LOCUS       sequence-495754-        7209 bp    DNA     circular SYN 18-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 0 CTG repeats in exon 15 located.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7209)
  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 7209)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 18, 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: pBItetDMPKSGFP
FEATURES             Location/Qualifiers
     source          1..7209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(1..19,20..42,43..61,62..84,85..103,104..126,127..145,
                     146..168,169..187,188..210,211..229,230..252,253..271)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        304..342
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      486..495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(492..494,495..497,498..1211)
                     /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(492..494,495..497,498..1208)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(537..558)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(798..817)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1145..1166
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          1297..1869
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2001..2020)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2066..2121
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2120..2139)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2519..2536)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2690..3278)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2770..2789)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3449..4240,4241..4309))
                     /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     4072..4091
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4310..4414)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4482..4500
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
       61 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
      121 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
      181 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
      241 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagctcg gtacccgggt
      301 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
      361 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
      421 agcctccgcg gccccgaatt cgagctcggt acccggggat cctctagtca gctgacgcgc
      481 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      541 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      601 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      661 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
      721 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      781 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      841 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
      901 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
      961 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1021 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1081 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1141 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1201 tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     1261 tttacttgct ttatctctag agctgagaac ttcagggtga gtttggggac ccttgattgt
     1321 tctttctttt tcgctattgt aaaattcatg ttatatggag ggggcaaagt tttcagggtg
     1381 ttgtttagaa tgggaagatg tcccttgtat caccatggac cctcatgata attttgtttc
     1441 tttcactttc tactctgttg acaaccattg tctcctctta ttttcttttc attttctgta
     1501 actttttcgt taaactttag cttgcatttg taacgaattt ttaaattcac ttttgtttat
     1561 ttgtcagatt gtaagtactt tctctaatca cttttttttc aaggcaatca gggtatatta
     1621 tattgtactt cagcacagtt ttagagaaca attgttataa ttaaatgata aggtagaata
     1681 tttctgcata taaattctgg ctggcgtgga aatattctta ttggtagaaa caactacacc
     1741 ctggtcatca tcctgccttt ctctttatgg ttacaatgat atacactgtt tgagatgagg
     1801 ataaaatact ctgagtccaa accgggcccc tctgctaacc atgttcatgc cttcttctct
     1861 ttcctacagc tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt tggcaaagaa
     1921 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     1981 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     2041 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     2101 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     2161 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     2221 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     2281 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     2341 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     2401 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgactgcatt
     2461 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2521 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2581 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2641 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2701 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2761 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2821 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2881 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2941 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3001 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3061 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3121 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3181 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3241 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3301 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3361 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3421 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3481 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3541 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3601 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3661 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3721 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3781 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3841 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3901 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3961 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4021 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     4081 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4141 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4201 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4261 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4321 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4381 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4441 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4501 cctttcgtct tcactcgacg cggccagctt ggtcgagctg gatacttccc gtccgccagg
     4561 gggacatgcc ggcgatgctg aaggtcgcgc gcattcccga tgaagaggcc ggttactcga
     4621 gcgcaggcgc agtggaagga ggatggccgc gcgcgctgcc agcccagccc cctcttctcg
     4681 acgctcggtg gcacagctgg gccacagctg ggcgggggcg gtgcctccgg gtggcccgct
     4741 cgccctccta ttggccggac gccaaagccc cgccccgtgg cttttcctcc cccaaccctg
     4801 attcggccgc ttcgcatccc gctagctcct cccagacctt cggccgcctc cacacgcctc
     4861 cggattggcc cgctgcggag cctccggccc acaacgcaaa ccgcggacac tgtggagtcc
     4921 agagctttgg gcagatggag ggccttttat tcgcgagggt cgggggtggg ggtcctaggt
     4981 ggggacagac aatagatacc gaggaatgtc ggggtctcag tgcatccaaa acgtggattg
     5041 gggttgttgg gggtcctgta gcctgtcagc gagtcggagg acgaggtcaa taaatatcca
     5101 aaccgccgaa gcgggcgggg ccggctgggg ctccgagagc agcgcaagtg aggagggggg
     5161 cgcgggatcc ccgaaaaagc gggtttggca aaagcaaatt tcccgagtaa gcaggcagag
     5221 atcgcgccag acgctcccca gagcagggcg tcatgcacaa gaaagctttg cactttgcga
     5281 accaacgata ggtgggggtg cgtggaggat ggaacacgga cggcccggct tgctgccttc
     5341 ccaggcctgc agtttgccca tccacgtcag ggcctcagcc tggccgaaag aaagaaatgg
     5401 tctgtgatcc cccgtcgaca ttcccggcta caaggaccct tcgagccccg ttcgccggcc
     5461 gcggacccgg cccctccctc cccggccgct agggggcggg cccggatcac aggactggag
     5521 ctgggcggag acccacgctc ggagcggttg tgaactggca ggcggtgggc gcggcttctg
     5581 tgccgtgccc cgggcactca gtcttccaac ggggccccgg agtcgaagac agttctaggg
     5641 ttcagggagc gcgggcggct cctgggcggc gccagactgc ggtgagttgg ccggcgtggg
     5701 ccaccaaccc aatgcagccc agggcggcgg cacgagacag aacaacggcg aacaggagca
     5761 gggaaagcgc ctccgatagg ccaggcctag ggacctgcgg ggagagggcg aggtcaacac
     5821 ccggcatggg cctctgattg gctcctggga ctcgccccgc ctacgcccat aggtgggccc
     5881 gcactcttcc ctgcgccccg cccccgcccc aacagcctac agctgttgtt agtccactcg
     5941 cacgcctcga atcccgtccg aactcgtcat tggctgcttc ctagcggcct gtgttgattg
     6001 gctgcccgaa gatccgccct cctgccgtgg gcccagcccc gcaaatgcgc agctaagcgg
     6061 gtggcacggg gcgggtggag cgcggggcgc gacggcggag gggggcgtgg gcagccggac
     6121 gtaccctggc agggagcagc aggtggcggc ggtgcatggg gcctggcccc accagcgggc
     6181 actggcccac agccacggcc ggggggccat ctagctggag agagaaggga caggtgaccc
     6241 gatcggagcc cagcccagcc ctcagcggtg gggcgagaga cagcgagggg aatcgaggtt
     6301 ggggaggtta tctagggaga tcccggaggg aatctggtga ggcctgaacg gagggagatc
     6361 tggggctgaa taaagggctt ctgccctcta aagtcgcaaa gacgtagggt gagccctata
     6421 tctggacggg gagaccagga gccagggagg ggatctgcag aatgggcagc aggtctgagg
     6481 caggggaaag agaggggtct tacatgggaa ggtggatccg tggcccgggg actggggacc
     6541 cccgtgacag ctggaaggag aagaaagagg catagggcgc gtggaggggc gaaggagggc
     6601 ggtggcgcgg cgtgccccag cgtgggtccc ttccctcctc caggtgtcta tacacgcccc
     6661 gcggagcaga cggcccacct cctcccggtc ctccggggaa ggggacacat gagggactca
     6721 cctgtggctc cctctgcctg cagcaactcc atccgctcct gcaactgccg gacgtgtgcc
     6781 tctaggtccc ggttccgagc ctctgcctcg cgtagttgac tgtggggagg taaggacggt
     6841 gagtccgtcc gggccggacg agaggggctg ccaagggttg ccaccggccc gcatcccggc
     6901 cccggccccg gccccgatcc cgacctggcg aagttctggt tgtccgtgcg gatggcctcc
     6961 atctcccggc tcaggctctg ccgggtgagc acctcctcct ccagggcttc ctggagctcc
     7021 cgcagcgtca cctcggcctc agcctgatct gagtccggta gctcgacgcg gccgctgcag
     7081 gaattcgggg ccgcggaggc tggatcggtc ccggtgtctt ctatggaggt caaaacagcg
     7141 tggatggcgt ctccaggcga tctgacggtt cactaaacga gctctgctta tataggtcga
     7201 gtttaccac
//
