LOCUS       40924_39927        9830 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pSG76, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9830)
  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 9830)
  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 9830)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9830)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            5, 5329 (2014)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-AUG-2014) Biological Engineering, MIT, 77 Massachusetts Avenue, 
            Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(17..35)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(53..69)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(77..93)
                     /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(101..131)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(146..167)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(455..1043)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1294..1797
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     repeat_region   1951..2505
     repeat_region   2506..3044
     gene            3052..4408
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     regulatory      complement(4456..5019)
                     /label=derived from P. falciparum HRPII 3' UTR
                     /note="derived from P. falciparum HRPII 3' UTR"
                     /regulatory_class="terminator"
     CDS             complement(5058..5990)
                     /label=Rluc
                     /note="luciferase from the anthozoan coelenterate Renilla 
                     reniformis (sea pansy)"
     regulatory      complement(6007..6611)
                     /label=5'UTR from the PcDT gene
                     /note="5'UTR from the PcDT gene"
                     /regulatory_class="promoter"
     regulatory      6612..7241
                     /note="contains promoter and 5' UTR from P. falciparum 
                     calmodulin gene"
                     /regulatory_class="promoter"
     misc_feature    7244..7290
                     /label=mutated TetR aptamer
                     /note="mutated TetR aptamer"
     CDS             7313..8962
                     /label=luciferase
                     /note="firefly luciferase"
     regulatory      8972..9807
                     /label=derived from P. falciparum hsp86 3' UTR
                     /note="derived from P. falciparum hsp86 3' UTR"
                     /regulatory_class="terminator"
ORIGIN
        1 ccccaagctt gagtattcta tagtgtcacc taaatagctt ggcgtaatca tggtcatagc
       61 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      121 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      181 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      241 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
      301 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      361 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      421 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      481 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      541 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      601 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      661 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      721 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      781 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      841 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
      901 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      961 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1021 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1081 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1141 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1201 cttggtctga cagttaccaa tgcttaatca gtgaggcagc ttcgtacgct gcaggtcgag
     1261 atttccccga aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa taattataat
     1321 ttaaattttt taatataaat atataaatta aaaatagaaa gtaaaaaaag aaattaaaga
     1381 aaaaatagtt tttgttttcc gaagatgtaa aagactctag ggggatcgcc aacaaatact
     1441 accttttatc ttgctcttcc tgctctcagg tattaatgcc gaattgtttc atcttgtctg
     1501 tgtagaagac cacacacgaa aatcctgtga ttttacattt tacttatcgt taatcgaatg
     1561 tatatctatt taatctgctt ttcttgtcta ataaatatat atgtaaagta cgctttttgt
     1621 tgaaattttt taaacctttg tttatttttt tttcttcatt ccgtaactct tctaccttct
     1681 ttatttactt tctaaaatcc aaatacaaaa cataaaaata aataaacaca gagtaaattc
     1741 ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa gttacaggca agcgatccgt
     1801 cctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggatct
     1861 gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggtcta tatttgttaa
     1921 attaagacca cccggccagc gacatggtta ttgttaaatt aagaccaaac ttagtacctt
     1981 aatgacttat gttagttacg ttaagaccta tataacttat gttaggacct atgttggtta
     2041 agttaaggcc tatagtactt atattaggac ctatactatt tatgttagga cctatattta
     2101 ttaagtcaag acctaaatta gttaccttaa gacctatata ttagttacct tacgacctaa
     2161 gttagtacct taattactta agtcagtaat attaagacct atattagtga aggtaagacc
     2221 taagttagtt aagttaagac ctacattact aatgttagga cttatgttag tcaaattagg
     2281 acctaagtta gtaatcataa gacctatatc agtgaatgta agaccttagt tagtcgaagt
     2341 atgaccttag ttactgaatt taagacctaa gttagtaaaa gtaataccta agttagtgaa
     2401 agtaagacat atatcagtaa aggtaagacc ttagtttagt taccataaga cttatatgag
     2461 tgaaagtcag acctacatta ctaatgttag gccagcgaca tgttattgtt aaattaagac
     2521 caaacttagt accttaatga cttatgttag ttacgttaag acctatataa cttatgttag
     2581 gacctatgtt ggttaagtta aggcctatag tacttatatt aggacctata ctatttatgt
     2641 taggacctat atttattaag tcaagaccta aattagttac cttaagacct atatattagt
     2701 taccttacga cctaagttag taccttaatt acttaagtca gtaatattaa gacctatatt
     2761 agtgaaggta agacctaagt tagttaagtt aagacctaca ttactaatgt taggacttat
     2821 gttagtcaaa ttaggaccta agttagtaat cataagacct atatcagtga atgtaagacc
     2881 ttagttagtc gaagtatgac cttagttact gaatttaaga cctaagttag taaaagtaat
     2941 acctaagtta gtgaaagtaa gacatatatc agtaaaggta agaccttagt ttagttacca
     3001 taagacttat atgagtgaaa gtcagaccta cattactaat gttaggccag cgacatggag
     3061 gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt
     3121 cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat acattttgat
     3181 ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc agacctgcga gcagggaaac
     3241 gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa
     3301 ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga
     3361 tacagttctc acatcacatc cgaacataaa caaccatggg taaggaaaag actcacgttt
     3421 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     3481 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
     3541 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     3601 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     3661 ctgatgatgc atggttactc accactgcga tccccggcaa aacagcattc caggtattag
     3721 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     3781 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     3841 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     3901 atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg
     3961 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     4021 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     4081 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     4141 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     4201 tctaatcagt actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt
     4261 tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg
     4321 cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa
     4381 tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc gccatccagt
     4441 gtcgaaaacg agctcggtac ctctagatgt aataaatatg ttcttatata taatgagaaa
     4501 taaatattta acatatgttt aaaaagaaaa atttaagatt tacatgatta ggaataaaaa
     4561 tattaaaagc tatatattaa tgttgtactt tatgttcata ttgtaaaaga taaatataat
     4621 ttttattaaa atttatatta ttattattat aagttcttag aaaatacatt ttgttttaaa
     4681 ataatactat aaaaagaaca ataataaaaa taaaataaat acaattaaaa cataattaaa
     4741 aaagaaaata cagacagtaa aaaaaatcgc tatcccataa attacaaaac atgaattaat
     4801 aaacatatat tgtttattaa taaaattgta agttattttt atatatacat aataaattaa
     4861 agatactgaa acaataattt taaatgtaaa taaatgaata atgatatgtt ttatatgatt
     4921 cattattcta tatttataag gaagattaca aaaaaaaaat tcatatgtat tttttttgta
     4981 atttctgtgt ttatgttttt atttttataa tatttttaat ctattattaa ataaatttaa
     5041 tggggtaccc gcggttattg ttcatttttg agaactcgct caacgaacga tttgatatat
     5101 tttcccattt catcaggtgc atcttcttgc gaaaaatgaa gaccttttac tttgacaaat
     5161 tcagtattag gaaacttctt ggcaccttca acaatagcat tggaaaagaa tcctgggtcc
     5221 gattcaataa acatttttgg taaatcatca cttgcacgta gataagcatt ataattccta
     5281 acaatttgta caacgtcagg tttaccacct tttactaacg ggatttcacg aggccatgat
     5341 aatgttggac gacgaacttc acctttctct ttgaatggtt caagatatgc tgcaaattct
     5401 tctggttcta actttctcat gatttttgat ggcaacatgg tttccacgaa gaagttattc
     5461 tccaaaacca ttttttctcc ttcttcagat ttgatcaacg caatatcttc ttcaatatca
     5521 ggccattcat cccatgattc aatcacatct actacacttt cagcgtgaac tattgctttg
     5581 atcttatctt gatgctcata gctataatga aatgccaaac aagcacccca atcatggccg
     5641 acaaaaatga tcttctttgg taaattaaga agttcaaacc atgcagtaag atatttgtaa
     5701 tgatcaagta acctataaga accattacca gatttgcctg atttgcccat accaataagg
     5761 tctggtataa tacaccgcgc tactggctca atatgtggca caacatgtcg ccataaataa
     5821 gaagaggccg cgttaccatg taaaaaaata acagcatttt ctgcatgttt ttctgaatca
     5881 taataattaa taaatgaatc aagaacattc atttgtttac atctggccca ccactgcgga
     5941 ccagttatca tccgtttcct ttgttctgga tcataaactt tcgaagtcat ggtggcccta
     6001 ggatactttg taagttttag gtgtgtgtat atatttaagc actacacaat tgttactagc
     6061 aaatgtagaa agaaatagcc agtatttaaa ctaaatatgt gaatattaaa atatagtgac
     6121 aaaaaaggaa agtgtatgta aaataaaatg aaaattgaaa agaatttata ttataatata
     6181 aaaaattgta gtgaacatat ataatcctgt gttatatata tttaatatta ttattgactt
     6241 ttttaaaaaa aatatttatt ataaaaattt tttaatttat gcactttttt attgtactta
     6301 tatatattcc attattttac ttacacctaa atttagtaga aaattttatt caacccaaat
     6361 aaaaattatg gcgctatata ggatgagatc atatattttt ttctcttttt caaatccttt
     6421 atattttatc atacaaactg ttttaaacac gagaaaaagg ggatgccatg cacatgctta
     6481 gtacacatgc aatagatata tatagtatat atattacata tatatatgtg tacttgttaa
     6541 aatgctatat caaatagttt tgcataaacg agagtacctc ttacaagcga attagctaac
     6601 ctagtgttta taagctttta tatgtgatta attttatata ttatcaattt atatattttt
     6661 aaatgcttac ttaattatct tttttttttt tttttttttt ttttcccctc tttttatatt
     6721 aatttatttt tgaaaaaatt gatatatata tatatatata atatatatat atacatgtag
     6781 tagtattaaa caatgtataa tatatataaa taatatattt atatatttca tttcaatttt
     6841 aatttttttt ggtttttttt ttttttcttt ttgtcatatt taaaaaaaat tatattcata
     6901 taagttatgc attttttata aacattattc aatatatgta taatataata tatatatata
     6961 tattaatgta ttattccaat gtgcatgata aaagaaaaaa ataatattta taaaaaaaaa
     7021 gaaaaataaa acaaaaaaag aaaaaaaaaa aaaaaaaaaa aaaaatacaa aaataaataa
     7081 tataatttat aattatatat tcttgtcaca ataaaaatat atatatatat atatatattt
     7141 ataatatgta tattttaaac tagaaaagga ataactaata ttttatttat tatcattcaa
     7201 gatttatatt ttataataat aaatacctaa tagaaatata tcaatccagg cagtgtaagg
     7261 tcgatacgga cggaatgtga tggcctggat aaaacacaaa actcgagaac atatggaaga
     7321 cgccaaaaac ataaagaaag gcccggcgcc attctatcca ctagaggatg gaaccgctgg
     7381 agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac
     7441 agatgcacat atcgaggtga acatcacgta cgcggaatac ttcgaaatgt ccgttcggtt
     7501 ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga
     7561 aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc
     7621 gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgaaca tttcgcagcc
     7681 taccgtagtg tttgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaaatt
     7741 accaataatc cagaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc
     7801 gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtacc
     7861 agagtccttt gatcgtgaca aaacaattgc actgataatg aattcctctg gatctactgg
     7921 gttacctaag ggtgtggccc ttccgcatag aactgcctgc gtcagattct cgcatgccag
     7981 agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt
     8041 ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt
     8101 cttaatgtat agatttgaag aagagctgtt tttacgatcc cttcaggatt acaaaattca
     8161 aagtgcgttg ctagtaccaa ccctattttc attcttcgcc aaaagcactc tgattgacaa
     8221 atacgattta tctaatttac acgaaattgc ttctgggggc gcacctcttt cgaaagaagt
     8281 cggggaagcg gttgcaaaac gcttccatct tccagggata cgacaaggat atgggctcac
     8341 tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg
     8401 taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg
     8461 cgttaatcag agaggcgaat tatgtgtcag aggacctatg attatgtccg gttatgtaaa
     8521 caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat
     8581 agcttactgg gacgaagacg aacacttctt catagttgac cgcttgaagt ctttaattaa
     8641 atacaaagga tatcaggtgg cccccgctga attggaatcg atattgttac aacaccccaa
     8701 catcttcgac gcgggcgtgg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc
     8761 cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc
     8821 cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc
     8881 gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa
     8941 gaagggcgga aagtccgagt tgtaactcga gttatataat atatttatgt actcacaatg
     9001 gggtctacaa aaaaaaaaaa ataaagatat atatatatat atattatctt tcattttatc
     9061 atcaccccaa tatacatata tatattttta tatttttata tatttatatt tttcttttat
     9121 ttatccataa aagaaaatta aatataaatt tatacctttt atttgaatta ccatttgtat
     9181 aaaatataaa aaaaaaaaaa aataataaaa agaagaaaaa acgaaaaaag taatatttaa
     9241 ataataaaga tcaatataca taataaaaaa aaaaaaaaaa aaaaaaatta aaatttaaaa
     9301 aattaaacat tgaaataatt tcatatacac acatatacat acatacatat atatatatat
     9361 atatatatat ttatacattt atgcatataa aatatattaa caacttatgt gtattcatat
     9421 atattttttt acatatgtat tgaaagactt tactgagaca tgaaatatct aataaccaaa
     9481 cgtttataaa ttctaattat atgtaaagca taatatagct tttgtctaat ttaattcctt
     9541 aaaacgtatg tattaaaaat atagtaagat ggggaaatta taattttttt tttaaaggtt
     9601 tttctttgat acagtaaaaa aaaaaaataa aaaaaaataa taataataaa aaaataaaaa
     9661 aataaaaaaa aataaaataa ttataaagat atatataatt cctcccaaat acaaatgatt
     9721 aagtttaaaa attaataaat aaatcaaaaa aaaatttaat aataattgta ttattttaag
     9781 tgtagttaat tcatcaaata gcatgcctgc aggtcgactc tagaggatcc
//
