LOCUS       40924_13395        6844 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Site-specific excision vector pCRE5, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6844)
  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 6844)
  AUTHORS   Choi K-H., Schweizer HP.
  TITLE     Genetic tools for Pseudomonas and its related bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6844)
  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 6844)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6844)
  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            "Unpublished"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..6844
                     /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(2415..3443)
                     /label=Cre
                     /note="site-specific recombinase"
     promoter        complement(3475..3593)
                     /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             3739..4572
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
     CDS             4649..5494
                     /label=rhaR
                     /note="transcriptional activator of rhaR and rhaS"
     primer_bind     complement(5531..5547)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5555..5571)
                     /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(5579..5609)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5624..5645)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     oriT            6029..6137
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(6209..6797)
                     /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 ctagtgatag atctgcaggt cgactatcaa ctaattatag caatcattta
     2401 cgcgttaatg gctaatcgcc atcttccagc aggcgcacca ttgcccctgt ttcactatcc
     2461 aggttacgga tatagttcat gacaatattt acattggtcc agccaccagc ttgcatgatc
     2521 tccggtattg aaactccagc gcgggccata tctcgcgcgg ctccgacacg ggcactgtgt
     2581 ccagaccagg ccaggtatct ctgaccagag tcatccttag cgccgtaaat caatcgatga
     2641 gttgcttcaa aaatcccttc cagggcgcga gttgatagct ggctggtggc agatggcgcg
     2701 gcaacaccat tttttctgac ccggcaaaac aggtagttat tcggatcatc agctacacca
     2761 gagacggaaa tccatcgctc gaccagttta gttaccccca ggctaagtgc cttctctaca
     2821 cctgcggtgc taaccagcgt tttcgttctg ccaatatgga ttaacattct cccaccgtca
     2881 gtacgtgaga tatctttaac cctgatcctg gcaatttcgg ctatacgtaa cagggtgtta
     2941 taagcaatcc ccagaaatgc cagattacgt atatcctggc agcgatcgct attttccatg
     3001 agtgaacgaa cctggtcgaa atcagtgcgt tcgaacgcta gagcctgttt tgcacgttca
     3061 ccggcatcaa cgttttcttt tcggatccgc cgcataacca gtgaaacagc attgctgtca
     3121 cttggtcgtg gcagcccgga ccgacgatga agcatgttta gctggcccaa atgttgctgg
     3181 atagttttta ctgccagacc gcgcgcctga agatatagaa gataatcgcg aacatcttca
     3241 ggttctgcgg gaaaccattt ccggttattc aacttgcacc atgccgccca cgaccggcaa
     3301 acggacagaa gcattttcca ggtatgctca gaaaacgcct ggcgatccct gaacatgtcc
     3361 atcaggttct tgcgaacctc atcactcgtt gcatcgaccg gtaatgcagg caaattttgg
     3421 tgtacggtca gtaaattgga catttaacac tcagataatg gtgctgaatt tcattacgac
     3481 cagtctaaaa agcgcctgaa ttcgcgacct tctcgttact gacaggaaaa tgggccattg
     3541 gcaaccaggg aaagatgaac gtgatgatgt tcacaatttg ctgaattgtg gtgatgtgat
     3601 gctcaccgca tttcctgaaa attcacgctg tatcttgaaa aatcgacgtt ttttacgtgg
     3661 ttttccgtcg aaaatttaag gtaagaacct gacctcgtga ttactatttc gccgtgttga
     3721 cgacatcagg aggccagtat gaccgtatta catagtgtgg atttttttcc gtctggtaac
     3781 gcgtccgtgg cgatagaacc ccggctcccg caggcggatt ttcctgaaca tcatcatgat
     3841 tttcatgaaa ttgtgattgt cgaacatggc acgggtattc atgtgtttaa tgggcagccc
     3901 tataccatca ccggtggcac ggtctgtttc gtacgcgatc atgatcggca tctgtatgaa
     3961 cataccgata atctgtgtct gaccaatgtg ctgtatcgct cgccggatcg atttcagttt
     4021 ctcgccgggc tgaatcagtt gctgccacaa gagctggatg ggcagtatcc gtctcactgg
     4081 cgcgttaacc acagcgtatt gcagcaggtg cgacagctgg ttgcacagat ggaacagcag
     4141 gaaggggaaa atgatttacc ctcgaccgcc agtcgcgaga tcttgtttat gcaattactg
     4201 ctcttgctgc gtaaaagcag tttgcaggag aacctggaaa acagcgcatc acgtctcaac
     4261 ttgcttctgg cctggctgga ggaccatttt gccgatgagg tgaattggga tgccgtggcg
     4321 gatcaatttt ctctttcact gcgtacgcta catcggcagc ttaagcagca aacgggactg
     4381 acgcctcagc gatacctgaa ccgcctgcga ctgatgaaag cccgacatct gctacgccac
     4441 agcgaggcca gcgttactga catcgcctat cgctgtggat tcagcgacag taaccacttt
     4501 tcgacgcttt ttcgccgaga gtttaactgg tcaccgcgtg atattcgcca gggacgggat
     4561 ggctttctgc aataacgcga atcttctcaa cgtatttgta cgccatattg cgaataatca
     4621 acttcgttct ctggccgagg tagccacggt ggcgcatcag ttaaaacttc tcaaagatga
     4681 tttttttgcc agcgaccagc aggcagtcgc tgtggctgac cgttatccgc aagatgtctt
     4741 tgctgaacat acacatgatt tttgtgagct ggtgattgtc tggcgcggta atggcctgca
     4801 tgtactcaac gatcgccctt atcgcattac ccgtggcgat ctcttttaca ttcatgctga
     4861 cgataaacac tcctacgctt ccgttaacga tctggttttg cagaatatta tttattgccc
     4921 ggagcgtctg aagctgaatc ttgactggca gggggcgatt ccgggattta acgccagcgc
     4981 agggcaacca cactggcgct taggtagcat ggggatggcg caggcgcggc aggttatcgg
     5041 tcagcttgag catgaaagta gtcagcatgt gccgtttgct aacgaaatgg ctgagttgct
     5101 gttcgggcag ttggtgatgt tgctgaatcg ccatcgttac accagtgatt cgttgccgcc
     5161 aacatccagc gaaacgttgc tggataagct gattacccgg ctggcggcta gcctgaaaag
     5221 tccctttgcg ctggataaat tttgtgatga ggcatcgtgc agtgagcgcg ttttgcgtca
     5281 gcaatttcgc cagcagactg gaatgaccat caatcaatat ctgcgacagg tcagagtgtg
     5341 tcatgcgcaa tatcttctcc agcatagccg cctgttaatc agtgatattt cgaccgaatg
     5401 tggctttgaa gatagtaact atttttcggt ggtgtttacc cgggaaaccg ggatgacgcc
     5461 cagccagtgg cgtcatctca attcgcagaa agattaaatg catatgatcg agctcgaatt
     5521 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5581 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5641 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5701 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5761 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5821 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgggga
     5881 ttccttaagg tatactttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt
     5941 cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact
     6001 ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga
     6061 gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg
     6121 ctctgcgagg ctggccgata agctagctta tgtgagcaaa aggccagcaa aaggccagga
     6181 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6241 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6301 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6361 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6421 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6481 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6541 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6601 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     6661 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6721 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6781 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6841 acga
//
