LOCUS       40924_20256        7072 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Site-specific excision vector pFLPe4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7072)
  AUTHORS   Choi KH, Mima T, Casart Y, Rholl D, Kumar A, Beacham IR, Schweizer 
            HP.
  TITLE     Genetic tools for select-agent-compliant manipulation of 
            Burkholderia pseudomallei
  JOURNAL   Appl. Environ. Microbiol. 74 (4), 1064-1075 (2008)
  PUBMED    18156318
REFERENCE   2  (bases 1 to 7072)
  AUTHORS   Choi K-H., Schweizer HP.
  TITLE     Genetic tools for Pseudomonas and its related bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 7072)
  AUTHORS   Choi K-H., Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-SEP-2007) Microbiology, Immunology and Pathology, 
            Colorado State University, 1682 Campus Delivery, Ft. Collins, CO 
            80523-1682, USA
REFERENCE   4  (bases 1 to 7072)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7072)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "4"; pages:
            "1064-1075"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (21-SEP-2007) Microbiology, Immunology and Pathology, Colorado State
            University, 1682 Campus Delivery, Ft. Collins, CO 80523-1682, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(37..867)
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     rep_origin      881..1232
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     regulatory      1286..1333
                     /label=ribosomal S12 promoter from B. thailandensis E264
                     /note="ribosomal S12 promoter from B. thailandensis E264"
                     /regulatory_class="promoter"
     CDS             1448..2260
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(2410..3690)
                     /label=FLPo
                     /note="nuclear-targeted site-specific recombinase"
     promoter        complement(3703..3821)
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             3967..4800
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
     CDS             4877..5722
                     /label=rhaR
                     /note="transcriptional activator of rhaR and rhaS"
     primer_bind     complement(5759..5775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5783..5799)
                     /label=lac operator
                     /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        complement(5807..5837)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5852..5873)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     oriT            6257..6365
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(6437..7025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cggtcgagcg gccgccagtg tgatggatgg cctctaggcc agatccagcg gcatctgggt
       61 tagtcgagcg cgggccgctt cccatgtctc accagggcga gcctgtttcg cgatctcagc
      121 atctgaaatc ttcctggcct tgcgcttcgc aggggcctta cccaccgcct tggcgggctt
      181 cttcggtcca aaactgaaca acagatgtgt gaccttgcgc ccggtctttc gctgcgccca
      241 ctccacctgt agcgggctgt gctcgttgat ctgcgtcacg gctggatcaa gcactcgcaa
      301 cttgaagtcc ttgatcgagg gataccggcc ttccagttga aaccactttc gcagctggtc
      361 aatttctatt tcacgctggc cgatgctgtc ccattgcgtg agcagctcgt aaagcctgat
      421 cgcgtgggtg ctgcccatct tggccacgtc agccaaggcg tatttggtga actgtttggt
      481 gagttccgtc agatacggca gcatgtcttt ggtgaacctg agttctacac ggccctcacc
      541 ctcccggtag atgattgttt gcacccagcc ggtaatcatc acactcggtc ttttcccctt
      601 gccattgggc tcttgggtta accggacttc ccgccgtttc aggcgcaggg ccgcttcttt
      661 gagctggttg taggaagatt cgatagggac acccgccatc gtcgctatgt cctccgccgt
      721 cactgaatac atcacttcat cggtgacagg ctcgctcctc ttcacctggc taatactggc
      781 cagaacgatc cgctgttcct gaacactgag gcaatacgcg gcctcgacca gggcattgct
      841 tttgtaaacc attgggggtg aggccacgtt cgacattcct tgtgtataag gggacactgt
      901 atcagcgtcc cacaatacaa caaatccgtc cctttacaac aacaaatccg tcccttctta
      961 acaacaaatc cgtcccttaa tggcaacaaa tccgtccctt tttaaactct acaggccacg
     1021 gattacgtgg cctgtagacg tcctaaaagg tttaaaaggg aaaaggaaga aaagggtgga
     1081 aacgcaaaaa acgcaccact acgtggcccc gttggggccg catttgtgcc cctgaagggg
     1141 cgggggaggc gtctgggcaa tccccgtttt accagtcccc tatcgccgcc tgagagggcg
     1201 caggaagcga gtaatcaggg tatcgaggcg gattcaccct tggcgtccaa ccagcggcac
     1261 cagcggcgaa ttcacgaaat ctgcagagct gttgactcgc ttgggatttt cggaatatca
     1321 tgccgggtgg gcccgggaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca
     1381 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1441 gggtgttatg agccatattc aacgggaaac gtcttgctcc aggccgcgat taaattccaa
     1501 catggatgct gatttatatg ggtataaatg ggcacgcgat aatgtcgggc aatcaggtgc
     1561 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     1621 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     1681 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     1741 cactgcgatc cccggcaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     1801 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     1861 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     1921 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     1981 ctggaaagaa atgcataagc tcttgccatt ctcaccggat tcagtcgtca ctcatggtga
     2041 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2101 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2161 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2221 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     2281 gttgtaacac tggcagagca ttacgctgac ttcccgggaa agccgaattc cagcacactg
     2341 gcggccgtta ctagtggatc cactagtaac ggccgccagt gtgctggaat tcggcttgat
     2401 gatggcctat atgcgtctat ttatgtagga tgaaaggtag tctagtacct cctgtgatat
     2461 tatcccattc catgcggggt atcgtatgct tccttcagca ctacccttta gctgttctat
     2521 atgctgccac tcctcaattg gattagtctc atccttcaat gctatcattt cctttgatat
     2581 tggatcatat gcatagtacc gagaaactag tgcgaagtag tgatcaggta ttgctgttat
     2641 ctgatgagta tacgttgtcc tggccacggc agaagcacgc ttatcgctcc aatttcccac
     2701 aacattagtc aactccgtta ggcccttcat tgaaagaaat gaggtcatca aatgtcttcc
     2761 aatgtgagat tttgggccat tttttatagc aaagatcgga taaggcgcat ttttcttcaa
     2821 agctttattg tacgatctga ctaagttatc ttttaataat tggtattcct gtttattgct
     2881 tgaagaattg ccggtcctat ttactcgttt taggactggt tcagaattcc tcaaaaattc
     2941 atccaaatat acaagtggat cgatcctacc ccttgcgcta aagaagtata tgtgcctact
     3001 aacgcttgtc tttgtctctg tcactaaaca ctggattatt actcccagat acttattttg
     3061 gactaattta aatgatttcg gatcaacgtt cttaatatcg ctgaatcttc cacaattgat
     3121 gaaagtagct aggaagagga attggtataa agtttttgtt tttgtaaatc tcgaagtata
     3181 ctcaaacgaa tttagtattt tctcagtgat ctcccagatg ctttcaccct cacttagaag
     3241 tgctttaagc atttttttac tgtggctatt tcccttatct gcttcttccg atgattcgaa
     3301 ctgtaattgc aaactactta caatatcagt gatatcagat tgatgttttt gtccgttgta
     3361 aggaataatt gtaaattccc aagcaggaat caatttcttt aatgaggctt ccagaattgt
     3421 tgctttttgc gtcttgtatt taaactggag tgatttattg acaatatcga aactcagcga
     3481 attgcttatg atagtattat agctcatgaa tgtggctctc ttgattgctg ttccgttatg
     3541 tgtaatcatc caacataaat aggttagttc agcagcacag gatgctattt tctcacctga
     3601 aggtctttca aacctttcca caaactgacg aacaagcacc ttaggcgggg ttttacacag
     3661 gatatcgaac tggctcatgt atatctcctt gctgaatttc attacgacca gtctaaaaag
     3721 cgcctgaatt cgcgaccttc tcgttactga caggaaaatg ggccattggc aaccagggaa
     3781 agatgaacgt gatgatgttc acaatttgct gaattgtggt gatgtgatgc tcaccgcatt
     3841 tcctgaaaat tcacgctgta tcttgaaaaa tcgacgtttt ttacgtggtt ttccgtcgaa
     3901 aatttaaggt aagaacctga cctcgtgatt actatttcgc cgtgttgacg acatcaggag
     3961 gccagtatga ccgtattaca tagtgtggat ttttttccgt ctggtaacgc gtccgtggcg
     4021 atagaacccc ggctcccgca ggcggatttt cctgaacatc atcatgattt tcatgaaatt
     4081 gtgattgtcg aacatggcac gggtattcat gtgtttaatg ggcagcccta taccatcacc
     4141 ggtggcacgg tctgtttcgt acgcgatcat gatcggcatc tgtatgaaca taccgataat
     4201 ctgtgtctga ccaatgtgct gtatcgctcg ccggatcgat ttcagtttct cgccgggctg
     4261 aatcagttgc tgccacaaga gctggatggg cagtatccgt ctcactggcg cgttaaccac
     4321 agcgtattgc agcaggtgcg acagctggtt gcacagatgg aacagcagga aggggaaaat
     4381 gatttaccct cgaccgccag tcgcgagatc ttgtttatgc aattactgct cttgctgcgt
     4441 aaaagcagtt tgcaggagaa cctggaaaac agcgcatcac gtctcaactt gcttctggcc
     4501 tggctggagg accattttgc cgatgaggtg aattgggatg ccgtggcgga tcaattttct
     4561 ctttcactgc gtacgctaca tcggcagctt aagcagcaaa cgggactgac gcctcagcga
     4621 tacctgaacc gcctgcgact gatgaaagcc cgacatctgc tacgccacag cgaggccagc
     4681 gttactgaca tcgcctatcg ctgtggattc agcgacagta accacttttc gacgcttttt
     4741 cgccgagagt ttaactggtc accgcgtgat attcgccagg gacgggatgg ctttctgcaa
     4801 taacgcgaat cttctcaacg tatttgtacg ccatattgcg aataatcaac ttcgttctct
     4861 ggccgaggta gccacggtgg cgcatcagtt aaaacttctc aaagatgatt tttttgccag
     4921 cgaccagcag gcagtcgctg tggctgaccg ttatccgcaa gatgtctttg ctgaacatac
     4981 acatgatttt tgtgagctgg tgattgtctg gcgcggtaat ggcctgcatg tactcaacga
     5041 tcgcccttat cgcattaccc gtggcgatct cttttacatt catgctgacg ataaacactc
     5101 ctacgcttcc gttaacgatc tggttttgca gaatattatt tattgcccgg agcgtctgaa
     5161 gctgaatctt gactggcagg gggcgattcc gggatttaac gccagcgcag ggcaaccaca
     5221 ctggcgctta ggtagcatgg ggatggcgca ggcgcggcag gttatcggtc agcttgagca
     5281 tgaaagtagt cagcatgtgc cgtttgctaa cgaaatggct gagttgctgt tcgggcagtt
     5341 ggtgatgttg ctgaatcgcc atcgttacac cagtgattcg ttgccgccaa catccagcga
     5401 aacgttgctg gataagctga ttacccggct ggcggctagc ctgaaaagtc cctttgcgct
     5461 ggataaattt tgtgatgagg catcgtgcag tgagcgcgtt ttgcgtcagc aatttcgcca
     5521 gcagactgga atgaccatca atcaatatct gcgacaggtc agagtgtgtc atgcgcaata
     5581 tcttctccag catagccgcc tgttaatcag tgatatttcg accgaatgtg gctttgaaga
     5641 tagtaactat ttttcggtgg tgtttacccg ggaaaccggg atgacgccca gccagtggcg
     5701 tcatctcaat tcgcagaaag attaaatgca tatgatcgag ctcgaattcg taatcatggt
     5761 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5821 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5881 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5941 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6001 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6061 tacggttatc cacagaatca ggggataacg caggaaagaa catggggatt ccttaaggta
     6121 tactttccgc tgcataaccc tgcttcgggg tcattatagc gattttttcg gtatatccat
     6181 cctttttcgc acgatataca ggattttgcc aaagggttcg tgtagacttt ccttggtgta
     6241 tccaacggcg tcagccgggc aggataggtg aagtaggccc acccgcgagc gggtgttcct
     6301 tcttcactgt cccttattcg cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct
     6361 ggccgataag ctagcttatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6421 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6481 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6541 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6601 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6661 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6721 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6781 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6841 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6901 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6961 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7021 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac ga
//
