LOCUS       Exported                5748 bp DNA     circular SYN 15-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5748)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5748)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          54..58
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      82..470
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     promoter        498..943
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             944..2362
                     /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(3207..3863)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIHSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3864..3966)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     oriT            4695..4804
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             4837..5205
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 cgcgaacgac atggagcggc acgcgggcgt ggaaagcctg gtcggctgga tcgatccggc
       61 cacgatgcgt ccggcgtaga ggatctgaag atcagcagtt caacctgttg atagtacgta
      121 ctaagctctc atgtttcacg tactaagctc tcatgtttaa cgtactaagc tctcatgttt
      181 aacgaactaa accctcatgg ctaacgtact aagctctcat ggctaacgta ctaagctctc
      241 atgtttcacg tactaagctc tcatgtttga acaataaaat taatataaat cagcaactta
      301 aatagcctct aaggttttaa gttttataag aaaaaaaaga atatataagg cttttaaagc
      361 ttttaaggtt taacggttgt ggacaacaag ccagggatgt aacgcactga gaagccctta
      421 gagcctctca aagcaatttt gagtgacaca ggaacactta acggctgaca tgggaattct
      481 gatccttttt aacccatcac atatacctgc cgttcactat tatttagtga aatgagatat
      541 tatgatattt tctgaattgt gattaaaaag gcaactttat gcccatgcaa cagaaactat
      601 aaaaaataca gagaatgaaa agaaacagat agatttttta gttctttagg cccgtagtct
      661 gcaaatcctt ttatgatttt ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa
      721 acgagagtct aatagaatga ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc
      781 aggcaagacc taaaatgtgt aaagggcaaa gtgtatactt tggcgtcacc ccttacatat
      841 tttaggtctt tttttattgt gcgtaactaa cttgccatct tcaaacagga gggctggaag
      901 aagcagaccg ctaacacagt acataaaaaa ggagacatga acgatgaaca tcaaaaagtt
      961 tgcaaaacaa gcaacagtat taacctttac taccgcactg ctggcaggag gcgcaactca
     1021 agcgtttgcg aaagaaacga accaaaagcc atataaggaa acatacggca tttcccatat
     1081 tacacgccat gatatgctgc aaatccctga acagcaaaaa aatgaaaaat atcaagttcc
     1141 tgagttcgat tcgtccacaa ttaaaaatat ctcttctgca aaaggcctgg acgtttggga
     1201 cagctggcca ttacaaaacg ctgacggcac tgtcgcaaac tatcacggct accacatcgt
     1261 ctttgcatta gccggagatc ctaaaaatgc ggatgacaca tcgatttaca tgttctatca
     1321 aaaagtcggc gaaacttcta ttgacagctg gaaaaacgct ggccgcgtct ttaaagacag
     1381 cgacaaattc gatgcaaatg attctatcct aaaagaccaa acacaagaat ggtcaggttc
     1441 agccacattt acatctgacg gaaaaatccg tttattctac actgatttct ccggtaaaca
     1501 ttacggcaaa caaacactga caactgcaca agttaacgta tcagcatcag acagctcttt
     1561 gaacatcaac ggtgtagagg attataaatc aatctttgac ggtgacggaa aaacgtatca
     1621 aaatgtacag cagttcatcg atgaaggcaa ctacagctca ggcgacaacc atacgctgag
     1681 agatcctcac tacgtagaag ataaaggcca caaatactta gtatttgaag caaacactgg
     1741 aactgaagat ggctaccaag gcgaagaatc tttatttaac aaagcatact atggcaaaag
     1801 cacatcattc ttccgtcaag aaagtcaaaa acttctgcaa agcgataaaa aacgcacggc
     1861 tgagttagca aacggcgctc tcggtatgat tgagctaaac gatgattaca cactgaaaaa
     1921 agtgatgaaa ccgctgattg catctaacac agtaacagat gaaattgaac gcgcgaacgt
     1981 ctttaaaatg aacggcaaat ggtatctgtt cactgactcc cgcggatcaa aaatgacgat
     2041 tgacggcatt acgtctaacg atatttacat gcttggttat gtttctaatt ctttaactgg
     2101 cccatacaag ccgctgaaca aaactggcct tgtgttaaaa atggatcttg atcctaacga
     2161 tgtaaccttt acttactcac acttcgctgt acctcaagcg aaaggaaaca atgtcgtgat
     2221 tacaagctat atgacaaaca gaggattcta cgcagacaaa caatcaacgt ttgcgcccag
     2281 cttcctgctg aacatcaaag gcaagaaaac atctgttgtc aaagacagca tccttgaaca
     2341 aggacaatta acagttaaca aataaaaacg caaaagaaaa tgccgatatc ctattggcat
     2401 tttcttttat ttcttatcaa cataaaggtg aatcccatat gaactatata aaagcaggca
     2461 aatggctaac cgtattccta accttttggt aatgactcca acttattgat agtgttttat
     2521 gttcagataa tgcccgatga ctttgtcatg cagctccacc gattttgaga acgacagcga
     2581 cttccgtccc agccgtgcca ggtgctgcct cagattcagg ttatgccgct caattcgctg
     2641 cgtatatcgc ttgctgatta cgtgcagctt tcccttcagg cgggattcat acagcggcca
     2701 gccatccgtc atccatatca ccacgtcaaa gggtgacagc aggctcataa gacgccccag
     2761 cgtcgccata gtgcgttcac cgaatacgtg cgcaacaacc gtcttccgga gactgtcata
     2821 cgcgtaaaac agccagcgct ggcgcgattt agccccgaca tagccccact gttcgtccat
     2881 ttccgcgcag acgatgacgt cactgcccgg ctgtatgcgc gaggttaccg actgcggcct
     2941 gagtttttta agtgacgtaa aatcgtgttg aggccaacgc ccataatgcg ggctgttgcc
     3001 cggcatccaa cgccattcat ggccatatca atgattttct ggtgcgtacc gggttgagaa
     3061 gcggtgtaag tgaactgcat gaattcccgg gagagctcga tatcgcatgc ggtacctcta
     3121 gaagaagctt gggatcgggc cctatcactt attcaggcgt agcaaccagg cgtttaaggg
     3181 caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt
     3241 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     3301 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     3361 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     3421 ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     3481 ccgtaacacg ccacatcttg cgaatgtatg tgtagaaact gccggaaatc gtcgtggtat
     3541 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     3601 acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca
     3661 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     3721 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     3781 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     3841 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     3901 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     3961 cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca
     4021 ccaggattta tttattctgc gaagtgatct tccgtcacag gtattagggc ccgatccttt
     4081 ttgtccggtg ttgggttgaa ggtgaagccg gtcggggccg cagcgggggc cggcttttca
     4141 gccttgcccc cctgcttcgg ccgccgtggc tccggcgtct tgggtgccgg cgcgggttcc
     4201 gcagccttgg cctgcggtgc gggcacatcg gcgggcttgg ccttgatgtg ccgcctggcg
     4261 tgcgagcgga acgtctcgta ggagaacttg accttccccg tttcccgcat gtgctcccaa
     4321 atggtgacga gcgcatagcc ggacgctaac gccgcctcga catccgccct caccgccagg
     4381 aacgcaaccg cagcctcatc acgccggcgc ttcttggccg cgcgggattc aacccactcg
     4441 gccagctcgt cggtgtagct ctttggcatc gtctctcgcc tgtcccctca gttcagtaat
     4501 ttcctgcatt tgcctgtttc cagtcggtag atattccaca aaacagcagg gaagcagcgc
     4561 ttttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt atatccatcc
     4621 tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc ttggtgtatc
     4681 caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc
     4741 ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct gcgaggctgg
     4801 ccggctaccg ccggcgtaac agatgagggc aagcggatgg ctgatgaaac caagccaacc
     4861 aggaagggca gcccacctat caaggtgtac tgccttccag acgaacgaag agcgattgag
     4921 gaaaaggcgg cggcggccgg catgagcctg tcggcctacc tgctggccgt cggccagggc
     4981 tacaaaatca cgggcgtcgt ggactatgag cacgtccgcg agctggcccg catcaatggc
     5041 gacctgggcc gcctgggcgg cctgctgaaa ctctggctca ccgacgaccc gcgcacggcg
     5101 cggttcggtg atgccacgat cctcgccctg ctggcgaaga tcgaagagaa gcaggacgag
     5161 cttggcaagg tcatgatggg cgtggtccgc ccgagggcag agccatgact tttttagccg
     5221 ctaaaacggc cggggggtgc gcgtgattgc caagcacgtc cccatgcgct ccatcaagaa
     5281 gagcgacttc gcggagctgg tgaagtacat caccgacgag caaggcaaga ccgagcgcct
     5341 gggtcacgtg cgcgtcacga actgcgaggc aaacaccctg cccgctgtca tggccgaggt
     5401 gatggcgacc cagcacggca acacccgttc cgaggccgac aagacctatc acctgctggt
     5461 tagcttccgc gcgggagaga agcccgacgc ggagacgttg cgcgcgattg aggaccgcat
     5521 ctgcgctggg cttggcttcg ccgagcatca gcgcgtcagt gccgtgcatc acgacaccga
     5581 caacctgcac atccatatcg ccatcaacaa gattcacccg acccgaaaca ccatccatga
     5641 gccgtatcgg gcctaccgcg ccctcgctga cctctgcgcg acgctcgaac gggactacgg
     5701 gcttgagcgt gacaatcacg aaacgcggca gcgcgtttcc gagaaccg
//
