LOCUS       sequence-437768-        5210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mScarlet regardless of TDP-43 knockdown. 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 5210)
  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 5210)
  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) matched positive cont...
FEATURES             Location/Qualifiers
     source          1..5210
                     /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             1351..1374
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1672..1793
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1683..1702)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1737..1756
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1908..1954
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1984..2120
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2165..2236
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2462..3050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2542..2561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3796..3864,3865..4656)
                     /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(4014..4033)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4669..4805
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4957..5028
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5033..5079)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5097..5145
                     /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 tccaaaggtg aggcagtcat aaaggagttt atgaggttca aggtgcacat ggaagggtca
      721 atgaacggac atgagttcga aattgaaggt gagggcgagg gccgccccta tgaagggaca
      781 caaactgcca agctcaaagt gaccaagggc gggcctctgc ccttctcttg ggatatcctg
      841 agcccgcagt ttatgtacgg cagccgggct ttcaccaaac accctgccga tatcccagac
      901 tactataaac agtcctttcc agaaggattt aagtgggagc gagtcatgaa tttcgaggac
      961 ggaggtgccg tgacggttac tcaggacacc agcctggagg acggcaccct gatctacaag
     1021 gtgaagctga ggggcaccaa cttccccccc gacggccccg tgatgcagaa gaagaccatg
     1081 ggctgggagg ccagcaccga gaggctgtac cccgaggacg gcgtgctgaa gggcgacatc
     1141 aagatggccc tgaggctgaa ggacggcggc aggtacctgg ccgacttcaa gaccacctac
     1201 aaggccaaga agcccgtgca gatgcccggc gcctacaacg tggacaggaa gctggacatc
     1261 accagccaca acgaggacta caccgtggtg gagcagtacg agaggagcga gggcaggcac
     1321 agcaccggcg gcatggacga gctgtacaag gactacaagg acgatgatga caagtgataa
     1381 acaaatggta aggaagggca catcaatctt tgcttaattg tcctttactc taaagatgta
     1441 ttttatcata ctgaatgcta aacttgatat ctccttttag gtcattgatg tccttcaccc
     1501 cgggaaggcg acagtgccta agacagaaat tcgggaaaaa ctagccaaaa tgtacaagac
     1561 cacaccggat gtcatctttg tatttggatt cagaactcag taaactggat ccgcaggcct
     1621 ctgctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
     1681 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1741 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1801 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     1861 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
     1921 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
     1981 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     2041 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     2101 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
     2161 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
     2221 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc
     2281 aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca
     2341 gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa
     2401 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2461 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2521 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2581 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2641 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2701 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2761 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2821 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2881 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2941 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3001 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3061 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3121 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3181 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3241 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga
     3301 actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct
     3361 acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg
     3421 cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga
     3481 gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg
     3541 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
     3601 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
     3661 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
     3721 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
     3781 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3841 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3901 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3961 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
     4021 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4081 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     4141 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4201 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4261 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4321 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
     4381 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
     4441 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4501 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4561 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4621 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
     4681 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     4741 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     4801 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
     4861 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
     4921 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
     4981 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
     5041 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
     5101 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
     5161 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
//
