LOCUS       sequence-222421-        1200 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Donor for precise CRISPR directed genomic integration using the 
            GeneWeld method.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1200)
  AUTHORS   Wierson WA, Welker JM, Almeida MP, Mann CM, Webster DA, Torrie ME, 
            Weiss TJ, Kambakam S, Vollbrecht MK, Lan M, McKeighan KC, Levey J, 
            Ming Z, Wehmeier A, Mikelson CS, Haltom JA, Kwan KM, Chien CB, 
            Balciunas D, Ekker SC, Clark KJ, Webber BR, Moriarity BS, Solin SL, 
            Carlson DF, Dobbs DL, McGrail M, Essner J
  TITLE     Efficient targeted integration directed by short homology in 
            zebrafish and mammalian cells.
  JOURNAL   Elife. 2020 May 15;9. pii: 53968. doi: 10.7554/eLife.53968.
  PUBMED    32412410
REFERENCE   2  (bases 1 to 1200)
  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: "Elife."; 
            date: "2020-05-15"; volume: "9. pii: 53968. doi"; pages: " 
            10.7554/eLife.53968"
COMMENT     Sequence Label: pGTag-TagRFP-CAAX-SV40
FEATURES             Location/Qualifiers
     source          1..1200
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             10..66
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             67..777
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    878..1012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(902..921)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     956..975
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(1075..1170)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggatccggag ccacgaactt ctctctgtta aagcaagcag gagacgtgga agaaaacccc
       61 ggtcctatgg tgtctaaggg cgaggaactg attaaggaga acatgcacat gaagctgtac
      121 atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc
      181 tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc
      241 ttcgacatcc tggctaccag cttcatgtac ggcagcagaa ccttcatcaa ccacacccag
      301 ggcatccccg acttctttaa gcagtccttc cctgagggct tcacatggga gagagtcacc
      361 acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc
      421 ctcatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc tgtgatgcag
      481 aagaaaacac tcggctggga ggccaacacc gagatgctgt accccgctga cggcggcctg
      541 gaaggcagaa gcgacatggc cctgaagctc gtgggcgggg gccacctgat ctgcaacttc
      601 aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg cgtctactat
      661 gtggaccaca gactggaaag aatcaaggag gccgacaaag agacctacgt cgagcagcac
      721 gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcacaa acttaataag
      781 ctgaaccctc ctgatgagag tggccccggc tgcatgagct gcaagtgtgt gctctcctga
      841 ggcgcgcctc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
      901 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
      961 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1021 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttccaactt
     1081 tattatacaa agttggcatt ataaaaaagc attgcttatc aatttgttgc aacgaacagg
     1141 tcactatcag tcaaaataaa atcattattt ggagctccat ggcgttgtct agcaaggaag
//
