pG18-STM vector (V006136)

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LOCUS       40924_20835        7657 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transposon mutagenesis vector pG18-STM, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7657)
  AUTHORS   Pobigaylo N, Wetter D, Szymczak S, Schiller U, Kurtz S, Meyer F, 
            Nattkemper TW, Becker A.
  TITLE     Construction of a large signature-tagged mini-Tn5 transposon library
            and its application to mutagenesis of Sinorhizobium meliloti
  JOURNAL   Appl. Environ. Microbiol. 72 (6), 4329-4337 (2006)
  PUBMED    16751548
REFERENCE   2  (bases 1 to 7657)
  AUTHORS   Pobigaylo N, Becker A.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-FEB-2006) Department of Genetics, University of 
            Bielefeld, Universitaetsstrasse 25, Bielefeld D-33615, Germany
REFERENCE   3  (bases 1 to 7657)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7657)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2006"; volume: "72"; issue: "6"; pages:
            "4329-4337"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-FEB-2006) Department of Genetics, University of Bielefeld, 
            Universitaetsstrasse 25, Bielefeld D-33615, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    145..468
                     /label=similar to transposase
                     /note="similar to transposase"
     CDS             complement(509..1039)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(1228..1256)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     oriT            complement(1479..1587)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      1799..2387
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2675..2696
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2711..2741
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2749..2765
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     repeat_region   2777..2795
                     /note="Tn5 outside end; required for transposition"
     CDS             2876..4684
                     /label=GUS
                     /note="beta-glucuronidase"
     promoter        4759..4787
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4795..4811
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             4863..5654
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    5791..6033
                     /note="artificial linker for cloning and amplification of 
                     signature tags"
     gap             5855..5878
                     /estimated_length=24
     misc_feature    5855..5878
                     /note="h-tag; variable region, signature tag"
     gap             5946..5969
                     /estimated_length=24
     misc_feature    5946..5969
                     /note="k-tag; variable region, signature tag"
     repeat_region   6034..6052
                     /note="Tn5 inside end; required for transposition"
     CDS             6139..7566
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
     primer_bind     complement(7636..7652)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"