LOCUS       40924_14850        8707 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pDM359, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8707)
  AUTHORS   Veltman DM, Keizer-Gunnink I, Haastert PJ.
  TITLE     An extrachromosomal, inducible expression system for Dictyostelium 
            discoideum
  JOURNAL   Plasmid 61 (2), 119-125 (2009)
  PUBMED    19046986
REFERENCE   2  (bases 1 to 8707)
  AUTHORS   Veltman DM, Keizer I, van Haastert PJM.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUL-2008) Cell Biochemistry, Rijks Universiteit 
            Groningen, Kerklaan 30, Haren, Groningen 9751 NN, The Netherlands
REFERENCE   3  (bases 1 to 8707)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8707)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2009"; volume: "61"; issue: "2"; pages: "119-125"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-JUL-2008) Cell Biochemistry, Rijks Universiteit Groningen, 
            Kerklaan 30, Haren, Groningen 9751 NN, The Netherlands"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    28..298
                     /label=tetracycline response element
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     regulatory      473..734
                     /label=act8 terminator
                     /note="act8 terminator"
                     /regulatory_class="terminator"
     regulatory      complement(741..995)
                     /label=mhcA terminator
                     /note="mhcA terminator"
                     /regulatory_class="terminator"
     CDS             complement(1005..2039)
                     /label=HygR
                     /note="hygromycin B phosphotransferase"
     regulatory      complement(2084..2305)
                     /label=act6 promoter
                     /note="act6 promoter"
                     /regulatory_class="promoter"
     promoter        2360..2464
                     /label=AmpR promoter
     CDS             2465..3322
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3496..4084
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      4297..4580
                     /label=act15 promoter
                     /note="act15 promoter"
                     /regulatory_class="promoter"
     CDS             4593..5213
                     /label=TetR
                     /note="tetracycline repressor TetR"
     CDS             5235..5468
                     /label=VP16 AD
                     /note="transcriptional activation domain of herpes simplex
                     virus protein VP16 (Triezenberg et al., 1988; Cousens et 
                     al., 1989)"
     regulatory      5510..5771
                     /label=act8 terminator
                     /note="act8 terminator"
                     /regulatory_class="terminator"
     rep_origin      complement(5896..6438)
                     /direction=LEFT
                     /label=Ddp1 ori
                     /note="Ddp1 ori"
     CDS             6348..6941
                     /codon_start=1
                     /product="G5"
                     /label=G5
                     /protein_id="ACG80839.1"
                     /translation="MFNHIKDSGYYATNEEIEIFLESCTLCKEITAQTKRNSYKKRNII
                     NKLPEEEEEEEEEEEEEEEEEEQEEEVEKPTISEEEEEETPAVSEEEKEEEEQEEDKEK
                     DKEKKIEEDTETGKKKDEVMNERQDGIEISEKTNDKPTEKTKVKKNVSRVKDINLHKKG
                     NPITPKKIKNFSQQQLRLASGVSNEKRAIITLKK"
     CDS             complement(6949..7713)
                     /codon_start=1
                     /product="D5"
                     /label=D5
                     /protein_id="ACG80840.1"
                     /translation="MYSTTLLFFLFFFIVSKSSVLQNDQIEVKYFQLLIVMYQKENCVE
                     KSMTGAVIYDECNIHGRVETNSTHALFYDDIETNNSRCNNFRNLTNLIKLNECINDEFG
                     ESILYKEYNETDDGYLFRVEDSFVEITSLSMDCTKNSKTIIEKFNICSKFENVYHITNI
                     TQEKSNRFTCTDPLCHYCKNENIQNNLDFKTTKCTPKYGASDSEFLSTIYNPKLDGSNN
                     GMEKSVTQEKNISNNLKINIYLIFFLIIFLIK"
     CDS             complement(8216..8707)
                     /codon_start=1
                     /product="D1'"
                     /label=D1'
                     /protein_id="ACG80841.1"
                     /translation="MDFEKVNSNYYKIQSESTQINSNEIADLKKFIKEEVNKTSSKIDF
                     FLVSSTDALSNPENYSLLEVKCINCHSLCQGKNLYISCTRDGCQNNICYNCLGININIY
                     NVVINSKLCPPCFNDSVINKKCAMCSKNGTKCNLNQECKLHLCAQCSKKCLYILRVKTN
                     "
ORIGIN
        1 aagcttccgg gggatcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
       61 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
      121 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
      181 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
      241 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
      301 agtgaaagtc gagctcggta cccaattttt tttttttttt tttttttttt tttttttttt
      361 ttttcagatt gcataaaaag attttttttt tttttttttc ttatttctta aaacaaataa
      421 attaaattaa ataaaaaata aaaatcagat ctagtactag ttaaataaat aaattattta
      481 ataaataata aaaaaacaaa ttgttgtaat aatctaatat tttctttttt ttttaatttt
      541 ttttttttaa atcttaataa ttattaagtt attttaattt tttttttttt tttttttttt
      601 tttttttttt tttttctatc aaaaaaatca aatatattta aaaaatttat tatttacaga
      661 tacattttga atggtgaaga taaatatatg cattagatgt aaaacagcca aagagtatga
      721 aaatcaaaaa gatactcgag attttattta atatactaaa taataaaaaa gttaaaaaat
      781 gatcattgga taaatttttt ataattataa ataaagataa taattttttt tttaacaaaa
      841 ctaaaaataa aaataataaa ataattgtta aaataggttt tttttttttt tttttttttt
      901 taataaatgg tatttattaa tttatttgtt gtgtgtgttt ttttttttat aatatttttt
      961 tttttagcat tgaattaaga agaaatcaaa ttgattctag tttagttagc ctcccccatc
     1021 tcccgatccg gacgagtgct ggggcgtcgg tttccactat cggcgagtac ttctacacag
     1081 ccatcggtcc agacggccgc gcttctgcgg gcgatttgtg tacgcccgac agtcccggct
     1141 ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc gaaattgccg
     1201 tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga gcatatacgc ccggagccgc
     1261 ggcgatcctg caagctccgg atgcctccgc tcgaagtagc gcgtctgctg ctccatacaa
     1321 gccaaccacg gcctccagaa gaagatgttg gcgacctcgt attgggaatc cccgaacatc
     1381 gcctcgctcc agtcaatgac cgctgttatg cggccattgt ccgtcaggac attgttggag
     1441 ccgaaatccg cgtgcacgag gtgccggact tcggggcagt cctcggccca aagcatcagc
     1501 tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt ttgccagtga
     1561 tacacatggg gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta ttgaccgatt
     1621 ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa gatcggccgc agcgatcgca
     1681 tccatggcct ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg caggtcttgc
     1741 aacgtgacac cctgtgcacg gcgggagatg caataggtca ggctctcgct aaattcccca
     1801 atgtcaagca cttccggaat cgggagcgcg gccgatgcaa agtgccgata aacataacga
     1861 tctttgtaga aaccatcggc gcagctattt acccgcagga catatccacg ccctcctaca
     1921 tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc ggagacgctg
     1981 tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg ctttttcatt
     2041 aggtcttgtt gagaaatgtt aaattgatcc attttggatc ttttatatta tatttattta
     2101 ttgattattt ttttgaatta attaaaaaaa aaaaaaattt cattttataa tctcagaaac
     2161 ctcaaaaaaa aaaaaataaa aaataaaaaa tataaaaaaa taaaaataaa atcccaattt
     2221 taaagcgaaa aaccacccat ggtttgaaaa tttcaatcaa tttcaaataa ctttacttaa
     2281 aaaaaaccca ttttttattt aaaaaatcta gtggatcccg ctacagggcg cgtcaggtgg
     2341 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2401 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2461 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2521 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2581 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2641 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2701 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2761 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2821 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2881 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2941 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3001 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3061 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3121 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3181 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3241 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3301 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3361 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3421 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3481 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3541 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3601 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3661 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3721 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3781 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3841 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3901 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3961 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4021 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4081 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4141 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4201 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4261 ggaagagcgc ccaatacgca aaccgcctct gccggctaaa aaaaattttt atttattttt
     4321 atttattttg aattaaatag attacaaatt aattaatccc atcaaatctt taaaaaaaaa
     4381 tggtttaaaa aaacttgggt tggttaatta ttatttgaaa attttaaaac ccaaattaaa
     4441 aaaaaaaaat gggattcaaa aatttttttt tttttttttt tttttttttt tttttttttt
     4501 tcagattgca taaaaagatt tttttttttt ttttttctta tttcttaaaa caaataaatt
     4561 aaattaaata aaaaataaaa atcagatcca aaatgtcaag attagataaa agtaaagtga
     4621 ttaacggcgc attagagctg cttaatgggg tcggaatcga aggtttaaca acccgtaaac
     4681 tcgcccagaa gctaggtgta gagcagccta cattgtattg gcatgtaaaa aataagcggg
     4741 ctttgctcga cgccttaccc attgagatgt tagataggca ccatactcac ttttgccctt
     4801 tagaagggga aagctggcaa gattttttac gtaataacgc taaaagtttt agatgtgctt
     4861 tactaagtca tcgcgatgga gcaaaagtac atttaggtac acggcctaca gaaaaacagt
     4921 atgaaactct cgaaaatcaa ttagcctttt tatgccaaca aggtttttca ctagagaatg
     4981 cattatatgc actcagcgct gtggggcact ttactttagg ttgcgtattg gaagaacaag
     5041 agcatcaagt cgctaaagaa gaaagggaaa cacctactac tgatagtatg ccgccattat
     5101 tacgacaagc tatcgaatta tttgatcgcc aaggtgcaga gccagccttc ttattcggcc
     5161 ttgaattgat catatgcgga ttagaaaaac aacttaaatg tgaaagtgga tcacacactc
     5221 gtagactctc aactgcccca ccaaccgatg tcagcctggg ggacgagctc cacttagacg
     5281 gcgaggacgt ggcgatggcg catgccgacg cgctagacga tttcgatctg gacatgttgg
     5341 gggacgggga ttccccgggc ccgggattta ccccccacga ctccgccccc tacggcgctc
     5401 tggatatggc cgacttcgag tttgagcaga tgtttaccga tgcccttgga attgacgagt
     5461 acggtggtta atctagtacg cgtaaattgt tcacagaaaa tgtttctaaa ttatttaata
     5521 aataataaaa aaacaaattg ttgtaataat ctaatatttt cttttttttt taattttttt
     5581 tttttaaatc ttaataatta ttaagttatt ttaatttttt tttttttttt tttttttttt
     5641 tttttttttt ttctatcaaa aaaatcaaat atatttaaaa aatttattat ttacagatac
     5701 attttgaatg gtgaagataa atatatgcat tagatgtaaa acagccaaag agtatgaaaa
     5761 tcaaaaagat agccggcctc gacatttgaa attacaatga atgtttttcg gtttttattt
     5821 tttacttttt atttgatttt tttttttttt tttttttttt tttttttttt aaaatttttt
     5881 tttttttttt ttcaaatttt ttttttttta aaaaaaaaaa aattcatttt tgtatttaaa
     5941 ttattttata aataaaaaat gatagtagta gaaattgttt tttggacaag aatcgtaaag
     6001 tcaaaatatt cagttgatga tgaaatcttt aaatatttgg ttaatagagc aaaagttgtg
     6061 tataacaatt cagattataa aacactatct caaactgatt tagagaaatg ggcgattcaa
     6121 aacactcaca gaacaataac ttcaattaaa aatcgttttt cagtcaaaaa gattggagat
     6181 gaagaaaagt tatttagaat ttcaaaaaat ggtaaaatcc atctttatac atgttcattt
     6241 taatttattt taactaattt tttaaattag gggagcttat tgttcttaat gaattagaat
     6301 tcgataattt tcatattaaa gaaggaaagc atttaagaaa atcaaaaatg tttaatcata
     6361 tcaaagacag tggttattat gcaacaaacg aagaaattga aatttttctt gaaagctgca
     6421 ctctatgtaa ggaaattact gcacaaacca aacgtaattc atataaaaaa agaaatataa
     6481 ttaataaatt accggaagaa gaagaagaag aagaagaaga agaagaagaa gaagaagaag
     6541 aagaagaaca agaagaagaa gtagaaaaac cgacaatatc ggaagaagaa gaagaagaaa
     6601 cacctgcagt atcagaagaa gaaaaagaag aggaagaaca agaagaagat aaagaaaaag
     6661 ataaagaaaa aaaaatagaa gaagatacag aaacaggaaa aaaaaaagat gaagttatga
     6721 atgaacgtca agatggtatt gaaattagtg aaaaaacaaa tgataaacca acagaaaaaa
     6781 caaaagttaa aaagaatgtt tcacgagtca aagatattaa tcttcataaa aaaggaaatc
     6841 ctattacccc aaaaaaaata aaaaatttca gtcaacaaca attacgttta gcttctggtg
     6901 tttcaaatga aaaaagagca ataattactc ttaaaaaata ataaaacttt atttaattaa
     6961 aaaaataatt aaaaagaaaa ttaaatatat attaattttt aaattatttg aaatgttttt
     7021 ttcttgagtt actgactttt ccataccgtt atttgagcca tcgagctttg gattgtaaat
     7081 tgttgataaa aattcagaat cagatgcacc atactttgga gtacactttg ttgttttaaa
     7141 atcaagattg ttttgaatgt tttcattctt acaatagtgg cacaatggat ctgtacatgt
     7201 aaatctattg gatttctctt gtgtaatgtt tgtaatatga tatacatttt caaattttga
     7261 acaaatgttg aatttttcaa taattgtttt actatttttt gtacaatcca ttgaaagaga
     7321 agtaatttca acaaagctgt cttccactct aaacaaataa ccatcatcag tttcattata
     7381 ttctttataa agaatagact ctccaaactc gtcattaata cattcattaa gtttaattaa
     7441 gtttgttaaa ttacgaaaat tgttacatct tgaattattt gtttcaatgt catcataaaa
     7501 aagcgcatga gtactatttg tttcaactct tccatgaata ttacattcat cataaattac
     7561 ggcgcctgtc attgactttt caacacagtt ttctttttga tacataacaa taagaagttg
     7621 aaaatatttt acttcaatct gatcattttg aagaacagaa gattttgata caataaaaaa
     7681 aaaaagaaaa aataataacg ttgttgagta cattatatac aatgaaaata ataaatgaac
     7741 aaaaaataat tttttattat aatatattaa accccacccc acaaccacac ctaaaaaaaa
     7801 gttaatactt ttttaaatat aattttttta atagttattt taatacgttt ttaacataaa
     7861 atatatttta attttttttt taatttttta gaaaattttt ttttcataat tattttaata
     7921 cttttttaca aaaaatattt ttaaaatttt aaaaattttt tttttataat tattttaata
     7981 gcttaattac aaaaacaatt taataatttt tttaattttt ttttcataat atttttaata
     8041 gcttttttac aaattatttt tacgaaaatt tttgttatat ttttttataa ttattttaat
     8101 acatttttaa cattaaataa ttatataatt ttttaataca ataattacag ttataaattc
     8161 atattattta aatattttta aatgagttta tttagaaatt aagtttatat tttatttaat
     8221 tagttttgac tctcagaatg tatagacact ttttagaaca ctgtgcacaa agatgaagtt
     8281 tacattcttg gttcaaatta catttagttc cgttcttact acacatggca cacttcttgt
     8341 tgattaccga atcattgaaa catggagggc aaagtttaga attaataaca acattatata
     8401 tgtttatgtt tattcctaaa caattatagc aaatattgtt ttgacatcca tctcttgtac
     8461 atgaaatata taaatttttt ccttgacaca aagaatgaca attaatacac tttacttcta
     8521 agagagaata attttctgga tttgaaaggg catctgttga agaaactaaa aagaaatcaa
     8581 ttttggaaga agttttattg acttcttctt taataaactt ttttaaatcg gcaatttcat
     8641 tggaatttat ttgggtagat tcggattgaa ttttgtaata attgctattt actttttcga
     8701 aatcgat
//
