LOCUS       sequence-306112-        8773 bp    DNA     circular SYN 14-APR-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 480 interrupted CTG repeats in exon 15.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8773)
  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 8773)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pBItetDT480GFP
FEATURES             Location/Qualifiers
     source          1..8773
                     /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 agctggatac
     4681 ttcccgtccg ccagggggac atgccggcga tgctgaaggt cgcgcgcatt cccgatgaag
     4741 aggccggtta ctcgagcgca ggcgcagtgg aaggaggatg gccgcgcgcg ctgccagccc
     4801 agccccctct tctcgacgct cggtggcaca gctgggccac agctgggcgg gggcggtgcc
     4861 tccgggtggc ccgctcgccc tcctattggc cggacgccaa agccccgccc cgtggctttt
     4921 cctcccccaa ccctgattcg gccgcttcgc atcccgctag ctcctcccag accttcggcc
     4981 gcctccacac gcctccggat tggcccgctg cggagcctcc ggcccacaac gcaaaccgcg
     5041 gacactgtgg agtccagagc tttgggcaga tggagggcct tttattcgcg agggtcgggg
     5101 gtgggggtcc taggtgggga cagacaatag ataccgagga atgtcggggt ctcagtgcat
     5161 ccaaaacgtg gattggggtt gttgggggtc ctgtagcctg tcagcgagtc ggaggacgag
     5221 gtcaataaat atccaaaccg ccgaagcggg cggggccggc tggggctccg agagcagcgc
     5281 aagtgaggag gggggcgcgg gatccccgaa aaagcgggtt tggcaaaagc aaatttcccg
     5341 agtaagcagg cagagatcgc gccagacgct ccccagagca gggcgtcatg cacaagaaag
     5401 ctttgcactt tgcgaaccaa cgataggtgg gggtgcgtgg aggatggaac acggacggcc
     5461 cggcttgctg ccttcccagg cctgcagttt gcccatccac gtcagggcct cagcctggcc
     5521 gaaagaaaga aatggtctgt gatcccccgt cgatcgagca gcagcagcag cagcagcagc
     5581 agcagcagca gcagcagcag cagcagcagc agcagcagtc gagcagcagc agcagcagca
     5641 gcagcagcag cagcagcagc agcagcagca gcagcagcag cagtcgagca gcagcagcag
     5701 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagtc gagcagcagc
     5761 agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagtcgagca
     5821 gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagtc
     5881 gagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
     5941 cagtcgagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc
     6001 agcagcagtc gagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
     6061 gcagcagcag cagtcgagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
     6121 cagcagcagc agcagcagtc gagcagcagc agcagcagca gcagcagcag cagcagcagc
     6181 agcagcagca gcagcagcag cagtcgagca gcagcagcag cagcagcagc agcagcagca
     6241 gcagcagcag cagcagcagc agcagcagtc gagcagcagc agcagcagca gcagcagcag
     6301 cagcagcagc agcagcagca gcagcagcag cagtcgagca gcagcagcag cagcagcagc
     6361 agcagcagca gcagcagcag cagcagcagc agcagcagtc gagcagcagc agcagcagca
     6421 gcagcagcag cagcagcagc agcagcagca gcagcagcag cagtcgagca gcagcagcag
     6481 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagtc gagcagcagc
     6541 agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagtcgagca
     6601 gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagyc
     6661 gagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
     6721 cagtcgagca gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc
     6781 agcagcagtc gagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
     6841 gcagcagcag cagtcgagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
     6901 cagcagcagc agcagcagtc gagcagcagc agcagcagca gcagcagcag cagcagcagc
     6961 agcagcagca gcagcagcag cagtcgagca gcagcagcag cagcagcagc agcagcagca
     7021 gcagcagcag cagcagcagc agcagcagtc gagcagcagc agcagcagca gcagcagcag
     7081 cagcagcagc agcagcagca gcagcagcag cagtcgacat tcccggctac aaggaccctt
     7141 cgagccccgt tcgccggccg cggacccggc ccctccctcc ccggccgcta gggggcgggc
     7201 ccggatcaca ggactggagc tgggcggaga cccacgctcg gagcggttgt gaactggcag
     7261 gcggtgggcg cggcttctgt gccgtgcccc gggcactcag tcttccaacg gggccccgga
     7321 gtcgaagaca gttctagggt tcagggagcg cgggcggctc ctgggcggcg ccagactgcg
     7381 gtgagttggc cggcgtgggc caccaaccca atgcagccca gggcggcggc acgagacaga
     7441 acaacggcga acaggagcag ggaaagcgcc tccgataggc caggcctagg gacctgcggg
     7501 gagagggcga ggtcaacacc cggcatgggc ctctgattgg ctcctgggac tcgccccgcc
     7561 tacgcccata ggtgggcccg cactcttccc tgcgccccgc ccccgcccca acagcctaca
     7621 gctgttgtta gtccactcgc acgcctcgaa tcccgtccga actcgtcatt ggctgcttcc
     7681 tagcggcctg tgttgattgg ctgcccgaag atccgccctc ctgccgtggg cccagccccg
     7741 caaatgcgca gctaagcggg tggcacgggg cgggtggagc gcggggcgcg acggcggagg
     7801 ggggcgtggg cagccggacg taccctggca gggagcagca ggtggcggcg gtgcatgggg
     7861 cctggcccca ccagcgggca ctggcccaca gccacggccg gggggccatc tagctggaga
     7921 gagaagggac aggtgacccg atcggagccc agcccagccc tcagcggtgg ggcgagagac
     7981 agcgagggga atcgaggttg gggaggttat ctagggagat cccggaggga atctggtgag
     8041 gcctgaacgg agggagatct ggggctgaat aaagggcttc tgccctctaa agtcgcaaag
     8101 acgtagggtg agccctatat ctggacgggg agaccaggag ccagggaggg gatctgcaga
     8161 atgggcagca ggtctgaggc aggggaaaga gaggggtctt acatgggaag gtggatccgt
     8221 ggcccgggga ctggggaccc ccgtgacagc tggaaggaga agaaagaggc atagggcgcg
     8281 tggaggggcg aaggagggcg gtggcgcggc gtgccccagc gtgggtccct tccctcctcc
     8341 aggtgtctat acacgccccg cggagcagac ggcccacctc ctcccggtcc tccggggaag
     8401 gggacacatg agggactcac ctgtggctcc ctctgcctgc agcaactcca tccgctcctg
     8461 caactgccgg acgtgtgcct ctaggtcccg gttccgagcc tctgcctcgc gtagttgact
     8521 gtggggaggt aaggacggtg agtccgtccg ggccggacga gaggggctgc caagggttgc
     8581 caccggcccg catcccggcc ccggccccgg ccccgatccc gacctggcga agttctggtt
     8641 gtccgtgcgg atggcctcca tctcccggct caggctctgc cgggtgagca cctcctcctc
     8701 cagggcttcc tggagctccc gcagcgtcac ctcggcctca gcctgatctg agtccggtag
     8761 ctcgacgcgg ccg
//
