LOCUS       sequence-222412-        1843 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 1843)
  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 1843)
  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-NLS-TagRFP-B-actin
FEATURES             Location/Qualifiers
     source          1..1843
                     /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             73..93
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             94..807
                     /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"
ORIGIN
        1 ggatccggag ccacgaactt ctctctgtta aagcaagcag gagacgtgga agaaaacccc
       61 ggtcctatgg ctccaaagaa gaagcgtaag gtaatggtgt ctaagggcga ggaactgatt
      121 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaacca ccacttcaag
      181 tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg
      241 gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt catgtacggc
      301 agcagaacct tcatcaacca cacccagggc atccccgact tctttaagca gtccttccct
      361 gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc
      421 caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac
      481 ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc caacaccgag
      541 atgctgtacc ccgctgacgg cggcctggaa ggcagaagcg acatggccct gaagctcgtg
      601 ggcgggggcc acctgatctg caacttcaag accacataca gatccaagaa acccgctaag
      661 aacctcaaga tgcccggcgt ctactatgtg gaccacagac tggaaagaat caaggaggcc
      721 gacaaagaga cctacgtcga gcagcacgag gtggctgtgg ccagatactg cgacctccct
      781 agcaaactgg ggcacaaact taattgaggc gcgccacgga ctgttaccac ttcacgccga
      841 ctcaaactgc gcagagaaac ttcaaacgac aacattggca tggcttttgt tatttttggc
      901 gcttgactca ggatctaaaa actggaacgg tgaaggtgac ggcaatgttt tttggcaaat
      961 aagcatcccc gaagttctac aatgcatctg aggactccat gttttgtttt ctttagtcat
     1021 tccaaatgtt tgttaaatgc attgttccga aacttatttg cctctatgaa ggctgcccag
     1081 taattgggag catacttaac attgtagtat tgtatgtaaa ttatgtaaca aaacaatgtc
     1141 tgggtttttg tacttttcag ccttaaaatc ttgggtttat ttttttttct ttgttccaaa
     1201 aaaacgaagc tttaccattc aagatgtaaa ggtttccatt ccccctgggc atattgtaaa
     1261 agctgtgtgg aacgtggcgg tgccagacat ttggtggggc caacctgtac actgactaat
     1321 tcaattccaa taaaagtgca catgtaagac atcctacccc tgtctcattt cctgtcttgg
     1381 ctctgagtgg cttcaataaa aatggctttt gtgaagttgc accatgttaa ttacatttca
     1441 gcactttcac accaggtttt tgactaattt gaatttatcg aagtttgacc tttttttttt
     1501 tttttcttca agctaatttg aggttatcaa accatttcat tggaggaaac ttaatgttta
     1561 aaagcataaa ttcgaaacat gattcattta tttaactgct taatggtgaa acaaacttca
     1621 agtaatacac caagtgttcc ctcatatgaa ggtagctaac tgcaaacggc cttaactttc
     1681 catcttaagt tgagctacat aaaggcttta acaggaagga actgccagtt taaaagtcaa
     1741 accaccatga ctgatataaa ttcttgtcca tataaccttg cgtttcacac aatatagcta
     1801 tctactgcta aataaattac catggcgttg tctagcaagg aag
//
