LOCUS       Exported                9983 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Tn7 transposase expression vector pTNS3, complete sequence.
ACCESSION   EU215432
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9983)
  AUTHORS   Choi KH, Mima T, Casart Y, Rholl D, Kumar A, Beacham IR, Schweizer 
            HP.
  TITLE     Genetic tools for select-agent-compliant manipulation of 
            Burkholderia pseudomallei
  JOURNAL   Appl. Environ. Microbiol. 74 (4), 1064-1075 (2008)
  PUBMED    18156318
REFERENCE   2  (bases 1 to 9983)
  AUTHORS   Choi K-H., Rholl D, Schweizer HP.
  TITLE     A Tn7-based broad-host bacterial cloning and expression system
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 9983)
  AUTHORS   Choi K-H., Rholl D, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-SEP-2007) Microbiology, Immunology and Pathology, 
            Colorado State University, 1682 Campus Delivery, Ft. Collins, CO 
            80523-1682, USA
REFERENCE   4  (bases 1 to 9983)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9983
                     /organism="Tn7 transposase expression vector pTNS3"
                     /mol_type="other DNA"
                     /db_xref="taxon:477962"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ABW35342.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            201..1061
                     /gene="bla"
                     /label=bla
     rep_origin      1465..1846
                     /label=R6K
                     /note="R6K"
     rep_origin      1470..1826
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     rep_origin      1870..2129
                     /label=oriT
                     /note="oriT"
     oriT            1877..1985
                     /note="incP origin of transfer"
     protein_bind    2361..2382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2397..2427
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2435..2451
                     /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)."
     primer_bind     2459..2475
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2496..2870
                     /regulatory_class="promoter"
                     /note="P1 integron promoter"
     promoter        2512..2540
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     mobile_element  3051..3275
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             3185..4006
                     /codon_start=1
                     /gene="tnsA"
                     /product="TnsA"
                     /label=tnsA
                     /note="Tn7 transposase subunit"
                     /protein_id="ABW35343.1"
                     /translation="MAKANSSFSEVQIARRIKEGRGQGHGKDYIPWLTVQEVPSSGRSH
                     RIYSHKTGRVHHLLSDLELAVFLSLEWESSVLDIREQFPLLPSDTRQIAIDSGIKHPVI
                     RGVDQVMSTDFLVDCKDGPFEQFAIQVKPAAALQDERTLEKLELERRYWQQKQIPWFIF
                     TDKEINPVVKENIEWLYSVKTEEVSAELLAQLSPLAHILQEKGDENIINVCKQVDIAYD
                     LELGKTLSEIRALTANGFIKFNIYKSFRANKCADLCISQVVNMEELRYVAN"
     gene            3185..4006
                     /gene="tnsA"
                     /label=tnsA
     CDS             3993..6101
                     /codon_start=1
                     /gene="tnsB"
                     /product="TnsB"
                     /label=tnsB
                     /note="Tn7 transposase subunit"
                     /protein_id="ABW35344.1"
                     /translation="MWQINEVVLFDNDPYRILAIEDGQVVWMQISADKGVPQARAELLL
                     MQYLDEGRLVRTDDPYVHLDLEEPSVDSVSFQKREEDYRKILPIINSKDRFDPKVRSEL
                     VEHVVQEHKVTKATVYKLLRRYWQRGQTPNALIPDYKNSGAPGERRSATGTAKIGRARE
                     YGKGEGTKVTPEIERLFRLTIEKHLLNQKGTKTTVAYRRFVDLFAQYFPRIPQEDYPTL
                     RQFRYFYDREYPKAQRLKSRVKAGVYKKDVRPLSSTATSQALGPGSRYEIDATIADIYL
                     VDHHDRQKIIGRPTLYIVIDVFSRMITGFYIGFENPSYVVAMQAFVNACSDKTAICAQH
                     DIEISSSDWPCVGLPDVLLADRGELMSHQVEALVSSFNVRVESAPPRRGDAKGIVESTF
                     RTLQAEFKSFAPGIVEGSRIKSHGETDYRLDASLSVFEFTQIILRTILFRNNHLVMDKY
                     DRDADFPTDLPSIPVQLWQWGMQHRTGSLRAVEQEQLRVALLPRRKVSISSFGVNLWGL
                     YYSGSEILREGWLQRSTDIARPQHLEAAYDPVLVDTIYLFPQVGSRVFWRCNLTERSRQ
                     FKGLSFWEVWDIQAQEKHNKANAKQDELTKRRELEAFIQQTIQKANKLTPSTTEPKSTR
                     IKQIKTNKKEAVTSERKKRAEHLKPSSSGDEAKVIPFNAVEADDQEDYSLPTYVPELFQ
                     DPPEKDES"
     gene            3993..6101
                     /gene="tnsB"
                     /label=tnsB
     CDS             6098..7765
                     /codon_start=1
                     /gene="tnsC"
                     /product="TnsC"
                     /label=tnsC
                     /note="Tn7 transposase subunit"
                     /protein_id="ABW35345.1"
                     /translation="MSATRIQAVYRDTGVEAYRDNPFIEALPPLQESVNSAASLKSSLQ
                     LTSSDLQKSRVIRAHTICRIPDDYFQPLGTHLLLSERISVMIRGGYVGRNPKTGDLQKH
                     LQNGYERVQTGELETFRFEEARSTAQSLLLIGCSGSGKTTSLHRILATYPQVIYHRELN
                     VEQVVYLKIDCSHNGSLKEICLNFFRALDRALGSNYERRYGLKRHGIETMLALMSQIAN
                     AHALGLLVIDEIQHLSRSRSGGSQEMLNFFVTMVNIIGVPVMLIGTPKAREIFEADLRS
                     ARRGAGFGAIFWDPIQQTQRGKPNQEWIAFTDNLWQLQLLQRKDALLSDEVRDVWYELS
                     QGVMDIVVKLFVLAQLRALALGNERITAGLLRQVYQDELKPVHPMLEALRSGIPERIAR
                     YSDLVVPEIDKRLIQLQLDIAAIQEQTPEEKALQELDTEDQRHLYLMLKEDYDSSLLIP
                     TIKKAFSQNPTMTRQKLLPLVLQWLMEGETVVSELEKPSKSKKVSAIKVVKPSDWDSLP
                     DTDLRYIYSQRQPEKTMHERLKGKGVIVDMASLFKQAG"
     gene            6098..7765
                     /gene="tnsC"
                     /label=tnsC
     CDS             7768..9294
                     /codon_start=1
                     /gene="tnsD"
                     /product="TnsD"
                     /label=tnsD
                     /note="Tn7 transposase subunit"
                     /protein_id="ABW35346.1"
                     /translation="MRNFPVPYSNELIYSTIARAGVYQGIVSPKQLLDEVYGNRKVVAT
                     LGLPSHLGVIARHLHQTGRYAVQQLIYEHTLFPLYAPFVGKERRDEAIRLMEYQAQGAV
                     HLMLGVAASRVKSDNRFRYCPDCVALQLNRYGEAFWQRDWYLPALPYCPKHGALVFFDR
                     AVDDHRHQFWALGHTELLSDYPKDSLSQLTALAAYIAPLLDAPRAQELSPSLEQWTLFY
                     QRLAQDLGLTKSKHIRHDLVAERVRQTFSDEALEKLDLKLAENKDTCWLKSIFRKHRKA
                     FSYLQHSIVWQALLPKLTVIEALQQASALTEHSITTRPVSQSVQPNSEDLSVKHKDWQQ
                     LVHKYQGIKAARQSLEGGVLYAWLYRHDRDWLVHWNQQHQQERLAPAPRVDWNQRDRIA
                     VRQLLRIIKRLDSSLDHPRATSSWLLKQTPNGTSLAKNLQKLPLVALCLKRYSESVEDY
                     QIRRISQAFIKLKQEDVELRRWRLLRSATLSKERITEEAQRFLEMVYGEE"
     gene            7768..9294
                     /gene="tnsD"
                     /label=tnsD
     primer_bind     complement(9589..9605)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"