RSFmobcat vector (V001600)

Basic Vector Information

      • Vector Name:
      • RSFmobcat
      • Antibiotic Resistance:
      • Chloramphenicol
      • Length:
      • 9363 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Katashkina JI, Kuvaeva TM, Skorokhodova AY, Biryukova IV, Golubeva LI, Mashko SV.

RSFmobcat vector Vector Map

RSFmobcat9363 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200strAstrBRSF1010 oriVcat promotercatlac 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 ℃.

RSFmobcat vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                9363 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector RSFmobcat, complete sequence.
ACCESSION   EF467358
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9363)
  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  JOURNAL   Unpublished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 9363)
  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 9363)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9363
                     /organism="Cloning vector RSFmobcat"
                     /mol_type="other DNA"
                     /note="RSF1010-based non-mobilizable cloning vector; all 
                     known DNA sequences involved in mobilization process were 
                     substituted"
                     /db_xref="taxon:437157"
     CDS             63..866
                     /codon_start=1
                     /gene="strA"
                     /product="streptomycin resistance protein A"
                     /label=strA
                     /protein_id="ABP62933.1"
                     /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="ABP62934.1"
                     /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        3046..3148
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3149..3808
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="ABP62935.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            3149..3808
                     /gene="cat"
                     /label=cat
     promoter        3874..3904
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     regulatory      3874..3879
                     /regulatory_class="minus_35_signal"
                     /note="PlacUV5 promoter -35 signal"
     regulatory      3898..3903
                     /regulatory_class="minus_10_signal"
                     /note="PlacUV5 promoter -10 signal"
     protein_bind    3912..3928
                     /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    3941..3982
                     /note="cloning/expression region of pET22b+ cloning vector 
                     (Novagen) fragment"
     RBS             3953..3975
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3983..5065
                     /codon_start=1
                     /gene="lacI"
                     /product="transcription repressor"
                     /label=lacI
                     /protein_id="ABP62936.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            3983..5065
                     /gene="lacI"
                     /label=lacI
     CDS             5087..6058
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B"
                     /label=repB
                     /protein_id="ABP62937.1"
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     gene            5087..6058
                     /gene="repB"
                     /label=repB
     CDS             6119..6331
                     /codon_start=1
                     /gene="orfE"
                     /product="unknown protein E"
                     /label=orfE
                     /note="hypothetical protein"
                     /protein_id="ABP62938.1"
                     /translation="MEYEKSASGSVYLIKSDKGYWLPGGFGYTSNKAEAGRFSVADMAS
                     LNLDGCTLSLFREDKPFGPGKFLGD"
     gene            6119..6331
                     /gene="orfE"
                     /label=orfE
     CDS             6333..6539
                     /codon_start=1
                     /gene="orfF"
                     /product="repressor protein F"
                     /label=orfF
                     /note="transcriptional regulator of repAC operon"
                     /protein_id="ABP62939.1"
                     /translation="MKDQKDKQTGDLLASPDAVRQARYAERMKAKGMRQRKFWLTDDEY
                     EALRECLEELRAAQGGGSDPASA"
     gene            6333..6539
                     /gene="orfF"
                     /label=orfF
     CDS             6569..7408
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A"
                     /label=repA
                     /note="hexameric replicative helicase; ssDNA-dependent 
                     ATPase"
                     /protein_id="ABP62940.1"
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     gene            6569..7408
                     /gene="repA"
                     /label=repA
     CDS             7395..8246
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C"
                     /label=repC
                     /note="required for origin recognition"
                     /protein_id="ABP62941.1"
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     gene            7395..8246
                     /gene="repC"
                     /label=repC
     CDS             8554..9342
                     /codon_start=1
                     /gene="sul"
                     /product="sulfonamide resistance protein"
                     /label=sul
                     /protein_id="ABP62942.1"
                     /translation="MNKSLIIFGIVNITSDSFSDGGRYLAPDAAIAQARKLMAEGADVI
                     DLVRHPAIPTPRLFRPTQKSRVCAGAGRAQADGIPVSLDSYQPATQAYALSRGVAYLND
                     IRGFPDAAFYPQLAKSSAKLVVMHSVQDGQADRREAPAGDIMDHIAAFFDARIAALTGA
                     GIKRNRLVLDPGMGFFLGAAPETSLSVLARFDELRLRFDLPVLLSVSRKSFLRALTGRG
                     PGVSGPRHSLQSLPPPQVELTSSAHTSRAPCATGWRYWRR"
     gene            8554..9342
                     /gene="sul"
                     /label=sul

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