LOCUS       V009287                 6722 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009287
VERSION     V009287
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6722)
  AUTHORS   Lopez CM, Rholl DA, Trunck LA, Schweizer HP.
  TITLE     Versatile dual-technology system for markerless allele replacement
            in Burkholderia pseudomallei
  JOURNAL   Appl. Environ. Microbiol. 75 (20), 6496-6503 (2009)
   PUBMED   19700544
REFERENCE   2  (bases 1 to 6722)
  AUTHORS   Lopez CM, Schweizer HP.
  TITLE     Homing Endonuclease-based System for Select Agent-compliant
            Markerless Gene Replacement in Burkholderia pseudomallei
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6722)
  AUTHORS   Lopez CM, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAR-2009) Microbiology, Immunology and Patholofy,
            Colorado State University, 3205 Rampart Rd., Fort Collins, CO 80523,
            USA
REFERENCE   4  (bases 1 to 6722)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6722)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2009"; volume: "75"; issue: "20";
            pages: "6496-6503"
            SGRef: number: 2; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (02-MAR-2009) Microbiology, Immunology and Patholofy, Colorado State
            University, 3205 Rampart Rd., Fort Collins, CO 80523, USA"
            SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      216..302
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      394..421
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        440..531
                     /label="AmpR promoter"
     CDS             complement(933..1304)
                     /label="BleoR"
                     /note="antibiotic-binding protein"
     promoter        complement(1323..1370)
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     rep_origin      complement(1490..1841)
                     /direction=LEFT
                     /label="pRO1600 oriV"
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             1855..2685
                     /label="pRO1600 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     rep_origin      2918..3373
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      3484..4072
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4772..5647)
                     /label="araC"
                     /note="L-arabinose regulatory protein"
     promoter        5674..5958
                     /label="araBAD promoter"
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite
                     direction (Guzman et al., 1995)"
     CDS             5999..6703
                     /gene="SCEI"
                     /label="Intron-encoded endonuclease I-SceI"
                     /note="Intron-encoded endonuclease I-SceI from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
                     Accession#: P03882"
ORIGIN
        1 ggcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
       61 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
      121 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
      181 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
      241 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
      301 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
      361 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
      421 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
      481 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
      541 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
      601 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
      661 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
      721 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
      781 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtct
      841 agatgcatgc tcgagcggcc gccagtgtga tggataattc ggcttatctc tgatgttaca
      901 ttgcacaaga taaaataata tcatcatgat cagtcctgct cctcggccac gaagtgcacg
      961 cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc acggctgctc gccgatctcg
     1021 gtcatggccg gcccggaggc gtcccggaag ttcgtggaca cgacctccga ccactcggcg
     1081 tacagctcgt ccaggccgcg cacccacacc caggccaggg tgttgtccgg caccacctgg
     1141 tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga ccacaccggc gaagtcgtcc
     1201 tccacgaagt cccgggagaa cccgagccgg tcggtccaga actcgaccgc tccggcgacg
     1261 tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg ccatggttta gttcctcacc
     1321 ttgtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac acgtgctgat
     1381 catgaccaaa atcccttaac gtgagttacg cgtcgaagcc gaattatctg cagaattcgg
     1441 cttttcgtga attcgccgct ggtgccgctg gttggacgcc aagggtgaat ccgcctcgat
     1501 accctgatta ctcgcttcct gcgccctctc aggcggcgat aggggactgg taaaacgggg
     1561 attgcccaga cgcctccccc gccccttcag gggcacaaat gcggccccaa cggggccacg
     1621 tagtggtgcg ttttttgcgt ttccaccctt ttcttccttt tcccttttaa accttttagg
     1681 acgtctacag gccacgtaat ccgtggcctg tagagtttaa aaagggacgg atttgttgcc
     1741 attaagggac ggatttgttg ttaagaaggg acggatttgt tgttgtaaag ggacggattt
     1801 gttgtattgt gggacgctga tacagtgtcc ccttatacac aaggaatgtc gaacgtggcc
     1861 tcacccccaa tggtttacaa aagcaatgcc ctggtcgagg ccgcgtattg cctcagtgtt
     1921 caggaacagc ggatcgttct ggccagtatt agccaggtga agaggagcga gcctgtcacc
     1981 gatgaagtga tgtattcagt gacggcggag gacatagcga cgatggcggg tgtccctatc
     2041 gaatcttcct acaaccagct caaagaagcg gccctgcgcc tgaaacggcg ggaagtccgg
     2101 ttaacccaag agcccaatgg caaggggaaa agaccgagtg tgatgattac cggctgggtg
     2161 caaacaatca tctaccggga gggtgagggc cgtgtagaac tcaggttcac caaagacatg
     2221 ctgccgtatc tgacggaact caccaaacag ttcaccaaat acgccttggc tgacgtggcc
     2281 aagatgggca gcacccacgc gatcaggctt tacgagctgc tcacgcaatg ggacagcatc
     2341 ggccagcgtg aaatagaaat tgaccagctg cgaaagtggt ttcaactgga aggccggtat
     2401 ccctcgatca aggacttcaa gttgcgagtg cttgatccag ccgtgacgca gatcaacgag
     2461 cacagcccgc tacaggtgga gtgggcgcag cgaaagaccg ggcgcaaggt cacacatctg
     2521 ttgttcagtt ttggaccgaa gaagcccgcc aaggcggtgg gtaaggcccc tgcgaagcgc
     2581 aaggccagga agatttcaga tgctgagatc gcgaaacagg ctcgccctgg tgagacatgg
     2641 gaagcggccc gcgctcgact aacccagatg ccgctggatc tggcctagag gccaagccga
     2701 attccagcac actggcggcc gttactagtg ggtctcgcgg tatcattgca gcactggggc
     2761 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2821 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2881 cagaccaagt ttactcatat atactttaga ttgatttacg cgccctgtag cggcgcatta
     2941 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3001 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3061 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3121 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3181 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aacttgaaca
     3241 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     3301 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3361 acgtttacaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3421 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3481 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3541 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3601 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     3661 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3721 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3781 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3841 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3901 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3961 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4021 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4081 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4141 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4201 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     4261 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag ggtcatggct
     4321 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4381 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4441 tcatcaccga aacgcgcgag gcagcaagga gatggcgccc aacagtcccc cggccacggg
     4501 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     4561 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     4621 gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atcatcgatg
     4681 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca
     4741 agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt
     4801 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
     4861 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
     4921 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
     4981 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat
     5041 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac
     5101 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc
     5161 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc
     5221 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat
     5281 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt
     5341 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac
     5401 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt
     5461 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt
     5521 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta
     5581 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt
     5641 cagccatact tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca
     5701 gacattgccg tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc
     5761 ttattaaaag cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa
     5821 gtgtctataa tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct
     5881 atgccatagc atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac
     5941 tctctactgt ttctccatac ccgttttttt gggctagcag gagggtacct atatgcatat
     6001 gaaaaacatc aaaaaaaacc aggtaatgaa cctgggtccg aactctaaac tgctgaaaga
     6061 atacaaatcc cagctgatcg aactgaacat cgaacagttc gaagcaggta tcggtctgat
     6121 cctgggtgat gcttacatcc gttctcgtga tgaaggtaaa acctactgta tgcagttcga
     6181 gtggaaaaac aaagcataca tggaccacgt atgtctgctg tacgatcagt gggtactgtc
     6241 cccgccgcac aaaaaagaac gtgttaacca cctgggtaac ctggtaatca cctggggcgc
     6301 ccagactttc aaacaccaag ctttcaacaa actggctaac ctgttcatcg ttaacaacaa
     6361 aaaaaccatc ccgaacaacc tggttgaaaa ctacctgacc ccgatgtctc tggcatactg
     6421 gttcatggat gatggtggta aatgggatta caacaaaaac tctaccaaca aatcgatcgt
     6481 actgaacacc cagtctttca ctttcgaaga agtagaatac ctggttaagg gtctgcgtaa
     6541 caaattccaa ctgaactgtt acgtaaaaat caacaaaaac aaaccgatca tctacatcga
     6601 ttctatgtct tacctgatct tctacaacct gatcaaaccg tacctgatcc cgcagatgat
     6661 gtacaaactg ccgaacacta tctcctccga aactttcctg aaataagggc ccgacgtctg
     6721 ca
//
