LOCUS       sequence-371989-        3946 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  vector for encoding a human codon-optimized dead RfxCas13d (dCas13d)
            driven by CBh promoter, guide RNAs compatible with RfxCas13d driven 
            by hU6, EGFP driven by SV40 promoter, and mCherry driven by SV40 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3946)
  AUTHORS   Tong H, Huang J, Xiao Q, He B, Dong X, Liu Y, Yang X, Han D, Wang Z,
            Wang X, Ying W, Zhang R, Wei Y, Xu C, Zhou Y, Li Y, Cai M, Wang Q, 
            Xue M, Li G, Fang K, Zhang H, Yang H
  TITLE     High-fidelity Cas13 variants for targeted RNA degradation with 
            minimal collateral effects.
  JOURNAL   Nat Biotechnol. 2022 Aug 11. pii: 10.1038/s41587-022-01419-7. doi: 
            10.1038/s41587-022-01419-7.
  PUBMED    35953673
REFERENCE   2  (bases 1 to 3946)
  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"; journalName: "Nat 
            Biotechnol. 2022 Aug 11. pii: 10.1038/s41587-022-01419-7. doi: 
            10.1038/s41587-022-01419-7."
COMMENT     Sequence Label: 
            pCBh-NLS_dCas13d_HA_NLS-pA-U6-DR-BpiI-BpiI-pSV40-EGFP-pA-pSV40-mCher
            ry-pA
FEATURES             Location/Qualifiers
     source          1..3946
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        1..286
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        288..565
                     /label=chicken beta-actin promoter
     intron          566..794
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             815..835
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3752..3778
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3779..3799
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3812..3946
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3849..3868)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3903..3922
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      121 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      181 cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag tacatgacct tatgggactt
      241 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      301 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      361 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccagg
      421 cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa
      481 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
      541 taaaaagcga agcgcgcggc gggcgggagt cgctgcgacg ctgccttcgc cccgtgcccc
      601 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
      661 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gctgagcaag aggtaagggt
      721 ttaagggatg gttggttggt ggggtattaa tgtttaatta cctggagcac ctgcctgaaa
      781 tcactttttt tcaggttgga ccggtgccac catgcctaaa aagaaaagaa aggtgggttc
      841 tggtatcgag aagaagaaga gcttcgccaa gggcatggga gtgaagagca ccctggtgtc
      901 cggctctaag gtgtacatga ccacatttgc tgagggaagc gacgccaggc tggagaagat
      961 cgtggagggc gatagcatca gatccgtgaa cgagggagag gctttcagcg ccgagatggc
     1021 tgacaagaac gctggctaca agatcggaaa cgccaagttt tcccacccaa agggctacgc
     1081 cgtggtggct aacaacccac tgtacaccgg accagtgcag caggacatgc tgggactgaa
     1141 ggagacactg gagaagaggt acttcggcga gtccgccgac ggaaacgata acatctgcat
     1201 ccaggtcatc cacaacatcc tggatatcga gaagatcctg gctgagtaca tcacaaacgc
     1261 cgcttacgcc gtgaacaaca tctccggcct ggacaaggat atcatcggct tcggaaagtt
     1321 ttctaccgtg tacacatacg acgagttcaa ggatccagag caccaccggg ccgcttttaa
     1381 caacaacgac aagctgatca acgccatcaa ggctcagtac gacgagttcg ataactttct
     1441 ggataacccc aggctgggct acttcggaca ggctttcttt tctaaggagg gcagaaacta
     1501 catcatcaac tacggaaacg agtgttacga catcctggcc ctgctgagcg gactggcgca
     1561 ctgggtggtg gctaacaacg aggaggagtc tcggatcagc cgcacctggc tgtacaacct
     1621 ggacaagaac ctggataacg agtacatctc cacactgaac tacctgtacg acaggatcac
     1681 caacgagctg acaaacagct tctccaagaa ctctgccgct aacgtgaact acatcgctga
     1741 gaccctgggc atcaacccag ctgagttcgc tgagcagtac ttcagatttt ccatcatgaa
     1801 ggagcagaag aacctgggct tcaacatcac aaagctgaga gaagtgatgc tggacagaaa
     1861 ggatatgtcc gagatcagga agaaccacaa ggtgttcgat tctatcagaa ccaaggtgta
     1921 cacaatgatg gactttgtga tctacaggta ctacatcgag gaggatgcca aggtggccgc
     1981 tgccaacaag agcctgcccg acaacgagaa gtctctgagc gagaaggata tcttcgtgat
     2041 caacctgaga ggctccttta acgacgatca gaaggacgct ctgtactacg atgaggccaa
     2101 caggatctgg agaaagctgg agaacatcat gcacaacatc aaggagttcc ggggaaacaa
     2161 gacccgcgag tacaagaaga aggacgctcc aaggctgcct aggatcctgc ctgctggaag
     2221 ggacgtgagc gccttcagca agctgatgta cgccctgaca atgtttctgg acggaaagga
     2281 gatcaacgat ctgctgacca cactgatcaa caagttcgac aacatccagt cttttctgaa
     2341 agtgatgcct ctgatcggcg tgaacgctaa gttcgtggag gagtacgcct tctttaagga
     2401 cagcgccaag atcgctgatg agctgcggct gatcaagtcc tttgccagga tgggagagcc
     2461 aatcgctgac gctaggagag ctatgtacat cgatgccatc cggatcctgg gaaccaacct
     2521 gtcttacgac gagctgaagg ctctggccga caccttcagc ctggatgaga acggcaacaa
     2581 gctgaagaag ggcaagcacg gaatgcgcaa cttcatcatc aacaacgtga tcagcaacaa
     2641 gcggtttcac tacctgatca gatacggcga cccagctcac ctgcacgaga tcgctaagaa
     2701 cgaggccgtg gtgaagttcg tgctgggacg gatcgccgat atccagaaga agcagggcca
     2761 gaacggaaag aaccagatcg accgctacta cgagacctgc atcggcaagg ataagggaaa
     2821 gtccgtgtct gagaaggtgg acgctctgac caagatcatc acaggcatga actacgacca
     2881 gttcgataag aagagatctg tgatcgagga caccggaagg gagaacgccg agagagagaa
     2941 gtttaagaag atcatcagcc tgtacctgac agtgatctac cacatcctga agaacatcgt
     3001 gaacatcaac gctagatacg tgatcggctt ccactgcgtg gagcgcgatg cccagctgta
     3061 caaggagaag ggatacgaca tcaacctgaa gaagctggag gagaagggct ttagctccgt
     3121 gaccaagctg tgcgctggaa tcgacgagac agcccccgac aagaggaagg atgtggagaa
     3181 ggagatggcc gagagagcta aggagagcat cgactccctg gagtctgcta accctaagct
     3241 gtacgccaac tacatcaagt actccgatga gaagaaggcc gaggagttca ccaggcagat
     3301 caacagagag aaggccaaga ccgctctgaa cgcctacctg aggaacacaa agtggaacgt
     3361 gatcatccgg gaggacctgc tgcgcatcga taacaagacc tgtacactgt tcgcaaacaa
     3421 ggctgtggcc ctggaggtgg ctcgctacgt gcacgcctac atcaacgaca tcgccgaggt
     3481 gaactcctac tttcagctgt accactacat catgcagagg atcatcatga acgagagata
     3541 cgagaagtct agcggcaagg tgtctgagta cttcgacgcc gtgaacgatg agaagaagta
     3601 caacgataga ctgctgaagc tgctgtgcgt gcctttcgga tactgtatcc cacggtttaa
     3661 gaacctgagc atcgaggccc tgttcgaccg caacgaggct gccaagtttg ataaggagaa
     3721 gaagaaggtg agcggcaact ccggttctgg ttacccatac gacgtaccag attacgctcc
     3781 caagaagaag cgcaaggtgg gataaaagct taacttgttt attgcagctt ataatggtta
     3841 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3901 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatc
//
