LOCUS       Exported                9251 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pN15E6, complete sequence.
ACCESSION   EF397941
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9251)
  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 9251)
  AUTHORS   Mardanov AV, Strakhova TS, Smagin VA, Ravin NV.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-JAN-2007) Lab of Molecular Cloning, Centre 
            Bioengineering RAS, Prosp. 60-let Oktiabria, bld. 7-1, Moscow 
            117312, Russia
REFERENCE   3  (bases 1 to 9251)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9251
                     /organism="Cloning vector pN15E6"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:424810"
     regulatory      272..444
                     /regulatory_class="terminator"
                     /note="terminator T1 of Escherichia coli rrnB gene"
     terminator      280..366
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      453..625
                     /regulatory_class="terminator"
                     /note="terminator T1 of Escherichia coli rrnB gene"
     terminator      461..547
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      634..806
                     /regulatory_class="terminator"
                     /note="terminator T1 of Escherichia coli rrnB gene"
     terminator      642..728
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      815..987
                     /regulatory_class="terminator"
                     /note="terminator T1 of Escherichia coli rrnB gene"
     terminator      823..909
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      1244..1324
                     /regulatory_class="promoter"
                     /note="phage T5 promoter/lac operator element"
     promoter        1247..1291
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1267..1283
                     /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)."
     protein_bind    1299..1315
                     /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)."
     RBS             1334..1345
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      1336..1342
                     /regulatory_class="ribosome_binding_site"
                     /note="optimized RBS"
     misc_feature    1350..1373
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      1374..1468
                     /regulatory_class="terminator"
                     /note="phage lambda T0 terminator"
     terminator      1374..1468
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2048..2863
                     /codon_start=1
                     /gene="kan"
                     /product="Kan"
                     /label=kan
                     /note="aminoglucoside phosphotransferase"
                     /protein_id="ABN12908.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            2048..2863
                     /gene="kan"
                     /label=kan
     CDS             3425..4033
                     /codon_start=1
                     /gene="cB"
                     /product="CB"
                     /label=cB
                     /note="main phage repressor"
                     /protein_id="ABN12909.1"
                     /translation="MIKGMKTRGERLKARRLELKLTLKQVAEAVGISLPGVQNLERGDV
                     MPSLEIGLSLAKCLRKPVQWILYGTESDPDRVPVIGTTETGPDRDWQPGEPANTERFLP
                     FVSQRNTVYALTVGNQIQRNYQPGDVILVDSSLTLVPGEDVLVCDNNGEITIQRLARYD
                     EQHYYLDSANSQRVIHDKSDLQFVHQVVGTIKSFMVEGR"
     gene            3425..4033
                     /gene="cB"
                     /label=cB
     CDS             4282..8256
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein"
                     /protein_id="ABN12910.1"
                     /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            4282..8256
                     /gene="repA"
                     /label=repA
     rep_origin      7916..8005
                     /label=N15 ori
                     /note="bidirectional replication origin of coliphage N15"
     CDS             8268..8516
                     /codon_start=1
                     /gene="36"
                     /product="gp36"
                     /label=36
                     /protein_id="ABN12911.1"
                     /translation="MGKQRECLVIVEGYDPYVVKVSHDATSYSDYKTGRELPFTIIRMN
                     GFSDSEGVYCCIASDKSMEDHTDEARALLKAYKVKPL"
     gene            8268..8516
                     /gene="36"
                     /label=36
     CDS             8513..8842
                     /codon_start=1
                     /gene="35"
                     /product="gp35"
                     /label=35
                     /protein_id="ABN12912.1"
                     /translation="MTSTPEFLKQLDYEQLKYCRELCDNRIRAIEAEEKKMAWAVTDGG
                     INYGWFRTEDYMKAVECLVATAAERWEESDKADPSGRGWLDLSIRGSRLPVSEYEALFA
                     DGQWG"
     gene            8513..8842
                     /gene="35"
                     /label=35
     CDS             8846..9175
                     /codon_start=1
                     /gene="34"
                     /product="gp34"
                     /label=34
                     /protein_id="ABN12913.1"
                     /translation="MAHDLQKMMDGELVRLQLEIKKEQERRLAEKMIPVFGVWQSPINQ
                     YEFADPEQAISCAAGLLDKVLREVKDDLSKEGGLQGWNGNLLRIYVQHVNESDFAVLQP
                     YLKDK"
     gene            8846..9175
                     /gene="34"
                     /label=34