LOCUS       Exported                6672 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pYMD2 DNA, complete sequence.
ACCESSION   AB477002
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6672
                     /organism="Suicide vector pYMD2"
                     /mol_type="other DNA"
                     /db_xref="taxon:589555"
     CDS             38..1459
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /protein_id="BAH11108.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            38..1459
                     /gene="sacB"
                     /label=sacB
     protein_bind    2678..2699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /bound_moiety="lac repressor encoded by lacI"
                     /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"
     CDS             2788..3207
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWGSIPSRVGSAAAWSWTTSRSLPAVQIR
                     RHPGNQQRLSAHPCPRTAGVGRHDGRFGRSCLARR"
     misc_feature    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(3309..3680)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            3713..3822
                     /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(5207..6067)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="BAH11109.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5207..6067)
                     /gene="bla"
                     /label=bla
     promoter        complement(6068..6172)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6204..6220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6264..37
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"