LOCUS       Exported                7572 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAH-mini-Mu(LER)-YS, complete sequence.
ACCESSION   MG014200
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7572)
  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  JOURNAL   Unpublished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 7572)
  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 7572)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7572
                     /organism="Cloning vector pAH-mini-Mu(LER)-YS"
                     /mol_type="other DNA"
                     /db_xref="taxon:2049510"
     CDS             complement(771..1565)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /note="KmR"
                     /protein_id="ATO98419.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    complement(1922..2039)
                     /label=Mu-R
                     /note="Mu-R"
     regulatory      complement(2043..2140)
                     /regulatory_class="terminator"
                     /note="deoT"
     misc_recomb     complement(2146..2179)
                     /label=lox71
                     /note="lox71"
     protein_bind    2146..2179
                     /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)."
     regulatory      complement(2186..2553)
                     /regulatory_class="enhancer"
                     /note="Mu phage E-element"
     CDS             complement(2565..3401)
                     /codon_start=1
                     /product="streptomycin resistance protein subunit B"
                     /label=streptomycin resistance protein subunit B
                     /note="strB"
                     /protein_id="ATO98420.1"
                     /translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
                     IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
                     AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
                     SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
                     QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
     CDS             complement(3401..4204)
                     /codon_start=1
                     /product="streptomycin resistance protein subunit A"
                     /label=streptomycin resistance protein subunit A
                     /note="strA"
                     /protein_id="ATO98421.1"
                     /translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
                     GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
                     MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
                     ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
                     NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
     CDS             complement(4274..4990)
                     /codon_start=1
                     /product="yECitrine"
                     /label=yECitrine
                     /protein_id="ATO98422.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      complement(4991..5134)
                     /regulatory_class="promoter"
                     /note="P17"
     misc_recomb     5162..5195
                     /label=lox66
                     /note="lox66"
     protein_bind    5162..5195
                     /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)."
     misc_feature    5246..5286
                     /label=MCS
                     /note="MCS"
     regulatory      5287..5433
                     /regulatory_class="terminator"
                     /note="hisT"
     misc_feature    complement(5434..5651)
                     /label=Mu-L
                     /note="Mu-L"
     rep_origin      5781..6169
                     /label=ori_R6k
                     /note="ori_R6k"
     regulatory      6207..6668
                     /regulatory_class="promoter"
                     /note="PsacB"
     promoter        6223..6668
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             6669..518
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"