QKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            1280..2032
                     /gene="aphA"
                     /label=aphA
                     /note="aminoglycoside 3'-phosphotransferase"
     misc_feature    2097..2350
                     /label=oriT origin of transfer
                     /note="oriT origin of transfer"
     oriT            2232..2340
                     /note="incP origin of transfer"
     rep_origin      2359..2750
                     /label=R6K
                     /note="R6K"
     rep_origin      2359..2747
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    2759..2924
                     /label=Tn7L transposase recognition site
                     /note="Tn7L transposase recognition site"
     mobile_element  2759..2924
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     regulatory      2925..3029
                     /regulatory_class="terminator"
                     /note="T1 transcriptional terminator"
     terminator      2938..3024
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3090..3623)
                     /codon_start=1
                     /gene="aacC1"
                     /product="3'-N-acetyltransferase"
                     /label=aacC1
                     /note="gentamicin resistance marker"
                     /protein_id="AKQ71013.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(3090..3623)
                     /gene="aacC1"
                     /label=aacC1
                     /note="3'-N-acetyltransferase"
     promoter        complement(3812..3840)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"

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