LOCUS       40924_46593        8261 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWM91 DNA, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8261)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "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/"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(59..415)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     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    complement(799..906)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     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    complement(982..998)
                     /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)."
     promoter        complement(1006..1036)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1051..1072)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     oriT            1778..1887
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(2113..2156)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     oriT            2208..2317
                     /label=incP origin of transfer
                     /note="incP origin of transfer"
     terminator      2565..2624
                     /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(2819..3407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4444..4889
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             4890..6308
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     CDS             complement(7199..8056)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8057..8161)
                     /label=AmpR promoter
ORIGIN
        1 gatcgctagt ttgttttgac tccatccatt agggcttcta aaacgccttc taaggccatg
       61 tcagccgtta agtgttcctg tgtcactgaa aattgctttg agaggctcta agggcttctc
      121 agtgcgttac atccctggct tgttgtccac aaccgttaaa ccttaaaagc tttaaaagcc
      181 ttatatattc ttttttttct tataaaactt aaaaccttag aggctattta agttgctgat
      241 ttatattaat tttattgttc aaacatgaga gcttagtacg tgaaacatga gagcttagta
      301 cgttagccat gagagcttag tacgttagcc atgagggttt agttcgttaa acatgagagc
      361 ttagtacgtt aaacatgaga gcttagtacg tgaaacatga gagcttagta cgtactatca
      421 acaggttgaa ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
      481 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
      541 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
      601 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      661 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      721 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
      781 tcactatagg gcgaattggg taccgggccc cccctcgagg tcgacggtat cgataagctt
      841 gatatcgaat tcctgcagcc cgggggatcc actagttcta gagcggccgc caccgcggtg
      901 gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
      961 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1021 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1081 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cactgcatta atgaatcggc
     1141 caacgcgcgg ggagagatcc tttttgtccg gtgttgggtt gaaggtgaag ccggtcgggg
     1201 ccgcagcggg ggccggcttt tcagccttgc ccccctgctt cggccgccgt ggctccggcg
     1261 tcttgggtgc cggcgcgggt tccgcagcct tggcctgcgg tgcgggcaca tcggcgggct
     1321 tggccttgat gtgccgcctg gcgtgcgagc ggaacgtctc gtaggagaac ttgaccttcc
     1381 ccgtttcccg catgtgctcc caaatggtga cgagcgcata gccggacgct aacgccgcct
     1441 cgacatccgc cctcaccgcc aggaacgcaa ccgcagcctc atcacgccgg cgcttcttgg
     1501 ccgcgcggga ttcaacccac tcggccagct cgtcggtgta gctctttggc atcgtctctc
     1561 gcctgtcccc tcagttcagt aatttcctgc atttgcctgt ttccagtcgg tagatattcc
     1621 acaaaacagc agggaagcag cgcttttccg ctgcataacc ctgcttcggg gtcattatag
     1681 cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc
     1741 gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc
     1801 cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg
     1861 ggaatcctgc tctgcgaggc tggccggcta ccgccggcgt aacagatgag ggcaagcgga
     1921 tggctgatga aaccaagcca accaggaagg gcagcccacc tatcaaggtg tactgccttc
     1981 cagacgaacg aagagcgatt gaggaaaagg cggcggcggc cggcatgagc ctgtcggcct
     2041 acccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     2101 tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctgga
     2161 attcatcctg aacttatcta gacttggtgt atccaacggc gtcagccggg caggataggt
     2221 gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg
     2281 gtgctcaacg ggaatcctgc tctgcnaggc tggccggcta ccgccggcgt aacagatgag
     2341 ggcaagcgga tggctgatga aaccaagcca accaggaagg gcagcccacc tatcacggaa
     2401 ttgatccccc tcgaattgac gcgtcaattc tcatgtttga cagcttatca tcgataagct
     2461 agctttaatg cggtagttta tcacagttaa attgctaacc agtcactatg gcgtgctgct
     2521 agcgctatat gcgttgatgc actagttaac ttcaggacct gcagtccggc aaaaaaacgg
     2581 gcaaggtgtc accaccctgc cctttttctt taaaaccgaa aagattactt cgcgttatgc
     2641 aggcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2701 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2761 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2821 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2881 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2941 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3001 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3061 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3121 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3181 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3241 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3301 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3361 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3421 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3481 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3541 caatctaaag tatatatgag tatgctggcc gtcggccagg gctacaaaat cacgggcgtc
     3601 gtggactatg agcacgtccg cgagctggcc cgcatcaatg gcgacctggg ccgcctgggc
     3661 ggcctgctga aactctggct caccgacgac ccgcgcacgg cgcggttcgg tgatgccacg
     3721 atcctcgccc tgctggcgaa gatcgaagag aagcaggacg agcttggcaa ggtcatgatg
     3781 ggcgtggtcc gcccgagggc agagccatga cttttttagc cgctaaaacg gccggggggt
     3841 gcgcgtgatt gccaagcacg tccccatgcg ctccatcaag aagagcgact tcgcggagct
     3901 ggtgaagtac atcaccgacg agcaaggcaa gaccgagcgc ctgggtcacg tgcgcgtcac
     3961 gaactgcgag gcaaacaccc tgcccgctgt catggccgag gtgatggcga cccagcacgg
     4021 caacacccgt tccgaggccg acaagaccta tcacctgctg gttagcttcc gcgcgggaga
     4081 gaagcccgac gcggagacgt tgcgcgcgat tgaggaccgc atctgcgctg ggcttggctt
     4141 cgccgagcat cagcgcgtca gtgccgtgca tcacgacacc gacaacctgc acatccatat
     4201 cgccatcaac aagattcacc cgacccgaaa caccatccat gagccgtatc gggcctaccg
     4261 cgccctcgct gacctctgcg cgacgctcga acgggactac gggcttgagc gtgacaatca
     4321 cgaaacgcgg cagcgcgttt ccgagaaccg cgcgaacgac atggagcggc acgcgggcgt
     4381 ggaaagcctg gtcggctggc gggactctgg ggttcgctag aggatcgatc ctttttaacc
     4441 catcacatat acctgccgtt cactattatt tagtgaaatg agatattatg atattttctg
     4501 aattgtgatt aaaaaggcaa ctttatgccc atgcaacaga aactataaaa aatacagaga
     4561 atgaaaagaa acagatagat tttttagttc tttaggcccg tagtctgcaa atccttttat
     4621 gattttctat caaacaaaag aggaaaatag accagttgca atccaaacga gagtctaata
     4681 gaatgaggtc gaaaagtaaa tcgcgcgggt ttgttactga taaagcaggc aagacctaaa
     4741 atgtgtaaag ggcaaagtgt atactttggc gtcacccctt acatatttta ggtctttttt
     4801 tattgtgcgt aactaacttg ccatcttcaa acaggagggc tggaagaagc agaccgctaa
     4861 cacagtacat aaaaaaggag acatgaacga tgaacatcaa aaagtttgca aaacaagcaa
     4921 cagtattaac ctttactacc gcactgctgg caggaggcgc aactcaagcg tttgcgaaag
     4981 aaacgaacca aaagccatat aaggaaacat acggcatttc ccatattaca cgccatgata
     5041 tgctgcaaat ccctgaacag caaaaaaatg aaaaatatca agtttctgaa tttgattcgt
     5101 ccacaattaa aaatatctct tctgcaaaag gcctggacgt ttgggacagc tggccattac
     5161 aaaacgctga cggcactgtc gcaaactatc acggctacca catcgtcttt gcattagccg
     5221 gagatcctaa aaatgcggat gacacatcga tttacatgtt ctatcaaaaa gtcggcgaaa
     5281 cttctattga cagctggaaa aacgctggcc gcgtctttaa agacagcgac aaattcgatg
     5341 caaatgattc tatcctaaaa gaccaaacac aagaatggtc aggttcagcc acatttacat
     5401 ctgacggaaa aatccgttta ttctacactg atttctccgg taaacattac ggcaaacaaa
     5461 cactgacaac tgcacaagtt aacgtatcag catcagacag ctctttgaac atcaacggtg
     5521 tagaggatta taaatcaatc tttgacggtg acggaaaaac gtatcaaaat gtacagcagt
     5581 tcatcgatga aggcaactac agctcaggcg acaaccatac gctgagagat cctcactacg
     5641 tagaagataa aggccacaaa tacttagtat ttgaagcaaa cactggaact gaagatggct
     5701 accaaggcga agaatcttta tttaacaaag catactatgg caaaagcaca tcattcttcc
     5761 gtcaagaaag tcaaaaactt ctgcaaagcg ataaaaaacg cacggctgag ttagcaaacg
     5821 gcgctctcgg tatgattgag ctaaacgatg attacacact gaaaaaagtg atgaaaccgc
     5881 tgattgcatc taacacagta acagatgaaa ttgaacgcgc gaacgtcttt aaaatgaacg
     5941 gcaaatggta cctgttcact gactcccgcg gatcaaaaat gacgattgac ggcattacgt
     6001 ctaacgatat ttacatgctt ggttatgttt ctaattcttt aactggccca tacaagccgc
     6061 tgaacaaaac tggccttgtg ttaaaaatgg atcttgatcc taacgatgta acctttactt
     6121 actcacactt cgctgtacct caagcgaaag gaaacaatgt cgtgattaca agctatatga
     6181 caaacagagg attctacgca gacaaacaat caacgtttgc gccgagcttc ctgctgaaca
     6241 tcaaaggcaa gaaaacatct gttgtcaaag acagcatcct tgaacaagga caattaacag
     6301 ttaacaaata aaaacgcaaa agaaaatgcc gatgggtacc gagcgaaatg accgaccaag
     6361 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6421 gcttcggaat cgttttccgg gacgccctcg cggacgtgct catagtccac gacgcccgtg
     6481 attttgtagc cctggccgac ggccagcagg taggccgaca ggctcatgcc ggccgccgcc
     6541 gccttttcct caatcgctct tcgttcgtct ggaaggcagt acaccttgat aggtgggctg
     6601 cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt tacgccggcg
     6661 gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataagga
     6721 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
     6781 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
     6841 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
     6901 ccttgctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     6961 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     7021 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     7081 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccaaaatt
     7141 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7201 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7261 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7321 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7381 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7441 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7501 ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7561 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7621 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7681 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7741 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7801 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7861 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7921 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7981 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8041 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8101 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8161 gcacatttcc ccgaaaagtg ccacctgcag atctgcaggt cgacggatcc caagcttctt
     8221 ctagaggtac cgcatgcgat atcgagctct cccgggaatt c
//
