LOCUS       Exported               10445 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Tn7 transposase expression vector pTNS2-ColE1, complete sequence.
ACCESSION   KM576903
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10445)
  AUTHORS   Lee KW, Periasamy S, Mukherjee M, Xie C, Kjelleberg S, Rice SA.
  TITLE     Biofilm development and enhanced stress resistance of a model, 
            mixed-species community biofilm
  JOURNAL   ISME J 8 (4), 894-907 (2014)
  PUBMED    24152718
REFERENCE   2  (bases 1 to 10445)
  AUTHORS   Lee KWK., Periasamy S, Mukherjee M, Xie C, Kjelleberg S, Rice SA.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-SEP-2014) Singapore Centre on Environmental Life 
            Sciences Engineering, Nanyang Technological University, 60 Nanyang 
            Drive, SBS-01N-27 637551, Singapore
REFERENCE   3  (bases 1 to 10445)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10445
                     /organism="Tn7 transposase expression vector pTNS2-ColE1"
                     /mol_type="other DNA"
                     /collection_date=2013
                     /db_xref="taxon:1582478"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(690..2216)
                     /codon_start=1
                     /gene="tnsD"
                     /product="TnsD"
                     /label=tnsD
                     /note="Tn7 transposition site selector"
                     /protein_id="AIZ73053.1"
                     /translation="MRNFPVPYSNELIYSTIARAGVYQGIVSPKQLLDEVYGNRKVVAT
                     LGLPSHLGVIARHLHQTGRYAVQQLIYEHTLFPLYAPFVGKERRDEAIRLMEYQAQGAV
                     HLMLGVAASRVKSDNRFRYCPDCVALQLNRYGEAFWQRDWYLPALPYCPKHGALVFFDR
                     AVDDHRHQFWALGHTELLSDYPKDSLSQLTALAAYIAPLLDAPRAQELSPSLEQWTLFY
                     QRLAQDLGLTKSKHIRHDLVAERVRQTFSDEALEKLDLKLAENKDTCWLKSIFRKHRKA
                     FSYLQHSIVWQALLPKLTVIEALQQASALTEHSITTRPVSQSVQPNSEDLSVKHKDWQQ
                     LVHKYQGIKAARQSLEGGVLYAWLYRHDRDWLVHWNQQHQQERLAPAPRVDWNQRDRIA
                     VRQLLRIIKRLDSSLDHPRATSSWLLKQTPNGTSLAKNLQKLPLVALCLKRYSESVEDY
                     QIRRISQAFIKLKQEDVELRRWRLLRSATLSKERITEEAQRFLEMVYGEE"
     gene            complement(690..2216)
                     /gene="tnsD"
                     /label=tnsD
     CDS             complement(2219..3886)
                     /codon_start=1
                     /gene="tnsC"
                     /product="TnsC"
                     /label=tnsC
                     /note="Tn7 transposition regulator"
                     /protein_id="AIZ73054.1"
                     /translation="MSATRIQAVYRDTGVEAYRDNPFIEALPPLQESVNSAASLKSSLQ
                     LTSSDLQKSRVIRAHTICRIPDDYFQPLGTHLLLSERISVMIRGGYVGRNPKTGDLQKH
                     LQNGYERVQTGELETFRFEEARSTAQSLLLIGCSGSGKTTSLHRILATYPQVIYHRELN
                     VEQVVYLKIDCSHNGSLKEICLNFFRALDRALGSNYERRYGLKRHGIETMLALMSQIAN
                     AHALGLLVIDEIQHLSRSRSGGSQEMLNFFVTMVNIIGVPVMLIGTPKAREIFEADLRS
                     ARRGAGFGAIFWDPIQQTQRGKPNQEWIAFTDNLWQLQLLQRKDALLSDEVRDVWYELS
                     QGVMDIVVKLFVLAQLRALALGNERITAGLLRQVYQDELKPVHPMLEALRSGIPERIAR
                     YSDLVVPEIDKRLIQLQLDIAAIQEQTPEEKALQELDTEDQRHLYLMLKEDYDSSLLIP
                     TIKKAFSQNPTMTRQKLLPLVLQWLMEGETVVSELEKPSKSKKVSAIKVVKPSDWDSLP
                     DTDLRYIYSQRQPEKTMHERLKGKGVIVDMASLFKQAG"
     gene            complement(2219..3886)
                     /gene="tnsC"
                     /label=tnsC
     CDS             complement(3883..5991)
                     /codon_start=1
                     /gene="tnsB"
                     /product="TnsB"
                     /label=tnsB
                     /note="Tn7 tansposase"
                     /protein_id="AIZ73055.1"
                     /translation="MWQINEVVLFDNDPYRILAIEDGQVVWMQISADKGVPQARAELLL
                     MQYLDEGRLVRTDDPYVHLDLEEPSVDSVSFQKREEDYRKILPIINSKDRFDPKVRSEL
                     VEHVVQEHKVTKATVYKLLRRYWQRGQTPNALIPDYKNSGAPGERRSATGTAKIGRARE
                     YGKGEGTKVTPEIERLFRLTIEKHLLNQKGTKTTVAYRRFVDLFAQYFPRIPQEDYPTL
                     RQFRYFYDREYPKAQRLKSRVKAGVYKKDVRPLSSTATSQALGPGSRYEIDATIADIYL
                     VDHHDRQKIIGRPTLYIVIDVFSRMITGFYIGFENPSYVVAMQAFVNACSDKTAICAQH
                     DIEISSSDWPCVGLPDVLLADRGELMSHQVEALVSSFNVRVESAPPRRGDAKGIVESTF
                     RTLQAEFKSFAPGIVEGSRIKSHGETDYRLDASLSVFEFTQIILRTILFRNNHLVMDKY
                     DRDADFPTDLPSIPVQLWQWGMQHRTGSLRAVEQEQLRVALLPRRKVSISSFGVNLWGL
                     YYSGSEILREGWLQRSTDIARPQHLEAAYDPVLVDTIYLFPQVGSRVFWRCNLTERSRQ
                     FKGLSFWEVWDIQAQEKHNKANAKQDELTKRRELEAFIQQTIQKANKLTPSTTEPKSTR
                     IKQIKTNKKEAVTSERKKRAEHLKPSSSGDEAKVIPFNAVEADDQEDYSLPTYVPELFQ
                     DPPEKDES"
     gene            complement(3883..5991)
                     /gene="tnsB"
                     /label=tnsB
     CDS             complement(5978..6799)
                     /codon_start=1
                     /gene="tnsA"
                     /product="TnsA"
                     /label=tnsA
                     /note="Tn7 transposase"
                     /protein_id="AIZ73056.1"
                     /translation="MAKANSSFSEVQIARRIKEGRGQGHGKDYIPWLTVQEVPSSGRSH
                     RIYSHKTGRVHHLLSDLELAVFLSLEWESSVLDIREQFPLLPSDTRQIAIDSGIKHPVI
                     RGVDQVMSTDFLVDCKDGPFEQFAIQVKPAAALQDERTLEKLELERRYWQQKQIPWFIF
                     TDKEINPVVKENIEWLYSVKTEEVSAELLAQLSPLAHILQEKGDENIINVCKQVDIAYD
                     LELGKTLSEIRALTANGFIKFNIYKSFRANKCADLCISQVVNMEELRYVAN"
     gene            complement(5978..6799)
                     /gene="tnsA"
                     /label=tnsA
     mobile_element  6709..6933
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     rep_origin      7120..7955
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(7197..7785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7971..7987)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7995..8011
                     /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)."
     promoter        complement(8019..8049)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8064..8085
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      8317..8576
                     /label=oriT
                     /note="oriT"
     oriT            8461..8569
                     /note="incP origin of transfer"
     rep_origin      8600..8981
                     /label=R6K
                     /note="R6K"
     rep_origin      8620..8976
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(9385..10245)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="AIZ73057.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(9385..10245)
                     /gene="bla"
                     /label=bla
     promoter        complement(10246..10350)
                     /gene="bla"
                     /label=AmpR promoter