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  JOURNAL   Unpublished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            "Unpublished"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"
ORIGIN
        1 tgcatgatag tttccttgca tgaatccata aaaggcgcct gtagtgccat ttacccctga
       61 agcggcgcac gaaaaacgcg aaagcgtttc acgataaatg cgaaaacttt agctttcgcg
      121 cttcaaatga aacagatgta ttaattactg ctttttattc attacatggg gatcttgaaa
      181 gctggagaga aaaatgaata gatactaaaa agccggagca gtctcccgct ccggcttcac
      241 aaggcaatcg ccttgcagcg aaacacaatt actcttatcg ctaccgttcg tatagcatac
      301 attatacgaa gttatgattc cagtgcgatt ttgatcatct gcatccttag ttcctattcc
      361 gaagttccta ttctctagaa agtataggaa cttcagagcg cttttgaagc tggggtgggc
      421 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
      481 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
      541 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
      601 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
      661 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
      721 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
      781 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
      841 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
      901 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
      961 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     1021 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     1081 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     1141 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc
     1201 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     1261 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     1321 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     1381 cgaaacgatc ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt
     1441 ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc
     1501 ccagctggca attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat
     1561 gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgttttccct tgtccagata
     1621 gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgtg
     1681 ttccgcttcc tttagcagcc cttgcgccct gagtgcttgc ggcagcgtgg gggatcttga
     1741 agttcctatt ccgaagttcc tattctctag aaagtatagg aacttcgaag cagctccagc
     1801 ctacacaatc gctcaagacg tgtaatgctg caagggacat ttaaaaaccc tcctaagttt
     1861 tgtaatctat aaagttagca atttaactaa gtgtaaaaac ttagctagct tactaaaaag
     1921 ataattactg aaaagctttt tgtaaagctg ctttttggta aattcctttg attactgatt
     1981 aacgcgttga taaataagga gtttaaattt tgaaaagtaa ctttatagaa aagaataata
     2041 ctgaaaagtc aatttggtgt tcgccgcaag aaattatggc tgctgatggt atgcctggat
     2101 ctgttgctgg tgttcactat cgagcaaatg ttcaaggctg gacgaagcaa aaaaaggaag
     2161 gtgtcaaggg ggggaaagct gttgaatacg atgtgatgtc gatgccccgg gccatctagt
     2221 atgacgtctg tcgcacctgc ttgatcgcgg ccgcgatagc tagatcgcgt tgctcctctt
     2281 ctccatccgc gttccaagct gcggaaaggc acccataagc gtacgcctgg tcgagcaggc
     2341 gacgcggatc gacgtccagc gcacgagaga atgcgtccgc catctgtgca atgcgtctag
     2401 gatcgagaca aaggtcgtct ctgtcagccg gatcgtagaa catattggcg gcgccaaagc
     2461 ccacttcacc gaccagaccg acgggatcta tcaccagcca gccgcgactg gagaacatga
     2521 tgttttcatg atgcagatcg ccatgtagcc cacgcagttc cgaggcattg ctcatcattt
     2581 gatcggctat aatcgccgcg tggacgtagt cagtttgaca acctgcgttt tgatcatcgc
     2641 gcgcccgctg aaacaaagct gcaaagcgat cccggatcgg gagaagggca gaaggcaggg
     2701 gttcctcaga tgcggcatac agcttcgcca ttagttccgc tgcaatttcg gtcgcctggt
     2761 agtcgccgtg ctcggcaacg atgtgagaga gcattcgctc cccggcatat tcgagcaaca
     2821 tcagattgtt ctcacgaccg agcaaccgga ctgctcccct cccattgcgc cataccagat
     2881 agtcggcccc gcgcagttca tcagcaatgt cttctatagg tttcaatccc ttgacgattg
     2941 caggagtccc gtctggcaat gaaactttcc aaacgaggct ggaaaaggtg tccgcaatga
     3001 gaacaggttg cgaaacgtgc caatgagcag gaaaaacagg cggcatgaac atcaacccca
     3061 agtcagaggg tccaatcgca gatagaaggc aaggcgttcg cggtcggggg cttcgatccc
     3121 caatacattg aataggacag cgaaggcgcg ctctgcttca tctggcgctg cccagttctc
     3181 ttcggcgtta gcaatcatga gtgccaaatc ggcatagcga tctgctgttc cgagccgccc
     3241 aaggtcgatc agacccgtgc attgaagagt tttagggtcc accatgaagt tcggcatgca
     3301 gggatcacca tggcaaacaa ccatatcggt gcgctcttgg tcgagccgca ccggtagctc
     3361 tcgttcgaca cgagccaaaa gatcgtgcag cggcgtactc ttgtcctcgt ccggtaagaa
     3421 gtcgggattg acggcattgc gggacaccac atcaacggcg cgtccgaaca ttcgcgacag
     3481 cctgcgctca aacggacatt gatcaaccga taggctgtga acagcgccaa gttgctgccc
     3541 cattgacggc cacgctttga gcaaatccgc tccagacaga tcagccgccg gtactcccgg
     3601 aattgccgtt atcaccaagc atgcaccctc ctgttcctcc tgccagttga tgacctcggg
     3661 gcaagccaca cctcgacctt tgagccaaat gaggcggtca cgctctccag cgagctcacc
     3721 gcggcgggaa gcaggtgcga ttttcgcgaa ggcatgcccg tcaccacgtc gaaaaacaaa
     3781 atcaccagat tctccgcctc tgacaggcaa ccagtcagaa tgcgattcac caaaaaaaat
     3841 attagttcga ttcaatggag gttccttcag ttttctgatg aagcgcgaat atagagaaat
     3901 atcccgaatg tgcagttcct cgagataact tcgtatagca tacattatac gaacggtatt
     3961 atttgtacaa ttcatccata ccatgggtaa taccagcagc agtaacaaat tctaacaaga
     4021 ccatgtggtc tctcttttcg tttggatctt tggataaggc agattgatag gataagtaat
     4081 ggttgtctgg taacaagact ggaccatcac caattggagt attttgttga taatggtcag
     4141 ctaattgaac agaaccatct tcaatgttgt gtctaatttt gaagttaact ttgataccat
     4201 tcttttgttt gtcagccatg atgtaaacat tgtgagagtt atagttgtat tccaatttgt
     4261 gacctaaaat gttaccatct tctttaaaat caataccttt taattcgatt ctattaacta
     4321 aggtatcacc ttcaaacttg acttcagctc tggtcttgta gttaccgtca tctttgaaaa
     4381 aaatagttct ttcttgaaca taaccttctg gcatggcaga cttgaaaaag tcatgttgtt
     4441 tcatatgatc tgggtatcta gcaaaacaca tcaaaccata acctaaagta gtgactaagg
     4501 ttggccatgg aactggcaat ttaccagtag tacaaataaa ttttaaggtc aatttaccgt
     4561 aagtagcatc accttcacct tcaccggaga cagaaaattt gtgaccatta acatcaccat
     4621 ctaattcaac caaaattggg acaacaccag tgaataattc ttcaccttta gacatgttaa
     4681 ttaacctcct atcgatgggg actgacatga tcgataatga tgacatcgct ggttagaggg
     4741 gaggtgccat aaaataggca acctttacat ttcgacagaa atcatagcaa aatagtgccc
     4801 ttacagatag caggaattcg aagcttctgc agacgcgtcg acgtcatatg gatccgatat
     4861 cgccgtggcg gccgctctag aactagttag tgagtcgtat tacaattcac tggccgtcgt
     4921 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     4981 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     5041 gttgcaattc ttaacgacgg taattgagaa cctgcgtaaa cggcatcagt gagttgcggg
     5101 gtaagcggat gcgatattgt tgccgcatcc ggcaaaaaaa cgggcaaggt gtcaccaccc
     5161 tgcccttttt ctttaaaacc gaaaagatta cttcgcgtga acgttttttg aagctgttat
     5221 tgaaatgatt tgcagtgttt ttgtttttcg tattttcaat gaaacaaaag caatttttta
     5281 ctatctttcg cgtttcattg attaacgact aaaatttgca ctacaggctt gcaagcccca
     5341 ccaaatctaa tcccatcaga tcccgaataa tccaatgtcc tcccggtttt tttcgtactt
     5401 caagtgaatc aatacaaccc ccattcactg ccagagccgt gagcgcagcg aactgaatgt
     5461 cacgaaaaag acagcgactc aggtgcctga tggtcggaga caaaaggaat attcagcgac
     5521 tagctagtta catggctaat tcccatgtca gccgttaagt gttcctgtgt cactgaaaat
     5581 tgctttgaga ggctctaagg gcttctcagt gcgttacatc cctggcttgt tgtccacaac
     5641 cgttaaacct taaaagcttt aaaagcctta tatattcttt tttttcttat aaaacttaaa
     5701 accttagagg ctatttaagt tgctgattta tattaatttt attgttcaaa catgagagct
     5761 tagtacgtga aacatgagag cttagtacgt tagccatgag agcttagtac gttagccatg
     5821 agggtttagt tcgttaaaca tgagagctta gtacgttaaa catgagagct tagtacgtga
     5881 aacatgagag cttagtacgt actatcaaca ggttgaactg ctgatcttca gatcctctac
     5941 gccggacgca tcgtggccgg atcttgcggc cgcaaaaatt aaaaatgaag ttttggaggc
     6001 ctcatttggt gacgaaataa ctaagcactt gtctcctgtt tactcccctg agcttgaggg
     6061 gtcaacatga aggtcattga tagcaggata ataatacagt aaaacgctaa accaataatc
     6121 caaatccagc catcccaaat tggtagtgaa tgattataaa taacagtaaa cagtaatggg
     6181 ccaataacac cggttgcatt ggtaaggctc accaataatc cctgtaaagc accttgctca
     6241 tgactctttg tttggataga catcactccc tgtaatgcag gtaaagcgat cccaccacca
     6301 gccaataaaa ttaaaacagg gaaatctaac caaccttcag atataaacgc taaaaaggca
     6361 aatgcactac tatctgcaat aaattcgagc agtactgccg ttttttcgcc ccatttagtg
     6421 gctattcttc ctgccacaaa ggcttggaat actgagtgta aaagaccaag acccgctaat
     6481 gaaaagccaa ccatcatgct attccatcca aaacgatttt cggtaaatag cacccacacc
     6541 gttgcgggaa tttggcctat caattgcgct gaaaaataaa taatcaacaa aatgggcatc
     6601 gttttaaata aagtgatgta caccgaattt gattgcgtct caacccctac ttcggtatct
     6661 gtattatcac gtgtattttt ggtttcacgg aaccaaaaca taaccacaag gaaagtgaca
     6721 atatttagca acgcagcgat aaaaaaggga ctatgcggtg aaatctctcc tgcaaaacca
     6781 ccaataatag gccccgctat taaaccaagc ccaaaacttg cccctaacca accgaaccac
     6841 ttcacgcgtt gagaagctga ggtggtatcg gcaatgaccg atgccgcgac agccccagta
     6901 gctcctgtga tccctgaaag caaacggcct aaatacagca tccaaagcgc acttgaaaaa
     6961 gccagcaata agtaatccag cgatgcgcct attaatgaca acaacagcac tgggcgccga
     7021 ccaaatcggt cagacatttt tccaagccaa ggagcaaaga taacctgcat taacgcataa
     7081 agtgcaagca atacgccaaa gtggttagcg atatcttccg aagcaataaa ttcacgtaat
     7141 aacgttggca agactggcat gataaggcca atccccatgg catcgagtaa cgtaattacc
     7201 aatgcgatct ttgtcgaact attcatttca cttttctcta tcactgatag ggagtgggaa
     7261 aataactcta tcaatgatag ggtgtcacc
//
