LOCUS       Exported               10870 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Tn7 transposase expression vector pSTNSK-Tp, complete sequence.
ACCESSION   JQ436538
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10870)
  AUTHORS   Crepin S, Harel J, Dozois CM.
  TITLE     Chromosomal complementation using Tn7 transposon vectors in 
            Enterobacteriaceae
  JOURNAL   Appl. Environ. Microbiol. 78 (17), 6001-6008 (2012)
  PUBMED    22706059
REFERENCE   2  (bases 1 to 10870)
  AUTHORS   Crepin S, Harel J, Dozois CM.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JAN-2012) Microbiology and Biothechnology, 
            INRS-Institut Armand-Frappier, 531, boul. des Prairies, Laval, 
            Quebec H7V 1B7, Canada
REFERENCE   3  (bases 1 to 10870)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10870
                     /organism="Tn7 transposase expression vector pSTNSK-Tp"
                     /mol_type="other DNA"
                     /db_xref="taxon:1201166"
     CDS             complement(281..1807)
                     /codon_start=1
                     /gene="tnsD"
                     /product="Tn7 transposase subunit"
                     /label=tnsD
                     /protein_id="AFM78104.1"
                     /translation="MRNFPVPYSNELIYSTIARAGVYQGIVSPKQLLDEVYGNRKVVAT
                     LGLPSHLGVIARHLHQTGRYAVQQLIYEHTLFPLYAPFVGKERRDEAIRLMEYQAQGAV
                     HLMLGVAASRVKSDNRFRYCPDCVALQLNRYGEAFWQRDWYLPALPYCPKHGALVFFDR
                     AVDDHRHQFWALGHTELLSDYPKDSLSQLTALAAYIAPLLDAPRAQELSPSLEQWTLFY
                     QRLAQDLGLTKSKHIRHDLVAERVRQTFSDEALEKLDLKLAENKDTCWLKSIFRKHRKA
                     FSYLQHSIVWQALLPKLTVIEALQQASALTEHSITTRPVSQSVQPNSEDLSVKHKDWQQ
                     LVHKYQGIKAARQSLEGGVLYAWLYRHDRDWLVHWNQQHQQERLAPAPRVDWNQRDRIA
                     VRQLLRIIKRLDSSLDHPRATSSWLLKQTPNGTSLAKNLQKLPLVALCLKRYSESVEDY
                     QIRRISQAFIKLKQEDVELRRWRLLRSATLSKERITEEAQRFLEMVYGEE"
     gene            complement(281..1807)
                     /gene="tnsD"
                     /label=tnsD
     CDS             complement(1810..3477)
                     /codon_start=1
                     /gene="tnsC"
                     /product="Tn7 transposase subunit"
                     /label=tnsC
                     /protein_id="AFM78105.1"
                     /translation="MSATRIQAVYRDTGVEAYRDNPFIEALPPLQESVNSAASLKSSLQ
                     LTSSDLQKSRVIRAHTICRIPDDYFQPLGTHLLLSERISVMIRGGYVGRNPKTGDLQKH
                     LQNGYERVQTGELETFRFEEARSTAQSLLLIGCSGSGKTTSLHRILATYPQVIYHRELN
                     VEQVVYLKIDCSHNGSLKEICLNFFRALDRALGSNYERRYGLKRHGIETMLALMSQIAN
                     AHALGLLVIDEIQHLSRSRSGGSQEMLNFFVTMVNIIGVPVMLIGTPKAREIFEADLRS
                     ARRGAGFGAIFWDPIQQTQRGKPNQEWIAFTDNLWQLQLLQRKDALLSDEVRDVWYELS
                     QGVMDIVVKLFVLAQLRALALGNERITAGLLRQVYQDELKPVHPMLEALRSGIPERIAR
                     YSDLVVPEIDKRLIQLQLDIAAIQEQTPEEKALQELDTEDQRHLYLMLKEDYDSSLLIP
                     TIKKAFSQNPTMTRQKLLPLVLQWLMEGETVVSELEKPSKSKKVSAIKVVKPSDWDSLP
                     DTDLRYIYSQRQPEKTMHERLKGKGVIVDMASLFKQAG"
     gene            complement(1810..3477)
                     /gene="tnsC"
                     /label=tnsC
     CDS             complement(3474..5582)
                     /codon_start=1
                     /gene="tnsB"
                     /product="Tn7 transposase subunit"
                     /label=tnsB
                     /protein_id="AFM78106.1"
                     /translation="MWQINEVVLFDNDPYRILAIEDGQVVWMQISADKGVPQARAELLL
                     MQYLDEGRLVRTDDPYVHLDLEEPSVDSVSFQKREEDYRKILPIINSKDRFDPKVRSEL
                     VEHVVQEHKVTKATVYKLLRRYWQRGQTPNALIPDYKNSGAPGERRSATGTAKIGRARE
                     YGKGEGTKVTPEIERLFRLTIEKHLLNQKGTKTTVAYRRFVDLFAQYFPRIPQEDYPTL
                     RQFRYFYDREYPKAQRLKSRVKAGVYKKDVRPLSSTATSQALGPGSRYEIDATIADIYL
                     VDHHDRQKIIGRPTLYIVIDVFSRMITGFYIGFENPSYVVAMQAFVNACSDKTAICAQH
                     DIEISSSDWPCVGLPDVLLADRGELMSHQVEALVSSFNVRVESAPPRRGDAKGIVESTF
                     RTLQAEFKSFAPGIVEGSRIKSHGETDYRLDASLSVFEFTQIILRTILFRNNHLVMDKY
                     DRDADFPTDLPSIPVQLWQWGMQHRTGSLRAVEQEQLRVALLPRRKVSISSFGVNLWGL
                     YYSGSEILREGWLQRSTDIARPQHLEAAYDPVLVDTIYLFPQVGSRVFWRCNLTERSRQ
                     FKGLSFWEVWDIQAQEKHNKANAKQDELTKRRELEAFIQQTIQKANKLTPSTTEPKSTR
                     IKQIKTNKKEAVTSERKKRAEHLKPSSSGDEAKVIPFNAVEADDQEDYSLPTYVPELFQ
                     DPPEKDES"
     gene            complement(3474..5582)
                     /gene="tnsB"
                     /label=tnsB
     gene            complement(5566..6390)
                     /gene="tnsA"
                     /label=tnsA
     CDS             complement(5569..6390)
                     /codon_start=1
                     /gene="tnsA"
                     /product="Tn7 transposase subunit"
                     /label=tnsA
                     /protein_id="AFM78107.1"
                     /translation="MAKANSSFSEVQIARRIKEGRGQGHGKDYIPWLTVQEVPSSGRSH
                     RIYSHKTGRVHHLLSDLELAVFLSLEWESSVLDIREQFPLLPSDTRQIAIDSGIKHPVI
                     RGVDQVMSTDFLVDCKDGPFEQFAIQVKPAAALQDERTLEKLELERRYWQQKQIPWFIF
                     TDKEINPVVKENIEWLYSVKTEEVSAELLAQLSPLAHILQEKGDENIINVCKQVDIAYD
                     LELGKTLSEIRALTANGFIKFNIYKSFRANKCADLCISQVVNMEELRYVAN"
     mobile_element  6300..6524
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     misc_recomb     6735..6782
                     /note="FRT; Flp recombinase target site"
     protein_bind    6735..6782
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(6844..7080)
                     /codon_start=1
                     /gene="dhfrII"
                     /product="dihydrofolate reductase"
                     /label=dhfrII
                     /note="trimethoprim resistant"
                     /protein_id="AFM78108.1"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     gene            complement(6844..7080)
                     /gene="dhfrII"
                     /label=dhfrII
     promoter        complement(7407..7435)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_recomb     7487..7534
                     /note="FRT; Flp recombinase target site"
     protein_bind    7487..7534
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        complement(7596..7614)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(7650..7683)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             7851..8645
                     /codon_start=1
                     /gene="neo"
                     /product="neomycin phosphotransferase"
                     /label=neo
                     /note="kanamycin resistance"
                     /protein_id="AFM78110.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            7851..8645
                     /gene="neo"
                     /label=neo
     rep_origin      9400..9780
                     /label=ts replication origin from plasmid pHS1
                     /note="ts replication origin from plasmid pHS1"
     rep_origin      9511..9733
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             9781..10731
                     /codon_start=1
                     /product="Rep ts replication protein"
                     /label=Rep ts replication protein
                     /note="from plasmid pHS1"
                     /protein_id="AFM78109.1"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"