LOCUS       sequence-99361-e        1090 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GFP reporter for CRISPR/CAS9 and gRNA_AAVS1-T2 activities.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1090)
  AUTHORS   Zhang JH, Pandey M, Kahler JF, Loshakov A, Harris B, Dagur PK, Mo Y,
            Simonds WF
  TITLE     Improving the specificity and efficacy of CRISPR/CAS9 and gRNA 
            through target specific DNA reporter.
  JOURNAL   J Biotechnol. 2014 Sep 2. pii: S0168-1656(14)00838-4. doi: 
            10.1016/j.jbiotec.2014.08.033.
  PUBMED    25193712
REFERENCE   2  (bases 1 to 1090)
  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: "J 
            Biotechnol. 2014 Sep 2. pii: S0168-1656(14)00838-4. doi: 
            10.1016/j.jbiotec.2014.08.033."
COMMENT     Sequence Label: psAAVS1-T2
FEATURES             Location/Qualifiers
     source          1..1090
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      101..110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(107..109,110..112,113..823)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(107..109,110..112,113..823)
                     /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(152..173)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(413..432)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     760..781
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    837..893
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(1054..1073)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ctatataagc agagctggtt tagtgaaccg tcagatcctg tatgagaccg gggccactag
       61 ggacaggatt ggggtctctg tttcgctagc gctaccggtc gccaccatgg tgagcaaggg
      121 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      181 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      241 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      301 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
      361 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
      421 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
      481 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
      541 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
      601 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
      661 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
      721 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
      781 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggcc ggactcagat
      841 ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat
      901 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
      961 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     1021 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1081 aagcattttt
//
