LOCUS       sequence-258172-        5299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insertional mutagenesis of GP transposon into chromosome of B. 
            cenocepacia. Trimethoprim selection for plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5299)
  AUTHORS   Flannagan RS, Aubert D, Kooi C, Sokol PA, Valvano MA
  TITLE     Burkholderia cenocepacia requires a periplasmic HtrA protease for 
            growth under thermal and osmotic stress and for survival in vivo.
  JOURNAL   Infect Immun. 2007 Apr;75(4):1679-89. Epub 2007 Jan 12.
  PUBMED    17220310
REFERENCE   2  (bases 1 to 5299)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Infect 
            Immun."; date: "2007-04"; volume: "75(4)"; pages: "1679-89. Epub 
            2007 Jan 12"
COMMENT     Sequence Label: pGP-Omega-Tp
FEATURES             Location/Qualifiers
     source          1..5299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(170..189)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     330..352
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     491..510
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     oriT            1191..1300
                     /note="incP origin of transfer"
     CDS             1333..1704
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      2322..2710
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2745..2761
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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        join(3503..3508,3509..3525,3526..3531)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3857..4093
                     /codon_start=1
                     /gene="dhfrIIb from plasmid R388"
                     /product="E. coli plasmid-associated dihydrofolate 
                     reductase"
                     /label=TpR
                     /note="confers resistance to trimethoprim"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
ORIGIN
        1 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
       61 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
      121 accaagttta ctcatatata ctttagattg atttatggtg cactctcagt acaatctgct
      181 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
      241 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
      301 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
      361 gtcatcaccg aaacgcgcga ggcagcaagg agatggcgcc caacagtccc ccggccacgg
      421 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
      481 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
      541 tgccggccac gatgcgtccg gcgtagagga tcctttttgt ccggtgttgg gttgaaggtg
      601 aagccggtcg gggccgcagc gggggccggc ttttcagcct tgcccccctg cttcggccgc
      661 cgtggctccg gcgtcttggg tgccggcgcg ggttccgcag ccttggcctg cggtgcgggc
      721 acatcggcgg gcttggcctt gatgtgccgc ctggcgtgcg agcggaacgt ctcgtaggag
      781 aacttgacct tccccgtttc ccgcatgtgc tcccaaatgg tgacgagcgc atagccggac
      841 gctaacgccg cctcgacatc cgccctcacc gccaggaacg caaccgcagc ctcatcacgc
      901 cggcgcttct tggccgcgcg ggattcaacc cactcggcca gctcgtcggt gtagctcttt
      961 ggcatcgtct ctcgcctgtc ccctcagttc agtaatttcc tgcatttgcc tgtttccagt
     1021 cggtagatat tccacaaaac agcagggaag cagcgctttt ccgctgcata accctgcttc
     1081 ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata tacaggattt
     1141 tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc gggcaggata
     1201 ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta ttcgcacctg
     1261 gcggtgctca acgggaatcc tgctctgcga ggctggccgg ctaccgccgg cgtaacagat
     1321 gagggcaagc ggatggctga tgaaaccaag ccaaccagga agggcagccc acctatcaag
     1381 gtgtactgcc ttccagacga acgaagagcg attgaggaaa aggcggcggc ggccggcatg
     1441 agcctgtcgg cctacctgct ggccgtcggc cagggctaca aaatcacggg cgtcgtggac
     1501 tatgagcacg tccgcgagct ggcccgcatc aatggcgacc tgggccgcct gggcggcctg
     1561 ctgaaactct ggctcaccga cgacccgcgc acggcgcggt tcggtgatgc cacgatcctc
     1621 gccctgctgg cgaagatcga agagaagcag gacgagcttg gcaaggtcat gatgggcgtg
     1681 gtccgcccga gggcagagcc atgacttttt tagccgctaa aacggccggg gggtgcgcgt
     1741 gattgccaag cacgtcccca tgcgctccat caagaagagc gacttcgcgg agctggtgaa
     1801 gtacatcacc gacgagcaag gcaagaccga gcgcctgggt cacgtgcgcg tcacgaactg
     1861 cgaggcaaac accctgcccg ctgtcatggc cgaggtgatg gcgacccagc acggcaacac
     1921 ccgttccgag gccgacaaga cctatcacct gctggttagc ttccgcgcgg gagagaagcc
     1981 cgacgcggag acgttgcgcg cgattgagga ccgcatctgc gctgggcttg gcttcgccga
     2041 gcatcagcgc gtcagtgccg tgcatcacga caccgacaac ctgcacatcc atatcgccat
     2101 caacaagatt cacccgaccc gaaacaccat ccatgagccg tatcgggcct accgcgccct
     2161 cgctgacctc tgcgcgacgc tcgaacggga ctacgggctt gagcgtgaca atcacgaaac
     2221 gcggcagcgc gtttccgaga accgcgcgaa cgacatggag cggcacgcgg gcgtggaaag
     2281 cctggtcggc tggatccggc cacgatgcgt ccggcgtaga ggatctgaag atcagcagtt
     2341 caacctgttg atagtacgta ctaagctctc atgtttcacg tactaagctc tcatgtttaa
     2401 cgtactaagc tctcatgttt aacgaactaa accctcatgg ctaacgtact aagctctcat
     2461 ggctaacgta ctaagctctc atgtttcacg tactaagctc tcatgtttga acaataaaat
     2521 taatataaat cagcaactta aatagcctct aaggttttaa gttttataag aaaaaaaaga
     2581 atatataagg cttttaaagc ttttaaggtt taacggttgt ggacaacaag ccagggatgt
     2641 aacgcactga gaagccctta gagcctctca aagcaatttt gagtgacaca ggaacactta
     2701 acggctgaca tgggaattcc acatgtggaa ttccacatgt ggaattgtga gcggataaca
     2761 atttgtggaa ttcccgggag agctcgatat cgcatgcggt acctctagaa gaagcttggg
     2821 atccgtcggg gatccggtga ttgattgagc aagctttatg cttgtaaacc gttttgtgaa
     2881 aaaattttta aaataaaaaa ggggacctct agggtcccca attaattagt aatataatct
     2941 attaaaggtc attcaaaagg tcatccaccg gatcagctta gtaaagccct cgctagattt
     3001 taatgcggat gttgcgatta cttcgccaac tattgcgata acaagaaaaa gccagccttt
     3061 catgatatat ctcccaattt gtgtagggct tattatgcac gcttaaaaat aataaaagca
     3121 gacttgacct gatagtttgg ctgtgagcaa ttatgtgctt agtgcatcta acgcttgagt
     3181 taagccgcgc cgcgaagcgg cgtcggcttg aacgaattgt tagacattat ttgccgacta
     3241 ccttggtgat ctcgcctttc acgtagtgga caaattcttc caactgatct agctttaatg
     3301 cggtagttta tcacagttaa attgctaacg cagtcaggca ccgtgtatga aatctaacaa
     3361 tgcgctcatc gtcatcctcg gcaccgtcac cctggatgct gtaggcatag gcttggttat
     3421 gccggtactg ccgggcctct tgcgggattc tacggggtct gacgctcagt ggaacgaaat
     3481 cgatgagctc gcacgaaccc agttgacata agcctgttcg gttcgtaaac tgtaatgcaa
     3541 gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg
     3601 cgcagtggcg gttttcatgg cttgttatga ctgttttttt gtacagtcta tgcctcgggc
     3661 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
     3721 tgttacgcag cagggcagtc gccctaaaac aaagttaggc agccgttgtg ctggtgcttt
     3781 ctgatagttg ttgtggggta ggcagtcaga gctcgatttg cttgtcgcca taatagattc
     3841 acaagaagga ttcgacatgg gtcaaagtag cgatgaagcc aacgctcccg ttgcagggca
     3901 gtttgcgctt cccctgagtg ccacctttgg cttaggggat cgcgtacgca agaaatctgg
     3961 tgccgcttgg cagggtcaag tcgtcggttg gtattgcaca aaactcactc ctgaaggcta
     4021 tgcggtcgag tccgaatccc acccaggctc agtgcaaatt tatcctgtgg ctgcacttga
     4081 acgtgtggcc taagcgatat cttaggatcc ccgacgcgag gctggatggc cttccccatt
     4141 atgattcttc tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg
     4201 caggtagatg acgaccatca gggacagctt caaggatcgc tcgcggctct taccagccta
     4261 acttcgatca ttggaccgct gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg
     4321 aacgggttgg catggattgt aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt
     4381 cgcggtgcat ggagccgggc cacctcgacc tgaatggaag ccggcggcac ctcgctaacg
     4441 gattcaccac tccaagaatt ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa
     4501 ccaacccttg gcagaacata tccatcgcgt ccgccatctc cagcagccgc acgcggcgca
     4561 tctcggcgga tcagtgaggg tttgcaactg cgggtcaagg atctggattt cgatcacggc
     4621 acgatcatcg tgcgggaggg caagggctcc aaggatcggg ccttgatgtt acccgagagc
     4681 ttggcaccca gcctgcgcga gcaggggaat tgatccggtg gatgaccttt tgaatgacct
     4741 ttaatagatt atattactaa ttaattgggg accctagagg tccccttttt tattttaaaa
     4801 attttttcac aaaacggttt acaagcataa agcttgctca atcaatcacc ggatccccac
     4861 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4921 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4981 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5041 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     5101 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5161 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5221 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5281 attgcagcac tggggccag
//
