LOCUS       sequence-222273-        3400 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 3400)
  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 3400)
  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-gcry-eGFP
FEATURES             Location/Qualifiers
     source          1..3400
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1909..1929
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1933..1935,1936..1938,1939..2652)
                     /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(1933..1935,1936..1938,1939..2649)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1978..1999)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2239..2258)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2586..2607
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2657..2881
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2951..3032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3156..3237
                     /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 cggcgttcct agttctttaa
     1261 actgttacaa agtgtttgga ccagcaactg aattgcactg taaattacca catcagggtc
     1321 tccggggtca acgctgctaa gcaatggact ggaaaatttg tgacattgag tttttggaat
     1381 aactgggaat gctggataac tctttgggta aagctatgga acaaaagtac cagcatgaac
     1441 taagaaatga ctgtcacagc acaacatcaa acacaatgca agacaaaggt tctctgcttt
     1501 tgccttactt gacagaaaaa gtaagctcta ggttcatcgc atgcagacag cagcaataca
     1561 tgacatatat aaaggctggc tcttttctcc cttccattga actgaaactt ccactcagtc
     1621 agacttgcgg gggatccaca gcctgactaa ctcgacaaac ccccccaaac ctaaggtgag
     1681 ttgatcttta agctttttac attttcagct cgcatatatc aattcgcgcg cattgtcaca
     1741 ctagcaccca ctagcggtca gactgcagat tgcagcacga aacaggaagc tgactccaca
     1801 tggtcacatg ctcactgaag tgttgacttc cctgacagct gtgcactttc taaaccggtt
     1861 ttctcattca tttacagttc agccggaatt cacctcgaga ccatggctcc aaagaagaag
     1921 cgtaaggtaa ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1981 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2041 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2101 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2161 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2221 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2281 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2341 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2401 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     2461 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2521 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     2581 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2641 ctgtacaagt aaagatctgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2701 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2761 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2821 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2881 gcttctgagg cggaaagaac caaagggcga attaattcat tgatgagttt ggacaaacca
     2941 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3001 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3061 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3121 gtcattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3181 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3241 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     3301 agcaagtaaa acctctacaa atgtggtcat tcagatcaat taaccctcac ccgggctagc
     3361 gatgcatggg agctctgacg tacgttgtct agcaaggaag
//
