p17GFP(lox-Sm-E-Kn)-Tc vector (V009781)

Basic Vector Information

      • Vector Name:
      • p17GFP(lox-Sm-E-Kn)-Tc
      • Antibiotic Resistance:
      • Kanamycin
      • Length:
      • 7272 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Gorshkova NV, Gak ER, Tokmakova IL, Mashko SV.

p17GFP(lox-Sm-E-Kn)-Tc vector Vector Map

p17GFP(lox-Sm-E-Kn)-Tc7272 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200ttRlox71FRT (minimal)NeoR/KanRFRTStrBStrAlox66ZsGreenM13 fwdhis operon terminatorttLR6K gamma oriTcRtet operator

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 ℃.

p17GFP(lox-Sm-E-Kn)-Tc vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_2433        7272 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p17GFP(lox-Sm-E-Kn)-Tc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7272)
  AUTHORS   Gorshkova NV, Gak ER, Tokmakova IL, Mashko SV.
  TITLE     Genetic tools for C. glutamicum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7272)
  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 7272)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7272)
  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            "Unpublished"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..7272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  complement(56..173)
                     /mobile_element_type="transposon:Mu bacteriophage attR"
                     /label=ttR
     protein_bind    complement(280..313)
                     /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    359..392
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             complement(586..1377)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     protein_bind    1738..1785
                     /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(1991..2138)
                     /regulatory_class="enhancer"
     CDS             complement(2217..3053)
                     /codon_start=1
                     /gene="strB"
                     /product="StrB"
                     /label=strB
                     /note="streptomycin phosphotransferase"
                     /protein_id="AKN19595.1"
                     /translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
                     IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
                     AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
                     SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
                     QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
     gene            complement(2217..3053)
                     /gene="strB"
                     /label=strB
     CDS             complement(3053..3856)
                     /codon_start=1
                     /gene="strA"
                     /product="StrA"
                     /label=strA
                     /note="streptomycin phosphotransferase"
                     /protein_id="AKN19596.1"
                     /translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
                     GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
                     MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
                     ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
                     NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
     gene            complement(3053..3856)
                     /gene="strA"
                     /label=strA
     protein_bind    complement(3926..3959)
                     /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..4657)
                     /label=ZsGreen
                     /note="Zoanthus green fluorescent protein (Matz et al.,
                     1999)"
     primer_bind     complement(4889..4905)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5108..5167
                     /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(5179..5396)
                     /mobile_element_type="transposon:Mu bacteriophage attL"
                     /label=ttL
     rep_origin      complement(5526..5918)
                     /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(6008..7210)
                     /label=TcR
                     /note="tetracycline efflux protein"
     protein_bind    complement(7219..7237)
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"

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