LOCUS       Exported                7272 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p17GFP(lox-Sm-E-Kn)-Tc, complete sequence.
ACCESSION   KP272131
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7272
                     /organism="Cloning vector p17GFP(lox-Sm-E-Kn)-Tc"
                     /mol_type="other DNA"
                     /db_xref="taxon:1655357"
     mobile_element  complement(56..173)
                     /mobile_element_type="transposon:Mu bacteriophage attR"
     misc_feature    280..313
                     /note="mutant loxP site; lox66"
     protein_bind    280..313
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    359..392
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(583..1377)
                     /codon_start=1
                     /gene="kmR(Tn5)"
                     /product="KmR"
                     /label=kmR(Tn5)
                     /note="amynoglycoside-3-o-phosphotransferase confers 
                     resistance to kanamycine"
                     /protein_id="AKN19594.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(583..1377)
                     /gene="kmR(Tn5)"
                     /label=kmR(Tn5)
     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
     misc_feature    3926..3959
                     /note="mutant loxP site; lox71"
     protein_bind    3926..3959
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(3962..4657)
                     /codon_start=1
                     /gene="ZsGreen"
                     /product="GFP"
                     /label=ZsGreen
                     /note="Zoanthus sp.green fluorescent protein"
                     /protein_id="AKN19597.1"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIADYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     gene            complement(3962..4657)
                     /gene="ZsGreen"
                     /label=ZsGreen
     primer_bind     complement(4889..4905)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     mobile_element  complement(5179..5396)
                     /mobile_element_type="transposon:Mu bacteriophage attL"
     rep_origin      5634..5918
     CDS             complement(6005..7210)
                     /codon_start=1
                     /gene="tetA"
                     /product="TetA"
                     /label=tetA
                     /note="confers tetracycline resistance"
                     /protein_id="AKN19598.1"
                     /translation="MNSSTKIALVITLLDAMGIGLIMPVLPTLLREFIASEDIANHFGV
                     LLALYALMQVIFAPWLGKMSDRFGRRPVLLLSLIGASLDYLLLAFSSALWMLYLGRLLS
                     GITGATGAVAASVIADTTSASQRVKWFGWLGASFGLGLIAGPIIGGFAGEISPHSPFFI
                     AALLNIVTFLVVMFWFRETKNTRDNTDTEVGVETQSNSVYITLFKTMPILLIIYFSAQL
                     IGQIPATVWVLFTENRFGWNSMMVGFSLAGLGLLHSVFQAFVAGRIATKWGEKTAVLLE
                     FIADSSAFAFLAFISEGWLDFPVLILLAGGGIALPALQGVMSIQTKSHEQGALQGLLVS
                     LTNATGVIGPLLFTVIYNHSLPIWDGWIWIIGLAFYCIIILLSMTFMLTPQAQGSKQET
                     SA"
     gene            complement(6005..7210)
                     /gene="tetA"
                     /label=tetA
     promoter        7218..7272
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    7219..7237
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"