LOCUS       Exported                6199 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pDMS197
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6199)
  AUTHORS   Edwards RA, Keller LH, Schifferli DM
  TITLE     Improved allelic exchange vectors and their use to analyze 987P 
            fimbria gene expression.
  JOURNAL   Gene. 1998 Jan 30;207(2):149-57.
  PUBMED    9511756
REFERENCE   2  (bases 1 to 6199)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6199
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             956..2377
                     /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"
     CDS             complement(3246..4436)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     4065..4084
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(4484..4512)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     oriT            5146..5255
                     /note="incP origin of transfer"
     CDS             5288..5659
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /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 ttgggatcca agggttggtt tgcgcattca cagttctccg
     3181 caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct
     3241 tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca
     3301 aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca
     3361 taaatcgccg tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc
     3421 gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac
     3481 gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct
     3541 cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg
     3601 gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg
     3661 tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg
     3721 tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag
     3781 aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact
     3841 gggttgaagg ctctcaaggg catcggtcga cgctctccct tatgcgactc ctgcattagg
     3901 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     3961 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     4021 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     4081 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     4141 aggatccaca ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc
     4201 gaagcgagca ggactgggcg gcggccaaag cggtcggaca gtgctccgag aacgggtgcg
     4261 catagaaatt gcatcaacgc atatagcgct agcagcacgc catagtgact ggcgatgctg
     4321 tcggaatgga cgatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct
     4381 acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac
     4441 ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg
     4501 ataagctgtc aaacatgaga attgatcctt tttgtccggt gttgggttga aggtgaagcc
     4561 ggtcggggcc gcagcggggg ccggcttttc agccttgccc ccctgcttcg gccgccgtgg
     4621 ctccggcgtc ttgggtgccg gcgcgggttc cgcagccttg gcctgcggtg cgggcacatc
     4681 ggcgggcttg gccttgatgt gccgcctggc gtgcgagcgg aacgtctcgt aggagaactt
     4741 gaccttcccc gtttcccgca tgtgctccca aatggtgacg agcgcatagc cggacgctaa
     4801 cgccgcctcg acatccgccc tcaccgccag gaacgcaacc gcagcctcat cacgccggcg
     4861 cttcttggcc gcgcgggatt caacccactc ggccagctcg tcggtgtagc tctttggcat
     4921 cgtctctcgc ctgtcccctc agttcagtaa tttcctgcat ttgcctgttt ccagtcggta
     4981 gatattccac aaaacagcag ggaagcagcg cttttccgct gcataaccct gcttcggggt
     5041 cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca
     5101 aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga
     5161 agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt
     5221 gctcaacggg aatcctgctc tgcgaggctg gccggctacc gccggcgtaa cagatgaggg
     5281 caagcggatg gctgatgaaa ccaagccaac caggaagggc agcccaccta tcaaggtgta
     5341 ctgccttcca gacgaacgaa gagcgattga ggaaaaggcg gcggcggccg gcatgagcct
     5401 gtcggcctac ctgctggccg tcggccaggg ctacaaaatc acgggcgtcg tggactatga
     5461 gcacgtccgc gagctggccc gcatcaatgg cgacctgggc cgcctgggcg gcctgctgaa
     5521 actctggctc accgacgacc cgcgcacggc gcggttcggt gatgccacga tcctcgccct
     5581 gctggcgaag atcgaagaga agcaggacga gcttggcaag gtcatgatgg gcgtggtccg
     5641 cccgagggca gagccatgac ttttttagcc gctaaaacgg ccggggggtg cgcgtgattg
     5701 ccaagcacgt ccccatgcgc tccatcaaga agagcgactt cgcggagctg gtgaagtaca
     5761 tcaccgacga gcaaggcaag accgagcgcc tgggtcacgt gcgcgtcacg aactgcgagg
     5821 caaacaccct gcccgctgtc atggccgagg tgatggcgac ccagcacggc aacacccgtt
     5881 ccgaggccga caagacctat cacctgctgg ttagcttccg cgcgggagag aagcccgacg
     5941 cggagacgtt gcgcgcgatt gaggaccgca tctgcgctgg gcttggcttc gccgagcatc
     6001 agcgcgtcag tgccgtgcat cacgacaccg acaacctgca catccatatc gccatcaaca
     6061 agattcaccc gacccgaaac accatccatg agccgtatcg ggcctaccgc gccctcgctg
     6121 acctctgcgc gacgctcgaa cgggactacg ggcttgagcg tgacaatcac gaaacgcggc
     6181 agcgcgtttc cgagaaccg
//
