LOCUS       sequence-423042-        3005 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CIRTS RNA targeting system for targeted knockdown of m6A-containing 
            Malat1 at the synapse. CIRTS-YTHDF2-Calm3 and Malat1 gRNA is 
            expressed under human Syn1 and U6 promoter, respectively..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3005)
  AUTHORS   Madugalle SU, Liau WS, Zhao Q, Li X, Gong H, Marshall PR, 
            Periyakaruppiah A, Zajaczkowski EL, Leighton LJ, Ren H, Musgrove 
            MRB, Davies JWA, Kim G, Rauch S, He C, Dickinson BC, Fulopova B, 
            Fletcher LN, Williams SR, Spitale RC, Bredy TW
  TITLE     Synapse-Enriched m(6)A-Modified Malat1 Interacts with the Novel 
            m(6)A Reader, DPYSL2, and Is Required for Fear-Extinction Memory.
  JOURNAL   J Neurosci. 2023 Oct 25;43(43):7084-7100. doi: 
            10.1523/JNEUROSCI.0943-23.2023. Epub 2023 Sep 5.
  PUBMED    37669863
REFERENCE   2  (bases 1 to 3005)
  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: 
            "10.1523/JNEUROSCI.0943-23.2023"; journalName: "J Neurosci"; date: 
            "2023-10-25- 25"; volume: "43"; issue: "43"; pages: "7084-7100"
COMMENT     Sequence Label: CIRTS-YTHDF2-Malat1
FEATURES             Location/Qualifiers
     source          1..3005
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             727..780
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1297..1323
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1957..1980
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
      721 tccggcgagg gcaggggaag tctactaaca tgcggggacg tggaggaaaa tcccggccca
      781 accggtatgg gaattatcaa cacccttcaa aaatactact gtcgcgtccg tgggggtcgt
      841 tgcgcagtcc tgtcgtgcct gcctaaagag gagcagatcg gtaaatgctc aacccgtgga
      901 cgtaagtgct gtcgtcgcaa gaaaggcggt tctggaggct caggggggag cggagggtcc
      961 ggtggatcag gaggctccat ggcagttccc gagacccgcc ctaaccacac tatttatatc
     1021 aacaacctca atagcaagat caagaaggat gagctaaaaa agtccctgta cgccatcttc
     1081 tcccagtttg gccagatcct ggatatcctg gtaccgcggc agcgtacccc gaggggccag
     1141 gcctttgtca tcttcaagga ggtcagcagc gccaccaacg ccctgcgctc catgcagggt
     1201 ttccctttct atgacaaacc tatgcgtatc cagtatgcca agaccgacaa acgtatcccg
     1261 gccaagatga aaggcacctt cgtgggatcc cttcaactgc ctccacttga aagactgaca
     1321 ctgggcggta gtggaggaag cggggggagc ggagggagtg gtggatccgg cggctccatg
     1381 tcggccagca gcctcttgga gcagagacca aaaggtcaag gaaacaaagt acaaaatgga
     1441 tctgtacatc aaaaggatgg attaaacgat gatgattttg aaccttactt gagtccacag
     1501 gcaaggccca ataatgcata tactgccatg tcagattcct acttacccag ttactacagt
     1561 ccctccattg gcttctccta ttctttgggt gaagctgctt ggtctacggg gggtgacaca
     1621 gccatgccct acttaacttc ttatggacag ctgagcaacg gagagcccca cttcctacca
     1681 gatgcaatgt ttgggcaacc aggagcccta ggtagcactc catttcttgg tcagcatggt
     1741 tttaatttct ttcccagtgg gattgacttc tcagcatggg gaaataacag ttctcaggga
     1801 cagtctactc agagctctgg atatagtagc aattatgctt atgcacctag ctccttaggt
     1861 ggagccatga ttgatggaca gtcagctttt gccaatgaga ccctcaataa ggctcctggc
     1921 atgaatacta tagaccaagg gatggcagca ggatccgatt acaaggacga cgatgacaag
     1981 taggctagcg ggagcctctg ccctcctcct gcagcccgga tggctctcct ctgtttttgg
     2041 tttgcttcct tttgtttgcc cttcttttgg gtgaccgggg tggccaccat tcctgtccct
     2101 gcccagtggg gaggggagca aggccttctg ccaagaggag gagcatgtcc cctgccactg
     2161 ttgcatgcac cccggccctg tgactctgtg tgcaagccca gcagcctgtg gggcagggtg
     2221 ccaagagggg acgggacgga cgttccagga cagagggagt cgaggatgtg atccaggacg
     2281 ggtggggggt ggaagggtgg gaaagggcag gaaggccttg gaagctaaca tgctttcact
     2341 actggagggg gacggataga gcggatggca ctcgggtctc actgacgctg ttctgaaacc
     2401 atctggctgg ctttctgagg tcaggctggg tagggggctg tcatcggcca tactgctgat
     2461 cacacttgcc ctcatccgtc cagcgatctc acgctgttct gtaaatacct ggtgctaaca
     2521 tcccatgccg ctcccccatg gcacccccag cctcctgccc agttcaggct ggtccaggaa
     2581 tggatgtggg ggatggtcgc tgtgtaacgt gctggtgatc tttttcccct acccctttta
     2641 tgacccttaa aactttgatt ttgtcctaaa catgccgggc caactaaagg gtggggagag
     2701 gagagacgag ccccaccatg ctcacaagaa ccaacctgca ataaacactt ctgtgggccg
     2761 ccgcgcagag cacgcagcgg gcacctcctg cccctccatg gcgacttggc atcgctttct
     2821 gcccgcctct gccctcgccc tgcctgcccc acccccagcg gagagcatga tccgcgccct
     2881 tgcttctgac tctagcctct gggaccatca gtcaacgtgg gcagttcagt ctgggttcgt
     2941 tcctttctct gttctcatct ggccccccag ggcgatcttg ttgctttttg ctgggacctg
     3001 cccag
//
