LOCUS       sequence-222409-        1167 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 1167)
  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 1167)
  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-SV40
FEATURES             Location/Qualifiers
     source          1..1167
                     /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"
     polyA_signal    845..979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(869..888)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     923..942
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(1042..1137)
                     /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 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 gcgcctctag aactatagtg agtcgtatta
      841 cgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
      901 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
      961 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     1021 ggaggtgtgg gaggtttttt ccaactttat tatacaaagt tggcattata aaaaagcatt
     1081 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgga
     1141 gctccatggc gttgtctagc aaggaag
//
