LOCUS       sequence-437759-        5421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of secreted Gaussia Luciferase specifically in cells with
            TDP-43 loss of function. 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 5421)
  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 5421)
  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(Gaussia Luciferase)#5 in pTwist-CMV
FEATURES             Location/Qualifiers
     source          1..5421
                     /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"
     polyA_signal    1883..2004
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1894..1913)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1948..1967
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2119..2165
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2195..2331
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2376..2447
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2673..3261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2753..2772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4007..4075,4076..4867)
                     /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(4225..4244)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4880..5016
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5168..5239
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5244..5290)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5308..5356
                     /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 accatgggag tcaaagttct gtttgccctg
      661 atctgcatcg ctgtggccga ggccaagccc accgagaaca acgaagactt caacatcgtg
      721 gccgtggcca gcaacttcgc gaccacggat ctcgatgctg accgcgggaa gttgcccggc
      781 aagaagctgc cgctggaggt gctcaaagag ttggaggcaa acgcgaggaa ggccggctgc
      841 accaggtaag aatgcacatc acttcttgag agtatggagg agtgaaatga cactcatgcc
      901 agagttactg tatgtctgca ctttaaaagt gtagctttta aaagatgatg aatgactgtc
      961 tgtttgtgtg tgtgtgtgtg aatgtgtgtg tgtgtgtgtg tcacccaggg gttgcctgat
     1021 ctgcttaagc cacatcaagt gtacgccaaa aatgaagaag ttcatcccag gtatgcatga
     1081 ctgcctgcat gcttgtttgt ttgtttgtat gtttgtatgg agtcggggtt tctgaatgta
     1141 tgcctgaggc tggttgcaga gtctcgctct ggatgtctac gctggatgtg cagtaacatg
     1201 agccactgtg cccggccaat cctaagaatt tcttttgcgg tggttgcaag tctgggcaga
     1261 actcttgtca ggggctgtaa ctggacttat ctttactcct ttgtcaggac ggtgtcatac
     1321 gtacgaaggc gataaagagt ccgcacaggg cggcataggc gaggcgatcg tcgacattcc
     1381 tgagattcct gggttcaagg acttggagcc cttggagcag ttcatcgcac aggtcgatct
     1441 gtgtgtggac tgcacaactg gctgcctcaa agggcttgcc aacgtgcagt gttctgacct
     1501 gctcaagaag tggctgccgc aacgctgtgc gacctttgcc agcaagatcc agggccaggt
     1561 ggacaagatc aagggggccg gtggtgacta aacaaatggt aaggaagggc acatcaatct
     1621 ttgcttaatt gtcctttact ctaaagatgt attttatcat actgaatgct aaacttgata
     1681 tctcctttta ggtcattgat gtccttcacc ccgggaaggc gacagtgcct aagacagaaa
     1741 ttcgggaaaa actagccaaa atgtacaaga ccacaccgga tgtcatcttt gtatttggat
     1801 tcagaactca gtaaactgga tccgcaggcc tctgctagct tgactgactg agatacagcg
     1861 taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga
     1921 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1981 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2041 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc
     2101 ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     2161 ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga
     2221 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     2281 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag
     2341 ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga
     2401 caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta
     2461 ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt
     2521 ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc
     2581 acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc
     2641 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2701 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2761 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2821 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2881 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2941 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3001 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3061 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3121 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3181 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3241 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3301 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3361 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3421 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3481 cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt
     3541 caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct
     3601 tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat
     3661 cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat
     3721 ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct
     3781 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     3841 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     3901 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     3961 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     4021 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4081 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     4141 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     4201 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4261 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     4321 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4381 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     4441 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4501 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4561 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     4621 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4681 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4741 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4801 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4861 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     4921 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     4981 ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac
     5041 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     5101 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     5161 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     5221 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     5281 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     5341 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     5401 ggggtcatta gttcatagcc c
//
