LOCUS       Exported                8335 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector RSFmob, complete sequence.
ACCESSION   EF467359
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8335)
  AUTHORS   Katashkina JI, Kuvaeva TM, Skorokhodova AY, Biryukova IV, Golubeva 
            LI, Mashko SV.
  TITLE     Construction of Stably Maintained mob- Derivatives of RSF1010 
            Plasmid Lacking All the Known DNA Sequences Involved in the 
            Mobilization Process
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8335)
  AUTHORS   Katashkina JI, Kuvaeva TM, Mashko SV.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAR-2007) Ajinomoto-Genetika Research Institute, 1st 
            Dorozhny Pr., Moscow 117545, Russia
REFERENCE   3  (bases 1 to 8335)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8335
                     /organism="Cloning vector RSFmob"
                     /mol_type="other DNA"
                     /note="non-mobilizable derivative of RSF1010; CmS 
                     derivative of RSFmobcat"
                     /db_xref="taxon:437158"
     CDS             63..866
                     /codon_start=1
                     /gene="strA"
                     /product="streptomycin resistance protein A"
                     /label=strA
                     /protein_id="ABP62943.1"
                     /translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
                     GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
                     MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
                     ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
                     NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
     gene            63..866
                     /gene="strA"
                     /label=strA
     CDS             866..1702
                     /codon_start=1
                     /gene="strB"
                     /product="streptomycin resistance protein B"
                     /label=strB
                     /protein_id="ABP62944.1"
                     /translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
                     IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
                     AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
                     SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
                     QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
     gene            866..1702
                     /gene="strB"
                     /label=strB
     rep_origin      2346..2740
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     rep_origin      2347..2771
                     /note="oriV; independent of host replication functions; 
                     requires RepA/B/C"
     promoter        2846..2876
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     regulatory      2846..2851
                     /regulatory_class="minus_35_signal"
                     /note="PlacUV5 promoter -35 signal"
     regulatory      2870..2875
                     /regulatory_class="minus_10_signal"
                     /note="PlacUV5 promoter -10 signal"
     protein_bind    2884..2900
                     /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)."
     misc_feature    2913..2954
                     /note="cloning/expression region of pET22b+ cloning vector 
                     (Novagen) fragment"
     RBS             2925..2947
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2955..4037
                     /codon_start=1
                     /gene="lacI"
                     /product="transcription repressor"
                     /label=lacI
                     /protein_id="ABP62945.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            2955..4037
                     /gene="lacI"
                     /label=lacI
     CDS             4059..5030
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B"
                     /label=repB
                     /protein_id="ABP62946.1"
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     gene            4059..5030
                     /gene="repB"
                     /label=repB
     CDS             5091..5303
                     /codon_start=1
                     /gene="orfE"
                     /product="unknown protein E"
                     /label=orfE
                     /note="hypothetical protein"
                     /protein_id="ABP62947.1"
                     /translation="MEYEKSASGSVYLIKSDKGYWLPGGFGYTSNKAEAGRFSVADMAS
                     LNLDGCTLSLFREDKPFGPGKFLGD"
     gene            5091..5303
                     /gene="orfE"
                     /label=orfE
     CDS             5305..5511
                     /codon_start=1
                     /gene="orfF"
                     /product="repressor protein F"
                     /label=orfF
                     /note="transcriptional regulator of repAC operon"
                     /protein_id="ABP62948.1"
                     /translation="MKDQKDKQTGDLLASPDAVRQARYAERMKAKGMRQRKFWLTDDEY
                     EALRECLEELRAAQGGGSDPASA"
     gene            5305..5511
                     /gene="orfF"
                     /label=orfF
     CDS             5541..6380
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A"
                     /label=repA
                     /note="hexameric replicative helicase; ssDNA-dependent 
                     ATPase"
                     /protein_id="ABP62949.1"
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     gene            5541..6380
                     /gene="repA"
                     /label=repA
     CDS             6367..7218
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C"
                     /label=repC
                     /note="required for origin recognition"
                     /protein_id="ABP62950.1"
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     gene            6367..7218
                     /gene="repC"
                     /label=repC
     CDS             7526..8314
                     /codon_start=1
                     /gene="sul"
                     /product="sulfonamide resistance protein"
                     /label=sul
                     /protein_id="ABP62951.1"
                     /translation="MNKSLIIFGIVNITSDSFSDGGRYLAPDAAIAQARKLMAEGADVI
                     DLVRHPAIPTPRLFRPTQKSRVCAGAGRAQADGIPVSLDSYQPATQAYALSRGVAYLND
                     IRGFPDAAFYPQLAKSSAKLVVMHSVQDGQADRREAPAGDIMDHIAAFFDARIAALTGA
                     GIKRNRLVLDPGMGFFLGAAPETSLSVLARFDELRLRFDLPVLLSVSRKSFLRALTGRG
                     PGVSGPRHSLQSLPPPQVELTSSAHTSRAPCATGWRYWRR"
     gene            7526..8314
                     /gene="sul"
                     /label=sul