LOCUS       sequence-161529-         926 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  Expresses GFP from the CBA promoter. Includes a Csy4 target hairpin 
            in the 3'UTR. 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 926)
  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 926)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-GFP-3'UTR-HP
FEATURES             Location/Qualifiers
     source          1..926
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      113..122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(119..121,122..124,125..838)
                     /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(119..121,122..124,125..835)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(164..185)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(425..444)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     772..793
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    869..888
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
ORIGIN
        1 taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt
       61 taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg tcgccaccat
      121 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      181 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      241 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      301 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
      361 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      421 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      481 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
      541 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
      601 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
      661 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
      721 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
      781 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaact
      841 cgagcggccg cttcgagcag acatgatagt tcactgccgt ataggcagct aagaaataag
      901 atacattgat gagtttggac aaacca
//
