LOCUS       sequence-437747-        5841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mCherry specifically in cells with TDP-43 knockdown. 
            Uses AARS1-derived cryptic exon to control mCherry expression. 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 5841)
  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 5841)
  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-REGv1(mCherry-FLAG) in pTwist-CMV
FEATURES             Location/Qualifiers
     source          1..5841
                     /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             1223..1279
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1985..2008
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2303..2424
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2314..2333)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2368..2387
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2539..2585
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2615..2751
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2796..2867
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3093..3681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3173..3192)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4427..4495,4496..5287)
                     /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(4645..4664)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5300..5436
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5588..5659
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5664..5710)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5728..5776
                     /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 gaaccatggt agccatggag
      661 accatggggc tcatgacaac agatctggca aaatttgggg taagaatgca catcacttct
      721 tgagagtatg gaggagtgaa atgacactca gtgccagagt tactgtatat ctacacttta
      781 aaagtgtagc ttttaaaaga taagcaagca caatcttttg tgtgtgtgtg tgtgaatgtg
      841 tgtgtgtgtg tgtgtcaccc aggctggagt gcagtggcat gatcacagct cactgcagcc
      901 tcaaacttcc tgggctcaag tgatcctctc ccgagtagct gggactacag gtatgcatca
      961 cccccccagc taattttttt ttgtattttt taccgagtcg gggtttcgca atgttgccca
     1021 ggctggtctc agagtctcgc tctgttgtct acgctggagt gcagtaacat gagccactgt
     1081 gcccggccaa tcctaagaat ttcttttgcg gtggttgcaa gtctgggcag aactcttgtc
     1141 aggggctgta actggactta tctttactcc tttgtcaggc tggatgccac caaaatcctc
     1201 ccaggcaaca tacggcagcg gcgccaccaa cttttccctg ctcaagcaag ccggcgacgt
     1261 ggaagagaat cccggccccg tcagcaaagg ggaagaggac aacatggcca tcattaagga
     1321 gtttatgcga ttcaaagtac acatggaggg atctgttaat ggccatgaat ttgagataga
     1381 gggggaaggt gagggtcgcc cttacgaagg cacgcagacg gctaagctga aggtcacgaa
     1441 agggggaccc ttgcccttcg catgggacat actctcccca cagtttatgt atggttctaa
     1501 ggcatatgtt aagcaccctg cagacatccc agactatctg aagctctcct ttcctgaggg
     1561 gtttaagtgg gaacgcgtta tgaactttga ggatggaggg gtcgtgactg ttacccagga
     1621 ttcttccctg caagatggag agttcatata caaagtgaaa cttcggggaa cgaatttccc
     1681 atcagacggg ccagtgatgc agaaaaagac gatggggtgg gaggcttcat ccgagaggat
     1741 gtatcccgag gacggagcat tgaaaggcga aataaaacaa aggctgaagt tgaaggatgg
     1801 gggccactac gacgcggagg ttaaaacaac gtataaagct aaaaagccag tacagctccc
     1861 aggcgcatat aacgtgaata taaagcttga cataacgagt cataacgagg attacacaat
     1921 cgtagaacag tacgaaagag ctgaaggacg gcactccacc ggtgggatgg atgaactcta
     1981 taaagactac aaggacgatg atgacaagta aacaaatggt aaggaagggc acatcaatct
     2041 ttgcttaatt gtcctttact ctaaagatgt attttatcat actgaatgct aaacttgata
     2101 tctcctttta ggtcattgat gtccttcacc ccgggaaggc gacagtgcct aagacagaaa
     2161 ttcgggaaaa actagccaaa atgtacaaga ccacaccgga tgtcatcttt gtatttggat
     2221 tcagaactca gtaaactgga tccgcaggcc tctgctagct tgactgactg agatacagcg
     2281 taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2341 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2401 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2461 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc
     2521 ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     2581 ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga
     2641 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     2701 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag
     2761 ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga
     2821 caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta
     2881 ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt
     2941 ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc
     3001 acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc
     3061 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3121 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3181 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3241 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3301 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3361 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3421 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3481 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3541 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3601 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3661 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3721 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3781 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3841 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3901 cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt
     3961 caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct
     4021 tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat
     4081 cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat
     4141 ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct
     4201 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     4261 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     4321 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     4381 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     4441 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4501 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     4561 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     4621 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4681 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     4741 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4801 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     4861 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4921 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4981 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     5041 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5101 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5161 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5221 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5281 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     5341 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     5401 ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac
     5461 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     5521 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     5581 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     5641 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     5701 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     5761 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     5821 ggggtcatta gttcatagcc c
//
