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"