LOCUS       40924_4155        7572 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAH-mini-Mu(LER)-YS, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7572)
  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            "Unpublished"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..7572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(774..1565)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    complement(2146..2179)
                     /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)."
     CDS             complement(4277..4990)
                     /codon_start=1
                     /label=Citrine
                     /note="enhanced variant of YFP (Heikal et al., 2001)"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     protein_bind    complement(5162..5195)
                     /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)."
     terminator      5363..5422
                     /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."
     rep_origin      complement(5781..6169)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     promoter        6223..6668
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(6669..7572,1..515)
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
ORIGIN
        1 aaaaacgcac ggctgagtta gcaaacggcg ctctcggtat gattgagcta aacgatgatt
       61 acacactgaa aaaagtgatg aaaccgctga ttgcatctaa cacagtaaca gatgaaattg
      121 aacgcgcgaa cgtctttaaa atgaacggca aatggtacct gttcactgac tcccgcggat
      181 caaaaatgac gattgacggc attacgtcta acgatattta catgcttggt tatgtttcta
      241 attctttaac tggcccatac aagccgctga acaaaactgg ccttgtgtta aaaatggatc
      301 ttgatcctaa cgatgtaacc tttacttact cacacttcgc tgtacctcaa gcgaaaggaa
      361 acaatgtcgt gattacaagc tatatgacaa acagaggatt ctacgcagac aaacaatcaa
      421 cgtttgcgcc aagcttcctg ctgaacatca aaggcaagaa aacatctgtt gtcaaagaca
      481 gcatccttga acaaggacaa ttaacagtta acaaataaaa acgcaaaaga aaatgccgat
      541 atcctattgg cattttcttt tatttcttat caacataaag gtgaatccca tactagtggg
      601 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
      661 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga
      721 tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac
      781 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
      841 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
      901 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
      961 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
     1021 tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     1081 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     1141 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     1201 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     1261 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     1321 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     1381 tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     1441 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     1501 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     1561 atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag
     1621 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     1681 ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat
     1741 cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc
     1801 cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc
     1861 tacgtgttcc gcttccttta gcagcccttg cgccctgagt gcttgcggca gcgtgtccgg
     1921 atgaagcggc gcacgaaaaa cgcgaaagcg tttcacgata aatgcgaaaa ctttagcttt
     1981 cgcgcttcaa atgaaacaga tgtattaatt actgcttttt attcattaca tggggatctt
     2041 gaaagctgga gagaaaaatg aatagatact aaaaagccgg agcagtctcc cgctccggct
     2101 tcacaaggca atcgccttgc agcgaaacac aattactctt atcgctaccg ttcgtatagc
     2161 atacattata cgaagttatg gtaccacatt taaaaaccct cctaagtttt gtaatctata
     2221 aagttagcaa tttaactaag tgtaaaaact tagctagctt actaaaaaga taattactga
     2281 aaagcttttt gtaaagctgc tttttggtaa attcctttga ttactgatta acgcgttgat
     2341 aaataaggag tttaaatttt gaaaagtaac tttatagaaa agaataatac tgaaaagtca
     2401 atttggtgtt cgccgcaaga aattatggct gctgatggta tgcctggatc tgttgctggt
     2461 gttcactatc gagcaaatgt tcaaggctgg acgaagcaaa aaaaggaagg tgtcaagggg
     2521 gggaaagctg ttgaatacga tgtgatgtcg atgccccggg ccatctagta tgacgtctgt
     2581 cgcacctgct tgatcgcggc cgcgatagct agatcgcgtt gctcctcttc tccatccgcg
     2641 ttccaagctg cggaaaggca cccataagcg tacgcctggt cgagcaggcg acgcggatcg
     2701 acgtccagcg cacgagagaa tgcgtccgcc atctgtgcaa tgcgtctagg atcgagacaa
     2761 aggtcgtctc tgtcagccgg atcgtagaac atattggcgg cgccaaagcc cacttcaccg
     2821 accagaccga cgggatctat caccagccag ccgcgactgg agaacatgat gttttcatga
     2881 tgcagatcgc catgtagccc acgcagttcc gaggcattgc tcatcatttg atcggctata
     2941 atcgccgcgt ggacgtagtc agtttgacaa cctgcgtttt gatcatcgcg cgcccgctga
     3001 aacaaagctg caaagcgatc ccggatcggg agaagggcag aaggcagggg ttcctcagat
     3061 gcggcataca gcttcgccat tagttccgct gcaatttcgg tcgcctggta gtcgccgtgc
     3121 tcggcaacga tgtgagagag cattcgctcc ccggcatatt cgagcaacat cagattgttc
     3181 tcacgaccga gcaaccggac tgctcccctc ccattgcgcc ataccagata gtcggccccg
     3241 cgcagttcat cagcaatgtc ttctataggt ttcaatccct tgacgattgc aggagtcccg
     3301 tctggcaatg aaactttcca aacgaggctg gaaaaggtgt ccgcaatgag aacaggttgc
     3361 gaaacgtgcc aatgagcagg aaaaacaggc ggcatgaaca tcaaccccaa gtcagagggt
     3421 ccaatcgcag atagaaggca aggcgttcgc ggtcgggggc ttcgatcccc aatacattga
     3481 ataggacagc gaaggcgcgc tctgcttcat ctggcgctgc ccagttctct tcggcgttag
     3541 caatcatgag tgccaaatcg gcatagcgat ctgctgttcc gagccgccca aggtcgatca
     3601 gacccgtgca ttgaagagtt ttagggtcca ccatgaagtt cggcatgcag ggatcaccat
     3661 ggcaaacaac catatcggtg cgctcttggt cgagccgcac cggtagctct cgttcgacac
     3721 gagccaaaag atcgtgcagc ggcgtactct tgtcctcgtc cggtaagaag tcgggattga
     3781 cggcattgcg ggacaccaca tcaacggcgc gtccgaacat tcgcgacagc ctgcgctcaa
     3841 acggacattg atcaaccgat aggctgtgaa cagcgccaag ttgctgcccc attgacggcc
     3901 acgctttgag caaatccgct ccagacagat cagccgccgg tactcccgga attgccgtta
     3961 tcaccaagca tgcaccctcc tgttcctcct gccagttgat gacctcgggg caagccacac
     4021 ctcgaccttt gagccaaatg aggcggtcac gctctccagc gagctcaccg cggcgggaag
     4081 caggtgcgat tttcgcgaag gcatgcccgt caccacgtcg aaaaacaaaa tcaccagatt
     4141 ctccgcctct gacaggcaac cagtcagaat gcgattcacc aaaaaaaata ttagttcgat
     4201 tcaatggagg ttccttcagt tttctgatga agcgcgaata tagagaaata tcccgaatgt
     4261 gcagttcctc gagttatttg tacaattcat ccataccatg ggtaatacca gcagcagtaa
     4321 caaattctaa caagaccatg tggtctctct tttcgtttgg atctttggat aaggcagatt
     4381 gataggataa gtaatggttg tctggtaaca agactggacc atcaccaatt ggagtatttt
     4441 gttgataatg gtcagctaat tgaacagaac catcttcaat gttgtgtcta attttgaagt
     4501 taactttgat accattcttt tgtttgtcag ccatgatgta aacattgtga gagttatagt
     4561 tgtattccaa tttgtgacct aaaatgttac catcttcttt aaaatcaata ccttttaatt
     4621 cgattctatt aactaaggta tcaccttcaa acttgacttc agctctggtc ttgtagttac
     4681 cgtcatcttt gaaaaaaata gttctttctt gaacataacc ttctggcatg gcagacttga
     4741 aaaagtcatg ttgtttcata tgatctgggt atctagcaaa acacatcaaa ccataaccta
     4801 aagtagtgac taaggttggc catggaactg gcaatttacc agtagtacaa ataaatttta
     4861 aggtcaattt accgtaagta gcatcacctt caccttcacc ggagacagaa aatttgtgac
     4921 cattaacatc accatctaat tcaaccaaaa ttgggacaac accagtgaat aattcttcac
     4981 ctttagacat gttaattaac ctactatcga tggggactga catgatcgat aatgatgaca
     5041 tcgctggtta gaggggaggt gccataaaat aggcaacctt tacatttcga cagaaatcat
     5101 agcaaaatag tgcccttaca gatagcagga attcgcgttg gccgattcat taatgcagct
     5161 gataacttcg tatagcatac attatacgaa cggtatcgac tacgtgcggg cagcgcgcgg
     5221 gcaaatcacc gctagtgaag ctggttctag aatggttgtc gacaacatca gtactatcat
     5281 ggatcctaac gacggtaatt gagaacctgc gtaaacggca tcagtgagtt gcggggtaag
     5341 cggatgcgat attgttgccg catccggcaa aaaaacgggc aaggtgtcac caccctgccc
     5401 tttttcttta aaaccgaaaa gattacttcg cgtgaacgtt ttttgaagct gttattgaaa
     5461 tgatttgcag tgtttttgtt tttcgtattt tcaatgaaac aaaagcaatt ttttactatc
     5521 tttcgcgttt cattgattaa cgactaaaat ttgcactaca ggcttgcaag ccccaccaaa
     5581 tctaatccca tcagatcccg aataatccaa tgtcctcccg gtttttttcg tacttcaagt
     5641 gaatcaatac aacccccatt cactgccaga gccgtgagcg cagcgaactg aatgtcacga
     5701 aaaagacagc gactcaggtg cctgatggtc ggagacaaaa ggaatattca gcgactagct
     5761 agttacatgg ctaattccca tgtcagccgt taagtgttcc tgtgtcactg aaaattgctt
     5821 tgagaggctc taagggcttc tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta
     5881 aaccttaaaa gctttaaaag ccttatatat tctttttttt cttataaaac ttaaaacctt
     5941 agaggctatt taagttgctg atttatatta attttattgt tcaaacatga gagcttagta
     6001 cgtgaaacat gagagcttag tacgttagcc atgagagctt agtacgttag ccatgagggt
     6061 ttagttcgtt aaacatgaga gcttagtacg ttaaacatga gagcttagta cgtgaaacat
     6121 gagagcttag tacgtactat caacaggttg aactgctgat cttcagatcc tctacgccgg
     6181 acgcatcgtg gccggatctt actagtatcc tttttaaccc atcacatata cctgccgttc
     6241 actattattt agtgaaatga gatattatga tattttctga attgtgatta aaaaggcaac
     6301 tttatgccca tgcaacagaa actataaaaa atacagagaa tgaaaagaaa cagatagatt
     6361 ttttagttct ttaggcccgt agtctgcaaa tccttttatg attttctatc aaacaaaaga
     6421 ggaaaataga ccagttgcaa tccaaacgag agtctaatag aatgaggtcg aaaagtaaat
     6481 cgcgcgggtt tgttactgat aaagcaggca agacctaaaa tgtgtaaagg gcaaagtgta
     6541 tactttggcg tcacccctta catattttag gtcttttttt attgtgcgta actaacttgc
     6601 catcttcaaa caggagggct ggaagaagca gaccgctaac acagtacata aaaaaggaga
     6661 catgaacgat gaacatcaaa aagtttgcaa aacaagcaac agtattaacc tttactaccg
     6721 cactgctggc aggaggcgca actcaagcgt ttgcgaaaga aacgaaccaa aagccatata
     6781 aggaaacata cggcatttcc catattacac gccatgatat gctgcaaatc cctgaacagc
     6841 aaaaaaatga aaaatatcaa gttcctgaat tcgattcgtc cacaattaaa aatatctctt
     6901 ctgcaaaagg cctggacgtt tgggacagct ggccattaca aaacgctgac ggcactgtcg
     6961 caaactatca cggctaccac atcgtctttg cattagccgg agatcctaaa aatgcggatg
     7021 acacatcgat ttacatgttc tatcaaaaag tcggcgaaac ttctattgac agctggaaaa
     7081 acgctggccg cgtctttaaa gacagcgaca aattcgatgc aaatgattct atcctaaaag
     7141 accaaacaca agaatggtca ggttcagcca catttacatc tgacggaaaa atccgtttat
     7201 tctacactga tttctccggt aaacattacg gcaaacaaac actgacaact gcacaagtta
     7261 acgtatcagc atcagacagc tctttgaaca tcaacggtgt agaggattat aaatcaatct
     7321 ttgacggtga cggaaaaacg tatcaaaatg tacagcagtt catcgatgaa ggcaactaca
     7381 gctcaggcga caaccatacg ctgagagatc ctcactacgt agaagataaa ggccacaaat
     7441 acttagtatt tgaagcaaac actggaactg aagatggcta ccaaggcgaa gaatctttat
     7501 ttaacaaagc atactatggc aaaagcacat cattcttccg tcaagaaagt caaaaacttc
     7561 tgcaaagcga ta
//
