LOCUS       sequence-222442-        1882 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 1882)
  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 1882)
  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-eGFP-CAAX-B-actin
FEATURES             Location/Qualifiers
     source          1..1882
                     /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             join(67..69,70..72,73..783)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(67..69,70..72,73..783)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(112..133)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(373..392)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     720..741
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ggatccggag ccacgaactt ctctctgtta aagcaagcag gagacgtgga agaaaacccc
       61 ggtcctatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      121 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      181 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      241 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
      301 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      361 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      421 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
      481 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
      541 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
      601 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
      661 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
      721 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
      781 aagaagctga accctcctga tgagagtggc cccggctgca tgagctgcaa gtgtgtgctc
      841 tcctgaggcg cgccacggac tgttaccact tcacgccgac tcaaactgcg cagagaaact
      901 tcaaacgaca acattggcat ggcttttgtt atttttggcg cttgactcag gatctaaaaa
      961 ctggaacggt gaaggtgacg gcaatgtttt ttggcaaata agcatccccg aagttctaca
     1021 atgcatctga ggactccatg ttttgttttc tttagtcatt ccaaatgttt gttaaatgca
     1081 ttgttccgaa acttatttgc ctctatgaag gctgcccagt aattgggagc atacttaaca
     1141 ttgtagtatt gtatgtaaat tatgtaacaa aacaatgtct gggtttttgt acttttcagc
     1201 cttaaaatct tgggtttatt tttttttctt tgttccaaaa aaacgaagct ttaccattca
     1261 agatgtaaag gtttccattc cccctgggca tattgtaaaa gctgtgtgga acgtggcggt
     1321 gccagacatt tggtggggcc aacctgtaca ctgactaatt caattccaat aaaagtgcac
     1381 atgtaagaca tcctacccct gtctcatttc ctgtcttggc tctgagtggc ttcaataaaa
     1441 atggcttttg tgaagttgca ccatgttaat tacatttcag cactttcaca ccaggttttt
     1501 gactaatttg aatttatcga agtttgacct tttttttttt ttttcttcaa gctaatttga
     1561 ggttatcaaa ccatttcatt ggaggaaact taatgtttaa aagcataaat tcgaaacatg
     1621 attcatttat ttaactgctt aatggtgaaa caaacttcaa gtaatacacc aagtgttccc
     1681 tcatatgaag gtagctaact gcaaacggcc ttaactttcc atcttaagtt gagctacata
     1741 aaggctttaa caggaaggaa ctgccagttt aaaagtcaaa ccaccatgac tgatataaat
     1801 tcttgtccat ataaccttgc gtttcacaca atatagctat ctactgctaa ataaattacc
     1861 atggcgttgt ctagcaagga ag
//
