LOCUS       40924_9361        7720 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Triparental mating vector pCD13SK-Flp-oriT, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7720)
  AUTHORS   Son MS, Nguyen DT, Kang Y, Hoang TT.
  TITLE     Engineering of FRT-lacZ fusion constructs: induction of the 
            Pseudomonas aeruginosa fadAB1 operon by medium and long chain-length
            fatty acids
  JOURNAL   Plasmid 59 (2), 111-118 (2008)
  PUBMED    18221997
REFERENCE   2  (bases 1 to 7720)
  AUTHORS   Son MS, Kang Y, Nguyen DT, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-JUL-2007) Microbiology, University of Hawaii Manoa, 
            2538 McCarthy Mall, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 7720)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7720)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2008"; volume: "59"; issue: "2"; pages: "111-118"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-JUL-2007) Microbiology, University of Hawaii Manoa, 2538 
            McCarthy Mall, Honolulu, HI 96822, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      25..385
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(678..909)
                     /label=lambda attP
                     /note="integrase from phage lambda"
     CDS             complement(1801..2589)
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      complement(3413..3841)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     3985..4001
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4011..4029
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4055..4071
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            4636..4744
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(4961..6229)
                     /codon_start=1
                     /label=FLP
                     /note="site-specific recombinase"
                     /translation="MPQFGILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     RBS             6233..6241
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             6330..7040
                     /codon_start=1
                     /label=lambda repressor (ts)
                     /note="temperature-sensitive variant of the phage lambda
                     repressor"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     promoter        complement(7329..7347)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7368..7384)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7392..7408)
                     /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(7416..7446)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7461..7482)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      7720
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 gatcagcagt tcaacctgtt gatagtacgt actaagctct catgtttcac gtactaagct
       61 ctcatgttta acgtactaag ctctcatgtt taacgaacta aaccctcatg gctaacgtac
      121 taagctctca tggctaacgt actaagctct catgtttcac gtactaagct ctcatgtttg
      181 aacaataaaa ttaatataaa tcagcaactt aaatagcctc taaggtttta agttttataa
      241 gaaaaaaaag aatatataag gcttttaaag ctagctttta aggtttaacg gttgtggaca
      301 acaagccagg gatgtaacgc actgagaagc ccttagagcc tctcaaagca attttcagtg
      361 acacaggaac acttaacggc tgacatgacg ctcagtggaa cgaaaactca cgttaaggga
      421 ttttggtcat gagacaataa ccctgataaa tgcttcaata atcggtgcat ggagccgggc
      481 cacctcgacc tgaatggaag ccggcggcac ctcgctaacg gattcaccgt ttttatcagg
      541 ctctgggagg cagaataaat gatccccaat tcagctttgc actggattgc gaggctttgt
      601 gcttctctgg agtgcgacag gtttgatgac aaaaaattag cgcaagaaga caaaaacacc
      661 ttgcgctaat gctctgttac aggtcactaa taccatctaa gtagttgatt catagtgact
      721 gcatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
      781 tattgatatt tatatcattt tacgtttctc gttcagcttt tttatactaa gttggcatta
      841 taaaaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
      901 tcattatttg atttcaattt tgtcccactc cctgcctctg tcatcacgat actgtgatgc
      961 catggtgtcc gacttatgcc cgagaagatg ttgagcaaac ttatcgctta tctgcttctc
     1021 atagagtctt gcagacaaac tgcgcaactc gtgaaaggta ggcagatcat atcgtcaatt
     1081 attacctcca cggggagagc ctgagcaaac tggcctcagg catttgagaa gcacacggtc
     1141 acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa
     1201 cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg ccattcatcc
     1261 gcttattatc acttattcag gcgtagcacc aggcgtttaa gggcaccaat aactgcctta
     1321 aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     1381 gccgacatgg aagccatcac agacggcatg atgaacctga atcgccagcg gcatcagcac
     1441 cttgtcgcct tgcgtataat aaaataaaaa aggggacctc tagggtcccc aattaattag
     1501 taatataatc tattaaaggt cattcaaaag gtcatccacc ggatcagctt agtaaagccc
     1561 tcgctagatt ttaatgcgga tgttgcgatt acttcgccaa ctattgcgat aacaagaaaa
     1621 agccagcctt tcatgatata tctcccaatt tgtgtagggc ttattatgca cgcttaaaaa
     1681 taataaaagc agacttgacc tgatagtttg gctgtgagca attatgtgct tagtgcatct
     1741 aacgcttgag ttaagccgcg ccgcgaagcg gcgtcggctt gaacgaattg ttagacatta
     1801 tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc
     1861 tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat
     1921 gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg
     1981 cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg
     2041 ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc
     2101 aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc
     2161 aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg
     2221 ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt
     2281 agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg
     2341 gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg
     2401 ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg
     2461 cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag
     2521 atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc
     2581 ttccctcatg atgtttaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
     2641 gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg
     2701 catagactgt accccaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     2761 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     2821 ttacagctta cgaaccgaac aggcttatgt ccactgggtt cgtgccttca tccgtttcca
     2881 cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gaggcatttc tgtcctggct
     2941 ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag gcattggcgg ccttgctgtt
     3001 cttctacggc aaggtgctgt gcacggatct gccctggctt caggagatcg gaagacctcg
     3061 gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa gtggttcgca tcctcggttt
     3121 tctggaaggc gagcatcgtt tgttcgccca gcttctgtat ggaacgggca tgcggatcag
     3181 tgagggtttg caactgcggg tcaaggatct ggatttcgat cacggcacga tcatcgtgcg
     3241 ggagggcaag ggctccaagg atcgggcctt gatgttaccc gagagcttgg cacccagcct
     3301 gcgcgagcag gggaattgat ccggtggatg accttttgaa tgacctttaa tagattatat
     3361 tactaattaa ttggggaccc tagaggtccc cttttttatt ttattttgtt aaaattcgcg
     3421 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3481 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3541 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3601 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3661 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3721 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3781 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3841 tcagccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     3901 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     3961 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact
     4021 cactataggg cgaattgggt accgggcccc ccctcgaggt cgacggtatc gataagcttg
     4081 atatcgaatt cctgcagccc tatggggatt ggtggcgacg actcctggag cccgtcagta
     4141 tcggcggatt tccagctgag cgccggtcgc taccattacc agttggtctg gtgtcaaaaa
     4201 taataataac cgggcaggcc atgtctgccc gtatttcgcg taaggaaatc cattatgtac
     4261 tatttaaaaa acacaaactt ttggatgttc ggtttattct ttttctttta cttttttatc
     4321 atgggagcct acttcccgtt tttcccgatt tggctacatg acatcaacca tatcagcaaa
     4381 agtgatacgg gtattatttt tgccgctatt tctctgttct cgctattatt ccaaccgctg
     4441 tttggtctgc tttctgacaa actcgggcag cgttgggtcc tgggggattc cttaaggtat
     4501 actttccgct gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc
     4561 ctttttcgca cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat
     4621 ccaacggcgt cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt
     4681 cttcactgtc ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg
     4741 gccgataagc tcctctcagg cggggggatc cccctagaga ataggaactt cggaatagga
     4801 acttcaaagc gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac
     4861 agctcactgt tcacgtcgca cctatatctg cgtgttgcct gtatatatat atacatgaga
     4921 agaacggcat agtgcgtgtt tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg
     4981 atgaaaggta gtctagtacc tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc
     5041 ttccttcagc actacccttt agctgttcta tatgctgcca ctcctcaatt ggattagtct
     5101 catccttcaa tgctatcatt tcctttgata ttggatcata tgcatagtac cgagaaacta
     5161 gtgcgaagta gtgatcaggt attgctgtta tctgatgagt atacgttgtc ctggccacgg
     5221 cagaagcacg cttatcgctc caatttccca caacattagt caactccgtt aggcccttca
     5281 ttgaaagaaa tgaggtcatc aaatgtcttc caatgtgaga ttttgggcca ttttttatag
     5341 caaagattga ataaggcgca tttttcttca aagctttatt gtacgatctg actaagttat
     5401 cttttaataa ttggtattcc tgtttattgc ttgaagaatt gccggtccta tttactcgtt
     5461 ttaggactgg ttcagaattc ctcaaaaatt catccaaata tacaagtgga tcgatcctac
     5521 cccttgcgct aaagaagtat atgtgcctac taacgcttgt ctttgtctct gtcactaaac
     5581 actggattat tactcccaga tacttatttt ggactaattt aaatgatttc ggatcaacgt
     5641 tcttaatatc gctgaatctt ccacaattga tgaaagtagc taggaagagg aattggtata
     5701 aagtttttgt ttttgtaaat ctcgaagtat actcaaacga atttagtatt ttctcagtga
     5761 tctcccagat gctttcaccc tcacttagaa gtgctttaag cattttttta ctgtggctat
     5821 ttcccttatc tgcttcttcc gatgattcga actgtaattg caaactactt acaatatcag
     5881 tgatatcaga ttgatgtttt tgtccatagt aaggaataat tgtaaattcc caagcaggaa
     5941 tcaatttctt taatgaggct tccagaattg ttgctttttg cgtcttgtat ttaaactgga
     6001 gtgatttatt gacaatatcg aaactcagcg aattgcttat gatagtatta tagctcatga
     6061 atgtggctct cttgattgct gttccgttat gtgtaatcat ccaacataaa taggttagtt
     6121 cagcagcaca taatgctatt ttctcacctg aaggtctttc aaacctttcc acaaactgac
     6181 gaacaagcac cttaggtggt gttttacata atataccaaa ttgtggcata caacctcctt
     6241 agtacatgca accattatca ccgccagagg taaaatagtc aacacgcacg gtgttagata
     6301 tttatccctt gcggtgatag atttaacgta tgagcacaaa aaagaaacca ttaacacaag
     6361 agcagcttga ggacgcacgt cgccttaaag caatttatga aaaaaagaaa aatgaacttg
     6421 gcttatccca ggaatctgtc gcagacaaga tggggatggg gcagtcaggc gttggtgctt
     6481 tatttaatgg catcaatgca ttaaatgctt ataacgccgc attgcttaca aaaattctca
     6541 aagttagcgt tgaagaattt agcccttcaa tcgccagaga aatctacgag atgtatgaag
     6601 cggttagtat gcagccgtca cttagaagtg agtatgagta ccctgttttt tctcatgttc
     6661 aggcagggat gttctcacct aagcttagaa cctttaccaa aggtgatgcg gagagatggg
     6721 taagcacaac caaaaaagcc agtgattctg cattctggct tgaggttgaa ggtaattcca
     6781 tgaccgcacc aacaggctcc aagccaagct ttcctgacgg aatgttaatt ctcgttgacc
     6841 ctgagcaggc tgttgagcca ggtgatttct gcatagccag acttgggggt gatgagttta
     6901 ccttcaagaa actgatcagg gatagcggtc aggtgttttt acaaccacta aacccacagt
     6961 acccaatgat cccatgcaat gagagttgtt ccgttgtggg gaaagttatc gctagtcagt
     7021 ggcctgaaga gacgtttggc tgatcggcaa ggtgttctgg tcggcgcata gctgataaca
     7081 attgagcaag aatcttcatc gaattagggg aattttcact cccctcagaa cataacatag
     7141 taaatggatt gaattatgaa gaatggtttt tatgcgactt accgcagcaa aaataaaggg
     7201 aaagataagc gctcaataaa cctgtctgtt ttccttaatt ctctgctggc tgataatcat
     7261 cacctgcagg ttggctccaa ttatttgtat attcataaaa tcggggtacc gagctccagc
     7321 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     7381 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     7441 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     7501 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     7561 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     7621 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7681 acagaatcag gggataacgc aggaaagaac atgtgagatc
//
