LOCUS       V003268                 9593 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003268
VERSION     V003268
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9593)
  AUTHORS   Goldfless SJ, Wagner JC, Niles JC.
  TITLE     Versatile control of Plasmodium falciparum gene expression with an
            inducible protein-RNA interaction
  JOURNAL   Nat Commun 5, 5329 (2014)
   PUBMED   25370483
REFERENCE   2  (bases 1 to 9593)
  AUTHORS   Goldfless SJ, Niles JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-AUG-2014) Biological Engineering, MIT, 77
            Massachusetts Avenue, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 9593)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9593)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun
            5, 5329 (2014)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (10-AUG-2014) Biological Engineering, MIT, 77 Massachusetts Avenue,
            Cambridge, MA 02139, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      9..428
                     /note="native upstream sequence from P. falciparum CRT
                     gene"
                     /regulatory_class="promoter"
     misc_feature    435..503
                     /label="TetR aptamer"
                     /note="TetR aptamer"
     CDS             504..1775
                     /gene="CG10"
                     /label="Chloroquine resistance transporter"
                     /note="Chloroquine resistance transporter from Plasmodium
                     falciparum (isolate 7G8). Accession#: W7FI62"
     regulatory      1787..2490
                     /label="derived from P. berghei CRT gene"
                     /note="derived from P. berghei CRT gene"
                     /regulatory_class="terminator"
     regulatory      2510..3499
                     /note="promoter and 5'UTR from P. falciparum calmodulin
                     gene"
                     /regulatory_class="promoter"
     CDS             3512..4072
                     /gene="DHFR"
                     /label="Dihydrofolate reductase"
                     /note="Dihydrofolate reductase from Homo sapiens.
                     Accession#: P00374"
     CDS             4088..4141
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             4142..4759
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     CDS             4772..4825
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label="T2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4826..5377
                     /label="hGLuc"
                     /note="secreted Gaussia luciferase"
     regulatory      5435..6006
                     /label="derived from P. falciparum HRPII gene"
                     /note="derived from P. falciparum HRPII gene"
                     /regulatory_class="terminator"
     regulatory      6013..7062
                     /label="derived from P. falciparum CRT gene"
                     /note="derived from P. falciparum CRT gene"
                     /regulatory_class="terminator"
     promoter        7425..7529
                     /label="AmpR promoter"
     CDS             7530..8387
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      8561..9149
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    9437..9458
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9473..9503
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    9511..9527
                     /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     9535..9551
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        9569..9587
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gcggccgctt ctatatattt gtggttccat ttttgtatat tatatataat atatttatat
       61 atatattgat atgtcaatat ttgtataaca catgaagttt ttgttttttt tttttttttt
      121 ttttaatgga gaatatttaa taatatatga gaaaaatttt atataaatat atatatatat
      181 atacatatat atatatatat atatgtatat atatttatat atacatgtat gttttttaaa
      241 aagttaaata attctataga ttattttcat tgtcttccac atatatgaca taaatatttt
      301 aaaatcgaca ttccgatata ttatattttt agactataat atccgttaat aataaataca
      361 cgcagtcata ttatttatta tacattcatt tattattttg ttttttttaa tttcttacat
      421 ataacaaaac cggtatccag gcagagaaag gtcgatacgg acggaatgtg atggcctgga
      481 taaaacacaa aactcgagaa catatgaaat ttgcatcaaa gaagaacaac caaaagaatt
      541 catcaaaaaa cgacgagagg tacagagagt tggacaacct tgtacaagag ggtaatggct
      601 cacgcctagg tggaggttct tgtcttggta aatgtgctca tgtgtttaaa cttattttta
      661 aagagattaa ggataatatt tttatttata ttttaagtat tatttattta agtgtatgtg
      721 taattgaaac aatttttgct aaaagaactt taaacaaaat tggtaactat agttttgtaa
      781 catccgaaac tcacaacttt atttgtatga ttatgttctt tattgtttat tccttatttg
      841 gaaataaaaa gggaaattca aaagaacgac accgaagctt taatttacaa ttttttgcta
      901 tatccatgtt agatgcctgt tcagtcattt tggccttcat aggtcttaca agaactactg
      961 gaaatatcca atcatttgtt cttcaattaa gtattcctat taatatgttc ttctgctttt
     1021 taatattaag atatagatat cacttataca attatctcgg agcagttatt attgttgtaa
     1081 caatagctct tgtagaaatg aaattatctt ttgaaacaca agaagaaaat tctatcatat
     1141 ttaatcttgt cttaattagt tccttaattc ctgtatgctt ttcaaacatg acaagggaaa
     1201 tagtttttaa aaaatataag attgacattt taagattaaa tgctatggta tcctttttcc
     1261 aattgttcac ttcttgtctt atattacctg tatacaccct tccattttta aaagaacttc
     1321 atttaccata taatgaaata tggacaaata taaaaaatgg tttcgcatgt ttattcttgg
     1381 gaagaaacac agtcgtagag aattgtggtc ttggtatggc taagttatgt gatgattgtg
     1441 acggagcatg gaaaaccttc gcattgtttt ccttctttag catttgtgat aatttaataa
     1501 ccagctatat tatcgacaaa ttttctacca tgacatatac tattgttagt tgtatacaag
     1561 gtccagcaac agcaattgct tattacttta aattcttagc cggtgatgtt gtaatagaac
     1621 caagattatt agatttcgta actttgtttg gctacctatt tggttctata atttaccgtg
     1681 taggaaatat tatcttagaa agaaaaaaaa tgagaaatga agaaaatgaa gattccgaag
     1741 gagaattaac caacgtcgat tcaattatta cacaataaat ccatggaaat gatttttgta
     1801 aatgtcacat aaagttataa agaagtatgt taacaattaa atatatgtat tatttttttt
     1861 ggattgtaat actttatttt tattcataaa tatttatatc ataaaattta tgtttttttt
     1921 ttttatgaaa atcatgtgtt acattttaaa tattcaaata catgaaatta atacatattt
     1981 tgtaaattat aacttatact gatgttctgc tattatatat aaattatgac cgtttgttcc
     2041 ttttgtgaat ttttcatttt ttaaaaaata ttttatatga attaatttat tttgttatat
     2101 gcctttatat tattcattaa aaaatcagat taccaaagat tttttaactt taatcatctt
     2161 tcttaagaaa aaggaaataa aataaaaaag tttttttaaa tgtgcacaca aggttcttaa
     2221 attttttttt ttttttcgga tatatataat gatgtatgtg ggcatacaaa aagacttaaa
     2281 aaatataaat gtagagaaat tattatttta tttttttata ttttcttttg tcttatatcc
     2341 ttatttgcct atgtatagag agcatgctcg ttgtttagaa gacattacaa ttatattttt
     2401 ttcttgtttt attattttaa taaaatataa tcaagatttt aactcctaat taaaaaatat
     2461 aagaatcctt atgctaagat ttttgaaata actagtccga agatctgtct taagcatttt
     2521 gtaaaaaaaa ttaaaatata tttatataat attattttat tttattatat attatattat
     2581 ttttattttt atttttattt ttttttctct acaaatttta tctattggtt tattataaaa
     2641 atatctattt ctaataataa ataattaaga tatcaattta tagaaacaaa atatatactt
     2701 gtataatttt atttttttat ataaatcatt acatatataa ttatacaata ttttttctaa
     2761 gagataatta tatattaata tatataaaaa aaggtgtttt tttttttttt ttttattttt
     2821 atttttattt tatggtaata ttttattttc cttattttat aaattatatt agtttatatg
     2881 tgattaattt tatatattat caatttatat atttttaaat gcttacttaa ttatcttttt
     2941 tttttttttt tttttttttt cccctctttt tatattaatt tatttttgaa aaaattgata
     3001 tatatatata tatataatat atatatatac atgtagtagt attaaacaat gtataatata
     3061 tataaataat atatttatat atttcatttc aattttaatt ttttttggtt tttttttttt
     3121 ttctttttgt catatttaaa aaaaattata ttcatataag ttatgcattt tttataaaca
     3181 ttattcaata tatgtataat ataatatata tatatatatt aatgtattat tccaatgtgc
     3241 atgataaaag aaaaaaataa tatttataaa aaaaaagaaa aataaaacaa aaaaagaaaa
     3301 aaaaaaaaaa aaaaaaaaaa atacaaaaat aaataatata atttataatt atatattctt
     3361 gtcacaataa aaatatatat atatatatat atatttataa tatgtatatt ttaaactaga
     3421 aaaggaataa ctaatatttt atttattatc attcaagatt tatattttat aataataaat
     3481 acctaataga aatatatcag gcgcgccaaa aatgcatggt tcgctaaact gcatcgtcgc
     3541 tgtgtcccag aacatgggca tcggcaagaa cggggacctg ccctggccac cgctcaggaa
     3601 cgaatttaga tatttccaga gaatgaccac aacctcttca gtagaaggta aacagaatct
     3661 ggtgattatg ggtaagaaga cctggttctc cattcctgag aagaatcgac ctttaaaggg
     3721 tagaattaat ttagttctca gcagagaact caaggaacct ccacaaggag ctcattttct
     3781 ttccagaagt ctagatgatg ccttaaaact tactgaacaa ccagaattag caaataaagt
     3841 agacatggtc tggatagttg gtggcagttc tgtttataag gaagccatga atcacccagg
     3901 ccatcttaaa ctatttgtga caaggatcat gcaagacttt gaaagtgaca cgttttttcc
     3961 agaaattgat ttggagaaat ataaacttct gccggaatac ccaggtgttc tctctgatgt
     4021 ccaggaggag aaaggcatta agtacaaatt tgaagtatat gagaagaatg atgccggcgg
     4081 aagtggagaa ggaagaggaa gtttattaac atgtggagat gtagaagaaa atccaggacc
     4141 atctagatta gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg
     4201 aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt
     4261 gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga
     4321 taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa
     4381 taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt
     4441 aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg
     4501 ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg ggcattttac
     4561 tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc
     4621 tactactgat agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg
     4681 tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact
     4741 taaatgtgaa agtgggtctc ccgggagtgg tgaaggtaga ggatcattat taacatgtgg
     4801 tgatgttgaa gaaaatccag gtccaggagt caaagttctg tttgccctga tctgcatcgc
     4861 tgtggccgag gccaagccca ccgagaacaa cgaagacttc aacatcgtgg ccgtggccag
     4921 caacttcgcg accacggatc tcgatgctga ccgcgggaag ttgcccggca agaagctgcc
     4981 gctggaggtg ctcaaagaga tggaagccaa tgcccggaaa gctggctgca ccaggggctg
     5041 tctgatctgc ctgtcccaca tcaagtgcac gcccaagatg aagaagttca tcccaggacg
     5101 ctgccacacc tacgaaggcg acaaagagtc cgcacagggc ggcataggcg aggcgatcgt
     5161 cgacattcct gagattcctg ggttcaagga cttggagccc atggagcagt tcatcgcaca
     5221 ggtcgatctg tgtgtggact gcacaactgg ctgcctcaaa gggcttgcca acgtgcagtg
     5281 ttctgacctg ctcaagaagt ggctgccgca acgctgtgcg acctttgcca gcaagatcca
     5341 gggccaggtg gacaagatca agggggccgg tggtgacagt gatgaattat aagctagcta
     5401 agtacaaatt tgaagtatat gagaagaatg attaagctta tttaataata gattaaaaat
     5461 attataaaaa taaaaacata aacacagaaa ttacaaaaaa aatacatatg aatttttttt
     5521 ttgtaatctt ccttataaat atagaataat gaatcatata aaacatatca ttattcattt
     5581 atttacattt aaaattattg tttcagtatc tttaatttat tatgtatata taaaaataac
     5641 ttacaatttt attaataaac aatatatgtt tattaattca tgttttgtaa tttatgggat
     5701 agcgattttt tttactgtct gtatttttct tttttaatta tgttttaatt gtattttatt
     5761 tttattattg ttctttttat agtattattt taaaacaaaa tgtattttct aagaacttat
     5821 aataataata aatataaatt ttaataaaaa ttatatttat cttttacaat atgaacataa
     5881 agtacaacat taatatatag cttttaatat ttttattcct aatcatgtaa atcttaaatt
     5941 tttcttttta aacatatgtt aaatatttat ttctcattat atataagaac atatttatta
     6001 aatctagaat tcaattatat catttatttt tttatatttt ttatattttt tttatttttc
     6061 atgttttttt ctttattttt tttttttttt ttttcctttt tttttttaat aaatcatttt
     6121 cctaatattt atatttgcta tcgcattaca ctttataagg aataatgttg aaggtataga
     6181 gatctcttta ttattattaa gatgtaaatt ttttatgaaa ttttgtactt ttttagtatt
     6241 tatattatat acacataaat atatatatat ttatttattg atatttgaaa aaagaataag
     6301 gaaataaaaa aagggaaaaa aaaaaaagtt aaaagtatat atatatatat atatatatat
     6361 atttatttat ttatatatat tattatgtat gtttataatt tatatatata tgtcataaaa
     6421 agaaatataa ttatatatat atatattata ttatatatat atataaattt ttttaaatta
     6481 taaactattg ttattgttaa atttaatttt taattcatta aaaatcagat taccaaagat
     6541 tttttccatt cttcaatcat gataatagaa atgagaagaa gcaaaaaaag aaaaaaaaaa
     6601 aaaattcata ctgtgtcaaa ggagcaaaca attttgtcca attgacaaat atgtatatat
     6661 ataaaatata tatatatata tatatatata tatatatata tatatatatt tattgattta
     6721 ttccttttct tttttttttc ccttttggtg ttttgataaa taggtaaaaa ataaaatata
     6781 taaaaattat ctctaacatt cgtaaagatt tattttattt tttaattttt tctattttat
     6841 caagtttgta aattatcatt tcattaatga aataataaga atacaaaatg ttttgtattt
     6901 gttcttgtta agaagatttt acaattatat tttttttttt aatttatttt ataaaatata
     6961 attaagatta tatcttctaa aagaaaaaaa aaaaagaaaa aagaaatatt ttgtataatg
     7021 aaatttgaaa tatctttcgc cacttatttc aagtttcagt tatcgataac tccatggaac
     7081 tcctaggctg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     7141 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     7201 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     7261 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     7321 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     7381 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     7441 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     7501 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     7561 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     7621 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     7681 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     7741 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     7801 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     7861 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     7921 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     7981 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     8041 taccaaacga cgagcgtgac accacgatgc ctgtagcaat gccaacaacg ttgcgcaaac
     8101 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     8161 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     8221 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     8281 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     8341 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     8401 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     8461 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     8521 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     8581 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     8641 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     8701 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     8761 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     8821 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     8881 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     8941 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     9001 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     9061 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     9121 gctcgtcagg ggggcggagc ctatcgaaaa acgccagcaa cgcggccttt ttacggttcc
     9181 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     9241 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     9301 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     9361 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     9421 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     9481 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     9541 acagctatga ccatgattac gccaagctat ttaggtgaca ctatagaata ctc
//
