LOCUS       40924_38793        5335 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSB890, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5335)
  AUTHORS   Klein K, Bliska J.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-DEC-2009) Microbiology and Molecular Genetics, Stony 
            Brook University, CMM 235, Stony Brook, NY 11794, USA
REFERENCE   2  (bases 1 to 5335)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5335)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (14-DEC-2009) Microbiology and Molecular Genetics, Stony Brook 
            University, CMM 235, Stony Brook, NY 11794, USA"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     88..104
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(830..2248)
                     /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"
     promoter        complement(2249..2694)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     rep_origin      complement(2749..3137)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     oriT            3314..3423
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             3456..3824
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     promoter        3993..4048
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     CDS             4056..5258
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MNSSTKIALVITLLDAMGIGLIMPVLPTLLREFIASEDIANHFGV
                     LLALYALMQVIFAPWLGKMSDRFGRRPVLLLSLIGASLDYLLLAFSSALWMLYLGRLLS
                     GITGATGAVAASVIADTTSASQRVKWFGWLGASFGLGLIAGPIIGGFAGEISPHSPFFI
                     AALLNIVTFLVVMFWFRETKNTRDNTDTEVGVETQSNSVYITLFKTMPILLIIYFSAQL
                     IGQIPATVWVLFTENRFGWNSMMVGFSLAGLGLLHSVFQAFVAGRIATKWGEKTAVLLG
                     FIADSSAFAFLAFISEGWLVFPVLILLAGGGIALPALQGVMSIQTKSHQQGALQGLLVS
                     LTNATXVIGPLLFAVIYNHSLPIWDGWIWIIGLAFYCIIILLSMTFMLTPQAQGSKQET
                     SA"
ORIGIN
        1 gataaagaag atttagcttc aaataaaacc tatctatttt atttatcttt caagctcaat
       61 aaaaagcccc accgcggtgg cggccgctct agaactagtg gatcccccgg gctgcagttc
      121 acttacaccg cttctcaacc cggtacgcac cagaaaatca ttgatatggc catgaatggc
      181 gttggatgcc gggcaacagc ccgcattatg ggcgttggcc tcaacacgat tttacgtcac
      241 ttaaaaaact caggccgcag tcggtaacct cgcgcataca gccgggcagt gacgtcatcg
      301 tctgcgcgga aatggacgaa cagtggggct atgtcggggc taaatcgcgc cagcgctggc
      361 tgttttacgc gtatgacagt ctccggaaga cggttgttgc gcacgtattc ggtgaamgca
      421 ctatggcgac gctggggcgt cttatgagcc tgctgtcacc ctttgacgtg gtgatatgga
      481 tgacggatgg ctggccgctg tatgaatccc gcctgaaggg aaagctgcac gtaatcagca
      541 agcgatatac gcagcgaatt gagcggcata acctgaatct gaggcagcac ctggcacggc
      601 tgggacggaa gtcgctgtcg ttctcaaaat cggtggagct gcatgacaaa gtcatcgggc
      661 attatctgaa cataaaacac tatcaataag ttggagtcat taccaaaagg ttaggaatac
      721 ggttagccat ttgcctgctt ttatatagtt catatgggat tcacctttat gttgataaga
      781 aataaaagaa aatgccaata ggatatcggc attttctttt gcgtttttat ttgttaactg
      841 ttaattgtcc ttgttcaagg atgctgtctt tgacaacaga tgttttcttg cctttgatgt
      901 tcagcaggaa gcttggcgca aacgttgatt gtttgtctgc gtagaatcct ctgtttgtca
      961 tatagcttgt aatcacgaca ttgtttcctt tcgcttgagg tacagcgaag tgtgagtaag
     1021 taaaggttac atcgttagga tcaagatcca tttttaacac aaggccagtt ttgttcagcg
     1081 gcttgtatgg gccagttaaa gaattagaaa cataaccaag catgtaaata tcgttagacg
     1141 taatgccgtc aatcgtcatt tttgatccgc gggagtcagt gaacaggtac catttgccgt
     1201 tcattttaaa gacgttcgcg cgttcaattt catctgttac tgtgttagat gcaatcagcg
     1261 gtttcatcac ttttttcagt gtgtaatcat cgtttagctc aatcataccg agagcgccgt
     1321 ttgctaactc agccgtgcgt tttttatcgc tttgcagaag tttttgactt tcttgacgga
     1381 agaatgatgt gcttttgcca tagtatgctt tgttaaataa agattcttcg ccttggtagc
     1441 catcttcagt tccagtgttt gcttcaaata ctaagtattt gtggccttta tcttctacgt
     1501 agtgaggatc tctcagcgta tggttgtcgc ctgagctgta gttgccttca tcgatgaact
     1561 gctgtacatt ttgatacgtt tttccgtcac cgtcaaagat tgatttataa tcctctacac
     1621 cgttgatgtt caaagagctg tctgatgctg atacgttaac ttgtgcagtt gtcagtgttt
     1681 gtttgccgta atgtttaccg gagaaatcag tgtagaataa acggattttt ccgtcagatg
     1741 taaatgtggc tgaacctgac cattcttgtg tttggtcttt taggatagaa tcatttgcat
     1801 cgaatttgtc gctgtcttta aagacgcggc cagcgttttt ccagctgtca atagaagttt
     1861 cgccgacttt ttgatagaac atgtaaatcg atgtgtcatc cgcattttta ggatctccgg
     1921 ctaatgcaaa gacgatgtgg tagccgtgat agtttgcgac agtgccgtca gcgttttgta
     1981 atggccagct gtcccaaacg tccaggcctt ttgcagaaga gatattttta attgtggacg
     2041 aatcgaattc aggaacttga tatttttcat ttttttgctg ttcagggatt tgcagcatat
     2101 catggcgtgt aatatgggaa atgccgtatg tttccttata tggcttttgg ttcgtttctt
     2161 tcgcaaacgc ttgagttgcg cctcctgcca gcagtgcggt agtaaaggtt aatactgttg
     2221 cttgttttgc aaactttttg atgttcatcg ttcatgtctc cttttttatg tactgtgtta
     2281 gcggtctgct tcttccagcc ctcctgtttg aagatggcaa gttagttacg cacaataaaa
     2341 aaagacctaa aatatgtaag gggtgacgcc aaagtataca ctttgccctt tacacatttt
     2401 aggtcttgcc tgctttatca gtaacaaacc cgcgcgattt acttttcgac ctcattctat
     2461 tagactctcg tttggattgc aactggtcta ttttcctctt ttgtttgata gaaaatcata
     2521 aaaggatttg cagactacgg gcctaaagaa ctaaaaaatc tatctgtttc ttttcattct
     2581 ctgtattttt tatagtttct gttgcatggg cataaagttg cctttttaat cacaattcag
     2641 aaaatatcat aatatctcat ttcactaaat aatagtgaac ggcaggtata tgtgatgggt
     2701 taaaaaggat cttaaggcct aggtctagct tggctgcagg aattcccatg tcagccgtta
     2761 agtgttcctg tgtcactcaa aattgctttg agaggctcta agggcttctc agtgcgttac
     2821 atccctggct tgttgtccac aaccgttaaa ccttaaaagc tttaaaagcc ttatatattc
     2881 ttttttttct tataaaactt aaaaccttag aggctattta agttgctgat ttatattaat
     2941 tttattgttc aaacatgaga gcttagtacg tgaaacatga gagcttagta cgttagccat
     3001 gagagcttag tacgttagcc atgagggttt agttcgttaa acatgagagc ttagtacgtt
     3061 aaacatgaga gcttagtacg tgaaacatga gagcttagta cgtactatca acaggttgaa
     3121 ctgctgatct tcagatcctc tacgccggac gcatcgtggc cggatctaga tgacggatct
     3181 tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct
     3241 ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc
     3301 aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct
     3361 tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc
     3421 cggctaccgc cggcgtaaca gatgaaggca agcggatggc tgatgaaacc aagccaacca
     3481 ggaagggcag cccacctatc aaggtgtact gccttccaga cgaacgaaga gcgattgagg
     3541 aaaaggcggc ggcggccggc atgagcctgt cggcctacct gctggccgtc ggccagggct
     3601 acaaaatcac gggcgtcgtg gactatgagc acgtccgcga gctggcccgc atcaatggcg
     3661 acctgggccg cctgggcggc ctgctgaaac tctggctcac cgacgacccg cgcacggcgc
     3721 ggttcggtga tgccacgatc ctcgccctgc tggcgaagat cgaagagaag caggacgagc
     3781 ttggcaaggt catgatgggc gtggtccgcc cgagggcaga gccatgactt ttttagccgc
     3841 taaaacggcc ggggggtgcg cgtgattgcc aagcacgtcc ccatgcgctc catcaagaag
     3901 agcgacttcg cggagctggt gaagtacatc accgacgagc aaggcaagac cgatccccga
     3961 cctcgactag acatcattaa ttcctaattt ttgttgaaac tctatcattg atagagttat
     4021 tttaccactc cctatcagtg atagagaaaa gtgaaatgaa tagttcgaca aagatcgcat
     4081 tggtaattac gttactcgat gccatgggga ttggccttat catgccagtc ttgccaacgt
     4141 tattacgtga atttattgct tcggaagata tcgctaacca ctttggcgta ttgcttgcac
     4201 tttatgcgtt aatgcaggtt atctttgctc cttggcttgg aaaaatgtct gaccgatttg
     4261 gtcggcgccc agtgctgttg ttgtcattaa taggcgcatc gctggattac ttattgctgg
     4321 ctttttcaag tgcgctttgg atgctgtatt taggccgttt gctttcaggg atcacaggag
     4381 ctactggggc tgtcgcggca tcggtcattg ccgataccac ctcagcttct caacgcgtga
     4441 agtggttcgg ttggttaggg gcaagttttg ggcttggttt aatagcgggg cctattattg
     4501 gtggttttgc aggagagatt tcaccgcata gtcccttttt tatcgctgcg ttgctaaata
     4561 ttgtcacttt ccttgtggtt atgttttggt tccgtgaaac caaaaataca cgtgataata
     4621 cagataccga agtaggggtt gagacgcaat cgaattcggt atacatcact ttatttaaaa
     4681 cgatgcccat tttgttgatt atttattttt cagcgcaatt gataggccaa attcccgcaa
     4741 cggtgtgggt gctatttacc gaaaatcgtt ttggatggaa tagcatgatg gttggctttt
     4801 cattagcggg tcttggtctt ttacactcag tattccaagc ctttgtggca ggaagaatag
     4861 ccactaaatg gggcgaaaaa acggcagtac tgctcggatt tattgcagat agtagtgcat
     4921 ttgccttttt agcgtttata tctgaaggtt ggttagtttt ccctgtttta attttattgg
     4981 ctggtggtgg gatcgcttta cctgcattac agggagtgat gtctatccaa acaaagagtc
     5041 atcagcaagg tgctttacag ggattattgg tgagccttac caatgcaacc gstgttattg
     5101 gcccattact gtttgctgtt atttataatc attcactacc aatttgggat ggctggattt
     5161 ggattattgg tttagcgttt tactgtatta ttatcctgct atcgatgacc ttcatgttaa
     5221 cccctcaagc tcaggggagt aaacaggaga caagtgctta gttatttcgt caccaaatga
     5281 tgttattccg cgaaatataa tgaccctctt gataacccaa gagggcattt tttac
//
