LOCUS       sequence-437762-        5483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mScarlet in cells with TDP-43 loss-of-function. This 
            plasmid has one of the best dynamic ranges of those tested 
            (~100-fold) in SK-N-BE2 cells. It uses a cryptic exon. [Code 'A11'] 
            Note: It is not recommended to produce lentiviruses containing 
            TDP-REG sequences – see Depositor Comments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5483)
  AUTHORS   Wilkins OG, Chien MZYJ, Wlaschin JJ, Barattucci S, Harley P, Mattedi
            F, Mehta PR, Pisliakova M, Ryadnov E, Keuss MJ, Thompson D, Digby H,
            Knez L, Simkin RL, Diaz JA, Zanovello M, Brown AL, Darbey A, Karda 
            R, Fisher EMC, Cunningham TJ, Le Pichon CE, Ule J, Fratta P
  TITLE     Creation of de novo cryptic splicing for ALS and FTD precision 
            medicine.
  JOURNAL   Science. 2024 Oct 4;386(6717):61-69. doi: 10.1126/science.adk2539. 
            Epub 2024 Oct 3.
  PUBMED    39361759
REFERENCE   2  (bases 1 to 5483)
  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: 
            "10.1126/science.adk2539"; journalName: "Science"; date: "2024-10-4-
            4"; volume: "386"; issue: "6717"; pages: "61-69"
COMMENT     Sequence Label: TDP-REGv2(mScarlet-FLAG)#7 in pTwist-CMV backbone
FEATURES             Location/Qualifiers
     source          1..5483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        321..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      628..637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1624..1647
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1945..2066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1956..1975)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2010..2029
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2181..2227
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2257..2393
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2438..2509
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2735..3323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2815..2834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4069..4137,4138..4929)
                     /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     complement(4287..4306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4942..5078
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5230..5301
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5306..5352)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5370..5418
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgcttcttgg
      601 tgccagctta tcatagcgct accggtcgcc accatggcga gaacaatggt tgctatggtg
      661 tccaagggtg aggcaggtaa gaatcgtagc atacaaaatt ataggagtgg ctgtgtgaat
      721 tggtcactgg caatgtccgt gcgtgagtgg tgcgatcagt ggtggttgaa tgcctggatg
      781 actgagtgtg tgtgtgtgct tcagtcatca aggagtttat gcgcttcaag gtgcacatgg
      841 aaggatcaat gaatggccac gagttcgaaa ttgaaggcga gggcgagggc cgcccctatg
      901 aagggacaca gactgccaag gtgtgtgtgt gtgtgtgagt gtgtggttga ttgtctgaca
      961 ggcaggtgat tagtgagtgc ttgaagacgt tatcaagcgt gattgttcct tgggagactg
     1021 aagtgtggtt ggaaaacgaa ttatcattgt tcttccccgc tacagctcaa ggtgacgaag
     1081 ggcgggcctc tgcccttctc ttgggatatc ctgagcccgc agtttatgta cggcagccgg
     1141 gctttcacca aacaccctgc cgatatccca gactactata aacagtcctt tccagaagga
     1201 tttaagtggg agcgagtcat gaatttcgag gacggaggtg ccgtgacggt tactcaggac
     1261 accagcctgg aggacggcac cctgatctac aaggtgaagc tgaggggcac caacttcccc
     1321 cccgacggcc ccgtgatgca gaagaagacc atgggctggg aggccagcac cgagaggctg
     1381 taccccgagg acggcgtgct gaagggcgac atcaagatgg ccctgaggct gaaggacggc
     1441 ggcaggtacc tggccgactt caagaccacc tacaaggcca agaagcccgt gcagatgccc
     1501 ggcgcctaca acgtggacag gaagctggac atcaccagcc acaacgagga ctacaccgtg
     1561 gtggagcagt acgagaggag cgagggcagg cacagcaccg gcggcatgga cgagctgtac
     1621 aaggactaca aggacgatga tgacaagtga taaacaaatg gtaaggaagg gcacatcaat
     1681 ctttgcttaa ttgtccttta ctctaaagat gtattttatc atactgaatg ctaaacttga
     1741 tatctccttt taggtcattg atgtccttca ccccgggaag gcgacagtgc ctaagacaga
     1801 aattcgggaa aaactagcca aaatgtacaa gaccacaccg gatgtcatct ttgtatttgg
     1861 attcagaact cagtaaactg gatccgcagg cctctgctag cttgactgac tgagatacag
     1921 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
     1981 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2041 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2101 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
     2161 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
     2221 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     2281 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     2341 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     2401 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     2461 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     2521 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     2581 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     2641 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     2701 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2761 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2821 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2881 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2941 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3001 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3061 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3121 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3181 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3241 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3301 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3361 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3421 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3481 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3541 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     3601 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     3661 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     3721 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     3781 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc
     3841 ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca
     3901 gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt
     3961 accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta
     4021 gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa
     4081 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4141 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac
     4201 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt
     4261 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4321 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca
     4381 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4441 ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag
     4501 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4561 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4621 gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag
     4681 ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4741 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4801 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4861 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4921 cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     4981 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     5041 ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg
     5101 actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt
     5161 tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc
     5221 ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa
     5281 cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa
     5341 ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg
     5401 atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt
     5461 acggggtcat tagttcatag ccc
//
