LOCUS       V002140                 5440 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002140
VERSION     V002140
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5440)
  AUTHORS   Barrett AR, Kang Y, Inamasu KS, Son MS, Vukovich JM, Hoang TT.
  TITLE     Genetic tools for allelic replacement in Burkholderia species
  JOURNAL   Appl. Environ. Microbiol. 74 (14), 4498-4508 (2008)
   PUBMED   18502918
REFERENCE   2  (bases 1 to 5440)
  AUTHORS   Barrett AR, Kang Y, Vukovich JM, Son MS, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-DEC-2007) Microbiology, University of Hawaii at Manoa,
            2538 McCarthy Mall Snyder Hall Rm. 310, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 5440)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5440)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "14";
            pages: "4498-4508"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (04-DEC-2007) Microbiology, University of Hawaii at Manoa, 2538
            McCarthy Mall Snyder Hall Rm. 310, Honolulu, HI 96822, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label="AmpR promoter"
     CDS             201..1058
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1232..1820
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2108..2129
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2206..2222
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    2289..2362
                     /label="Flip recognition target"
                     /note="Flip recognition target"
     protein_bind    complement(2306..2353)
                     /label="FRT"
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      2393..2434
                     /regulatory_class="promoter"
     RBS             2445..2467
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2474..2821
                     /gene="merT"
                     /label="Mercuric transport protein MerT"
                     /note="Mercuric transport protein MerT from Pseudomonas
                     aeruginosa. Accession#: P04140"
     CDS             2837..3109
                     /gene="merP"
                     /label="Mercuric transport protein periplasmic component"
                     /note="Mercuric transport protein periplasmic component
                     from Pseudomonas aeruginosa. Accession#: P04131"
     CDS             3184..4866
                     /gene="merA"
                     /label="Mercuric reductase"
                     /note="Mercuric reductase from Pseudomonas aeruginosa.
                     Accession#: P00392"
     protein_bind    complement(4932..4979)
                     /label="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(5046..5062)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcatgcctgc aggtcgactc taggatcccc gggtaccgag
     2281 ctcgaattag cttcaaaagc gctctgaagt tcctatactt tctagagaat aggaacttcg
     2341 gaataggaac ttcaagatcc cctgattccc tttgtcaaca gcaatggata tcagccgttg
     2401 acttagttgg tatttccgga atatcatgct gggttccgaa taattttgtt taactttaag
     2461 aaggagatat accatgtctg aaccaaaaac cgggcgcggc gcgctcttca ctggagggct
     2521 tgccgccatc ctcgcctcgg cttgctgcct cgggccgttg gttctgatcg ccttggggtt
     2581 cagcggcgct tggatcggca acttggcggt gttggaaccc tatcgcccca tctttatcgg
     2641 cgtggcgctg gtggcgttgt tcttcgcctg gcggcgcatc taccggcagg cagcggcctg
     2701 caaaccgggt gaggtctgcg cgattcccca agtgcgagct acttacaagc tcattttctg
     2761 gatcgtggcc gcgctggttc tggtcgcgct cggatttccc tacgtcatgc catttttcta
     2821 ctgatcggag ttcaccatga agaaactgtt tgcctccctc gccctcgccg ccgttgttgc
     2881 ccccgtctgg gccgccaccc agaccgtcac gctgtccgta ccgggcatga cctgctccgc
     2941 ctgcccgatc actgtcaaga aggcgatttc cgaggtcgaa ggcgtcagca aagttgacgt
     3001 gactttcgag acacgccaag cggtcgtcac cttcgacgat gccaagacca gcgtgcagaa
     3061 gctgaccaag gcaaccgcag acgcgggcta tccgtccagc gtcaagcagt gagtcactga
     3121 aaacggcacc gcagcacaac ggacgtcatt gtctggcgcc acaaacgata aaggatctgt
     3181 tgcatgaccc atctaaaaat caccggcatg acttgcgact cgtgcgcggc gcacgtcaag
     3241 gaagcgctgg aaaaagtgcc aggcgtgcag tcggcgctgg tgtcctatcc gaagggcaca
     3301 gcgcaactcg ccatcgtgcc gggcacatcg ccggacgcgc tgactgccgc cgtggccgga
     3361 ctgggctaca aggcaacgct agccgatgcg ccactggcgg acaaccgcgt cggactgctc
     3421 gacaaggtgc ggggatggat ggccgccgcc gaaaagcaca gtggcaacga gcccccggtg
     3481 caggtagcgg tcattggcag cggtggagcc gcgatggcgg cggcgctgaa ggccgtcgag
     3541 caaggcgcgc aggtcacgct gatcgagcgc ggcaccatcg gcggcacctg cgtcaatgtc
     3601 ggctgtgtgc cgtccaagat catgatccgc gccgcccaca tcgcccatct gcgccgggaa
     3661 agcccgttcg atggcggtat tgcggcaact gtgcctacga ttgaccgcag taagctgctg
     3721 gcccagcagc aggcccgcgt ggacgaactg cggcacgcca agtacgaagg catcctgggc
     3781 ggtaatccgg ccatcaccgt tgtgcacggt gaggcgcgct tcaaggacga ccagagcctt
     3841 accgtccgtt tgaacgaggg tggcgagcgc gtcgtgatgt tcgaccgctg cctggtcgcc
     3901 acgggtgcca gcccggcggt cccgccgatt ccggggttga aagagtcacc ctactggact
     3961 tccaccgagg ccctggcgag cgacaccatt cccgaacgcc ttgccgtaat cggctcgtcg
     4021 gtggtggcgc tggagctggc gcaagccttt gcccggctgg gcagcaaggt cacggtcctg
     4081 gcgcgcaata ccttgttctt ccgtgaagac ccggccatcg gcgaggcggt gacagccgct
     4141 ttccgtgccg agggcatcga ggtgctggag cacacgcaag ccagccaggt cgcccatatg
     4201 gacggtgagt tcgtgctgac caccacgcac ggtgaattgc gcgccgacaa actgctggtt
     4261 gccaccggtc ggacaccgaa cacgcgcagc ctcgcgctgg acgcagcggg ggtcactgtc
     4321 aatgcgcaag gtgccatcgt catcgaccaa ggtatgcgca cgagcaaccc gaacatctac
     4381 gcggccggcg actgcaccga ccagccgcag ttcgtctatg tggcggcagc ggccggcacc
     4441 cgtgccgcga tcaacatgac cggcggcgat gcggcgctcg acctgaccgc aatgccggcc
     4501 gtggtgttca ccgatccgca agtggcgacc gtgggctaca gcgaggcgga agcccaccac
     4561 gacgggatcg agaccgacag ccgcaccttg accttggaca acgtgccgcg tgcgctcgcc
     4621 aacttcgaca cacgcggctt catcaagttg gttatcgagg aaggcagcca tcggctgatc
     4681 ggcgtacagg cggtcgcgcc ggaagcgggt gaactgatcc agacggcggc tctggccatt
     4741 cgcaaccgca tgacggtgca ggaactggcc gaccagttgt tcccctacct gacgatggtc
     4801 gaggggttga agctcgcggc gcagaccttc aacaaggatg tgaagcagct ttcctgctgc
     4861 gccgggtgag aaaaagctcg agctaattcc attgagtaag tttttaagca catcagcttc
     4921 aaaagcgctc tgaagttcct atactttcta gagaatagga acttcggaat aggaacttca
     4981 agatccccaa ttcgagctcg gtacccgggg atcctagagt cgacctgcag gcatgcaagc
     5041 ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5101 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5161 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     5221 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     5281 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     5341 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5401 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
