Basic Vector Information
RSFmobcat vector Vector Map
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
LOCUS 40924_48728 9363 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector RSFmobcat, complete sequence. ACCESSION . 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mob- Derivatives of RSF1010 Plasmid Lacking All the Known DNA Sequences Involved in the Mobilization Process JOURNAL Unpublished REFERENCE 2 (bases 1 to 9363) AUTHORS Katashkina JI, Kuvaeva TM, Mashko SV.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) TITLE Direct Submission REFERENCE 4 (bases 1 to 9363) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (02-MAR-2007) Ajinomoto-Genetika Research Institute, 1st Dorozhny Pr., Moscow 117545, Russia" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..9363 /mol_type="other DNA" /organism="synthetic DNA construct" 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 complement(2346..2740) /direction=LEFT /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)" promoter 3046..3148 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 3149..3805 /codon_start=1 /label=CmR /note="chloramphenicol acetyltransferase" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" protein_bind 3838..3859 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 3874..3904 /label=lac UV5 promoter /note="E. coli lac promoter with an 'up' mutation" protein_bind 3912..3928 /label=lac operator /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 3953..3975 /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 3983..5062 /codon_start=1 /label=lacI /note="lac repressor" /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" CDS 5087..6055 /codon_start=1 /label=RSF1010 RepB /note="replication protein B of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)" /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM" 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..7405 /codon_start=1 /label=RSF1010 RepA /note="replication protein A of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)" /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA" CDS 7395..8243 /codon_start=1 /label=RSF1010 RepC /note="replication protein C of the broad-host-range plasmid RSF1010 (Scholz et al., 1989)" /translation="VVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA AG" 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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