LOCUS       Exported                9218 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pN15E4, complete sequence.
ACCESSION   EF392865
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9218)
  AUTHORS   Mardanov AV, Strakhova TS, Smagin VA, Ravin NV.
  TITLE     Tightly regulated, high-level expression from controlled copy number
            vectors based on the replicon of temperate phage N15
  JOURNAL   Gene 395 (1-2), 15-21 (2007)
  PUBMED    17433573
REFERENCE   2  (bases 1 to 9218)
  AUTHORS   Mardanov AV, Strakhova TS, Smagin VA, Ravin NV.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JAN-2007) Lab of Molecular Cloning, Centre 
            Bioengineering RAS, Prosp. 60-let Oktiabria, bld. 7-1, Moscow 
            117312, Russia
REFERENCE   3  (bases 1 to 9218)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9218
                     /organism="Cloning vector pN15E4"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:424811"
     CDS             complement(94..972)
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /protein_id="ABN12901.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            complement(94..972)
                     /gene="araC"
                     /label=araC
     promoter        999..1283
                     /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      1248..1275
                     /regulatory_class="promoter"
                     /note="araPBAD promoter"
     regulatory      1304..1310
                     /regulatory_class="ribosome_binding_site"
     misc_feature    1311..1340
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      1341..1435
                     /regulatory_class="terminator"
                     /note="phage lambda T0 terminator"
     terminator      1341..1435
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2015..2830
                     /codon_start=1
                     /gene="kan"
                     /product="Kan"
                     /label=kan
                     /note="aminoglucoside phosphotransferase"
                     /protein_id="ABN12902.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            2015..2830
                     /gene="kan"
                     /label=kan
     CDS             3392..4000
                     /codon_start=1
                     /gene="cB"
                     /product="CB"
                     /label=cB
                     /note="main phage repressor"
                     /protein_id="ABN12903.1"
                     /translation="MIKGMKTRGERLKARRLELKLTLKQVAEAVGISLPGVQNLERGDV
                     MPSLEIGLSLAKCLRKPVQWILYGTESDPDRVPVIGTTETGPDRDWQPGEPANTERFLP
                     FVSQRNTVYALTVGNQIQRNYQPGDVILVDSSLTLVPGEDVLVCDNNGEITIQRLARYD
                     EQHYYLDSANSQRVIHDKSDLQFVHQVVGTIKSFMVEGR"
     gene            3392..4000
                     /gene="cB"
                     /label=cB
     CDS             4249..8223
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein"
                     /protein_id="ABN12904.1"
                     /translation="MTLQEFYAERFGSDPFSLLDAARDELTELAKMAGINWPACADKIQ
                     LNPRGGVERYTTYNNSSPEALQKSLKGRVEIYSRLEQSKDGISYPFVNFVEKAHDAGSW
                     SGFSFLFSEYRREQQRNGATVVAQPEEERARMERQAEARRLRAEQQRVNDLKNNQMEQE
                     RLLGWLAFHRAWEHSPAEDGSWPYAVKKGIRDVFGACDIRRVTSHDSAKWSRGPTTYMA
                     IPLSHLDGRKDGRIVGWQRIDLNGGKFQTSAITNGDFVGACFVIGDLQGAQKIATAEGF
                     ATGASIWLATRNDPKKRFDAVVIAVSANNMIHVVEQLVNMYPAAQITCALDNDRKSSAE
                     GKGNTGLRTGFDIMEKFSGVKCVYPTFEDDPELECSDFNDLHSLRGLKEVARQLTRNHL
                     SRATDLLSITLNKLRTLPRLNRRTFAKELLRAVDIGMLTCPVPNSPKELMRLFSSTLRD
                     MGIAEIYNGTVKDHITRRLNRKCRAAQTSRSFSERITNPNLRPSHITYKRFETSRMTDE
                     VMTYAAQLQGIVIVRAGMGSGKSTGLLRPLMLQSTRGVSVAHRVSLIGGLHEMMTEGKG
                     AKADILHYQDPGYQEMAPYANKLTICINSILKGCWQPLMRQHDFFGFDEATQGLRAILA
                     GRAMENPVGVFNTLIDALARTEEHAIMVDADANDLLVDLAELAMKRREELGLPAWLQIH
                     VIELPVDVRNRETNKPIRVFYTEKNRIMTEVIAAVQRGERIMLATDSSTFAEDVTMQLR
                     LQFPDKKFLCVNQKNKQEKEVDDFTNQPKVMVKKYDGLIYSPSISSGVSIEEKHFHRHF
                     GMFCGEVVPSDAIQMLRRDRTAQEYIIGFDKLRGKRETDPEKIKRAYAQALLETAGHSG
                     LLTDVVFDGDRISLGVANSSFMQLKIKAAALEASARNDYASNMICIMHDDGYQVAPMAT
                     DALANSIGKDLRKEARELVFEQLMERHLSVDTPDQAEHDELIKKRTLSLDEQAQLVRWD
                     IEKELQLDVDEVALKFYFDGGLKKVRLFETMQLDEITARRLDREEALIHFTYAYRVAGR
                     WQQFVTTAMTREQADAEFQAKFPAITDYRVKSTPAVEIGMRGFYTLKSATLQQYFRDCG
                     IDPKTLEGEADMDALKRARDNLLTPERRDLLNNVLRIGGFNTEKGKKKAPADLCIGILE
                     SMGLSSKTRRARDGDARPTMRSIDPDSVEFLMNIVEKRREAGLSIHARKVEKTTIEVDR
                     DLDLNIDIHGNPRSKTEHVPDAPQSVIIQALEAIPVAVPEAWAENALPATEMEAVRLWP
                     VASIARTFASLYMTEFMDLLSVREIRLLKAFLSQRQAVAI"
     gene            4249..8223
                     /gene="repA"
                     /label=repA
     rep_origin      7883..7972
                     /label=N15 ori
                     /note="bidirectional replication origin of coliphage N15"
     CDS             8235..8483
                     /codon_start=1
                     /gene="36"
                     /product="gp36"
                     /label=36
                     /protein_id="ABN12905.1"
                     /translation="MGKQRECLVIVEGYDPYVVKVSHDATSYSDYKTGRELPFTIIRMN
                     GFSDSEGVYCCIASDKSMEDHTDEARALLKAYKVKPL"
     gene            8235..8483
                     /gene="36"
                     /label=36
     CDS             8480..8809
                     /codon_start=1
                     /gene="35"
                     /product="gp35"
                     /label=35
                     /protein_id="ABN12906.1"
                     /translation="MTSTPEFLKQLDYEQLKYCRELCDNRIRAIEAEEKKMAWAVTDGG
                     INYGWFRTEDYMKAVECLVATAAERWEESDKADPSGRGWLDLSIRGSRLPVSEYEALFA
                     DGQWG"
     gene            8480..8809
                     /gene="35"
                     /label=35
     CDS             8813..9142
                     /codon_start=1
                     /gene="34"
                     /product="gp34"
                     /label=34
                     /protein_id="ABN12907.1"
                     /translation="MAHDLQKMMDGELVRLQLEIKKEQERRLAEKMIPVFGVWQSPINQ
                     YEFADPEQAISCAAGLLDKVLREVKDDLSKEGGLQGWNGNLLRIYVQHVNESDFAVLQP
                     YLKDK"
     gene            8813..9142
                     /gene="34"
                     /label=34