LOCUS       V003640                 5540 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003640
VERSION     V003640
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5540)
  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 5540)
  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 5540)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5540)
  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..5540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(22..714)
                     /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(773..1459)
                     /label="AmCyan"
                     /note="Anemonia majano cyan fluorescent protein"
     regulatory      complement(1486..1491)
                     /label="PFP1"
                     /note="PFP1"
                     /regulatory_class="minus_10_signal"
     regulatory      complement(1509..1514)
                     /label="PFP1"
                     /note="PFP1"
                     /regulatory_class="minus_35_signal"
     CDS             complement(1524..2315)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(2617..2635)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2656..2672)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(2680..2696)
                     /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(2704..2734)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2749..2770)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3058..3646)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            3984..4092
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             complement(4279..5280)
                     /label="repB"
                     /note="RepB replication protein"
ORIGIN
        1 gtcgaggtcg actcactatt atttcaggaa agtttcggag gagatagtgt tcggcagttt
       61 gtacatcatc tgcgggatca ggtacggttt gatcaggttg tagaagatca ggtaagacat
      121 agaatcgatg tagatgatcg gtttgttttt gttgattttt acgtaacagt tcagttggaa
      181 tttgttacgc agacccttaa ccaggtattc tacttcttcg aaagtgaaag actgggtgtt
      241 cagtacgatc gatttgttgg tagagttttt gttgtaatcc catttaccac catcatccat
      301 gaaccagtat gccagagaca tcggggtcag gtagttttca accaggttgt tcgggatggt
      361 ttttttgttg ttaacgatga acaggttagc cagtttgttg aaagcttggt gtttgaaagt
      421 ctgggcgccc caggtgatta ccaggttacc caggtggtta acacgttctt ttttgtgcgg
      481 cggggacagt acccactgat cgtacagcag acatacgtgg tccatgtatg ctttgttttt
      541 ccactcgaac tgcatacagt aggttttacc ttcatcacga gaacggatgt aagcatcacc
      601 caggatcaga ccgatacctg cttcgaactg ttcgatgttc agttcgatca gctgggattt
      661 gtattctttc agcagtttag agttcggacc caggttcatt acctggtttt tttgatgcat
      721 atgcgttctc ctttcattgg atcccccggg agatctcgat atcgaattct cagaaaggga
      781 caacagaggt tatatgtgca acagcgtgct ccgtcagctg aacactgttg ccacctttgt
      841 caaggtcggt cctcgcaatg cgatgttcca ccgcatggtt tggtggcatc gtcaccggtt
      901 tttttgtctt gtaagaagtg tggaattggc atctgtaatt gccacctcct tgcagcatga
      961 ggaacgcggt gacatcaccc ttcaatattc catcgcagac agtcattttc tcaaaagatg
     1021 ggtcccaacc agttgtcatc ttcgccatca caggtccatc agcaggaaag ttcactccat
     1081 gaaacgtgga tttgtgctca aagcagttgc ctttaaggct tatttcccaa ctggctgtag
     1141 caactcctcc atcttcatag gtaaaagtcc tttcatatga cattccgtca ggaaatgctt
     1201 gtttgaaata gtcgggcata ctggtaggat acgcagtaaa gcatcgattt ccatacatga
     1261 acactgtaga tagtatgtca aaggagaatg caaggggccc accgttggcc atggtgactt
     1321 taaaagtcga ggtctgcgtc ccttcgtatg gcttcccgct gccttcacct ttgacggtaa
     1381 agtaatgccc attgacacag ccatccatat ggtaggtcat tttcatgtca tctccgataa
     1441 acttgtttga aagagccatc ataattccct ccaattaaac ttatcattat aagtttctaa
     1501 ctttatacta tcaacaaaca tcaaaacaat tcatccagta aaatataata ttttattttc
     1561 tcccaatcag gcttgatccc cagtaagtca aaaaatagct cgacatactg ttcttccccg
     1621 atatcctccc tgatcgaccg gacgcagaag gcaatgtcat accacttgtc cgccctgccg
     1681 cttctcccaa gatcaataaa gccacttact ttgccatctt tcacaaagat gttgctgtct
     1741 cccaggtcgc cgtgggaaaa gacaagttcc tcttcgggct tttccgtctt taaaaaatca
     1801 tacagctcgc gcggatcttt aaatggagtg tcttcttccc agttttcgca atccacatcg
     1861 gccagatcgt tattcagtaa gtaatccaat tcggctaagc ggctgtctaa gctattcgta
     1921 tagggacaat ccgatatgtc gatggagtga aagagcctga tgcactccgc atacagctcg
     1981 ataatctttt cagggctttg ttcatcttca tactcttccg agcaaaggac gccatcggcc
     2041 tcactcatga gcagattgct ccagccatca tgccgttcaa agtgcaggac ctttggaaca
     2101 ggcagctttc cttccagcca tagcatcatg tccttttccc gttctacatc ataggtggtc
     2161 cctttatacc ggctgtccgt catttttaaa tataggtttt cattttctcc caccagctta
     2221 tataccttag caggagacat tccttccgta tcttttacgc agcggtattt ttcgatcagt
     2281 tttttcaatt ccggtgatat tctcatttta gccatttatt atttccttcc tcttttctac
     2341 agtatttaaa gataccccaa gaagctaatt ataacaagac gaactccaat tcactgttcc
     2401 ttgcattcta aaaccttaaa taccagaaaa cagctttttc aaagttgttt tcaaagttgg
     2461 cgtataacat agtatcgacg gagccgattt tgaaaccaca attatgatag aatttacaag
     2521 ctatagggaa tcatttgaag gttggtacta tataaaaata atatgcattt aatactagcg
     2581 acgccatcta tgtggcgcgg ccgccagctt ttgttccctt tagtgagggt taattgcgcg
     2641 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2701 acacaacata cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2761 actcacatta attgcgttgc gctcactgcc cgctttccag tcaggaaacc tgtcgtgcca
     2821 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2881 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2941 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3001 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3061 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3121 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3181 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3241 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3301 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3361 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3421 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3481 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3541 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3601 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3661 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3721 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3781 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagggtacc
     3841 ggatcgcgct tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta
     3901 tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct
     3961 tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg
     4021 tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg
     4081 cgaggctggc cgcctaggaa taatgagggc agacgtagta atgatacgct tccctctttt
     4141 aattgaaccc tgttacattc attacacttc ataattaatt cctcctaaac ttgattaaaa
     4201 cattttacca catataaact aagttttaaa ttcagtattt catcacttat acaacaatat
     4261 ggcccgtttg ttgaactact ctttaataaa ataatttttc cgttcccaat tccacattgc
     4321 aataatagaa aatccatctt catcggcttt ttcgtcatca tctgtatgaa tcaaatcgcc
     4381 ttcttctgtg tcatcaaggt ttaatttttt atgtatttct tttaacaaac caccatagga
     4441 gattaacctt ttacggtgta aaccttcctc caaatcagac aaacgtttca aattcttttc
     4501 ttcatcatcg gtcataaaat ccgtatcctt tacaggatat tttgcagttt cgtcaattgc
     4561 cgattgtata tccgatttat atttattttt cggtcgaatc atttgaactt ttacatttgg
     4621 atcatagtct aatttcattg cctttttcca aaattgaatc cattgttttt gattcacgta
     4681 gttttctgta ttcttaaaat aagttggttc cacacatacc aatacatgca tgtgctgatt
     4741 ataagaatta tctttattat ttattgtcac ttccgttgca cgcataaaac caacaagatt
     4801 tttattaatt tttttatatt gcatcattcg gcgaaatcct tgagccatat ctgacaaact
     4861 cttatttaat tcttcgccat cataaacatt tttaactgtt aatgtgagaa acaaccaacg
     4921 aactgttggc ttttgtttaa taacttcagc aacaaccttt tgtgactgaa tgccatgttt
     4981 cattgctctc ctccagttgc acattggaca aagcctggat ttacaaaacc acactcgata
     5041 caactttctt tcgcctgttt cacgattttg tttatactct aatatttcag cacaatcttt
     5101 tactctttca gcctttttaa attcaagaat atgcagaagt tcaaagtaat caacattagc
     5161 gattttcttt tctctccatg gtctcacttt tccacttttt gtcttgtcca ctaaaaccct
     5221 tgatttttca tctgaataaa tgctactatt aggacacata atattaaaag aaacccccat
     5281 ctatttagtt atttgtttgg tcacttataa ctttaacaga tggggttttt ctgtgcaacc
     5341 aattttaagg gttttcaata ctttaaaaca catacatacc aacacttcaa cgcacctttc
     5401 agcaactaaa ataaaaatga cgttatttct atatgtatca agataagaaa gaacaagttc
     5461 aaaaccatca aaaaaagaca ccttttcagg tgcttttttt attttataaa ctcattccct
     5521 gatctcgact tcgttctttt
//
