LOCUS       Exported                4833 bp ds-DNA     circular SYN 18-MAR-2019
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4833)
  AUTHORS   SynthSys Center
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 18, 2019 from SnapGene 4.3.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4833
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     CDS             215..1225
                     /codon_start=1
                     /label=SpecR
                     /label=Sm/Sp no DraIII
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    1385..1999
                     /label=ori (pMB1)
                     /label=rep (pMB1)
     misc_feature    2232..2235
                     /label=AATG
     promoter        2236..2681
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             2682..4103
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     misc_feature    4818..4821
                     /label=AGGT
ORIGIN
        1 gtggctcttc agtggacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
       61 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      181 ataaatgctt caataatatt gaaaaaggaa gagtatgcgc tcacgcaact ggtccagaac
      241 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
      301 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
      361 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
      421 aaagttaaac atcatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
      481 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
      541 cgcagtggat ggcggcctga agccacacag cgatattgat ttgctggtta cggtgaccgt
      601 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
      661 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
      721 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
      781 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
      841 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
      901 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
      961 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     1021 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     1081 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     1141 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccattacg taaaaggcga
     1201 gatcaccaag gtagtcggca aataactgtc agaccaagtt tactcatata tactttagat
     1261 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1321 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1381 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1441 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1501 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1561 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1621 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1681 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1741 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1801 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1861 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1921 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1981 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2041 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2101 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2161 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatcact
     2221 ctgtggtctc aaatgcacat atacctgccg ttcactatta tttagtgaaa tgagatatta
     2281 tgatattttc tgaattgtga ttaaaaaggc aactttatgc ccatgcaaca gaaactataa
     2341 aaaatacaga gaatgaaaag aaacagatag attttttagt tctttaggcc cgtagtctgc
     2401 aaatcctttt atgattttct atcaaacaaa agaggaaaat agaccagttg caatccaaac
     2461 gagagtctaa tagaatgagg tcgaaaagta aatcgcgcgg gtttgttact gataaagcag
     2521 gcaagaccta aaatgtgtaa agggcaaagt gtatactttg gcgtcacccc ttacatattt
     2581 taggtctttt tttattgtgc gtaactaact tgccatcttc aaacaggagg gctggaagaa
     2641 gcagaccgct aacacagtac ataaaaaagg agacatgaac gatgaacatc aaaaagtttg
     2701 caaaacaagc aacagtatta acctttacta ccgcactgct ggcaggaggc gcaactcaag
     2761 cgtttgcgaa agaaacgaac caaaagccat ataaggaaac atacggcatt tcccatatta
     2821 cacgccatga tatgctgcaa atccctgaac agcaaaaaaa tgaaaaatat caagttcctg
     2881 aattcgattc gtccacaatt aaaaatatct cttctgcaaa aggcctggac gtttgggaca
     2941 gctggccatt acaaaacgct gacggcactg tcgcaaacta tcacggctac cacatcgtct
     3001 ttgcattagc cggagatcct aaaaatgcgg atgacacatc gatttacatg ttctatcaaa
     3061 aagtcggcga aacttctatt gacagctgga aaaacgctgg ccgcgtcttt aaagacagcg
     3121 acaaattcga tgcaaatgat tctatcctaa aagaccaaac acaagaatgg tcaggttcag
     3181 ccacatttac atctgacgga aaaatccgtt tattctacac tgatttctcc ggtaaacatt
     3241 acggcaaaca aacactgaca actgcacaag ttaacgtatc agcatcagac agctctttga
     3301 acatcaacgg tgtagaggat tataaatcaa tctttgacgg tgacggaaaa acgtatcaaa
     3361 atgtacagca gttcatcgat gaaggcaact acagctcagg cgacaaccat acgctgagag
     3421 atcctcacta cgtagaagat aaaggccaca aatacttagt atttgaagca aacactggaa
     3481 ctgaagatgg ctaccaaggc gaagaatctt tatttaacaa agcatactat ggcaaaagca
     3541 catcattctt ccgtcaagaa agtcaaaaac ttctgcaaag cgataaaaaa cgcacggctg
     3601 agttagcaaa cggcgctctc ggtatgattg agctaaacga tgattacaca ctgaaaaaag
     3661 tgatgaaacc gctgattgca tctaacacag taacagatga aattgaacgc gcgaacgtct
     3721 ttaaaatgaa cggcaaatgg tacctgttca ctgactcccg cggatcaaaa atgacgattg
     3781 acggcattac gtctaacgat atttacatgc ttggttatgt ttctaattct ttaactggcc
     3841 catacaagcc gctgaacaaa actggccttg tgttaaaaat ggatcttgat cctaacgatg
     3901 taacctttac ttactcacac ttcgctgtac ctcaagcgaa aggaaacaat gtcgtgatta
     3961 caagctatat gacaaacaga ggattctacg cagacaaaca atcaacgttt gcgccaagct
     4021 tcctgctgaa catcaaaggc aagaaaacat ctgttgtcaa agacagcatc cttgaacaag
     4081 gacaattaac agttaacaaa taaaaacgca aaagaaaatg ccgatatcct attggcattt
     4141 tcttttattt cttatcaaca taaaggtgaa tcccatatga actatataaa agcaggcaaa
     4201 tggctaaccg tattcctaac cttttggtaa tgactccaac ttattgatag tgttttatgt
     4261 tcagataatg cccgatgact ttgtcatgca gctccaccga ttttgagaac gacagcgact
     4321 tccgtcccag ccgtgccagg tgctgcctca gattcaggtt atgccgctca attcgctgcg
     4381 tatatcgctt gctgattacg tgcagctttc ccttcaggcg ggattcatac agcggccagc
     4441 catccgtcat ccatatcacc acgtcaaagg gtgacagcag gctcataaga cgccccagcg
     4501 tcgccatagt gcgttcaccg aatacgtgcg caacaaccgt ctaccggaga ctgtcatacg
     4561 cgtaaaacag ccagcgctgg cgcgatttag ccccgacata gccccactgt tcgtccattt
     4621 ccgcgcagac gatgacgtca ctgcccggct gtatgcgcga ggttaccgac tgcggcctga
     4681 gttttttaag tgacgtaaaa tcgtgttgag gccaacgccc ataatgcggg ctgttgcccg
     4741 gcatccaacg ccattcatgg ccatatcaat gattttctgg tgcgtaccgg gttgagaagc
     4801 ggtgtaagtg aactgcaagg ttgagaccac gaa
//
