LOCUS       sequence-212087-        7390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible allelic exchange vector for Burkholderia sp..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7390)
  AUTHORS   Fazli M, Harrison JJ, Gambino M, Givskov M, Tolker-Nielsen T
  TITLE     In-Frame and Unmarked Gene Deletions in Burkholderia cenocepacia via
            an Allelic Exchange System Compatible with Gateway Technology.
  JOURNAL   Appl Environ Microbiol. 2015 Jun;81(11):3623-30. doi: 
            10.1128/AEM.03909-14. Epub 2015 Mar 20.
  PUBMED    25795676
REFERENCE   2  (bases 1 to 7390)
  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"; doi: 
            "10.1128/AEM.03909-14"; journalName: "Appl Environ Microbiol"; date:
            "2015-06"; volume: "81"; issue: "11"; pages: "3623-30"
COMMENT     Sequence Label: pDONRPEX18Gm-SceI-pheS
FEATURES             Location/Qualifiers
     source          1..7390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    214..235
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(250..255,256..273,274..280)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    288..304
                     /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)."
     primer_bind     293..315
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     312..328
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     401..417
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     422..441
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        422..440
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    459..690
                     /gene="mutant version of attP"
                     /label=attP2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        835..937
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             938..1621
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     complement(1004..1023)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1941..2246
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2125..2144
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2642..2873)
                     /gene="mutant version of attP"
                     /label=attP1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2890..2906)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2963..2979)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3159..3176)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3329..3415
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3507..3534
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     oriT            3977..4086
                     /note="incP origin of transfer"
     primer_bind     complement(5357..5376)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5476..5498
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5536..5554)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5622..5726
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5744..5749,5750..5766,5767..5772)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5942..6475
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      6728..7316
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7217..7236
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
       61 cagccgaacg accgagcgca gcgggtcagt gagcgaggaa gcggaagagc gcccaatacg
      121 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
      181 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
      241 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
      301 acaatttcac acaggaaaca gctatgacat gattacgaat tcgagctcgg tacccgggga
      361 tcctctagat cagagatttt gagacacggg ccagagctgc caggaaacag ctatgaccat
      421 gtaatacgac tcactatagg ggatatcagc tggatggcaa ataatgattt tattttgact
      481 gatagtgacc tgttcgttgc aacaaattga taagcaatgc tttcttataa tgccaacttt
      541 gtacaagaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat
      601 tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt cactatgaat
      661 caactactta gatggtatta gtgacctgta gtcgactaag ttggcagcat cacccgacgc
      721 actttgcgcc gaataaatac ctgtgacgga agatcacttc gcagaataaa taaatcctgg
      781 tgtccctgtt gataccggga agccctgggc caacttttgg cgaaaatgag acgttgatcg
      841 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
      901 tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
      961 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     1021 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     1081 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     1141 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     1201 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     1261 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     1321 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     1381 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     1441 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     1501 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     1561 acaacagtac tgcgatgagt ggcagggggg gcgtaaacgc cgcgtggatc cggcttacta
     1621 aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact
     1681 gatatgtata cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagtga
     1741 cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct
     1801 ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga
     1861 aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga
     1921 cgagaacagg ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt
     1981 atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga
     2041 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     2101 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     2161 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     2221 ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag
     2281 gtcgatacag tagaaattac agaaacttta tcacgtttag taagtataga ggctgaaaat
     2341 ccagatgaag ccgaacgact tgtaagagaa aagtataaga gttgtgaaat tgttcttgat
     2401 gcagatgatt ttcaggacta tgacactagc gtatatgaat aggtagatgt ttttattttg
     2461 tcacacaaaa aagaggctcg cacctctttt tcttatttct ttttatgatt taatacggca
     2521 ttgaggacaa tagcgagtag gctggatacg acgattccgt ttgagaagaa catttggaag
     2581 gctgtcggtc gactaagttg gcagcatcac ccgaagaaca tttggaaggc tgtcggtcga
     2641 ctacaggtca ctaataccat ctaagtagtt gattcatagt gactggatat gttgtgtttt
     2701 acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     2761 attttacgtt tctcgttcag cttttttgta caaagttggc attataaaaa agcattgctc
     2821 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttggggccc
     2881 gagcttaaga ctggccgtcg ttttacaacg tcgtgactgg gaaaacatcc atgctattac
     2941 cctgttatcc ctaaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     3001 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     3061 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     3121 gagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
     3181 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
     3241 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
     3301 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     3361 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     3421 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     3481 ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt
     3541 tctacaaact cttttgttta tttttctaaa tacattcaaa tatgcgcctg atgcggtatt
     3601 ttctccttac gcatctgtgc ggtatttcac accgcatatc gacatccgcc ctcaccgcca
     3661 ggaacgcaac cgcagcctca tcacgccggc gcttcttggc cgcgcgggat tcaacccact
     3721 cggccagctc gtcggtgtag ctctttggca tcgcctctcg cctgtcccct cagttcagta
     3781 atttcttgca tttgcctgtt tccagtcggt agatattcca caaaacagca gggaagcagc
     3841 gcttttccgc tgcataaccc tgcttcgggg tcattatagc gattttttcg gtatatccat
     3901 cctttttcgc acgatataca ggattttgcc aaagggttcg tgtagacttt ccttggtgta
     3961 tccaacggcg tcagccgggc aggataggtg aagtaggccc acccgcgagc gggtgttcct
     4021 tcttcactgt cccttattcg cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct
     4081 ggccggctac cgccggcgta acagtactga gggcaagcac tagctgttga ctcgcttggg
     4141 attttcggaa tatcatgccg ggtctgacaa tcggatcgag cttcgaaagg acaagcatat
     4201 ggacctcgat cagatcgtgg cggatgccca gcagagcttt gagggcgcgg ccgacattac
     4261 gaccttggaa aatgaaaagg cccgcttcct gggcaagagc ggtgccctca cggaactctt
     4321 gaaggggctg ggtaagctgg acccggaaac ccggaagacc gagggtgccc ggattaacgt
     4381 ggccaagcag caggtggagg cggccttgaa tgcccggcgg caggccttgg cggatgccct
     4441 tctcaatcag cgcctggcgg ctgaagccat tgatgtgacc ctccccggtc gcggggcggg
     4501 cgccggctcg ctgcaccccg tcatgcgaac ctgggaacgt gtggagcaga ttttccgtag
     4561 cattggcttc gatgtcgcgg atggaccgga aattgagacg gattggtata atttcacggc
     4621 gctcaactcg cccgaaaacc accccgcacg gtccatgcag gacaccttct atgtggatgg
     4681 taaggacgcc gatgggcggc ccctcctcct ccggacccat acctcgccca tgcaggtgcg
     4741 gtacgcccgc atgaatcgac ccccgattaa ggtgattgct cccggtcgga cctatcgggt
     4801 cgattcggac gccacccata gccccatgtt taatcaggtg gaggggctct ggattgatga
     4861 aaacgtgtcg tttgccgatc tcaagggtgc ctacaccgac tttctcaaga aatttttcga
     4921 acgagatgac atcttggtcc gtttccgccc gagctacttc cccttcacgg aacccagcgc
     4981 ggaaattgat atgatgtttg aacatgggaa gaatgctggt aagtggttgg aaatcagcgg
     5041 cagcggtcag gtccacccca cggtcattcg gaatatgggc ctcgacccgg agcgttacat
     5101 tggcttcggc ttcggttccg ggttggaacg ccttaccatg ctccggtatg gtgtccagga
     5161 cctccggctc tttttcgaaa atgacctccg gttcctccgc cagttcgcct aaagatccaa
     5221 cgtttgcgcc tagcttcctg ctgaacatca aaggcaagaa aacatctgtt gtcaaagaca
     5281 gcatccttga acaaggacaa ttaacagtta acaaataaaa acgcaaaaga aaatgccgat
     5341 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     5401 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     5461 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     5521 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5581 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5641 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5701 ttcaataata ttgaaaaagg aagagtatga gtattcaacc gaattgacat aagcctgttc
     5761 ggttcgtaaa ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
     5821 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     5881 tgtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     5941 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttagg
     6001 tggctcaagt atgggcatca ttcgcacatg taggctcggc cctgaccaag tcaaatccat
     6061 gcgggctgct cttgatcttt tcggtcgtga gttcggagac gtagccacct actcccaaca
     6121 tcagccggac tccgattacc tcgggaactt gctccgtagt aagacattca tcgcgcttgc
     6181 tgccttcgac caagaagcgg ttgttggcgc tctcgcggct tacgttctgc ccaagtttga
     6241 gcagccgcgt agtgagatct atatctatga tctcgcagtc tccggcgagc accggaggca
     6301 gggcattgcc accgcgctca tcaatctcct caagcatgag gccaacgcgc ttggtgctta
     6361 tgtgatctac gtgcaagcag attacggtga cgatcccgca gtggctctct atacaaagtt
     6421 gggcatacgg gaagaagtga tgcactttga tatcgaccca agtaccgcca cctaacaatt
     6481 cgttcaagcc gagatcggct tcccggccgc ggagttgttc ggtaaattgt cacaacgccg
     6541 cggccaattc gcatcgattg tcagaccaag tttactcata tatactttag attgatttaa
     6601 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6661 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6721 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6781 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6841 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6901 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6961 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7021 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7081 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7141 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7201 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7261 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7321 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7381 ttcctgcgtt
//
