LOCUS       sequence-161527-        1016 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses GFP from the CBA promoter. Includes a Csy4 target hairpin 
            in the 5'UTR for Csy4-mediated regulation. Cloned into an 
            Adeno-Associated Virus backbone for packaging into AAV..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1016)
  AUTHORS   Borchardt EK, Vandoros LA, Huang M, Lackey PE, Marzluff WF, Asokan A
  TITLE     Controlling mRNA stability and translation with the CRISPR 
            endoribonuclease Csy4.
  JOURNAL   RNA. 2015 Nov;21(11):1921-30. doi: 10.1261/rna.051227.115. Epub 2015
            Sep 9.
  PUBMED    26354771
REFERENCE   2  (bases 1 to 1016)
  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: 
            "10.1261/rna.051227.115"; journalName: "RNA"; date: "2015-11"; 
            volume: "21"; issue: "11"; pages: "1921-30"
COMMENT     Sequence Label: TR-5'UTR-HP-GFP
FEATURES             Location/Qualifiers
     source          1..1016
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    98..117
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     regulatory      144..153
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(150..152,153..155,156..869)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(150..152,153..155,156..866)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(195..216)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(456..475)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     803..824
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(909..928)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     963..982
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gtaattagct gagcaagagg taagggttta agggatggtt ggttggtggg gtattaatgt
       61 ttaattacct ggagcacctg cctgaaatca ctttttagtt cactgccgta taggcagcta
      121 agaaatttca ggttggaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
      181 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      241 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      301 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
      361 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      421 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
      481 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
      541 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
      601 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
      661 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
      721 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
      781 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
      841 ctctcggcat ggacgagctg tacaagtaac tcgagcggcc gcttcgagca gacatgataa
      901 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
      961 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaa
//
