RSFmob vector (V001601)

Basic Vector Information

      • Vector Name:
      • RSFmob
      • Length:
      • 8335 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Katashkina JI, Kuvaeva TM, Skorokhodova AY, Biryukova IV, Golubeva LI, Mashko SV.

RSFmob vector Vector Map

RSFmob8335 bp400800120016002000240028003200360040004400480052005600600064006800720076008000strAstrBRSF1010 oriVlac UV5 promoterlac operatorcloning/expression region of pET22b+ cloning vector (Novagen) fragmentlacIrepBorfEorfFrepAsul

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

RSFmob vector Sequence

Copy Sequence

Download GeneBank File(.gb)

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  JOURNAL   Unpublished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

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