LOCUS       Exported                8031 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pI-SceI, complete sequence.
ACCESSION   EU372689
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8031)
  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) In press
  PUBMED    18341581
REFERENCE   2  (bases 1 to 8031)
  AUTHORS   Flannagan RS, Linn T, Valvano MA.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JAN-2008) Microbiology & Immunology, University of 
            Western Ontario, Dental Sciences Building, 3014, Canada, Ontario N6A
            5C1, Canada
REFERENCE   3  (bases 1 to 8031)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8031
                     /organism="Expression vector pI-SceI"
                     /mol_type="other DNA"
                     /note="used to express the I-SceI restriction endonuclease 
                     under the control of the dhfr promoter; not a cloning 
                     vector but it is part of a mutagenesis system for 
                     Burkholderia strains"
                     /db_xref="taxon:511658"
     CDS             complement(51..728)
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /label=tetR
                     /protein_id="ACC63377.1"
                     /translation="MFISDKVSSMTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGV
                     QQPALYWHFRNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRD
                     GARIHAGTRPGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSD
                     AGERGGTVEQAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRK
                     GDD"
     gene            complement(51..728)
                     /gene="tetR"
                     /label=tetR
     CDS             complement(51..701)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             732..2006
                     /codon_start=1
                     /gene="tetA"
                     /product="TetA"
                     /label=tetA
                     /note="tetracycline resistance"
                     /protein_id="ACC63378.1"
                     /translation="MSTNLSVIKNPRVQSDQRRLVRRPDVKPNIPLIVILSTVALDAVG
                     IGLIMPVLPGLLRDLVHSNDVTAHYGILLALYALVQFACAPVLGALSDRFGRRPILLVS
                     LAGATVDYAIMATAPFLWVLYIGRIVAGITGATGAVAGAYIADITDGDERARHFGFMSA
                     CFGFGMVAGPVLGGLMGGFSPHAPFFAAAALNGLNFLTGCFLLPESHKGERRPLRREAL
                     NPLASFRWARGMTVVAALMAVFFIMQLVGQVPAALWVIFGEDRFHWDATTIGISLAAFG
                     ILHSLAQAMITGPVAARLGERRALMLGMIADGTGYILLAFATRGWMAFPIMVLLASGGI
                     GMPALQAMLSRQVDEERQGQLQGSLAALTSLTSIVGPLLFTAIYAASITTWNGWAWIAG
                     AALYLLCLPALRRGLWSGAGQRADR"
     gene            732..2006
                     /gene="tetA"
                     /label=tetA
     CDS             807..2006
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     promoter        2243..2271
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     regulatory      2243..2248
                     /regulatory_class="minus_35_signal"
     regulatory      2266..2271
                     /regulatory_class="minus_10_signal"
     CDS             2606..3307
                     /codon_start=1
                     /gene="I-SceI"
                     /product="I-SceI-FLAG"
                     /label=I-SceI
                     /note="endonuclease; I-SceI-protein has a C-terminal FLAG 
                     oligopeptide"
                     /protein_id="ACC63379.1"
                     /translation="MHQKNQVMNLGPNSKLLKEYKSQLIELNIEQFEAGIGLILGDAYI
                     RSRDEGKTYCMQFEWKNKAYMDHVCLLYDQWVLSPPHKKERVNHLGNLVITWGAQTFKH
                     QAFNKLANLFIVNNKKTIPNNLVENYLTPMSLAYWFMDDGGKWDYNKNSTNKSIVLNTQ
                     SFTFEEVEYLVKGLRNKFQLNCYVKINKNKPIIYIDSMSYLIFYNLIKPYLIPQMMYKL
                     PNTISSETFLK"
     gene            2606..3307
                     /gene="I-SceI"
                     /label=I-SceI
     CDS             3326..3340
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     terminator      3568..3654
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3746..3773
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        complement(4628..4730)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(4880..6364)
                     /codon_start=1
                     /gene="mob"
                     /product="Mob"
                     /label=mob
                     /protein_id="ACC63380.1"
                     /translation="MSNRQAKRAGRRPEAFPQRKLKKLMGQGRRPRSWSRWVCGRRLGS
                     RWAIPVVKLVGRRSLSISLSSRVVHGPSEASQPVAARGHRPHIHGLARERSDRAGRSPE
                     SKPVGRRSRRRRSRLCEAKSSEYTQALSGPPEAPPCAAPVEPVGHQKGGAGMAAYAIMR
                     CKKLAKMGNVAASLKHAYRERETPNADASRTPENEHWAASSTDEAMGRLRELLPEKRRK
                     DAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLADKYGADRIVTASIHRDETSPHM
                     TAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVADLGLQRGIEGSKARHTRIQAFY
                     EALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKRVETPEAVADRLTKAVRQGYEPA
                     LQAAAGAREMRKKADQAQETARDLRERLKPVLDALGPLNRDMQAKAAAIIKAVGEKLLT
                     EQREVQRQKQAQRQQERGRAHFPEKCHLTSKKPLLS"
     gene            complement(4880..6364)
                     /gene="mob"
                     /label=mob
     rep_origin      6123..6892
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     rep_origin      6695..6706
     CDS             6881..7555
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="replication protein"
                     /protein_id="ACC63381.1"
                     /translation="MEISMATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAA
                     MLLHHLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVS
                     AYVVNDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTG
                     PGEEPPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLP
                     RR"
     gene            6881..7555
                     /gene="rep"
                     /label=rep
     CDS             6893..7555
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"