LOCUS       40924_48223        6672 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pYMD2 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6672)
  AUTHORS   Yamada K, Kaneko J, Itoh Y.
  TITLE     Role sharing among a population of Pectobacterium carotovorum subsp.
            carotovorum Er
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6672)
  AUTHORS   Yamada K, Kaneko J.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JAN-2009) Contact:Kazuteru Yamada Tohoku University, 
            Department of Microbial Biotechnology; Tsutsumidori Amamiya-machi 
            1-1, Aoba-ku, Sendai, Miyagi 981-8555, Japan
REFERENCE   3  (bases 1 to 6672)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6672)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-JAN-2009) Contact:Kazuteru Yamada Tohoku University, Department 
            of Microbial Biotechnology; Tsutsumidori Amamiya-machi 1-1, Aoba-ku,
            Sendai, Miyagi 981-8555, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             38..1456
                     /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"
     protein_bind    2678..2699
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2714..2744
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2752..2768
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2776..2792
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(2805..2861)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2862..2878)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3312..3680)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(3713..3822)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      3982..4370
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(5210..6067)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6068..6172)
                     /label=AmpR promoter
     primer_bind     6204..6220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6264..6672
                     /label=sacB promoter
                     /note="sacB promoter and control region"
ORIGIN
        1 accgctaaca cagtacataa aaaaggagac atgaacgatg aacatcaaaa agtttgcaaa
       61 acaagcaaca gtattaacct ttactaccgc actgctggca ggaggcgcaa ctcaagcgtt
      121 tgcgaaagaa acgaaccaaa agccatataa ggaaacatac ggcatttccc atattacacg
      181 ccatgatatg ctgcaaatcc ctgaacagca aaaaaatgaa aaatatcaag ttcctgaatt
      241 cgattcgtcc acaattaaaa atatctcttc tgcaaaaggc ctggacgttt gggacagctg
      301 gccattacaa aacgctgacg gcactgtcgc aaactatcac ggctaccaca tcgtctttgc
      361 attagccgga gatcctaaaa atgcggatga cacatcgatt tacatgttct atcaaaaagt
      421 cggcgaaact tctattgaca gctggaaaaa cgctggccgc gtctttaaag acagcgacaa
      481 attcgatgca aatgattcta tcctaaaaga ccaaacacaa gaatggtcag gttcagccac
      541 atttacatct gacggaaaaa tccgtttatt ctacactgat ttctccggta aacattacgg
      601 caaacaaaca ctgacaactg cacaagttaa cgtatcagca tcagacagct ctttgaacat
      661 caacggtgta gaggattata aatcaatctt tgacggtgac ggaaaaacgt atcaaaatgt
      721 acagcagttc atcgatgaag gcaactacag ctcaggcgac aaccatacgc tgagagatcc
      781 tcactacgta gaagataaag gccacaaata cttagtattt gaagcaaaca ctggaactga
      841 agatggctac caaggcgaag aatctttatt taacaaagca tactatggca aaagcacatc
      901 attcttccgt caagaaagtc aaaaacttct gcaaagcgat aaaaaacgca cggctgagtt
      961 agcaaacggc gctctcggta tgattgagct aaacgatgat tacacactga aaaaagtgat
     1021 gaaaccgctg attgcatcta acacagtaac agatgaaatt gaacgcgcga acgtctttaa
     1081 aatgaacggc aaatggtacc tgttcactga ctcccgcgga tcaaaaatga cgattgacgg
     1141 cattacgtct aacgatattt acatgcttgg ttatgtttct aattctttaa ctggcccata
     1201 caagccgctg aacaaaactg gccttgtgtt aaaaatggat cttgatccta acgatgtaac
     1261 ctttacttac tcacacttcg ctgtacctca agcgaaagga aacaatgtcg tgattacaag
     1321 ctatatgaca aacagaggat tctacgcaga caaacaatca acgtttgcgc caagcttcct
     1381 gctgaacatc aaaggcaaga aaacatctgt tgtcaaagac agcatccttg aacaaggaca
     1441 attaacagtt aacaaataaa aacgcaaaag aaaatgccga tatcctattg gcattttctt
     1501 ttatttctta tcaacataaa ggtgaatccc atatgaacta tataaaagca ggcaaatggc
     1561 taaccgtatt cctaaccttt tggtaatgac tccaacttat tgatagtgtt ttatgttcag
     1621 ataatgcccg atgactttgt catgcagctc caccgatttt gagaacgaca gcgacttccg
     1681 tcccagccgt gccaggtgct gcctcagatt caggttatgc cgctcaattc gctgcgtata
     1741 tcgcttgctg attacgtgca gctttccctt caggcgggat tcatacagcg gccagccatc
     1801 cgtcatccat atcaccacgt caaagggtga cagcaggctc ataagacgcc ccagcgtcgc
     1861 catagtgcgt tcaccgaata cgtgcgcaac aaccgtcttc cggagactgt catacgcgta
     1921 aaacagccag cgctggcgcg atttagcccc gacatagccc cactgttcgt ccatttccgc
     1981 gcagacgatg acgtcactgc ccggctgtat gcgcgaggtt accgactgcg gcctgagttt
     2041 tttaagtgac gtaaaatcgt gttgaggcca acgcccataa tgcgggctgt tgcccggcat
     2101 ccaacgccat tcatggccat atcaatgatt ttctggtgcg taccgggttg agaagcggtg
     2161 taagtgaacc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     2221 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     2281 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     2341 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     2401 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     2461 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     2521 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccccaa
     2581 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2641 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     2701 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     2761 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt gcatgcctgc
     2821 aggtcgactc tagaggatcc ccgggtaccg agctcgaatt cactggccgt cgttttacaa
     2881 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     2941 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     3001 agcctgaatg gcgaatgggg ctcgatcccc tcgcgagttg gttcagctgc tgcctggagc
     3061 tggacgacct cgcggagtct accggcagtg caaatccgtc ggcatccagg aaaccagcag
     3121 cggctatccg cgcatccatg cccccgaact gcaggagtgg ggaggcacga tggccgcttt
     3181 ggtcggtctt gccttgctcg tcggtgatgt acttcaccag ctccgcgaag tcgctcttct
     3241 tgatggagcg catggggacg tgcttggcaa tcacgcgcac cccccggccg ttttagcggc
     3301 taaaaaagtc atggctctgc cctcgggcgg accacgccca tcatgacctt gccaagctcg
     3361 tcctgcttct cttcgatctt cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc
     3421 gtgcgcgggt cgtcggtgag ccagagtttc agcaggccgc ccaggcggcc caggtcgcca
     3481 ttgatgcggg ccagctcgcg gacgtgctca tagtccacga cgcccgtgat tttgtagccc
     3541 tggccgacgg ccagcaggta ggccgacagg ctcatgccgg ccgccgccgc cttttcctca
     3601 atcgctcttc gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt
     3661 ggcttggttt catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag
     3721 cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag
     3781 gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata
     3841 caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat
     3901 ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaacggc
     3961 cacgatgcgt ccggcgtaga ggatctgaag atcagcagtt caacctgttg atagtacgta
     4021 ctaagctctc atgtttcacg tactaagctc tcatgtttaa cgtactaagc tctcatgttt
     4081 aacgaactaa accctcatgg ctaacgtact aagctctcat ggctaacgta ctaagctctc
     4141 atgtttcacg tactaagctc tcatgtttga acaataaaat taatataaat cagcaactta
     4201 aatagcctct aaggttttaa gttttataag aaaaaaaaga atatataagg cttttaaagc
     4261 ttttaaggtt taacggttgt ggacaacaag ccagggatgt aacgcactga gaagccctta
     4321 gagcctctca aagcaatttt gagtgacaca ggaacactta acggctgaca tgggaattag
     4381 cttcacgctg ccgcaagcac tcagggcgca agggctgcta aaggaagcgg aacacgtaga
     4441 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga
     4501 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat
     4561 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct
     4621 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
     4681 gatggcgcag gggatcaaga tcggccgcaa tcgggcaaat cgctgaatat tccttttgtc
     4741 tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac
     4801 ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag
     4861 gaaactcaaa ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg
     4921 tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc
     4981 agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta
     5041 aggcagcggt atcatcaacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5101 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5161 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5221 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5281 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5341 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5401 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5461 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5521 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5581 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5641 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5701 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5761 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5821 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5881 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5941 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6001 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6061 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6121 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6181 cccgaaaagt gccacttgca tcgcaggaaa cagctatgac catgattacg aattcgctag
     6241 aggatcgatc ctttttaacc catcacatat acctgccgtt cactattatt tagtgaaatg
     6301 agatattatg atattttctg aattgtgatt aaaaaggcaa ctttatgccc atgcaacaga
     6361 aactataaaa aatacagaga atgaaaagaa acagatagat tttttagttc tttaggcccg
     6421 tagtctgcaa atccttttat gattttctat caaacaaaag aggaaaatag accagttgca
     6481 atccaaacga gagtctaata gaatgaggtc gaaaagtaaa tcgcgcgggt ttgttactga
     6541 taaagcaggc aagacctaaa atgtgtaaag ggcaaagtgt atactttggc gtcacccctt
     6601 acatatttta ggtctttttt tattgtgcgt aactaacttg ccatcttcaa acaggagggc
     6661 tggaagaagc ag
//
