LOCUS       sequence-212135-        9145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast homing endonuclease I - SceI expression vector. Used for 
            resolution of merodiploids during allelic exchange in Burkholderia 
            sp..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9145)
  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 9145)
  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: pDAI-SceI-pheS
FEATURES             Location/Qualifiers
     source          1..9145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(988..993,994..1010,1011..1016))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     complement(1135..1154)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(1255..1905)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             2011..3210
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             complement(3738..4400)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQRRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      4401..5172
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     primer_bind     6430..6448
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6565..6667
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6734..6753)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(7521..7548)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(7640..7726)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     7879..7896
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 atctgcggga tcaggtacgg tttgatcagg ttgtagaaga tcaggtaaga catagaatcg
       61 atgtagatga tcggtttgtt tttgttgatt tttacgtaac agttcagttg gaatttgtta
      121 cgcagaccct taaccaggta ttctacttct tcgaaagtga aagactgggt gttcagtacg
      181 atcgatttgt tggtagagtt tttgttgtaa tcccatttac caccatcatc catgaaccag
      241 tatgccagag acatcggggt caggtagttt tcaaccaggt tgttcgggat ggtttttttg
      301 ttgttaacga tgaacaggtt agccagtttg ttgaaagctt ggtgtttgaa agtctgggcg
      361 ccccaggtga ttaccaggtt acccaggtgg ttaacacgtt cttttttgtg cggcggggac
      421 agtacccact gatcgtacag cagacatacg tggtccatgt atgctttgtt tttccactcg
      481 aactgcatac agtaggtttt accttcatca cgagaacgga tgtaagcatc acccaggatc
      541 agaccgatac ctgcttcgaa ctgttcgatg ttcagttcga tcagctggga tttgtattct
      601 ttcagcagtt tagagttcgg acccaggttc attacctggt ttttttgatg catatgtaga
      661 attccgaatc cttcttgtga atctattatg gcgacaagca aatcgagctc tgactgccta
      721 ccccacaaca actatcagaa agcaccagca caacggctgc ctaactttgt tttagggcga
      781 ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa
      841 cgcgcttgct gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc
      901 atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg
      961 cgtgagcgca tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaactgg
     1021 gttcgtgcga gctcatcgct agcatactcg agtaagctgt caaacatgag cagatcctct
     1081 acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata
     1141 tcgccgacat caccgatggg agatcctaag atatcgctta ggccacacgt tcaagtgcag
     1201 atcgatcgat ggctctgctg tagtgagtgg gttgcgctcc ggcagcggtc ctgatcaatc
     1261 gtcacccttt ctcggtcctt caacgttcct gacaacgagc ctccttttcg ccaatccatc
     1321 gacaatcacc gcgagtccct gctcgaacgc tgcgtccgga ccggcttcgt cgaaggcgtc
     1381 tatcgcggcc cgcaacagcg gcgagagcgg agcctgttca acggtgccgc cgcgctcgcc
     1441 ggcatcgctg tcgccggcct gctcctcaag cacggcccca acagtgaagt agctgattgt
     1501 catcagcgca ttgacggcgt ccccggccga aaaacccgcc tcgcagagga agcgaagctg
     1561 cgcgtcggcc gtttccatct gcggtgcgcc cggtcgcgtg ccggcatgga tgcgcgcgcc
     1621 atcgcggtag gcgagcagcg cctgcctgaa gctgcgggca ttcccgatca gaaatgagcg
     1681 ccagtcgtcg tcggctctcg gcaccgaatg cgtatgattc tccgccagca tggcttcggc
     1741 cagtgcgtcg agcagcgccc gcttgttcct gaagtgccag taaagcgccg gctgctgaac
     1801 ccccaaccgt tccgccagtt tgcgtgtcgt cagaccgtct acgccgacct cgttcaacag
     1861 gtccagggcg gcacggatca ctgtattcgg ctgcaacttt gtcatgcttg acactttatc
     1921 actgataaac ataatatgtc caccaactta tcagtgataa agaatccgcg cgttcaatcg
     1981 gaccagcgga ggctggtccg gaggccagac gtgaaaccca acatacccct gatcgtaatt
     2041 ctgagcactg tcgcgctcga cgctgtcggc atcggcctga ttatgccggt gctgccgggc
     2101 ctcctgcgcg atctggttca ctcgaacgac gtcaccgccc actatggcat tctgctggcg
     2161 ctgtatgcgt tggtgcaatt tgcctgcgca cctgtgctgg gcgcgctgtc ggatcgtttc
     2221 gggcggcggc caatcttgct cgtctcgctg gccggcgcca ctgtcgacta cgccatcatg
     2281 gcgacagcgc ctttcctttg ggttctctat atcgggcgga tcgtggccgg catcaccggg
     2341 gcgactgggg cggtagccgg cgcttatatt gccgatatca ctgatggcga tgagcgcgcg
     2401 cggcacttcg gcttcatgag cgcctgtttc gggttcggga tggtcgcggg acctgtgctc
     2461 ggtgggctga tgggcggttt ctccccccac gctccgttct tcgccgcggc agccttgaac
     2521 ggcctcaatt tcctgacggg ctgtttcctt ttgccggagt cgcacaaagg cgaacgccgg
     2581 ccgttacgcc gggaggctct caacccgctc gcttcgttcc ggtgggcccg gggcatgacc
     2641 gtcgtcgccg ccctgatggc ggtcttcttc atcatgcaac ttgtcggaca ggtgccggcc
     2701 gcgctttggg tcattttcgg cgaggatcgc tttcactggg acgcgaccac gatcggcatt
     2761 tcgcttgccg catttggcat tctgcattca ctcgcccagg caatgatcac cggccctgta
     2821 gccgcccggc tcggcgaaag gcgggcactc atgctcggaa tgattgccga cggcacaggc
     2881 tacatcctgc ttgccttcgc gacacgggga tggatggcgt tcccgatcat ggtcctgctt
     2941 gcttcgggtg gcatcggaat gccggcgctg caagcaatgt tgtccaggca ggtggatgag
     3001 gaacgtcagg ggcagctgca aggctcactg gcggcgctca ccagcctgac ctcgatcgtc
     3061 ggacccctcc tcttcacggc gatctatgcg gcttctataa caacgtggaa cgggtgggca
     3121 tggattgcag gcgctgccct ctacttgctc tgcctgccgg cgctgcgtcg cgggctttgg
     3181 agcggcgcag ggcaacgagc cgatcgctga tcgtggaaac gataggaaac gtggccaata
     3241 tggacaactt cttcgcatcg atttcgttcc actgagcgtc agaccccgta ggttattggt
     3301 cgacttaaac gcctggtgct acgcctgaat aagtgataat aagcggatga atggcagaaa
     3361 ttcgaaagca aattcgaccc ggtcgtcggt tcagggcagg gtcgttaaat agccgcttat
     3421 gtctattgct ggtttaccgg tttattgact accggaagca gtgtgaccgt gtgcttctca
     3481 aatgcctgag gccagtttgc tcaggctctc cccgtggagg taataattga cgatatgatc
     3541 atttattctg cctcccagag cctgataaaa acggtgaatc cgttagcgag gtgccgccgg
     3601 cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga
     3661 caaggtatag ggcggcgagg cggctacagc cgatagtctg gaacagcgca cttacgggtt
     3721 gctgcgcaac ccaagtgcta ccggcgcggc agcgtgaccc gtgtcggcgg ctccaacggc
     3781 tcgccatcgt ccagaaaaca cggctcatcg ggcatcggca ggcgccgctg cccgcgccgt
     3841 tcccattcct ccgtttcggt caaggctggc aggtctggtt ccatgcccgg aatgccgggc
     3901 tggctgggcg gctcctcgcc ggggccggtc ggtagttgct gctcgcccgg atacagggtc
     3961 gggatgcggc gcaggtcgcc atgccccaac agcgattcgt cctggtcgtc gtgatcaacc
     4021 accacggcgg cactgaacac cgacaggcgc aactggtcgc ggggctggcc ccacgccacg
     4081 cggtcattga ccacgtaggc cgacacggtg ccggggccgt tgagcttcac gacggagatc
     4141 cagcgctcgg ccaccaagtc cttgactgcg tattggaccg tccgcaaaga acgtccgatg
     4201 agcttggaaa gtgtcttctg gctgaccacc acggcgttct ggtggcccat ctgcgccacg
     4261 aggtgatgca gcagcattgc cgccgtgggt ttcctcgcaa taagcccggc ccacgcctca
     4321 tgcgctttgc gttccgtttg cacccagtga ccgggcttgt tcttggcttg aatgccgatt
     4381 tctctggact gcgtggccat gcttatctcc atgcggtagg ggtgccgcac ggttgcggca
     4441 ccatgcgcaa tcagctgcaa cttttcggca gcgcgacaac aattatgcgt tgcgtaaaag
     4501 tggcagtcaa ttacagattt tctttaacct acgcaatgag ctattgcggg gggtgccgca
     4561 atgagctgtt gcgtaccccc cttttttaag ttgttgattt ttaagtcttt cgcatttcgc
     4621 cctatatcta gttctttggt gcccaaagaa gggcacccct gcggggttcc cccacgcctt
     4681 cggcgcggct ccccctccgg caaaaagtgg cccctccggg gcttgttgat cgactgcgcg
     4741 gccttcggcc ttgcccaagg tggcgctgcc cccttggaac ccccgcactc gccgccgtga
     4801 ggctcggggg gcaggcgggc gggcttcgcc cttcgactgc ccccactcgc ataggcttgg
     4861 gtcgttccag gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc cttgacccgc
     4921 cttccacttg gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg aggcttttcc
     4981 ccagagaaaa ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt ggagccggtg
     5041 ggtatgtggt cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc tcgtgggcag
     5101 gcgcagcctg tccatcagct tgtccagcag ggttgtccac gggccgagcg aagcgagcca
     5161 gccggtggcc gctcgcggcc atcgtccaca tatccacggg ctggcaaggg agcgcagcga
     5221 ccgcgcaggg cgaagcccgg agagcaagcc cgtagggcgc cgcagccgcc gtaggcggtc
     5281 acgactttgc gaagcaaagt ctagtgagta tactcaagca ttgagtggcc cgccggaggc
     5341 accgccttgc gctgcccccg tcgagccggt tggacaccaa aagggagggg caggcatggc
     5401 ggcatacgcg atcatgcgat gcaagaagct ggcgaaaatg ggcaacgtgg cggccagtct
     5461 caagcacgcc taccgcgagc gcgagacgcc caacgctgac gccagcagga cgccagagaa
     5521 cgagcactgg gcggccagca gcaccgatga agcgatgggc cgactgcgcg agttgctgcc
     5581 agagaagcgg cgcaaggacg ctgtgttggc ggtcgagtac gtcatgacgg ccagcccgga
     5641 atggtggaag tcggccagcc aagaacagca ggcggcgttc ttcgagaagg cgcacaagtg
     5701 gctggcggac aagtacgggg cggatcgcat cgtgacggcc agcatccacc gtgacgaaac
     5761 cagcccgcac atgaccgcgt tcgtggtgcc gctgacgcag gacggcaggc tgtcggccaa
     5821 ggagttcatc ggcaacaaag cgcagatgac ccgcgaccag accacgtttg cggccgctgt
     5881 ggccgatcta gggctgcaac ggggcatcga gggcagcaag gcacgtcaca cgcgcattca
     5941 ggcgttctac gaggccctgg agcggccacc agtgggccac gtcaccatca gcccgcaagc
     6001 ggtcgagcca cgcgcctatg caccgcaggg attggccgaa aagctgggaa tctcaaagcg
     6061 cgttgagacg ccggaagccg tggccgaccg gctgacaaaa gcggttcggc aggggtatga
     6121 gcctgcccta caggccgccg caggagcgcg tgagatgcgc aagaaggccg atcaagccca
     6181 agagacggcc cgagaccttc gggagcgcct gaagcccgtt ctggacgccc tggggccgtt
     6241 gaatcgggat atgcaggcca aggccgccgc gatcatcaag gccgtgggcg aaaagctgct
     6301 gacggaacag cgggaagtcc agcgccagaa acaggcccag cgccagcagg aacgcgggcg
     6361 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     6421 acctataaaa ataggcgtat cacgaggccc tttcgtcttc gaataaatac ctgtgacgga
     6481 agatcacttc gcagaataaa taaatcctgg tgtccctgtt gataccggga agccctgggc
     6541 caacttttgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact ttcaccataa
     6601 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
     6661 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     6721 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     6781 gctggatatt acggcctttt tagcttttgc cattctcacc ggattcagtc gtcactcatg
     6841 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     6901 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     6961 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atacggtatt gataatcctg
     7021 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     7081 attggttgta acactggcag agcattacgc tgacttgacg ggacggcggc tttgttgaat
     7141 aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg
     7201 ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca aagctctcat
     7261 caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggcct ggtatgagtc
     7321 agcaacacct tcttcacgag gcagacctca gcgccagaag gccgccagag aggccgagcg
     7381 cggccgtgag gcttggacgc tagggcaggg catgaaaaag cccgtagcgg gctgctacgg
     7441 gcgtctgacg cggtggaaag ggggagggga tgttgtttaa atggctctgt gtagtgagtg
     7501 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
     7561 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcagcgt
     7621 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     7681 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
     7741 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
     7801 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
     7861 agaccgcttc tgcgttctga tttaatctgt atcaggctga aaatcttctc tcatccgcca
     7921 aaacagccaa gcttgcatgc aggcctctgc agtcgacggg cccgggatcc gatccaatag
     7981 tactagctgt tgactcgctt gggattttcg gaatatcatg ccgggtctga caatcggatc
     8041 gagcttcgaa aggacaagca tatggacctc gatcagatcg tggcggatgc ccagcagagc
     8101 tttgagggcg cggccgacat tacgaccttg gaaaatgaaa aggcccgctt cctgggcaag
     8161 agcggtgccc tcacggaact cttgaagggg ctgggtaagc tggacccgga aacccggaag
     8221 accgagggtg cccggattaa cgtggccaag cagcaggtgg aggcggcctt gaatgcccgg
     8281 cggcaggcct tggcggatgc ccttctcaat cagcgcctgg cggctgaagc cattgatgtg
     8341 accctccccg gtcgcggggc gggcgccggc tcgctgcacc ccgtcatgcg aacctgggaa
     8401 cgtgtggagc agattttccg tagcattggc ttcgatgtcg cggatggacc ggaaattgag
     8461 acggattggt ataatttcac ggcgctcaac tcgcccgaaa accaccccgc acggtccatg
     8521 caggacacct tctatgtgga tggtaaggac gccgatgggc ggcccctcct cctccggacc
     8581 catacctcgc ccatgcaggt gcggtacgcc cgcatgaatc gacccccgat taaggtgatt
     8641 gctcccggtc ggacctatcg ggtcgattcg gacgccaccc atagccccat gtttaatcag
     8701 gtggaggggc tctggattga tgaaaacgtg tcgtttgccg atctcaaggg tgcctacacc
     8761 gactttctca agaaattttt cgaacgagat gacatcttgg tccgtttccg cccgagctac
     8821 ttccccttca cggaacccag cgcggaaatt gatatgatgt ttgaacatgg gaagaatgct
     8881 ggtaagtggt tggaaatcag cggcagcggt caggtccacc ccacggtcat tcggaatatg
     8941 ggcctcgacc cggagcgtta cattggcttc ggcttcggtt ccgggttgga acgccttacc
     9001 atgctccggt atggtgtcca ggacctccgg ctctttttcg aaaatgacct ccggttcctc
     9061 cgccagttcg cctaaagatc tttcccaata tctagattat ttcaggaaag tttcggagga
     9121 gatagtgttc ggcagtttgt acatc
//
