LOCUS       Exported                7657 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Transposon mutagenesis vector pG18-STM, complete sequence.
ACCESSION   DQ408591
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7657
                     /organism="Transposon mutagenesis vector pG18-STM"
                     /mol_type="other DNA"
                     /note="derived from pG18Mob2 plasmid in GenBank Accession 
                     Number AY222820"
                     /db_xref="taxon:375452"
     misc_feature    145..468
                     /label=similar to transposase
                     /note="similar to transposase"
     CDS             complement(506..1039)
                     /codon_start=1
                     /gene="accC1"
                     /product="AacC1"
                     /label=accC1
                     /note="gentamicin acetyltransferase"
                     /protein_id="ABD73286.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(506..1039)
                     /gene="accC1"
                     /label=accC1
     promoter        complement(1228..1256)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     oriT            1479..1587
                     /note="incP origin of transfer"
     misc_feature    1523..1621
                     /label=RP4-mob region
                     /note="RP4-mob region"
     rep_origin      1799..2387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2386..2388
                     /label=oriV
                     /note="oriV"
     protein_bind    2675..2696
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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)."
     mobile_element  2777..6052
                     /mobile_element_type="transposon:mTn5-STM"
                     /note="modified mTn5-GNm"
     repeat_region   2777..2795
                     /note="Tn5 outside end; required for transposition"
     CDS             2876..4687
                     /codon_start=1
                     /gene="gusA"
                     /product="beta-glucuronidase"
                     /label=gusA
                     /note="promoterless coding region; GusA"
                     /protein_id="ABD73288.1"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            2876..4687
                     /gene="gusA"
                     /label=gusA
     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..5657
                     /codon_start=1
                     /gene="nptII"
                     /product="NptII"
                     /label=nptII
                     /note="confers resistance to neomycin and kanamycin"
                     /protein_id="ABD73289.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            4863..5657
                     /gene="nptII"
                     /label=nptII
     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..7569
                     /codon_start=1
                     /gene="tnpA**"
                     /product="TnpA**"
                     /label=tnpA**
                     /note="HindIII restriction site has been removed"
                     /protein_id="ABD73287.1"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSEAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTLPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     gene            6139..7569
                     /gene="tnpA**"
                     /label=tnpA**
     primer_bind     complement(7636..7652)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"