LOCUS       Exported                1016 bp ds-DNA     linear   SYN 25-MAY-2021
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    TR-5'UTR-HP-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
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 May 25, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..1016
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    103..117
                     /label=Csy4 site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease 
                     (Haurwitz et al., 2010)"
     regulatory      144..153
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             150..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             150..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
//
