LOCUS       40924_14810        8596 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pDM309, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8596)
  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 8596)
  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 8596)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8596)
  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..8596
                     /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..1796)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      complement(1844..2065)
                     /label=act6 promoter
                     /note="act6 promoter"
                     /regulatory_class="promoter"
     promoter        2120..2224
                     /label=AmpR promoter
     CDS             2225..3082
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3256..3844
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      4057..4340
                     /label=act15 promoter
                     /note="act15 promoter"
                     /regulatory_class="promoter"
     CDS             4353..5357
                     /label=tTA
                     /note="tetracycline-controlled transactivator, comprising a
                     fusion of the tetracycline repressor TetR with the 
                     C-terminal activation domain of herpes simplex virus VP16"
     regulatory      5399..5660
                     /label=act8 terminator
                     /note="act8 terminator"
                     /regulatory_class="terminator"
     rep_origin      complement(5785..6327)
                     /direction=LEFT
                     /label=Ddp1 ori
                     /note="Ddp1 ori"
     CDS             6237..6830
                     /codon_start=1
                     /product="G5"
                     /label=G5
                     /protein_id="ACG80851.1"
                     /translation="MFNHIKDSGYYATNEEIEIFLESCTLCKEITAQTKRNSYKKRNII
                     NKLPEEEEEEEEEEEEEEEEEEQEEEVEKPTISEEEEEETPAVSEEEKEEEEQEEDKEK
                     DKEKKIEEDTETGKKKDEVMNERQDGIEISEKTNDKPTEKTKVKKNVSRVKDINLHKKG
                     NPITPKKIKNFSQQQLRLASGVSNEKRAIITLKK"
     CDS             complement(6838..7602)
                     /codon_start=1
                     /product="D5"
                     /label=D5
                     /protein_id="ACG80852.1"
                     /translation="MYSTTLLFFLFFFIVSKSSVLQNDQIEVKYFQLLIVMYQKENCVE
                     KSMTGAVIYDECNIHGRVETNSTHALFYDDIETNNSRCNNFRNLTNLIKLNECINDEFG
                     ESILYKEYNETDDGYLFRVEDSFVEITSLSMDCTKNSKTIIEKFNICSKFENVYHITNI
                     TQEKSNRFTCTDPLCHYCKNENIQNNLDFKTTKCTPKYGASDSEFLSTIYNPKLDGSNN
                     GMEKSVTQEKNISNNLKINIYLIFFLIIFLIK"
     CDS             complement(8105..8596)
                     /codon_start=1
                     /product="D1'"
                     /label=D1'
                     /protein_id="ACG80853.1"
                     /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 ttcagaagaa ctcgtcaaga
     1021 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     1081 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     1141 tgatagcggt ccgccacacc cagccgtcca cagtcgatga atccagaaaa gcggccattt
     1201 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     1261 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     1321 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     1381 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     1441 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     1501 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     1561 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
     1621 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     1681 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     1741 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     1801 aacgatccag cttgaacatc ttcaccatcc attttggatc ttttatatta tatttattta
     1861 ttgattattt ttttgaatta attaaaaaaa aaaaaaattt cattttataa tctcagaaac
     1921 ctcaaaaaaa aaaaaataaa aaataaaaaa tataaaaaaa taaaaataaa atcccaattt
     1981 taaagcgaaa aaccacccat ggtttgaaaa tttcaatcaa tttcaaataa ctttacttaa
     2041 aaaaaaccca ttttttattt aaaaaatcta gtggatcccg ctacagggcg cgtcaggtgg
     2101 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2161 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2221 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2281 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2341 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2401 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2461 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2521 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2581 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2641 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2701 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     2761 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2821 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2881 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2941 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3001 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3061 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3121 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3181 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3241 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3301 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3361 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3421 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3481 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3541 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3601 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3661 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3721 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3781 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3841 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3901 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3961 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4021 ggaagagcgc ccaatacgca aaccgcctct gccggctaaa aaaaattttt atttattttt
     4081 atttattttg aattaaatag attacaaatt aattaatccc atcaaatctt taaaaaaaaa
     4141 tggtttaaaa aaacttgggt tggttaatta ttatttgaaa attttaaaac ccaaattaaa
     4201 aaaaaaaaat gggattcaaa aatttttttt tttttttttt tttttttttt tttttttttt
     4261 tcagattgca taaaaagatt tttttttttt ttttttctta tttcttaaaa caaataaatt
     4321 aaattaaata aaaaataaaa atcagatcca aaatgtcaag attagataaa agtaaagtga
     4381 ttaacagcgc attagagctg cttaatgagg tcggaatcga aggtttaaca acccgtaaac
     4441 tcgcccagaa gctaggtgta gagcagccta cattgtattg gcatgtaaaa aataagcggg
     4501 ctttgctcga cgccttagcc attgagatgt tagataggca ccatactcac ttttgccctt
     4561 tagaagggga aagctggcaa gattttttac gtaataacgc taaaagtttt agatgtgctt
     4621 tactaagtca tcgcgatgga gcaaaagtac atttaggtac acggcctaca gaaaaacagt
     4681 atgaaactct cgaaaatcaa ttagcctttt tatgccaaca aggtttttca ctagagaatg
     4741 cattatatgc actcagcgct gtggggcatt ttactttagg ttgcgtattg gaagatcaag
     4801 agcatcaagt cgctaaagaa gaaagggaaa cacctactac tgatagtatg ccgccattat
     4861 tacgacaagc tatcgaatta tttgatcacc aaggtgcaga gccagccttc ttattcggcc
     4921 ttgaattgat catatgcgga ttagaaaaac aacttaaatg tgaaagtggg tccgcgtaca
     4981 gccgcgcgcg tacgaaaaac aattacgggt ctaccatcga gggcctgctc gatctcccgg
     5041 acgacgacgc ccccgaagag gcggggctgg cggctccgcg cctgtccttt ctccccgcgg
     5101 gacacacgcg cagactgtcg acggcccccc cgaccgatgt cagcctgggg gacgagctcc
     5161 acttagacgg cgaggacgtg gcgatggcgc atgccgacgc gctagacgat ttcgatctgg
     5221 acatgttggg ggacggggat tccccgggtc cgggatttac cccccacgac tccgccccct
     5281 acggcgctct ggatatggcc gacttcgagt ttgagcagat gtttaccgat gcccttggaa
     5341 ttgacgagta cggtggttaa tctagtacgc gtaaattgtt cacagaaaat gtttctaaat
     5401 tatttaataa ataataaaaa aacaaattgt tgtaataatc taatattttc tttttttttt
     5461 aatttttttt ttttaaatct taataattat taagttattt taattttttt tttttttttt
     5521 tttttttttt tttttttttt tctatcaaaa aaatcaaata tatttaaaaa atttattatt
     5581 tacagataca ttttgaatgg tgaagataaa tatatgcatt agatgtaaaa cagccaaaga
     5641 gtatgaaaat caaaaagata gccggcctcg acatttgaaa ttacaatgaa tgtttttcgg
     5701 tttttatttt ttacttttta tttgattttt tttttttttt tttttttttt ttttttttta
     5761 aaattttttt tttttttttt tcaaattttt ttttttttaa aaaaaaaaaa attcattttt
     5821 gtatttaaat tattttataa ataaaaaatg atagtagtag aaattgtttt ttggacaaga
     5881 atcgtaaagt caaaatattc agttgatgat gaaatcttta aatatttggt taatagagca
     5941 aaagttgtgt ataacaattc agattataaa acactatctc aaactgattt agagaaatgg
     6001 gcgattcaaa acactcacag aacaataact tcaattaaaa atcgtttttc agtcaaaaag
     6061 attggagatg aagaaaagtt atttagaatt tcaaaaaatg gtaaaatcca tctttataca
     6121 tgttcatttt aatttatttt aactaatttt ttaaattagg ggagcttatt gttcttaatg
     6181 aattagaatt cgataatttt catattaaag aaggaaagca tttaagaaaa tcaaaaatgt
     6241 ttaatcatat caaagacagt ggttattatg caacaaacga agaaattgaa atttttcttg
     6301 aaagctgcac tctatgtaag gaaattactg cacaaaccaa acgtaattca tataaaaaaa
     6361 gaaatataat taataaatta ccggaagaag aagaagaaga agaagaagaa gaagaagaag
     6421 aagaagaaga agaagaacaa gaagaagaag tagaaaaacc gacaatatcg gaagaagaag
     6481 aagaagaaac acctgcagta tcagaagaag aaaaagaaga ggaagaacaa gaagaagata
     6541 aagaaaaaga taaagaaaaa aaaatagaag aagatacaga aacaggaaaa aaaaaagatg
     6601 aagttatgaa tgaacgtcaa gatggtattg aaattagtga aaaaacaaat gataaaccaa
     6661 cagaaaaaac aaaagttaaa aagaatgttt cacgagtcaa agatattaat cttcataaaa
     6721 aaggaaatcc tattacccca aaaaaaataa aaaatttcag tcaacaacaa ttacgtttag
     6781 cttctggtgt ttcaaatgaa aaaagagcaa taattactct taaaaaataa taaaacttta
     6841 tttaattaaa aaaataatta aaaagaaaat taaatatata ttaattttta aattatttga
     6901 aatgtttttt tcttgagtta ctgacttttc cataccgtta tttgagccat cgagctttgg
     6961 attgtaaatt gttgataaaa attcagaatc agatgcacca tactttggag tacactttgt
     7021 tgttttaaaa tcaagattgt tttgaatgtt ttcattctta caatagtggc acaatggatc
     7081 tgtacatgta aatctattgg atttctcttg tgtaatgttt gtaatatgat atacattttc
     7141 aaattttgaa caaatgttga atttttcaat aattgtttta ctattttttg tacaatccat
     7201 tgaaagagaa gtaatttcaa caaagctgtc ttccactcta aacaaataac catcatcagt
     7261 ttcattatat tctttataaa gaatagactc tccaaactcg tcattaatac attcattaag
     7321 tttaattaag tttgttaaat tacgaaaatt gttacatctt gaattatttg tttcaatgtc
     7381 atcataaaaa agcgcatgag tactatttgt ttcaactctt ccatgaatat tacattcatc
     7441 ataaattacg gcgcctgtca ttgacttttc aacacagttt tctttttgat acataacaat
     7501 aagaagttga aaatatttta cttcaatctg atcattttga agaacagaag attttgatac
     7561 aataaaaaaa aaaagaaaaa ataataacgt tgttgagtac attatataca atgaaaataa
     7621 taaatgaaca aaaaataatt ttttattata atatattaaa ccccacccca caaccacacc
     7681 taaaaaaaag ttaatacttt tttaaatata atttttttaa tagttatttt aatacgtttt
     7741 taacataaaa tatattttaa tttttttttt aattttttag aaaatttttt tttcataatt
     7801 attttaatac ttttttacaa aaaatatttt taaaatttta aaaatttttt ttttataatt
     7861 attttaatag cttaattaca aaaacaattt aataattttt ttaatttttt tttcataata
     7921 tttttaatag cttttttaca aattattttt acgaaaattt ttgttatatt tttttataat
     7981 tattttaata catttttaac attaaataat tatataattt tttaatacaa taattacagt
     8041 tataaattca tattatttaa atatttttaa atgagtttat ttagaaatta agtttatatt
     8101 ttatttaatt agttttgact ctcagaatgt atagacactt tttagaacac tgtgcacaaa
     8161 gatgaagttt acattcttgg ttcaaattac atttagttcc gttcttacta cacatggcac
     8221 acttcttgtt gattaccgaa tcattgaaac atggagggca aagtttagaa ttaataacaa
     8281 cattatatat gtttatgttt attcctaaac aattatagca aatattgttt tgacatccat
     8341 ctcttgtaca tgaaatatat aaattttttc cttgacacaa agaatgacaa ttaatacact
     8401 ttacttctaa gagagaataa ttttctggat ttgaaagggc atctgttgaa gaaactaaaa
     8461 agaaatcaat tttggaagaa gttttattga cttcttcttt aataaacttt tttaaatcgg
     8521 caatttcatt ggaatttatt tgggtagatt cggattgaat tttgtaataa ttgctattta
     8581 ctttttcgaa atcgat
//
