LOCUS       40924_46533        3728 bp DNA     circular SYN 18-DEC-2018
DEFINITION  FRT-Tp-FRT expression vector pwFRT-PCS12-Tp, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3728)
  AUTHORS   Barrett AR, Kang Y, Inamasu KS, Son MS, Vukovich JM, Hoang TT.
  TITLE     Genetic tools for allelic replacement in Burkholderia species
  JOURNAL   Appl. Environ. Microbiol. 74 (14), 4498-4508 (2008)
  PUBMED    18502918
REFERENCE   2  (bases 1 to 3728)
  AUTHORS   Barrett AR, Kang Y, Vukovich JM, Son MS, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-DEC-2007) Microbiology, University of Hawaii at Manoa,
            2538 McCarthy Mall Snyder Hall Rm 310, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 3728)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3728)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "14"; 
            pages: "4498-4508"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-DEC-2007) Microbiology, University of Hawaii at Manoa, 2538 
            McCarthy Mall Snyder Hall Rm 310, Honolulu, HI 96822, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     2289..2362
                     /note="FRT; Flp-recombination-target"
     protein_bind    complement(2306..2353)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      2393..2434
                     /label=Burkholderia cenocepacia S12 promoter
                     /note="Burkholderia cenocepacia S12 promoter"
                     /regulatory_class="enhancer"
     RBS             2445..2467
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        2565..2593
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2920..3153
                     /label=TpR
                     /note="E. coli plasmid-associated dihydrofolate reductase"
     protein_bind    complement(3220..3267)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(3334..3350)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcatgcctgc aggtcgactc taggatcccc gggtaccgag
     2281 ctcgaattag cttcaaaagc gctctgaagt tcctatactt tctagagaat aggaacttcg
     2341 gaataggaac ttcaagatcc cctgattccc tttgtcaaca gcaatggata tcagccgttg
     2401 acttagttgg tatttccgga atatcatgct gggttccgaa taattttgtt taactttaag
     2461 aaggagatat accatgtctg aaccaaaaac cgggcgcggc gcgctcttca ctggagggct
     2521 tgacgcacta gtggaacgaa atcgatgagc tcgcacgaac ccagttgaca taagcctgtt
     2581 cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
     2641 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
     2701 ttgtacagtc tagcctcggg catccaagct agctaagcgc gttacgccgt gggtcgatgt
     2761 ttgatgttat ggaacagcaa cgatgttacg cagcagggta gtcgccctaa aacaaagtta
     2821 ggcagccgtt gtgctggtgc tttctagtag ttgttgtggg gtaggcagtc agagctcgat
     2881 ttgcttgtcg ccataataga ttcacaagaa ggattcgaca tgggtcaaag tagcgatgaa
     2941 gccaacgctc ccgttgcagg gcagtttgcg cttcccctga gtgccacctt tggcttaggg
     3001 gatcgcgtac gcaagaaatc tggtgccgct tggcagggtc aagtcgtcgg ttggtattgc
     3061 acaaaactca ctcctgaagg ctatgcggtc gagtccgaat cccacccagg ctcagtgcaa
     3121 atttatcctg tggctgcact tgaacgtgtg gcctaagcga taagctcgag ctaattccat
     3181 tgagtaagtt tttaagcaca tcagcttcaa aagcgctctg aagttcctat actttctaga
     3241 gaataggaac ttcggaatag gaacttcaag atccccaatt cgagctcggt acccggggat
     3301 cctagagtcg acctgcaggc atgcaagctt ggcactggcc gtcgttttac aacgtcgtga
     3361 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     3421 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3481 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3541 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3601 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3661 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3721 acgcgcga
//
