LOCUS       Exported                8261 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWM91 DNA, complete sequence.
ACCESSION   LC144618
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8261)
  AUTHORS   Okamoto S, Niki H.
  TITLE     cloning vector pWM91 sequence
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8261)
  AUTHORS   Okamoto S, Niki H.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-APR-2016) Contact:Sho Okamoto National Institute of 
            Genetics, Microbial Genetics Laboratory, Genetics Strains Research 
            Center; 1111 Yata, Mishima, Shizuoka 411-8540, Japan URL 
            :https://www.nig.ac.jp/labs/MicroGen/
REFERENCE   3  (bases 1 to 8261)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8261
                     /organism="Cloning vector pWM91"
                     /mol_type="other DNA"
                     /db_xref="taxon:1825573"
     rep_origin      59..415
                     /label=R6K gamma ori
                     /note="R6K gamma ori"
     CDS             complement(405..962)
                     /codon_start=1
                     /gene="lacZ alpha"
                     /product="LacZ alpha fragment of beta-galactosidase"
                     /label=lacZ alpha
                     /protein_id="BAV13342.1"
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
                     FPRQALNRGLPLGFLQPVDSTY"
     gene            complement(405..962)
                     /gene="lacZ alpha"
                     /label=lacZ alpha
     primer_bind     746..762
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        772..790
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    799..906
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     816..832
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(866..882)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(919..937)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(958..974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    982..998
                     /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)."
     promoter        complement(1006..1036)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1051..1072
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     oriT            1778..1887
                     /note="incP origin of transfer"
     terminator      2113..2156
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     oriT            2208..2317
                     /label=incP origin of transfer
                     /note="incP origin of transfer"
     rep_origin      complement(2819..3407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4444..4889
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             4890..6311
                     /codon_start=1
                     /gene="sacB"
                     /product="secreted levansucrase"
                     /function="renders bacterial growth sensitive to sucrose"
                     /label=sacB
                     /protein_id="BAV13343.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            4890..6311
                     /gene="sacB"
                     /label=sacB
     CDS             complement(7196..8056)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="BAV13344.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7196..8056)
                     /gene="bla"
                     /label=bla
     promoter        complement(8057..8161)
                     /gene="bla"
                     /label=AmpR promoter