LOCUS       sequence-222282-        3289 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 3289)
  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 3289)
  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: pPRISM-Stop-cmlc2-eGFP
FEATURES             Location/Qualifiers
     source          1..3289
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1798..1818
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1822..1824,1825..1827,1828..2541)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1822..1824,1825..1827,1828..2538)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1867..1888)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2128..2147)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2475..2496
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2546..2770
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2840..2921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3045..3126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cttctataat gataggtagt aatgattagt aatgagacta gtaattaagt ctcagccacc
       61 gttaactgaa catgtcaaaa cctgtggaga ctgttgagat ttgatgttct gaaaagataa
      121 agcctataaa taaaatgttg cccaaatttc ctgcctgatg tttttctttg tctttgctac
      181 atggctttgc tgctcggatc ggctcactct gtgtatgcca cgttcacttt gtactctcct
      241 tctcacggta ggtttattat ttttagatgt gcagttagtt tctgtgaaat aacacaccac
      301 acactgatat tgtctgtgca ttgacttggt gagtgcacat tgtttttgat cttgacatat
      361 ttatatttga ttgatcaggt gaactgtgtg aatctaaagt gctccataca gatgttctgc
      421 attgaaaata ttctcatttt attagtggaa gtgagtgtat gccacatcca atcaatttca
      481 gcaaacaccc cagtatgatt taatgcaaaa aaatgaaggt atcaaacacg cattactact
      541 ttgcagttaa atatttaaca tttattccaa cacgaaaaaa agcagtaaat aacactttga
      601 caaacacgtc aggacatctt atttttgtca ccctcacagg caatttagta taatatatta
      661 tatatatata tatatcatat aataatattc agtataatat atatatatat atcatattat
      721 aatattcagt ataatataaa acacaaacac atatatgtat aatataatat aacattttta
      781 tttattgaga tgcctctatg gaccgtgtta taagaagtaa agatcaggag aagtaaacat
      841 gaagtgtaat tatgaatact gatgttaaat taagctatga tgagttttca ctgttaattt
      901 accatctcaa ttaaatgttg atgcctccat gaccaagtta agcagatgag actgagacaa
      961 ctgtagaaga caagatgttc actttgctga atatagctgg cttgacagtt atctatgact
     1021 ctataaatat atatatattt ttttttttat aaaatgattt atttataact atatatccat
     1081 ttctcagaca ggtgcttcat atccctcact cccgtagctg tccatgctgg atctgtcccc
     1141 gttgttttta aaaagctaaa taagttatta acatgactgc atccagcgag ccaaacctgt
     1201 ctggtgtaca gctaccagag aagcttgaga tcctagtcac cggcggcatt catccatcct
     1261 tttcatccct caaatctctc attcacgtcc ccctccccat ctgcacactt tatctcattt
     1321 tccaccctgc tggaatctga gcacttgtgc agttatcagg gctcctgtat ttaggaggct
     1381 ctgggtgtcc atgtagggga cgaacagaaa cactgcagac ctttatagaa gaacaaatga
     1441 taagagtcct catacataaa gactccatta gaaacgtcag tgacccagga gcccagacca
     1501 acagcaaagc agacagtgaa ggatccacag cctgactaac tcgacaaacc cccccaaacc
     1561 taaggtgagt tgatctttaa gctttttaca ttttcagctc gcatatatca attcgcgcgc
     1621 attgtcacac tagcacccac tagcggtcag actgcagatt gcagcacgaa acaggaagct
     1681 gactccacat ggtcacatgc tcactgaagt gttgacttcc ctgacagctg tgcactttct
     1741 aaaccggttt tctcattcat ttacagttca gccggaattc acctcgagac catggctcca
     1801 aagaagaagc gtaaggtaac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1861 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1921 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1981 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2041 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2101 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2161 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2221 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2281 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2341 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2401 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2461 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2521 atggacgagc tgtacaagta aagatctgtg ccttctagtt gccagccatc tgttgtttgc
     2581 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2641 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2701 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2761 ggctctatgg cttctgaggc ggaaagaacc aaagggcgaa ttaattcatt gatgagtttg
     2821 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2881 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2941 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     3001 acaaatgtgg tcattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     3061 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     3121 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     3181 gttttttaaa gcaagtaaaa cctctacaaa tgtggtcatt cagatcaatt aaccctcacc
     3241 cgggctagcg atgcatggga gctctgacgt acgttgtcta gcaaggaag
//
