LOCUS       40924_14815        8467 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pDM310, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8467)
  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 8467)
  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 8467)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8467)
  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..8467
                     /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..4973
                     /label=TetR
                     /note="tetracycline repressor TetR"
     CDS             4995..5228
                     /label=VP16 AD
                     /note="transcriptional activation domain of herpes simplex
                     virus protein VP16 (Triezenberg et al., 1988; Cousens et 
                     al., 1989)"
     regulatory      5270..5531
                     /label=act8 terminator
                     /note="act8 terminator"
                     /regulatory_class="terminator"
     rep_origin      complement(5656..6198)
                     /direction=LEFT
                     /label=Ddp1 ori
                     /note="Ddp1 ori"
     CDS             6108..6701
                     /codon_start=1
                     /product="G5"
                     /label=G5
                     /protein_id="ACG80833.1"
                     /translation="MFNHIKDSGYYATNEEIEIFLESCTLCKEITAQTKRNSYKKRNII
                     NKLPEEEEEEEEEEEEEEEEEEQEEEVEKPTISEEEEEETPAVSEEEKEEEEQEEDKEK
                     DKEKKIEEDTETGKKKDEVMNERQDGIEISEKTNDKPTEKTKVKKNVSRVKDINLHKKG
                     NPITPKKIKNFSQQQLRLASGVSNEKRAIITLKK"
     CDS             complement(6709..7473)
                     /codon_start=1
                     /product="D5"
                     /label=D5
                     /protein_id="ACG80834.1"
                     /translation="MYSTTLLFFLFFFIVSKSSVLQNDQIEVKYFQLLIVMYQKENCVE
                     KSMTGAVIYDECNIHGRVETNSTHALFYDDIETNNSRCNNFRNLTNLIKLNECINDEFG
                     ESILYKEYNETDDGYLFRVEDSFVEITSLSMDCTKNSKTIIEKFNICSKFENVYHITNI
                     TQEKSNRFTCTDPLCHYCKNENIQNNLDFKTTKCTPKYGASDSEFLSTIYNPKLDGSNN
                     GMEKSVTQEKNISNNLKINIYLIFFLIIFLIK"
     CDS             complement(7976..8467)
                     /codon_start=1
                     /product="D1'"
                     /label=D1'
                     /protein_id="ACG80835.1"
                     /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 ttaacggcgc attagagctg cttaatgggg tcggaatcga aggtttaaca acccgtaaac
     4441 tcgcccagaa gctaggtgta gagcagccta cattgtattg gcatgtaaaa aataagcggg
     4501 ctttgctcga cgccttaccc 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 gtggggcact ttactttagg ttgcgtattg gaagaacaag
     4801 agcatcaagt cgctaaagaa gaaagggaaa cacctactac tgatagtatg ccgccattat
     4861 tacgacaagc tatcgaatta tttgatcgcc aaggtgcaga gccagccttc ttattcggcc
     4921 ttgaattgat catatgcgga ttagaaaaac aacttaaatg tgaaagtgga tcacacactc
     4981 gtagactctc aactgcccca ccaaccgatg tcagcctggg ggacgagctc cacttagacg
     5041 gcgaggacgt ggcgatggcg catgccgacg cgctagacga tttcgatctg gacatgttgg
     5101 gggacgggga ttccccgggc ccgggattta ccccccacga ctccgccccc tacggcgctc
     5161 tggatatggc cgacttcgag tttgagcaga tgtttaccga tgcccttgga attgacgagt
     5221 acggtggtta atctagtacg cgtaaattgt tcacagaaaa tgtttctaaa ttatttaata
     5281 aataataaaa aaacaaattg ttgtaataat ctaatatttt cttttttttt taattttttt
     5341 tttttaaatc ttaataatta ttaagttatt ttaatttttt tttttttttt tttttttttt
     5401 tttttttttt ttctatcaaa aaaatcaaat atatttaaaa aatttattat ttacagatac
     5461 attttgaatg gtgaagataa atatatgcat tagatgtaaa acagccaaag agtatgaaaa
     5521 tcaaaaagat agccggcctc gacatttgaa attacaatga atgtttttcg gtttttattt
     5581 tttacttttt atttgatttt tttttttttt tttttttttt tttttttttt aaaatttttt
     5641 tttttttttt ttcaaatttt ttttttttta aaaaaaaaaa aattcatttt tgtatttaaa
     5701 ttattttata aataaaaaat gatagtagta gaaattgttt tttggacaag aatcgtaaag
     5761 tcaaaatatt cagttgatga tgaaatcttt aaatatttgg ttaatagagc aaaagttgtg
     5821 tataacaatt cagattataa aacactatct caaactgatt tagagaaatg ggcgattcaa
     5881 aacactcaca gaacaataac ttcaattaaa aatcgttttt cagtcaaaaa gattggagat
     5941 gaagaaaagt tatttagaat ttcaaaaaat ggtaaaatcc atctttatac atgttcattt
     6001 taatttattt taactaattt tttaaattag gggagcttat tgttcttaat gaattagaat
     6061 tcgataattt tcatattaaa gaaggaaagc atttaagaaa atcaaaaatg tttaatcata
     6121 tcaaagacag tggttattat gcaacaaacg aagaaattga aatttttctt gaaagctgca
     6181 ctctatgtaa ggaaattact gcacaaacca aacgtaattc atataaaaaa agaaatataa
     6241 ttaataaatt accggaagaa gaagaagaag aagaagaaga agaagaagaa gaagaagaag
     6301 aagaagaaca agaagaagaa gtagaaaaac cgacaatatc ggaagaagaa gaagaagaaa
     6361 cacctgcagt atcagaagaa gaaaaagaag aggaagaaca agaagaagat aaagaaaaag
     6421 ataaagaaaa aaaaatagaa gaagatacag aaacaggaaa aaaaaaagat gaagttatga
     6481 atgaacgtca agatggtatt gaaattagtg aaaaaacaaa tgataaacca acagaaaaaa
     6541 caaaagttaa aaagaatgtt tcacgagtca aagatattaa tcttcataaa aaaggaaatc
     6601 ctattacccc aaaaaaaata aaaaatttca gtcaacaaca attacgttta gcttctggtg
     6661 tttcaaatga aaaaagagca ataattactc ttaaaaaata ataaaacttt atttaattaa
     6721 aaaaataatt aaaaagaaaa ttaaatatat attaattttt aaattatttg aaatgttttt
     6781 ttcttgagtt actgactttt ccataccgtt atttgagcca tcgagctttg gattgtaaat
     6841 tgttgataaa aattcagaat cagatgcacc atactttgga gtacactttg ttgttttaaa
     6901 atcaagattg ttttgaatgt tttcattctt acaatagtgg cacaatggat ctgtacatgt
     6961 aaatctattg gatttctctt gtgtaatgtt tgtaatatga tatacatttt caaattttga
     7021 acaaatgttg aatttttcaa taattgtttt actatttttt gtacaatcca ttgaaagaga
     7081 agtaatttca acaaagctgt cttccactct aaacaaataa ccatcatcag tttcattata
     7141 ttctttataa agaatagact ctccaaactc gtcattaata cattcattaa gtttaattaa
     7201 gtttgttaaa ttacgaaaat tgttacatct tgaattattt gtttcaatgt catcataaaa
     7261 aagcgcatga gtactatttg tttcaactct tccatgaata ttacattcat cataaattac
     7321 ggcgcctgtc attgactttt caacacagtt ttctttttga tacataacaa taagaagttg
     7381 aaaatatttt acttcaatct gatcattttg aagaacagaa gattttgata caataaaaaa
     7441 aaaaagaaaa aataataacg ttgttgagta cattatatac aatgaaaata ataaatgaac
     7501 aaaaaataat tttttattat aatatattaa accccacccc acaaccacac ctaaaaaaaa
     7561 gttaatactt ttttaaatat aattttttta atagttattt taatacgttt ttaacataaa
     7621 atatatttta attttttttt taatttttta gaaaattttt ttttcataat tattttaata
     7681 cttttttaca aaaaatattt ttaaaatttt aaaaattttt tttttataat tattttaata
     7741 gcttaattac aaaaacaatt taataatttt tttaattttt ttttcataat atttttaata
     7801 gcttttttac aaattatttt tacgaaaatt tttgttatat ttttttataa ttattttaat
     7861 acatttttaa cattaaataa ttatataatt ttttaataca ataattacag ttataaattc
     7921 atattattta aatattttta aatgagttta tttagaaatt aagtttatat tttatttaat
     7981 tagttttgac tctcagaatg tatagacact ttttagaaca ctgtgcacaa agatgaagtt
     8041 tacattcttg gttcaaatta catttagttc cgttcttact acacatggca cacttcttgt
     8101 tgattaccga atcattgaaa catggagggc aaagtttaga attaataaca acattatata
     8161 tgtttatgtt tattcctaaa caattatagc aaatattgtt ttgacatcca tctcttgtac
     8221 atgaaatata taaatttttt ccttgacaca aagaatgaca attaatacac tttacttcta
     8281 agagagaata attttctgga tttgaaaggg catctgttga agaaactaaa aagaaatcaa
     8341 ttttggaaga agttttattg acttcttctt taataaactt ttttaaatcg gcaatttcat
     8401 tggaatttat ttgggtagat tcggattgaa ttttgtaata attgctattt actttttcga
     8461 aatcgat
//
