LOCUS       sequence-99989-e         834 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Drosophila, p3xP3-EGFP.vas-int.NLS.attB was digested SpeI to get rid
            of the attB site. used for RMCE. CRISPR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 834)
  AUTHORS   Zhang X, Koolhaas WH, Schnorrer F
  TITLE     A Versatile Two-Step CRISPR- and RMCE-Based Strategy for Efficient 
            Genome Engineering in Drosophila.
  JOURNAL   G3 (Bethesda). 2014 Oct 15. pii: g3.114.013979. doi: 
            10.1534/g3.114.013979.
  PUBMED    25324299
REFERENCE   2  (bases 1 to 834)
  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: "G3 
            (Bethesda). 2014 Oct 15. pii: g3.114.013979. doi: 
            10.1534/g3.114.013979."
COMMENT     Sequence Label: p3xP3-EGFP.vas-int.NLS
FEATURES             Location/Qualifiers
     source          1..834
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      237..246
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(243..245,246..248,249..761)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(288..309)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(549..568)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 gggtacgtac cgggcccgga tctaattcaa ttagagacta attcaattag agctaattca
       61 attaggatcc aagcttatcg atttcgaacc ctcgaccgcc ggagtataaa tagaggcgct
      121 tcgtctacgg agcgacaatt caattcaaac aagcaaagtg aacacgtcgc taagcgaaag
      181 ctaagcaaat aaacaagcgc agctgaacaa gctaaacaat cggctcgaga ccggtcgcca
      241 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      301 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      361 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      421 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
      481 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      541 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      601 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
      661 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
      721 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
      781 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccg
//
