LOCUS       V003045                 6668 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003045
VERSION     V003045
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6668)
  AUTHORS   Plaut RD, Stibitz S.
  TITLE     Improvements to a Markerless Allelic Exchange System for Bacillus
            anthracis
  JOURNAL   PLoS ONE 10 (12), E0142758 (2015)
   PUBMED   26624016
REFERENCE   2  (bases 1 to 6668)
  AUTHORS   Plaut RD, Stibitz S.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-SEP-2015) Center for Biologics Evaluation and
            Research, OVRR, DBPAP, Food and Drug Administration, 10903 New
            Hampshire Ave, Bldg 52/72, Rm 3310, Silver Spring, MD 20993, USA
REFERENCE   3  (bases 1 to 6668)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6668)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
            date: "2015"; volume: "10"; issue: "12"; pages: "E0142758"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (24-SEP-2015) Center for Biologics Evaluation and Research, OVRR,
            DBPAP, Food and Drug Administration, 10903 New Hampshire Ave, Bldg
            52/72, Rm 3310, Silver Spring, MD 20993, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(16..708)
                     /gene="SCEI"
                     /label="Intron-encoded endonuclease I-SceI"
                     /note="Intron-encoded endonuclease I-SceI from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
                     Accession#: P03882"
     CDS             complement(767..1453)
                     /label="AmCyan"
                     /note="Anemonia majano cyan fluorescent protein"
     regulatory      complement(1480..1485)
                     /label="PFP1"
                     /note="PFP1"
                     /regulatory_class="minus_10_signal"
     regulatory      complement(1503..1508)
                     /label="PFP1"
                     /note="PFP1"
                     /regulatory_class="minus_35_signal"
     CDS             complement(1629..2315)
                     /label="AmCyan"
                     /note="Anemonia majano cyan fluorescent protein"
     regulatory      complement(2342..2347)
                     /label="PFP1"
                     /note="PFP1"
                     /regulatory_class="minus_10_signal"
     regulatory      complement(2365..2370)
                     /label="PFP1"
                     /note="PFP1"
                     /regulatory_class="minus_35_signal"
     CDS             complement(2380..3171)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(3473..3491)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3512..3528)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(3536..3552)
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3560..3590)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3605..3626)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3914..4502)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            4840..4949
                     /label="oriT"
                     /note="incP origin of transfer"
     oriT            4955..5206
                     /note="from D2BpUC9Cam; tamdem repeat"
     oriT            5098..5207
                     /label="incP origin of transfer"
                     /note="incP origin of transfer"
     CDS             complement(5407..6408)
                     /label="repB"
                     /note="RepB replication protein"
ORIGIN
        1 gtcgactcac tattatttca ggaaagtttc ggaggagata gtgttcggca gtttgtacat
       61 catctgcggg atcaggtacg gtttgatcag gttgtagaag atcaggtaag acatagaatc
      121 gatgtagatg atcggtttgt ttttgttgat ttttacgtaa cagttcagtt ggaatttgtt
      181 acgcagaccc ttaaccaggt attctacttc ttcgaaagtg aaagactggg tgttcagtac
      241 gatcgatttg ttggtagagt ttttgttgta atcccattta ccaccatcat ccatgaacca
      301 gtatgccaga gacatcgggg tcaggtagtt ttcaaccagg ttgttcggga tggttttttt
      361 gttgttaacg atgaacaggt tagccagttt gttgaaagct tggtgtttga aagtctgggc
      421 gccccaggtg attaccaggt tacccaggtg gttaacacgt tcttttttgt gcggcgggga
      481 cagtacccac tgatcgtaca gcagacatac gtggtccatg tatgctttgt ttttccactc
      541 gaactgcata cagtaggttt taccttcatc acgagaacgg atgtaagcat cacccaggat
      601 cagaccgata cctgcttcga actgttcgat gttcagttcg atcagctggg atttgtattc
      661 tttcagcagt ttagagttcg gacccaggtt cattacctgg tttttttgat gcatatgcgt
      721 tctcctttca ttggatcccc cgggagatct cgatatcgaa ttctcagaaa gggacaacag
      781 aggttatatg tgcaacagcg tgctccgtca gctgaacact gttgccacct ttgtcaaggt
      841 cggtcctcgc aatgcgatgt tccaccgcat ggtttggtgg catcgtcacc ggtttttttg
      901 tcttgtaaga agtgtggaat tggcatctgt aattgccacc tccttgcagc atgaggaacg
      961 cggtgacatc acccttcaat attccatcgc agacagtcat tttctcaaaa gatgggtccc
     1021 aaccagttgt catcttcgcc atcacaggtc catcagcagg aaagttcact ccatgaaacg
     1081 tggatttgtg ctcaaagcag ttgcctttaa ggcttatttc ccaactggct gtagcaactc
     1141 ctccatcttc ataggtaaaa gtcctttcat atgacattcc gtcaggaaat gcttgtttga
     1201 aatagtcggg catactggta ggatacgcag taaagcatcg atttccatac atgaacactg
     1261 tagatagtat gtcaaaggag aatgcaaggg gcccaccgtt ggccatggtg actttaaaag
     1321 tcgaggtctg cgtcccttcg tatggcttcc cgctgccttc acctttgacg gtaaagtaat
     1381 gcccattgac acagccatcc atatggtagg tcattttcat gtcatctccg ataaacttgt
     1441 ttgaaagagc catcataatt ccctccaatt aaacttatca ttataagttt ctaactttat
     1501 actatcaaca aacatcaaaa caattcatcc agtaaaatat aatattttat tttctcccaa
     1561 tcaggcttga tccccagtaa gtcaaaaaat agctcgacat actgttcttc cccgatatcg
     1621 aattctcaga aagggacaac agaggttata tgtgcaacag cgtgctccgt cagctgaaca
     1681 ctgttgccac ctttgtcaag gtcggtcctc gcaatgcgat gttccaccgc atggtttggt
     1741 ggcatcgtca ccggtttttt tgtcttgtaa gaagtgtgga attggcatct gtaattgcca
     1801 cctccttgca gcatgaggaa cgcggtgaca tcacccttca atattccatc gcagacagtc
     1861 attttctcaa aagatgggtc ccaaccagtt gtcatcttcg ccatcacagg tccatcagca
     1921 ggaaagttca ctccatgaaa cgtggatttg tgctcaaagc agttgccttt aaggcttatt
     1981 tcccaactgg ctgtagcaac tcctccatct tcataggtaa aagtcctttc atatgacatt
     2041 ccgtcaggaa atgcttgttt gaaatagtcg ggcatactgg taggatacgc agtaaagcat
     2101 cgatttccat acatgaacac tgtagatagt atgtcaaagg agaatgcaag gggcccaccg
     2161 ttggccatgg tgactttaaa agtcgaggtc tgcgtccctt cgtatggctt cccgctgcct
     2221 tcacctttga cggtaaagta atgcccattg acacagccat ccatatggta ggtcattttc
     2281 atgtcatctc cgataaactt gtttgaaaga gccatcataa ttccctccaa ttaaacttat
     2341 cattataagt ttctaacttt atactatcaa caaacatcaa aacaattcat ccagtaaaat
     2401 ataatatttt attttctccc aatcaggctt gatccccagt aagtcaaaaa atagctcgac
     2461 atactgttct tccccgatat cctccctgat cgaccggacg cagaaggcaa tgtcatacca
     2521 cttgtccgcc ctgccgcttc tcccaagatc aataaagcca cttactttgc catctttcac
     2581 aaagatgttg ctgtctccca ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc
     2641 cgtctttaaa aaatcataca gctcgcgcgg atctttaaat ggagtgtctt cttcccagtt
     2701 ttcgcaatcc acatcggcca gatcgttatt cagtaagtaa tccaattcgg ctaagcggct
     2761 gtctaagcta ttcgtatagg gacaatccga tatgtcgatg gagtgaaaga gcctgatgca
     2821 ctccgcatac agctcgataa tcttttcagg gctttgttca tcttcatact cttccgagca
     2881 aaggacgcca tcggcctcac tcatgagcag attgctccag ccatcatgcc gttcaaagtg
     2941 caggaccttt ggaacaggca gctttccttc cagccatagc atcatgtcct tttcccgttc
     3001 cacatcatag gtggtccctt tataccggct gtccgtcatt tttaaatata ggttttcatt
     3061 ttctcccacc agcttatata ccttagcagg agacattcct tccgtatctt ttacgcagcg
     3121 gtatttttcg atcagttttt tcaattccgg tgatattctc attttagcca tttattattt
     3181 ccttcctctt ttctacagta tttaaagata ccccaagaag ctaattataa caagacgaac
     3241 tccaattcac tgttccttgc attctaaaac cttaaatacc agaaaacagc tttttcaaag
     3301 ttgttttcaa agttggcgta taacatagta tcgacggagc cgattttgaa accacaatta
     3361 tgatagaatt tacaagctat agggaatcat ttgaaggttg gtactatata aaaataatat
     3421 gcatttaata ctagcgacgc catctatgtg gcgcggccgc cagcttttgt tccctttagt
     3481 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3541 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3601 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3661 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3721 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3781 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3841 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3901 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3961 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4021 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4081 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4141 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4201 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4261 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4321 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4381 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4441 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4501 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4561 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4621 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4681 gcttaatcag ggtaccggat cgcgcttttc cgctgcataa ccctgcttcg gggtcattat
     4741 agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt
     4801 tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg
     4861 cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa
     4921 cgggaatcct gctctgcgag gctggccggg taccggatcg cgcttttccg ctgcataacc
     4981 ctgcttcggg gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac
     5041 aggattttgc caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg
     5101 caggataggt gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc
     5161 gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc tggccgggta cctaggaata
     5221 atgagggcag acgtagttta tagggttaat gatacgcttc cctcttttaa ttgaaccctg
     5281 ttacattcat tacacttcat aattaattcc tcctaaactt gattaaaaca ttttaccaca
     5341 tataaactaa gttttaaatt cagtatttca tcacttatac aacaatatgg cccgtttgtt
     5401 gaactactct ttaataaaat aatttttccg ttcccaattc cacattgcaa taatagaaaa
     5461 tccatcttca tcggcttttt cgtcatcatc tgtatgaatc aaatcgcctt cttctgtgtc
     5521 atcaaggttt aattttttat gtatttcttt taacaaacca ccataggaga ttaacctttt
     5581 acggtgtaaa ccttcctcca aatcagacaa acgtttcaaa ttcttttctt catcatcggt
     5641 cataaaatcc gtatccttta caggatattt tgcagtttcg tcaattgccg attgtatatc
     5701 cgatttatat ttatttttcg gtcgaatcat ttgaactttt acatttggat catagtctaa
     5761 tttcattgcc tttttccaaa attgaatcca ttgtttttga ttcacgtagt tttctgtatt
     5821 cttaaaataa gttggttcca cacataccaa tacatgcatg tgctgattat aagaattatc
     5881 tttattattt attgtcactt ccgttgcacg cataaaacca acaagatttt tattaatttt
     5941 tttatattgc atcattcggc gaaatccttg agccatatct gacaaactct tatttaattc
     6001 ttcgccatca taaacatttt taactgttaa tgtgagaaac aaccaacgaa ctgttggctt
     6061 ttgtttaata acttcagcaa caaccttttg tgactgaatg ccatgtttca ttgctctcct
     6121 ccagttgcac attggacaaa gcctggattt acaaaaccac actcgataca actttctttc
     6181 gcctgtttca cgattttgtt tatactctaa tatttcagca caatctttta ctctttcagc
     6241 ctttttaaat tcaagaatat gcagaagttc aaagtaatca acattagcga ttttcttttc
     6301 tctccatggt ctcacttttc cactttttgt cttgtccact aaaacccttg atttttcatc
     6361 tgaataaatg ctactattag gacacataat attaaaagaa acccccatct atttagttat
     6421 ttgtttggtc acttataact ttaacagatg gggtttttct gtgcaaccaa ttttaagggt
     6481 tttcaatact ttaaaacaca tacataccaa cacttcaacg cacctttcag caactaaaat
     6541 aaaaatgacg ttatttctat atgtatcaag ataagaaaga acaagttcaa aaccatcaaa
     6601 aaaagacacc ttttcaggtg ctttttttat tttataaact cattccctga tctcgacttc
     6661 gttctttt
//
