LOCUS       Exported               13352 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Tn7 delivery vector pMRE101_Ptac_eCFP, complete sequence.
ACCESSION   KU973693
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13352)
  AUTHORS   Remus-Emsermann MNP., Gisler P, Drissner D.
  TITLE     MiniTn7-transposon delivery vectors for inducible or constitutive 
            fluorescent protein expression in Enterobacteriaceae
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13352)
  AUTHORS   Remus-Emsermann MNP., Drissner D.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-MAR-2016) Institute for Food Sciences, Agroscope, 
            Schloss 1, Waedenswil, Zurich 8820, Switzerland
REFERENCE   3  (bases 1 to 13352)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13352
                     /organism="Tn7 delivery vector pMRE101_Ptac_eCFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1834226"
     CDS             complement(146..1075)
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /note="arabinose operon regulator"
                     /protein_id="AMY99976.1"
                     /translation="MQYGQLVSSLNGGSMKSMAEAQNDPLLPGYSFNAHLVAGLTPIEA
                     NGYLDFFIDRPLGMKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPE
                     AREWYHQWVYFRPRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGR
                     YSELLAINLLEQLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVC
                     LSPSRLSHLFRQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRV
                     FKKCTGASPSEFRAGCEEKVNDVAVKLS"
     gene            complement(146..1075)
                     /gene="araC"
                     /label=araC
     CDS             complement(146..1024)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1051..1335
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     regulatory      1174..1327
                     /regulatory_class="promoter"
     CDS             1453..2274
                     /codon_start=1
                     /gene="tnsA"
                     /product="tnsA"
                     /label=tnsA
                     /note="Tn7 transposase"
                     /protein_id="AMY99977.1"
                     /translation="MAKANSSFSEVQIARRIKEGRGQGHGKDYIPWLTVQEVPSSGRSH
                     RIYSHKTGRVHHLLSDLELAVFLSLEWESSVLDIREQFPLLPSDTRQIAIDSGIKHPVI
                     RGVDQVMSTDFLVDCKDGPFEQFAIQVKPAAALQDERTLEKLELERRYWQQKQIPWFIF
                     TDKEINPVVKENIEWLYSVKTEEVSAELLAQLSPLAHILQEKGDENIINVCKQVDIAYD
                     LELGKTLSEIRALTANGFIKFNIYKSFRANKCADLCISQVVNMEELRYVAN"
     gene            1453..2274
                     /gene="tnsA"
                     /label=tnsA
     CDS             2261..4369
                     /codon_start=1
                     /gene="tnsB"
                     /product="TnsB"
                     /label=tnsB
                     /note="Tn7 transposase"
                     /protein_id="AMY99978.1"
                     /translation="MWQINEVVLFDNDPYRILAIEDGQVVWMQISADKGVPQARAELLL
                     MQYLDEGRLVRTDDPYVHLDLEEPSVDSVSFQKREEDYRKILPIINSKDRFDPKVRSEL
                     VEHVVQEHKVTKATVYKLLRRYWQRGQTPNALIPDYKNSGAPGERRSATGTAKIGRARE
                     YGKGEGTKVTPEIERLFRLTIEKHLLNQKGTKTTVAYRRFVDLFAQYFPRIPQEDYPTL
                     RQFRYFYDREYPKAQRLKSRVKAGVYKKDVRPLSSTATSQALGPGSRYEIDATIADIYL
                     VDHHDRQKIIGRPTLYIVIDVFSRMITGFYIGFENPSYVVAMQAFVNACSDKTAICAQH
                     DIEISSSDWPCVGLPDVLLADRGELMSHQVEALVSSFNVRVESAPPRRGDAKGIVESTF
                     RTLQAEFKSFAPGIVEGSRIKSHGETDYRLDASLSVFEFTQIILRTILFRNNHLVMDKY
                     DRDADFPTDLPSIPVQLWQWGMQHRTGSLRAVEQEQLRVALLPRRKVSISSFGVNLWGL
                     YYSGSEILREGWLQRSTDIARPQHLEAAYDPVLVDTIYLFPQVGSRVFWRCNLTERSRQ
                     FKGLSFWEVWDIQAQEKHNKANAKQDELTKRRELEAFIQQTIQKANKLTPSTTEPKSTR
                     IKQIKTNKKEAVTSERKKRAEHLKPSSSGDEAKVIPFNAVEADDQEDYSLPTYVPELFQ
                     DPPEKDES"
     gene            2261..4369
                     /gene="tnsB"
                     /label=tnsB
     CDS             4366..6033
                     /codon_start=1
                     /gene="tnsC"
                     /product="TnsC"
                     /label=tnsC
                     /note="Tn7 transposition regulator"
                     /protein_id="AMY99979.1"
                     /translation="MSATRIQAVYRDTGVEAYRDNPFIEALPPLQESVNSAASLKSSLQ
                     LTSSDLQKSRVIRAHTICRIPDDYFQPLGTHLLLSERISVMIRGGYVGRNPKTGDLQKH
                     LQNGYERVQTGELETFRFEEARSTAQSLLLIGCSGSGKTTSLHRILATYPQVIYHRELN
                     VEQVVYLKIDCSHNGSLKEICLNFFRALDRALGSNYERRYGLKRHGIETMLALMSQIAN
                     AHALGLLVIDEIQHLSRSRSGGSQEMLNFFVTMVNIIGVPVMLIGTPKAREIFEADLRS
                     ARRGAGFGAIFWDPIQQTQRGKPNQEWIAFTDNLWQLQLLQRKDALLSDEVRDVWYELS
                     QGVMDIVVKLFVLAQLRALALGNERITAGLLRQVYQDELKPVHPMLEALRSGIPERIAR
                     YSDLVVPEIDKRLIQLQLDIAAIQEQTPEEKALQELDTEDQRHLYLMLKEDYDSSLLIP
                     TIKKAFSQNPTMTRQKLLPLVLQWLMEGETVVSELEKPSKSKKVSAIKVVKPSDWDSLP
                     DTDLRYIYSQRQPEKTMHERLKGKGVIVDMASLFKQAG"
     gene            4366..6033
                     /gene="tnsC"
                     /label=tnsC
     CDS             6036..7562
                     /codon_start=1
                     /gene="tnsD"
                     /product="TnsD"
                     /label=tnsD
                     /note="Tn7 transposition site-selector"
                     /protein_id="AMY99980.1"
                     /translation="MRNFPVPYSNELIYSTIARAGVYQGIVSPKQLLDEVYGNRKVVAT
                     LGLPSHLGVIARHLHQTGRYAVQQLIYEHTLFPLYAPFVGKERRDEAIRLMEYQAQGAV
                     HLMLGVAASRVKSDNRFRYCPDCVALQLNRYGEAFWQRDWYLPALPYCPKHGALVFFDR
                     AVDDHRHQFWALGHTELLSDYPKDSLSQLTALAAYIAPLLDAPRAQELSPSLEQWTLFY
                     QRLAQDLGLTKSKHIRHDLVAERVRQTFSDEALEKLDLKLAENKDTCWLKSIFRKHRKA
                     FSYLQHSIVWQALLPKLTVIEALQQASALTEHSITTRPVSQSVQPNSEDLSVKHKDWQQ
                     LVHKYQGIKAARQSLEGGVLYAWLYRHDRDWLVHWNQQHQQERLAPAPRVDWNQRDRIA
                     VRQLLRIIKRLDSSLDHPRATSSWLLKQTPNGTSLAKNLQKLPLVALCLKRYSESVEDY
                     QIRRISQAFIKLKQEDVELRRWRLLRSATLSKERITEEAQRFLEMVYGEE"
     gene            6036..7562
                     /gene="tnsD"
                     /label=tnsD
     regulatory      7994..8419
                     /regulatory_class="terminator"
     terminator      8196..8282
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8374..8401
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        8420..8511
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8512..9372
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance determinant"
                     /protein_id="AMY99981.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            8512..9372
                     /gene="bla"
                     /label=bla
     oriT            9543..9652
                     /note="incP origin of transfer"
     misc_feature    9554..9642
                     /note="oriT origin of transfer recognized by RP4 
                     conjugatable plasmid"
     rep_origin      10392..10614
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     rep_origin      10661..12031
                     /label=pSC101 temperature-sensitive origin of replication
                     /note="pSC101 temperature-sensitive origin of replication"
     CDS             10662..11612
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     repeat_region   12045..12276
                     /label=Tn7 left end
                     /note="Tn7 left end"
     mobile_element  12045..12210
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     regulatory      12294..12376
                     /regulatory_class="promoter"
                     /note="tac promoter"
     promoter        12302..12330
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    12338..12354
                     /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             12377..13096
                     /codon_start=1
                     /gene="eCFP"
                     /product="eCFP"
                     /label=eCFP
                     /note="cyan fluorescent protein"
                     /protein_id="AMY99982.1"
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            12377..13096
                     /gene="eCFP"
                     /label=eCFP
     repeat_region   13141..13352
                     /label=Tn7 right end
                     /note="Tn7 right end"