LOCUS       sequence-220096-        4559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construction of unmarked gene deletions. Suicide cloning vector 
            designed to introduce a targeted I-SceI restriction site in the 
            genome of Burkholderia.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4559)
  AUTHORS   Flannagan RS, Linn T, Valvano MA
  TITLE     A system for the construction of targeted unmarked gene deletions in
            the genus Burkholderia.
  JOURNAL   Environ Microbiol. 2008 Jun;10(6):1652-60. Epub 2008 Mar 13.
  PUBMED    18341581
REFERENCE   2  (bases 1 to 4559)
  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: "Environ 
            Microbiol."; date: "2008-06"; volume: "10(6)"; pages: "1652-60. Epub
            2008 Mar 13"
COMMENT     Sequence Label: pGPI-SceI
FEATURES             Location/Qualifiers
     source          1..4559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..237
                     /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"
     primer_bind     complement(873..892)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1033..1055
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1194..1213
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     oriT            1894..2003
                     /note="incP origin of transfer"
     CDS             2036..2407
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      3025..3413
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    3448..3464
                     /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(4206..4211,4212..4228,4229..4234)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 atgggtcaaa gtagcgatga agccaacgct cccgttgcag ggcagtttgc gcttcccctg
       61 agtgccacct ttggcttagg ggatcgcgta cgcaagaaat ctggtgccgc ttggcagggt
      121 caagtcgtcg gttggtattg cacaaaactc actcctgaag gctatgcggt cgagtccgaa
      181 tcccacccag gctcagtgca aatttatcct gtggctgcac ttgaacgtgt ggcctaagca
      241 ttaccctgtt atccctaact gcactcacca gtcacagaaa agcatcttac ggatggcatg
      301 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
      361 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
      421 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
      481 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
      541 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
      601 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
      661 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
      721 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
      781 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
      841 atactttaga ttgatttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
      901 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
      961 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     1021 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     1081 cgaggcagca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     1141 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     1201 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     1261 ccggcgtaga ggatcctttt tgtccggtgt tgggttgaag gtgaagccgg tcggggccgc
     1321 agcgggggcc ggcttttcag ccttgccccc ctgcttcggc cgccgtggct ccggcgtctt
     1381 gggtgccggc gcgggttccg cagccttggc ctgcggtgcg ggcacatcgg cgggcttggc
     1441 cttgatgtgc cgcctggcgt gcgagcggaa cgtctcgtag gagaacttga ccttccccgt
     1501 ttcccgcatg tgctcccaaa tggtgacgag cgcatagccg gacgctaacg ccgcctcgac
     1561 atccgccctc accgccagga acgcaaccgc agcctcatca cgccggcgct tcttggccgc
     1621 gcgggattca acccactcgg ccagctcgtc ggtgtagctc tttggcatcg tctctcgcct
     1681 gtcccctcag ttcagtaatt tcctgcattt gcctgtttcc agtcggtaga tattccacaa
     1741 aacagcaggg aagcagcgct tttccgctgc ataaccctgc ttcggggtca ttatagcgat
     1801 tttttcggta tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt
     1861 agactttcct tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc
     1921 cgcgagcggg tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa
     1981 tcctgctctg cgaggctggc cggctaccgc cggcgtaaca gatgagggca agcggatggc
     2041 tgatgaaacc aagccaacca ggaagggcag cccacctatc aaggtgtact gccttccaga
     2101 cgaacgaaga gcgattgagg aaaaggcggc ggcggccggc atgagcctgt cggcctacct
     2161 gctggccgtc ggccagggct acaaaatcac gggcgtcgtg gactatgagc acgtccgcga
     2221 gctggcccgc atcaatggcg acctgggccg cctgggcggc ctgctgaaac tctggctcac
     2281 cgacgacccg cgcacggcgc ggttcggtga tgccacgatc ctcgccctgc tggcgaagat
     2341 cgaagagaag caggacgagc ttggcaaggt catgatgggc gtggtccgcc cgagggcaga
     2401 gccatgactt ttttagccgc taaaacggcc ggggggtgcg cgtgattgcc aagcacgtcc
     2461 ccatgcgctc catcaagaag agcgacttcg cggagctggt gaagtacatc accgacgagc
     2521 aaggcaagac cgagcgcctg ggtcacgtgc gcgtcacgaa ctgcgaggca aacaccctgc
     2581 ccgctgtcat ggccgaggtg atggcgaccc agcacggcaa cacccgttcc gaggccgaca
     2641 agacctatca cctgctggtt agcttccgcg cgggagagaa gcccgacgcg gagacgttgc
     2701 gcgcgattga ggaccgcatc tgcgctgggc ttggcttcgc cgagcatcag cgcgtcagtg
     2761 ccgtgcatca cgacaccgac aacctgcaca tccatatcgc catcaacaag attcacccga
     2821 cccgaaacac catccatgag ccgtatcggg cctaccgcgc cctcgctgac ctctgcgcga
     2881 cgctcgaacg ggactacggg cttgagcgtg acaatcacga aacgcggcag cgcgtttccg
     2941 agaaccgcgc gaacgacatg gagcggcacg cgggcgtgga aagcctggtc ggctggatcc
     3001 ggccacgatg cgtccggcgt agaggatctg aagatcagca gttcaacctg ttgatagtac
     3061 gtactaagct ctcatgtttc acgtactaag ctctcatgtt taacgtacta agctctcatg
     3121 tttaacgaac taaaccctca tggctaacgt actaagctct catggctaac gtactaagct
     3181 ctcatgtttc acgtactaag ctctcatgtt tgaacaataa aattaatata aatcagcaac
     3241 ttaaatagcc tctaaggttt taagttttat aagaaaaaaa agaatatata aggcttttaa
     3301 agcttttaag gtttaacggt tgtggacaac aagccaggga tgtaacgcac tgagaagccc
     3361 ttagagcctc tcaaagcaat tttgagtgac acaggaacac ttaacggctg acatgggaat
     3421 tccacatgtg gaattccaca tgtggaattg tgagcggata acaatttgtg gaattcccgg
     3481 gagagctcga tatcgcatgc ggtacctcta gaagaagctt gggatccgtc ggggatccgg
     3541 tgattgattg agcaagcttt atgcttgtaa accgttttgt gaaaaaattt ttaaaataaa
     3601 aaaggggacc tctagggtcc ccaattaatt agtaatataa tctattaaag gtcattcaaa
     3661 aggtcatcca ccggatcagc ttagtaaagc cctcgctaga ttttaatgcg gatgttgcga
     3721 ttacttcgcc aactattgcg ataacaagaa aaagccagcc tttcatgata tatctcccaa
     3781 tttgtgtagg gcttattatg cacgcttaaa aataataaaa gcagacttga cctgatagtt
     3841 tggctgtgag caattatgtg cttagtgcat ctaacgcttg agttaagccg cgccgcgaag
     3901 cggcgtcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt gatctcgcct
     3961 ttcacgtagt ggacaaattc ttccaactga tctagcttta atgcggtagt ttatcacagt
     4021 taaattgcta acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc
     4081 tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc
     4141 tcttgcggga ttctacgggg tctgacgctc agtggaacga aatcgatgag ctcgcacgaa
     4201 cccagttgac ataagcctgt tcggttcgta aactgtaatg caagtagcgt atgcgctcac
     4261 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
     4321 tggcttgtta tgactgtttt tttgtacagt ctatgcctcg ggcatccaag cagcaagcgc
     4381 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     4441 gtcgccctaa aacaaagtta ggcagccgtt gtgctggtgc tttctgatag ttgttgtggg
     4501 gtaggcagtc agagctcgat ttgcttgtcg ccataataga ttcacaagaa ggattcgac
//
