LOCUS       sequence-151410-        7144 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 12 CTG repeats in exon 15 located.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7144)
  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 7144)
  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: pBItetDT12nGFP
FEATURES             Location/Qualifiers
     source          1..7144
                     /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..476
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      620..629
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(626..628,629..631,632..1345)
                     /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(626..628,629..631,632..1342)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(671..692)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(932..951)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1279..1300
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          1430..2002
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2134..2153)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2199..2254
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2253..2272)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2645..2662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2816..3404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2896..2915)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3575..4366,4367..4435))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4198..4217
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4436..4540)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4608..4626
                     /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 cgggtcgagt aggcgtgtac ggtgggaggc ctatataagc agagctcgtt
      481 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
      541 accgggaccg atccagcctc cgcggccccg aattcgagct cggtacccgg ggatcctcta
      601 gtcagctgac gcgccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
      661 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      721 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
      781 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
      841 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
      901 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
      961 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1021 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     1081 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     1141 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1201 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     1261 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1321 cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc
     1381 acatttgtag aggttttact tgctttatct ctagactgag aacttcaggg tgagtttggg
     1441 gacccttgat tgttctttct ttttcgctat tgaaaaattc atgttatatg gagggggcaa
     1501 agttttcagg gtgttgttta gaatgggaag atgtcccttg tatcaccatg gaccctcatg
     1561 ataattttgt ttctttcact ttctactctg ttgacaacca ttgtctcctc ttattttctt
     1621 ttcattttct gtaacttttt tcgttaaact ttagcttgca tttgtaacga atttttaaat
     1681 tcactttcgt ttatttgtca gattgtaagt actttctcta atcacttttt tttcaaggca
     1741 atcagggtaa ttatattgta cttcagcaca gttttagaga acaattgtta taattaaatg
     1801 ataaggtaga atatttctgc atataaattc tggctggcgt ggaaatattc ttattggtag
     1861 aaacaactac atcctggtaa tcatcctgcc tttctcttta tggttacaat gatatacact
     1921 gtttgagatg aggataaaat actctgagtc caaaccgggc ccctctgcta accatgttca
     1981 tgccttcttc tttttcctac agctcctggg caacgtgctg gttgttgtgc tgtctcatca
     2041 ttttggcaaa gaattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     2101 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     2161 ggacatcatg aagccccttg agcatctgac ttctgggtaa taaaggaaat ttattttcat
     2221 tgcaatagtg tgtgggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     2281 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg ccatatgctg
     2341 gctgccatga acaaaggtgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     2401 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     2461 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     2521 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tgaactcgac
     2581 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2641 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2701 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2761 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2821 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2881 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2941 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3001 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3061 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3121 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3181 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3241 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3301 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3361 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3421 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3481 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3541 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3601 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3661 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3721 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3781 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3841 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3901 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3961 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4021 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4081 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4141 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4201 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4261 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4321 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4381 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4441 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4501 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4561 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     4621 cgaggccctt tcgtcttcac tcgacgctcg agcgcaggcg cagtggaagg aggatggccg
     4681 cgcgcgctgc cagcccagcc ccctcttctc gacgctcggt ggcacagctg ggccacagct
     4741 gggcgggggc ggtgcctccg ggtggcccgc tcgccctcct attggccgga cgccaaagcc
     4801 ccgccccgtg gcttttcctc ccccaaccct gattcggccg cttcgcatcc cgctagctcc
     4861 tcccagacct tcggccgcct ccacacgcct ccggattggc ccgctgcgga gctccggccc
     4921 acaacgcaaa ccgcggacac tgtggagtcc agagctttgg gcagatggag ggccttttat
     4981 tcgcgagggt cgggggtggg ggtcctaggt ggggacagac aataaatacc gaggaatgtc
     5041 ggggtctcag tgcatccaaa acgtggattg gggttgttgg gggtcctgta gcctgtcagc
     5101 gagtcggagg acgaggtcaa taaatatcca aaccgccgaa gcgggcggag ccggctgggg
     5161 ctccgagagc agcgcaagtg aggagggggg cgcgggatcc ccgaaaaagc gggtttggca
     5221 aaagcaaatt tcccgagtaa gcaggcagag atcgcgccag acgctcccca gagcagggcg
     5281 tcatgcacaa gaaagctttg cactttgcga accaacgata ggtgggggtg cgtggaggat
     5341 ggaacacgga cggcccggct tgctgccttc ccaggcctgc agtttgccca tccacgtcag
     5401 ggcctcagcc tggccgaaag aaagaaatgg tctgtgatcc cccgtcgaca gcagcagcag
     5461 cagcagcagc agcagcagca gcagtcgaca ttcccggcta caaggaccct tcgagccccg
     5521 ttcgccggcc gcggacccgg cccctccctc cccggccgct agggggcggg cccggatcac
     5581 aggactggag ctgggcggag acccacgctc ggagcggttg tgaactggca ggcggtgggc
     5641 gcggcttctg tgccgtgccc cgggcactca gtcttccaac ggggccccgg agtcgaagac
     5701 agttctaggg ttcagggagc gcgggcggct cctgggcggc gccagactgc ggtgagttgg
     5761 ccggcgtggg ccaccaaccc aatgcagccc agggcggcgg cacgagacag aacaacggcg
     5821 aacaggagca gggaaagcgc ctccgatagg ccaggcctag ggacctgcgg ggagagggcg
     5881 aggtcaacac ccggcatggg cctctgattg gctcctggga ctcgccccgc ctacgcccat
     5941 aggtgggccc gcactcttcc ctgcgccccg cccccgcccc aacagcctac agctgttgtt
     6001 agtccactcg cacgcctcga atcccgtccg aactcgtcat tggctgcttc ctagcggcct
     6061 gtgttgattg gctgcccgaa gatccgccct cctgccgtgg gcccagcccc gcaaatgcgc
     6121 agctaagcgg gtggcacggg gcgggtggag cgcggggcgc gacggcggag gggggcgtgg
     6181 gcagccggac gtaccctggc agggagcagc aggtggcggc ggtgcatggg gcctggcccc
     6241 accagcgggc actggcccac agccacggcc ggggggccat ctagctggag agagaaggga
     6301 caggtgaccc gatcggagcc cagcccagcc ctcagcggtg gggcgagaga cagcgagcgg
     6361 aatcgaggtt ggggaggtta tctagggaga tcccggaggg aatctggtga ggcctgaacg
     6421 gagggagatc tggggctgaa taaagggctt ctgccctcta aagtcgcaaa gacgtagggt
     6481 gagccctata tctggacggg gagaccagga gccagggagg ggatctgcag aatgggcagc
     6541 aggtctgagg caggggaaag agaggggtct tacatgggaa ggtggatccg tggcccgggg
     6601 actggggacc cccgtgacag ctggaaggag aagaaagagg catagggcgc gtggaggggc
     6661 gaaggagggc ggtggcgcgg cgtgccccag cgtgggtccc ttccctcctc caggtgtcta
     6721 tacacgcccc gcggagcaga cggcccacct cctcccggtc ctccggggaa ggggacacat
     6781 gagggactca cctgtggctc cctctgcctg cagcaactcc atccgctcct gcaactgccg
     6841 gacgtgtgcc tctaggtccc ggttccgagc ctctgcctcg cgtagttgac tgtggggagg
     6901 taaggacggt gagtccgtcc gggccggacg agaggggctg ccaagggttg ccaccggccc
     6961 gcatcccggc cccggccccg gccccgatcc cgacctggcg aagttctggt tgtccgtgcg
     7021 gatggcctcc atctcccggc tcaggctctg ccgggtgagc acctcctcct ccagggcttc
     7081 ctggagctcc cgcagcgtca cctcggcctc agcctgatct gagtccggta gctcgacgcg
     7141 gccg
//
