LOCUS       sequence-222448-        1298 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 1298)
  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 1298)
  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: Control-cx43.4-48H-pGTag
FEATURES             Location/Qualifiers
     source          1..1298
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             60..116
                     /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             117..827
                     /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    928..1062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(952..971)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1006..1025
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(1125..1220)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gttttcttac gcggttgttg gatgaaatct ccaaccactc caccttcgtg ggatccggag
       61 ccacgaactt ctctctgtta aagcaagcag gagacgtgga agaaaacccc ggtcctatgg
      121 tgtctaaggg cgaggaactg attaaggaga acatgcacat gaagctgtac atggagggca
      181 ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca
      241 cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc
      301 tggctaccag cttcatgtac ggcagcagaa ccttcatcaa ccacacccag ggcatccccg
      361 acttctttaa gcagtccttc cctgagggct tcacatggga gagagtcacc acatacgaag
      421 acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca
      481 acgtcaagat cagaggggtg aacttcccat ccaacggccc tgtgatgcag aagaaaacac
      541 tcggctggga ggccaacacc gagatgctgt accccgctga cggcggcctg gaaggcagaa
      601 gcgacatggc cctgaagctc gtgggcgggg gccacctgat ctgcaacttc aagaccacat
      661 acagatccaa gaaacccgct aagaacctca agatgcccgg cgtctactat gtggaccaca
      721 gactggaaag aatcaaggag gccgacaaag agacctacgt cgagcagcac gaggtggctg
      781 tggccagata ctgcgacctc cctagcaaac tggggcacaa acttaataag ctgaaccctc
      841 ctgatgagag tggccccggc tgcatgagct gcaagtgtgt gctctcctga ggcgcgcctc
      901 tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt
      961 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1021 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1081 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttccaactt tattatacaa
     1141 agttggcatt ataaaaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1201 tcaaaataaa atcattattt ggagctccat ggcgttgtct agcaaggaag tgggcaagat
     1261 atggctcacg ttattcatca tcttccgcat tgttttga
//
