LOCUS       V006226                 7680 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006226
VERSION     V006226
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7680)
  AUTHORS   Norris MH, Kang Y, Barrett AR, Lu D, Hoang TT.
  TITLE     Glyphosate resistance as a novel non-antibiotic selectable marker in
            the select agent Burkholderia pseudomallei
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7680)
  AUTHORS   Norris MH, Kang Y, Barrett AR, Lu D, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-OCT-2008) Molecular Biology and Bio-Engineering,
            University of Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308,
            Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 7680)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7680)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (14-OCT-2008) Molecular Biology and Bio-Engineering, University of
            Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308, Honolulu, HI 96822,
            USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(232..583)
                     /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             597..1427
                     /label="pRO1600 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     primer_bind     1591..1607
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      1662..1703
                     /note="PCS12; rpsL promoter from Burkholderia pseudomallei"
                     /regulatory_class="promoter"
     CDS             1739..2749
                     /gene="pheS"
                     /label="Phenylalanine--tRNA ligase alpha subunit"
                     /note="Phenylalanine--tRNA ligase alpha subunit from
                     Burkholderia pseudomallei (strain 1710b). Accession#:
                     Q3JT08"
     CDS             complement(2964..4232)
                     /label="FLP"
                     /note="site-specific recombinase"
     RBS             4236..4244
                     /label="Shine-Dalgarno sequence"
                     /note="full consensus sequence for ribosome-binding sites
                     upstream of start codons in E. coli; complementary to a
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             4333..5043
                     /label="lambda repressor (ts)"
                     /note="temperature-sensitive variant of the phage lambda
                     repressor"
     primer_bind     complement(5335..5351)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5359..5375)
                     /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(5383..5413)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5428..5449)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     oriT            5833..5941
                     /label="oriT"
                     /note="incP origin of transfer"
     rep_origin      complement(6013..6601)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6921..7361)
                     /codon_start=1
                     /gene="gat"
                     /product="Gat"
                     /label="gat"
                     /note="glyphosate acetyl transferase; confers resistance to
                     glyphosate"
                     /protein_id="ACJ70061.1"
                     /translation="MIEVKPINAEDTYDLRHRVLRPNQPIEACMFESDLTRSAFHLGGF
                     YGGKLISVASFHQAEHSELQGKKQYQLRGVATLEGYREQKAGSSLVKHAEEILRKRGAD
                     MIWCNARTSASGYYRKLGFSEQGEVFDTPPVGPHILMYKRIT"
     gene            complement(6921..7361)
                     /gene="gat"
                     /label="gat"
     RBS             complement(7397..7419)
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      complement(7430..7471)
                     /note="PCS12; rpsL promoter from Burkholderia cenocepacia"
                     /regulatory_class="promoter"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatacctc tcaggcgccg ctggtgccgc tggttggacg ccaagggtga atccgcctcg
      241 ataccctgat tactcgcttc ctgcgccctc tcaggcggcg ataggggact ggtaaaacgg
      301 ggattgccca gacgcctccc ccgccccttc aggggcacaa atgcggcccc aacggggcca
      361 cgtagtggtg cgttttttgc gtttccaccc ttttcttcct tttccctttt aaacctttta
      421 ggacgtctac aggccacgta atccgtggcc tgtagagttt aaaaagggac ggatttgttg
      481 ccattaaggg acggatttgt tgttaagaag ggacggattt gttgttgtaa agggacggat
      541 ttgttgtatt gtgggacgca gatacagtgt ccccttatac acaaggaatg tcgaacgtgg
      601 cctcaccccc aatggtttac aaaagcaatg ccctggtcga ggccgcgtat cgcctcagtg
      661 ttcaggaaca gcggatcgtt ctggcctgta ttagccaggt gaagaggagc gagcctgtca
      721 ccgatgaagt gatgtattca gtgacggcgg aggacatagc gacgatggcg ggtgtcccta
      781 tcgaatcttc ctacaaccag ctcaaagaag cggccctgcg cctgaaacgg cgggaagtcc
      841 ggttaaccca agagcccaat ggcaagggga aaagaccgag tgtgatgatt accggctggg
      901 tgcaaacaat catctaccgg gagggtgagg gccgtgtaga actcaggttc accaaagaca
      961 tgctgccgta cctgacggaa ctcaccaaac agttcaccaa atacgccttg gctgacgtgg
     1021 ccaagatgga cagcacccac gcgatcaggc tttacgagct gctcatgcaa tgggacagca
     1081 tcggccagcg cgaaatagaa attgaccagc tgcgaaagtg gtttcaactg gaaggccggt
     1141 atccctcgat caaggacttc aagttgcgag tgcttgatcc agccgtgacg cagatcaacg
     1201 agcacagccc gctacaggtg gagtgggcgc agcgaaagac cgggcgcaag gtcacacatc
     1261 tgttgttcag ttttggaccg aagaagcccg ccaaggcggt gggtaaggcc ccagcgaagc
     1321 gcaaggccgg gaagatttca gatgctgaga tcgcgaaaca ggctcgccct ggtgagacat
     1381 gggaagcggc ccgcgctcga ctaacccaga tgccgctgga tctggcctag aggccgtggc
     1441 caccacggcc cggcctgcct ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1501 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1561 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgcat
     1621 gcaggcctct gcagtcgacg ggcccgggat ccgatagtac tagctgttga ctcgcttggg
     1681 attttcggaa tatcatgccg ggtctgacaa tcggatcgag cttcgaaagg acaagcatat
     1741 ggacctcgat cagatcgtgg cggatgccca gcagagcttt gagggcgcgg ccgacattac
     1801 gaccttggaa aatgaaaagg cccgcttcct gggcaagagc ggtgccctca cggaactctt
     1861 gaaggggctg ggtaagctgg acccggaaac ccggaagacc gagggtgccc ggattaacgt
     1921 ggccaagcag caggtggagg cggccttgaa tgcccggcgg caggccttgg cggatgccct
     1981 tctcaatcag cgcctggcgg ctgaagccat tgatgtgacc ctccccggtc gcggggcggg
     2041 cgccggctcg ctgcaccccg tcatgcgaac ctgggaacgt gtggagcaga ttttccgtag
     2101 cattggcttc gatgtcgcgg atggaccgga aattgagacg gattggtata atttcacggc
     2161 gctcaactcg cccgaaaacc accccgcacg gtccatgcag gacaccttct atgtggatgg
     2221 taaggacgcc gatgggcggc ccctcctcct ccggacccat acctcgccca tgcaggtgcg
     2281 gtacgcccgc atgaatcgac ccccgattaa ggtgattgct cccggtcgga cctatcgggt
     2341 cgattcggac gccacccata gccccatgtt taatcaggtg gaggggctct ggattgatga
     2401 aaacgtgtcg tttgccgatc tcaagggtgc ctacaccgac tttctcaaga aatttttcga
     2461 acgagatgac atcttggtcc gtttccgccc gagctacttc cccttcacgg aacccagcgc
     2521 ggaaattgat atgatgtttg aacatgggaa gaatgctggt aagtggttgg aaatcagcgg
     2581 cagcggtcag gtccacccca cggtcattcg gaatatgggc ctcgacccgg agcgttacat
     2641 tggcttcggc ttcggttccg ggttggaacg ccttaccatg ctccggtatg gtgtccagga
     2701 cctccggctc tttttcgaaa atgacctccg gttcctccgc cagttcgcct aaagatcttt
     2761 catctagagg atccccctag agaataggaa cttcggaata ggaacttcaa agcgtttccg
     2821 aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac tgttcacgtc
     2881 gcacctatat ctgcgtgttg cctgtatata tatatacatg agaagaacgg catagtgcgt
     2941 gtttatgctt aaatgcgtac ttatatgcgt ctatttatgt aggatgaaag gtagtctagt
     3001 acctcctgtg atattatccc attccatgcg gggtatcgta tgcttccttc agcactaccc
     3061 tttagctgtt ctatatgctg ccactcctca attggattag tctcatcctt caatgctatc
     3121 atttcctttg atattggatc atatgcatag taccgagaaa ctagtgcgaa gtagtgatca
     3181 ggtattgctg ttatctgatg agtatacgtt gtcctggcca cggcagaagc acgcttatcg
     3241 ctccaatttc ccacaacatt agtcaactcc gttaggccct tcattgaaag aaatgaggtc
     3301 atcaaatgtc ttccaatgtg agattttggg ccatttttta tagcaaagat tgaataaggc
     3361 gcatttttct tcaaagcttt attgtacgat ctgactaagt tatcttttaa taattggtat
     3421 tcctgtttat tgcttgaaga attgccggtc ctatttactc gttttaggac tggttcagaa
     3481 ttcctcaaaa attcatccaa atatacaagt ggatcgatcc taccccttgc gctaaagaag
     3541 tatatgtgcc tactaacgct tgtctttgtc tctgtcacta aacactggat tattactccc
     3601 agatacttat tttggactaa tttaaatgat ttcggatcaa cgttcttaat atcgctgaat
     3661 cttccacaat tgatgaaagt agctaggaag aggaattggt ataaagtttt tgtttttgta
     3721 aatctcgaag tatactcaaa cgaatttagt attttctcag tgatctccca gatgctttca
     3781 ccctcactta gaagtgcttt aagcattttt ttactgtggc tatttccctt atctgcttct
     3841 tccgatgatt cgaactgtaa ttgcaaacta cttacaatat cagtgatatc agattgatgt
     3901 ttttgtccat agtaaggaat aattgtaaat tcccaagcag gaatcaattt ctttaatgag
     3961 gcttccagaa ttgttgcttt ttgcgtcttg tatttaaact ggagtgattt attgacaata
     4021 tcgaaactca gcgaattgct tatgatagta ttatagctca tgaatgtggc tctcttgatt
     4081 gctgttccgt tatgtgtaat catccaacat aaataggtta gttcagcagc acataatgct
     4141 attttctcac ctgaaggtct ttcaaacctt tccacaaact gacgaacaag caccttaggt
     4201 ggtgttttac ataatatacc aaattgtggc atacaacctc cttagtacat gcaaccatta
     4261 tcaccgccag aggtaaaata gtcaacacgc acggtgttag atatttatcc cttgcggtga
     4321 tagatttaac gtatgagcac aaaaaagaaa ccattaacac aagagcagct tgaggacgca
     4381 cgtcgcctta aagcaattta tgaaaaaaag aaaaatgaac ttggcttatc ccaggaatct
     4441 gtcgcagaca agatggggat ggggcagtca ggcgttggtg ctttatttaa tggcatcaat
     4501 gcattaaatg cttataacgc cgcattgctt acaaaaattc tcaaagttag cgttgaagaa
     4561 tttagccctt caatcgccag agaaatctac gagatgtatg aagcggttag tatgcagccg
     4621 tcacttagaa gtgagtatga gtaccctgtt ttttctcatg ttcaggcagg gatgttctca
     4681 cctaagctta gaacctttac caaaggtgat gcggagagat gggtaagcac aaccaaaaaa
     4741 gccagtgatt ctgcattctg gcttgaggtt gaaggtaatt ccatgaccgc accaacaggc
     4801 tccaagccaa gctttcctga cggaatgtta attctcgttg accctgagca ggctgttgag
     4861 ccaggtgatt tctgcatagc cagacttggg ggtgatgagt ttaccttcaa gaaactgatc
     4921 agggatagcg gtcaggtgtt tttacaacca ctaaacccac agtacccaat gatcccatgc
     4981 aatgagagtt gttccgttgt ggggaaagtt atcgctagtc agtggcctga agagacgttt
     5041 ggctgatcgg caaggtgttc tggtcggcgc atagctgata acaattgagc aagaatcttc
     5101 atcgaattag gggaattttc actcccctca gaacataaca tagtaaatgg attgaattat
     5161 gaagaatggt ttttatgcga cttaccgcag caaaaataaa gggaaagata agcgctcaat
     5221 aaacctgtct gttttcctta attctctgct ggctgataat catcacctgc aggttggctc
     5281 caattatttg tatattcata aaatcggggt accgagctcg aattcgtaat catggtcata
     5341 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5401 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5461 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5521 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5581 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5641 gttatccaca gaatcagggg ataacgcagg aaagaacatg gggattcctt aaggtatact
     5701 ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt
     5761 tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca
     5821 acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt
     5881 cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc
     5941 gataagctag cttatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6001 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6061 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6121 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6181 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6241 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6301 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6361 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6421 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6481 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6541 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6601 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6661 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6721 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6781 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6841 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6901 aatgataccg cctcctcgag ttaggtgatc cgcttgtaca tcaggatgtg gggccccacg
     6961 ggcggggtgt cgaagacctc cccctgctcg ctgaacccca gcttccggta gtaccccgag
     7021 gccgaggtcc gggcgttgca ccagatcatg tcggcccccc gcttccgcag gatctcttcg
     7081 gcgtgcttga ccagcgacga cccggccttc tgctcccggt agccctccag ggtggccaca
     7141 ccgcgcagct ggtactgctt cttcccctgg agctccgagt gctcggcctg gtggaacgaa
     7201 gcgaccgaaa tcagcttgcc cccgtagaac ccgcccaggt ggaaggcgct ccgcgtgaga
     7261 tcgctttcga acatgcacgc ctcgatcggc tggttgggcc ggaggacccg atgccgcagg
     7321 tcgtaggtgt cctcggcgtt gatgggcttg acctcgatca tatgcttgtc ctttcgaagc
     7381 tcgatcccat ggtatatctc cttcttaaag ttaaacaaaa ttattcggaa cccagcatga
     7441 tattccggaa ataccaacta agtcaacggc tgatcatggt atatctcctt cttaaagatt
     7501 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7561 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     7621 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
