LOCUS       sequence-134274-         816 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral introduction of BFP reporter (Point mutation H66 in mEGFP
            results in BFP expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 816)
  AUTHORS   Richardson CD, Ray GJ, DeWitt MA, Curie GL, Corn JE
  TITLE     Enhancing homology-directed genome editing by catalytically active 
            and inactive CRISPR-Cas9 using asymmetric donor DNA.
  JOURNAL   Nat Biotechnol. 2016 Jan 20. doi: 10.1038/nbt.3481.
  PUBMED    26789497
REFERENCE   2  (bases 1 to 816)
  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. 2016 Jan 20. doi: 10.1038/nbt.3481."
COMMENT     Sequence Label: BFP dest clone
FEATURES             Location/Qualifiers
     source          1..816
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    46..70
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(85..798,799..801,802..804))
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(130..151)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     479..498
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     738..759
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      801..810
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atccagaggt tgattgtcga gcggccgcca ctgtgctgga tatcaaccac tttgtacaag
       61 aaagctgggt ctagatatct cgagttactt gtacagctcg tccatgccga gagtgatccc
      121 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
      181 cagcttggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
      241 gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg
      301 gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg
      361 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
      421 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
      481 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
      541 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
      601 gccatgggtc agggtggtca cgagggtggg ccagggcacc ggcagcttgc cggtggtgca
      661 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
      721 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
      781 cagctcctcg cccttgctca ccatggtggc ggtacc
//
