Basic Vector Information
p17YFP(lox-Sm-E-Kn)-Tc 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 ℃.
p17YFP(lox-Sm-E-Kn)-Tc vector Sequence
LOCUS 40924_2438 7277 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector p17YFP(lox-Sm-E-Kn)-Tc, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7277) AUTHORS Gorshkova NV, Gak ER, Tokmakova IL, Mashko SV. TITLE Genetic tools for C. glutamicum JOURNAL Unpublished REFERENCE 2 (bases 1 to 7277) AUTHORS Gorshkova NV, Gak ER, Tokmakova IL, Mashko SV. TITLE Direct Submission JOURNAL Submitted (12-DEC-2014) Lab3, AGRI, 1st Dorozhny Proezd, 1-1, Moscow 117545, Russia REFERENCE 3 (bases 1 to 7277) TITLE Direct Submission REFERENCE 4 (bases 1 to 7277) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (12-DEC-2014) Lab3, AGRI, 1st Dorozhny Proezd, 1-1, Moscow 117545, Russia" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..7277 /mol_type="other DNA" /organism="synthetic DNA construct" mobile_element complement(58..175) /mobile_element_type="transposon:Mu bacteriophage attR" /label=ttR protein_bind complement(282..315) /label=lox71 /note="Left element (LE) mutant of loxP (Araki et al., 2010). Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." protein_bind 361..394 /label=FRT (minimal) /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" CDS complement(588..1379) /label=NeoR/KanR /note="aminoglycoside phosphotransferase" protein_bind 1740..1787 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." regulatory complement(1993..2140) /regulatory_class="enhancer" CDS complement(2219..3055) /codon_start=1 /gene="strB" /product="StrB" /label=strB /note="streptomycin phosphotransferase" /protein_id="AKN19590.1" /translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY" gene complement(2219..3055) /gene="strB" /label=strB CDS complement(3055..3858) /codon_start=1 /gene="strB" /product="StrA" /label=strB /note="streptomycin phosphotransferase" /protein_id="AKN19591.1" /translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG" gene complement(3055..3858) /gene="strB" /label=strB protein_bind complement(3928..3961) /label=lox66 /note="Right element (RE) mutant of loxP (Araki et al., 2010). Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." CDS complement(3965..4678) /label=Citrine /note="enhanced variant of YFP (Heikal et al., 2001)" primer_bind complement(4893..4909) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" terminator 5112..5171 /label=his operon terminator /note="This putative transcriptin terminator from the E. coli his operon has a 2-bp deletion introduced during synthesis. Its efficiency has not been determined." mobile_element complement(5183..5400) /mobile_element_type="transposon:Mu bacteriophage attL" /label=ttL rep_origin complement(5530..5922) /direction=LEFT /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" CDS complement(6012..7214) /label=TcR /note="tetracycline efflux protein" protein_bind complement(7223..7241) /label=tet operator /note="bacterial operator O2 for the tetR and tetA genes"
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