LOCUS       62056_18775        5760 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5760)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      93..482
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     promoter        510..955
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             956..2374
                     /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"
     CDS             complement(3219..3875)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3876..3978)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     oriT            4707..4816
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             4849..5217
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 cgcgtttccg agaaccgcgc gaacgacatg gagcggcacg cgggcgtgga aagcctggtc
       61 ggctggatcg gcacgatgcg tccggcgtag aggatctgaa gatccagcag ttcaacctgt
      121 tgatagtacg tactaagctc tcatgtttca cgtactaagc tctcatgttt aacgtactaa
      181 gctctcatgt ttaacgaact aaaccctcat ggctaacgta ctaagctctc atggctaacg
      241 tactaagctc tcatgtttca cgtactaagc tctcatgttt gaacaataaa attaatataa
      301 atcagcaact taaatagcct ctaaggtttt aagttttata agaaaaaaaa gaatatataa
      361 ggcttttaaa gcttttaagg tttaacggtt gtggacaaca agccagggat gtaacgcact
      421 gagaagccct tagagcctct caaagcaatt ttcagtgaca caggaacact taacggctga
      481 catgggaatt ctgatccttt ttaacccatc acatatacct gccgttcact attatttagt
      541 gaaatgagat attatgatat tttctgaatt gtgattaaaa aggcaacttt atgcccatgc
      601 aacagaaact ataaaaaata cagagaatga aaagaaacag atagattttt tagttcttta
      661 ggcccgtagt ctgcaaatcc ttttatgatt ttctatcaaa caaaagagga aaatagacca
      721 gttgcaatcc aaacgagagt ctaatagaat gaggtcgaaa agtaaatcgc gcgggtttgt
      781 tactgataaa gcaggcaaga cctaaaatgt gtaaagggca aagtgtatac tttggcgtca
      841 ccccttacat attttaggtc tttttttatt gtgcgtaact aacttgccat cttcaaacag
      901 gagggctgga agaagcagac cgctaacaca gtacataaaa aaggagacat gaacgatgaa
      961 catcaaaaag tttgcaaaac aagcaacagt attaaccttt actaccgcac tgctggcagg
     1021 aggcgcaact caagcgtttg cgaaagaaac gaaccaaaag ccatataagg aaacatacgg
     1081 catttcccat attacacgcc atgatatgct gcaaatccct gaacagcaaa aaaatgaaaa
     1141 atatcaagtt cctgagttcg attcgtccac aattaaaaat atctcttctg caaaaggcct
     1201 ggacgtttgg gacagctggc cattacaaaa cgctgacggc actgtcgcaa actatcacgg
     1261 ctaccacatc gtctttgcat tagccggaga tcctaaaaat gcggatgaca catcgattta
     1321 catgttctat caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg ctggccgcgt
     1381 ctttaaagac agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc aaacacaaga
     1441 atggtcaggt tcagccacat ttacatctga cggaaaaatc cgtttattct acactgattt
     1501 ctccggtaaa cattacggca aacaaacact gacaactgca caagttaacg tatcagcatc
     1561 agacagctct ttgaacatca acggtgtaga ggattataaa tcaatctttg acggtgacgg
     1621 aaaaacgtat caaaatgtac agcagttcat cgatgaaggc aactacagct caggcgacaa
     1681 ccatacgctg agagatcctc actacgtaga agataaaggc cacaaatact tagtatttga
     1741 agcaaacact ggaactgaag atggctacca aggcgaagaa tctttattta acaaagcata
     1801 ctatggcaaa agcacatcat tcttccgtca agaaagtcaa aaacttctgc aaagcgataa
     1861 aaaacgcacg gctgagttag caaacggcgc tctcggtatg attgagctaa acgatgatta
     1921 cacactgaaa aaagtgatga aaccgctgat tgcatctaac acagtaacag atgaaattga
     1981 acgcgcgaac gtctttaaaa tgaacggcaa atggtatctg ttcactgact cccgcggatc
     2041 aaaaatgacg attgacggca ttacgtctaa cgatatttac atgcttggtt atgtttctaa
     2101 ttctttaact ggcccataca agccgctgaa caaaactggc cttgtgttaa aaatggatct
     2161 tgatcctaac gatgtaacct ttacttactc acacttcgct gtacctcaag cgaaaggaaa
     2221 caatgtcgtg attacaagct atatgacaaa cagaggattc tacgcagaca aacaatcaac
     2281 gtttgcgccc agcttcctgc tgaacatcaa aggcaagaaa acatctgttg tcaaagacag
     2341 catccttgaa caaggacaat taacagttaa caaataaaaa cgcaaaagaa aatgccgata
     2401 tcctattggc attttctttt atttcttatc aacataaagg tgaatcccat atgaactata
     2461 taaaagcagg caaatggcta accgtattcc taaccttttg gtaatgactc caacttattg
     2521 atagtgtttt atgttcagat aatgcccgat gactttgtca tgcagctcca ccgattttga
     2581 gaacgacagc gacttccgtc ccagccgtgc caggtgctgc ctcagattca ggttatgccg
     2641 ctcaattcgc tgcgtatatc gcttgctgat tacgtgcagc tttcccttca ggcgggattc
     2701 atacagcggc cagccatccg tcatccatat caccacgtca aagggtgaca gcaggctcat
     2761 aagacgcccc agcgtcgcca tagtgcgttc accgaatacg tgcgcaacaa ccgtcttccg
     2821 gagactgtca tacgcgtaaa acagccagcg ctggcgcgat ttagccccga catagcccca
     2881 ctgttcgtcc atttccgcgc agacgatgac gtcactgccc ggctgtatgc gcgaggttac
     2941 cgactgcggc ctgagttttt taagtgacgt aaaatcgtgt tgaggccaac gcccataatg
     3001 cgggctgttg cccggcatcc aacgccattc atggccatat caatgatttt ctggtgcgta
     3061 ccgggttgag aagcggtgta agtgaactgc atgaattccc gggagagctc gatatcgcat
     3121 gcggtacctc tagaagaagc ttgggatcgg gccctatcac ttattcaggc gtagcaacca
     3181 ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg
     3241 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca gacggcatga
     3301 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
     3361 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
     3421 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
     3481 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
     3541 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
     3601 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat
     3661 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
     3721 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
     3781 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     3841 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     3901 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     3961 cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta
     4021 tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac aggtattagg
     4081 gcccgatcct ttttgtccgg tgttgggttg aaggtgaagc cggtcggggc cgcagcgggg
     4141 gccggctttt cagccttgcc cccctgcttc ggccgccgtg gctccggcgt cttgggtgcc
     4201 ggcgcgggtt ccgcagcctt ggcctgcggt gcgggcacat cggcgggctt ggccttgatg
     4261 tgccgcctgg cgtgcgagcg gaacgtctcg taggagaact tgaccttccc cgtttcccgc
     4321 atgtgctccc aaatggtgac gagcgcatag ccggacgcta acgccgcctc gacatccgcc
     4381 ctcaccgcca ggaacgcaac cgcagcctca tcacgccggc gcttcttggc cgcgcgggat
     4441 tcaacccact cggccagctc gtcggtgtag ctctttggca tcgtctctcg cctgtcccct
     4501 cagttcagta atttcctgca tttgcctgtt tccagtcggt agatattcca caaaacagca
     4561 gggaagcagc gcttttccgc tgcataaccc tgcttcgggg tcattatagc gattttttcg
     4621 gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg tgtagacttt
     4681 ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc acccgcgagc
     4741 gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg gaatcctgct
     4801 ctgcgaggct ggccggctac cgccggcgta acagatgagg gcaagcggat ggctgatgaa
     4861 accaagccaa ccaggaaggg cagcccacct atcaaggtgt actgccttcc agacgaacga
     4921 agagcgattg aggaaaaggc ggcggcggcc ggcatgagcc tgtcggccta cctgctggcc
     4981 gtcggccagg gctacaaaat cacgggcgtc gtggactatg agcacgtccg cgagctggcc
     5041 cgcatcaatg gcgacctggg ccgcctgggc ggcctgctga aactctggct caccgacgac
     5101 ccgcgcacgg cgcggttcgg tgatgccacg atcctcgccc tgctggcgaa gatcgaagag
     5161 aagcaggacg agcttggcaa ggtcatgatg ggcgtggtcc gcccgagggc agagccatga
     5221 cttttttagc cgctaaaacg gccggggggt gcgcgtgatt gccaagcacg tccccatgcg
     5281 ctccatcaag aagagcgact tcgcggagct ggtgaagtac atcaccgacg agcaaggcaa
     5341 gaccgagcgc ctgggtcacg tgcgcgtcac gaactgcgag gcaaacaccc tgcccgctgt
     5401 catggccgag gtgatggcga cccagcacgg caacacccgt tccgaggccg acaagaccta
     5461 tcacctgctg gttagcttcc gcgcgggaga gaagcccgac gcggagacgt tgcgcgcgat
     5521 tgaggaccgc atctgcgctg ggcttggctt cgccgagcat cagcgcgtca gtgccgtgca
     5581 tcacgacacc gacaacctgc acatccatat cgccatcaac aagattcacc cgacccgaaa
     5641 caccatccat gagccgtatc gggcctaccg cgccctcgct gacctctgcg cgacgctcga
     5701 acgggactac gggcttgagc gtgacaatca cgaaacgcgg cagcgcgttt ccgagaaccg
//
