pAH-mini-Mu(LER)-YK vector (V009562)

LOCUS       40924_4150        7289 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAH-mini-Mu(LER)-YK, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7289)
  AUTHORS   Gorshkova NV, Lobanova JS, Tokmakova IL, Smirnov SV, Akhverdyan VZ, 
            Krylov AA, Mashko SV.
  TITLE     Mu-driven transposition of recombinant mini-Mu unit DNA in the 
            Corynebacterium glutamicum chromosome
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7289)
  AUTHORS   Gorshkova NV, Lobanova JS, Tokmakova IL, Smirnov SV, Akhverdyan VZ, 
            Krylov AA, Mashko SV.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-SEP-2017) Ajinomoto-Genetika Research Institute, 1st 
            Dorozhny pr. 1-1, Moscow 117545, Russian Federation
REFERENCE   3  (bases 1 to 7289)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7289)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-SEP-2017) Ajinomoto-Genetika Research Institute, 1st Dorozhny 
            pr. 1-1, Moscow 117545, Russian Federation"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     complement(58..174)
                     /label=Mu-R
                     /note="Mu-R"
     regulatory      complement(179..276)
                     /label=deoT
                     /note="deoT"
                     /regulatory_class="terminator"
     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    1739..1786
                     /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(1837..2204)
                     /label=Mu phage E-element
                     /note="Mu phage E-element"
                     /regulatory_class="enhancer"
     CDS             complement(2216..3052)
                     /codon_start=1
                     /product="streptomycin resistance protein subunit B"
                     /label=streptomycin resistance protein subunit B
                     /note="strB"
                     /protein_id="ATO98411.1"
                     /translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
                     IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
                     AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
                     SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
                     QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
     CDS             complement(3052..3855)
                     /codon_start=1
                     /product="streptomycin resistance protein subunit A"
                     /label=streptomycin resistance protein subunit A
                     /note="strA"
                     /protein_id="ATO98412.1"
                     /translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
                     GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
                     MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
                     ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
                     NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
     protein_bind    complement(3925..3958)
                     /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(3962..4675)
                     /label=Citrine
                     /note="enhanced variant of YFP (Heikal et al., 2001)"
     regulatory      complement(4716..4837)
                     /label=P17
                     /note="P17"
                     /regulatory_class="promoter"
     primer_bind     complement(4909..4925)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5128..5187
                     /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."
     misc_recomb     complement(5199..5416)
                     /label=Mu-L
                     /note="Mu-L"
     rep_origin      complement(5546..5934)
                     /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(6024..7226)
                     /label=TcR
                     /note="tetracycline efflux protein"
     protein_bind    complement(7235..7253)
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"