LOCUS       sequence-193222-        7899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tet inducible bidirectional plasmid expressing GFP in one direction 
            and in the other a human DMPK genomic segment containing exons 11-15
            with 240 interrupted CTG repeats in exon 15 located.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7899)
  AUTHORS   Lee JE, Bennett CF, Cooper TA
  TITLE     RNase H-mediated degradation of toxic RNA in myotonic dystrophy type
            1.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Mar 13;109(11):4221-6. doi: 
            10.1073/pnas.1117019109. Epub 2012 Feb 27.
  PUBMED    22371589
REFERENCE   2  (bases 1 to 7899)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.1117019109"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2012-03-13- 13"; volume: "109"; issue: "11"; pages: "4221-6"
COMMENT     Sequence Label: pBItetDT240GFP
FEATURES             Location/Qualifiers
     source          1..7899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(136..154,155..177,178..196,197..219,220..238,239..261,
                     262..280,281..303,304..322,323..345,346..364,365..387,
                     388..406)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        439..477
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      621..630
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(627..629,630..632,633..1346)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(627..629,630..632,633..1343)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(672..693)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(933..952)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1280..1301
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          1432..2004
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2136..2155)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2201..2256
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2255..2274)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2654..2671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2825..3413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2905..2924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3584..4375,4376..4444))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4207..4226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4445..4549)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4617..4635
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctgcaggaat tcggggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa
       61 acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata
      121 ggtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
      181 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
      241 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
      301 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
      361 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
      421 gctcggtacc cgggtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt
      481 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      541 caccgggacc gatccagcct ccgcggcccc gaattcgagc tcggtacccg gggatcctct
      601 agtcagctga cgcgccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
      661 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      721 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      781 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
      841 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
      901 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
      961 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1021 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1081 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1141 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1201 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     1261 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1321 tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac
     1381 cacatttgta gaggttttac ttgctttatc tctagagctg agaacttcag ggtgagtttg
     1441 gggacccttg attgttcttt ctttttcgct attgtaaaat tcatgttata tggagggggc
     1501 aaagttttca gggtgttgtt tagaatggga agatgtccct tgtatcacca tggaccctca
     1561 tgataatttt gtttctttca ctttctactc tgttgacaac cattgtctcc tcttattttc
     1621 ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc atttgtaacg aatttttaaa
     1681 ttcacttttg tttatttgtc agattgtaag tactttctct aatcactttt ttttcaaggc
     1741 aatcagggta tattatattg tacttcagca cagttttaga gaacaattgt tataattaaa
     1801 tgataaggta gaatatttct gcatataaat tctggctggc gtggaaatat tcttattggt
     1861 agaaacaact acaccctggt catcatcctg cctttctctt tatggttaca atgatataca
     1921 ctgtttgaga tgaggataaa atactctgag tccaaaccgg gcccctctgc taaccatgtt
     1981 catgccttct tctctttcct acagctcctg ggcaacgtgc tggttgttgt gctgtctcat
     2041 cattttggca aagaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     2101 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     2161 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     2221 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     2281 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     2341 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     2401 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     2461 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     2521 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     2581 tccctcgact gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2641 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2701 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2761 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2821 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2881 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2941 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3001 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3061 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3121 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3181 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3241 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3301 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3361 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3421 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3481 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3541 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3601 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3661 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3721 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3781 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3841 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3901 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3961 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4021 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4081 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4141 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4201 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4261 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4321 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4381 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4441 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4501 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4561 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     4621 ggcgtatcac gaggcccttt cgtcttcact cgacgcggcc agcttggtcg agctggatrg
     4681 aggatggccg cgcgcgctgc cagcccagcc ccctcttctc gacgctcggt ggcacagctg
     4741 ggccacagct gggcgggggc ggtgcctccg ggtggcccgc tcgccctcct attggccgga
     4801 cgccaaagcc ccgccccgtg gcttttcctc ccccaaccct gattcggccg cttcgcatcc
     4861 cgctagctcc tcccagacct tcggccgcct ccacacgcct ccggattggc ccgctgcgga
     4921 gcctccggcc cacaacgcaa accgcggaca ctgtggagtc cagagctttg ggcagatgga
     4981 gggcctttta ttcgcgaggg tcgggggtgg gggtcctagg tggggacaga caatagatac
     5041 cgaggaatgt cggggtctca gtgcatccaa aacgtggatt ggggttgttg ggggtcctgt
     5101 agcctgtcag cgagtcggag gacgaggtca ataaatatcc aaaccgccga agcgggcggg
     5161 gccggctggg gctccgagag cagcgcaagt gaggaggggg gcgcgggatc cccgaaaaag
     5221 cgggtttggc aaaagcaaat ttcccgagta agcaggcaga gatcgcgcca gacgctcccc
     5281 agagcagggc gtcatgcaca agaaagcttt gcactttgcg aaccaacgat aggtgggggt
     5341 gcgtggagga tggaacacgg acggcccggc ttgctgcctt cccaggcctg cagtttgccc
     5401 atccacgtca gggcctcagc ctggccgaaa gaaagaaatg gtctgtgatc ccccgtcgat
     5461 cgagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
     5521 gcagtcgagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
     5581 cagcagcagt cgagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc
     5641 agcagcagca gcagtcgagc agcagcagca gcagcagcag cagcagcagc agcagcagca
     5701 gcagcagcag cagcagcagt cgagcagcag cagcagcagc agcagcagca gcagcagcag
     5761 cagcagcagc agcagcagca gcagtcgagc agcagcagca gcagcagcag cagcagcagc
     5821 agcagcagca gcagcagcag cagcagcagt cgagcagcag cagcagcagc agcagcagca
     5881 gcagcagcag cagcagcagc agcagcagca gcagtcgagc agcagcagca gcagcagcag
     5941 cagcagcagc agcagcagca gcagcagcag cagcagcagt cgagcagcag cagcagcagc
     6001 agcagcagca gcagcagcag cagcagcagc agcagcagca gcagtcgagc agcagcagca
     6061 gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagt cgagcagcag
     6121 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagtcgagc
     6181 agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagt
     6241 cgacattccc ggctacaagg acccttcgag ccccgttcgc cggccgcgga cccggcccct
     6301 ccctccccgg ccgctagggg gcgggcccgg atcacaggac tggagctggg cggagaccca
     6361 cgctcggagc ggttgtgaac tggcaggcgg tgggcgcggc ttctgtgccg tgccccgggc
     6421 actcagtctt ccaacggggc cccggagtcg aagacagttc tagggttcag ggagcgcggg
     6481 cggctcctgg gcggcgccag actgcggtga gttggccggc gtgggccacc aacccaatgc
     6541 agcccagggc ggcggcacga gacagaacaa cggcgaacag gagcagggaa agcgcctccg
     6601 ataggccagg cctagggacc tgcggggaga gggcgaggtc aacacccggc atgggcctct
     6661 gattggctcc tgggactcgc cccgcctacg cccataggtg ggcccgcact cttccctgcg
     6721 ccccgccccc gccccaacag cctacagctg ttgttagtcc actcgcacgc ctcgaatccc
     6781 gtccgaactc gtcattggct gcttcctagc ggcctgtgtt gattggctgc ccgaagatcc
     6841 gccctcctgc cgtgggccca gccccgcaaa tgcgcagcta agcgggtggc acggggcggg
     6901 tggagcgcgg ggcgcgacgg cggagggggg cgtgggcagc cggacgtacc ctggcaggga
     6961 gcagcaggtg gcggcggtgc atggggcctg gccccaccag cgggcactgg cccacagcca
     7021 cggccggggg gccatctagc tggagagaga agggacaggt gacccgatcg gagcccagcc
     7081 cagccctcag cggtggggcg agagacagcg aggggaatcg aggttgggga ggttatctag
     7141 ggagatcccg gagggaatct ggtgaggcct gaacggaggg agatctgggg ctgaataaag
     7201 ggcttctgcc ctctaaagtc gcaaagacgt agggtgagcc ctatatctgg acggggagac
     7261 caggagccag ggaggggatc tgcagaatgg gcagcaggtc tgaggcaggg gaaagagagg
     7321 ggtcttacat gggaaggtgg atccgtggcc cggggactgg ggacccccgt gacagctgga
     7381 aggagaagaa agaggcatag ggcgcgtgga ggggcgaagg agggcggtgg cgcggcgtgc
     7441 cccagcgtgg gtcccttccc tcctccaggt gtctatacac gccccgcgga gcagacggcc
     7501 cacctcctcc cggtcctccg gggaagggga cacatgaggg actcacctgt ggctccctct
     7561 gcctgcagca actccatccg ctcctgcaac tgccggacgt gtgcctctag gtcccggttc
     7621 cgagcctctg cctcgcgtag ttgactgtgg ggaggtaagg acggtgagtc cgtccgggcc
     7681 ggacgagagg ggctgccaag ggttgccacc ggcccgcatc ccggccccgg ccccggcccc
     7741 gatcccgacc tggcgaagtt ctggttgtcc gtgcggatgg cctccatctc ccggctcagg
     7801 ctctgccggg tgagcacctc ctcctccagg gcttcctgga gctcccgcag cgtcacctcg
     7861 gcctcagcct gatctgagtc cggtagctcg acgcggccg
//
