LOCUS       sequence-222285-        4075 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 4075)
  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 4075)
  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-gal4-VP16-gcry-TagBFP
FEATURES             Location/Qualifiers
     source          1..4075
                     /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..510
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             2597..2617
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2621..3322
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    3332..3556
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    3626..3707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3831..3912
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cttcccggag ccacgaactt ctctctgtta aagcaagcag gagacgtgga agaaaacccc
       61 ggtccttcta gaaagctact gtcttctatc gaacaagcat gcgatatttg ccgacttaaa
      121 aagctcaagt gctccaaaga aaaaccgaag tgcgccaagt gtctgaagaa caactgggag
      181 tgtcgctact ctcccaaaac caaaaggtct ccgctgacta gggcacatct gacagaagtg
      241 gaatcaaggc tagaaagact ggaacagcta tttctactga tttttcctcg agaagacctt
      301 gacatgattt tgaaaatgga ttctttacag gatataaaag cattgttaac aggattattt
      361 gtacaagata atgtgaataa agatgccgtc acagatagat tggcttcagt ggagactgat
      421 atgcctctaa cattgagaca gcatagaata agtgcgacat catcatcgga agagagtagt
      481 aacaaaggtc aaagacagtt gactgtatcg tcgaggtcga ccccgggaat acagatctct
      541 cgagccgccc ccccgaccga tgtcagcctg ggggacgagc tccacttaga cggcgaggac
      601 gtggcgatgg cgcatgccga cgcgctagac gatttcgatc tggacatgtt gggggacggg
      661 gattccccgg gtccgggatt taccccccac gactccgccc cctacggcgc tctggatatg
      721 taaaactagt aattaagtct cagccaccgt taactgaaca tgtcaaaacc tgtggagact
      781 gttgagattt gatgttctga aaagataaag cctataaata aaatgttgcc caaatttcct
      841 gcctgatgtt tttctttgtc tttgctacat ggctttgctg ctcggatcgg ctcactctgt
      901 gtatgccacg ttcactttgt actctccttc tcacggtagg tttattattt ttagatgtgc
      961 agttagtttc tgtgaaataa cacaccacac actgatattg tctgtgcatt gacttggtga
     1021 gtgcacattg tttttgatct tgacatattt atatttgatt gatcaggtga actgtgtgaa
     1081 tctaaagtgc tccatacaga tgttctgcat tgaaaatatt ctcattttat tagtggaagt
     1141 gagtgtatgc cacatccaat caatttcagc aaacacccca gtatgattta atgcaaaaaa
     1201 atgaaggtat caaacacgca ttactacttt gcagttaaat atttaacatt tattccaaca
     1261 cgaaaaaaag cagtaaataa cactttgaca aacacgtcag gacatcttat ttttgtcacc
     1321 ctcacaggca atttagtata atatattata tatatatata tatcatataa taatattcag
     1381 tataatatat atatatatat catattataa tattcagtat aatataaaac acaaacacat
     1441 atatgtataa tataatataa catttttatt tattgagatg cctctatgga ccgtgttata
     1501 agaagtaaag atcaggagaa gtaaacatga agtgtaatta tgaatactga tgttaaatta
     1561 agctatgatg agttttcact gttaatttac catctcaatt aaatgttgat gcctccatga
     1621 ccaagttaag cagatgagac tgagacaact gtagaagaca agatgttcac tttgctgaat
     1681 atagctggct tgacagttat ctatgactct ataaatatat atatattttt ttttttataa
     1741 aatgatttat ttataactat atatccattt ctcagacagg tgcttcatat ccctcactcc
     1801 cgtagctgtc catgctggat ctgtccccgt tgtttttaaa aagctaaata agttattaac
     1861 atgactgcat ccagcgagcc aaacctgtct ggtgtacagc taccagagaa gcttgagatc
     1921 ctagtcaccg gcgttcctag ttctttaaac tgttacaaag tgtttggacc agcaactgaa
     1981 ttgcactgta aattaccaca tcagggtctc cggggtcaac gctgctaagc aatggactgg
     2041 aaaatttgtg acattgagtt tttggaataa ctgggaatgc tggataactc tttgggtaaa
     2101 gctatggaac aaaagtacca gcatgaacta agaaatgact gtcacagcac aacatcaaac
     2161 acaatgcaag acaaaggttc tctgcttttg ccttacttga cagaaaaagt aagctctagg
     2221 ttcatcgcat gcagacagca gcaatacatg acatatataa aggctggctc ttttctccct
     2281 tccattgaac tgaaacttcc actcagtcag acttgcgggg gatccacagc ctgactaact
     2341 cgacaaaccc ccccaaacct aaggtgagtt gatctttaag ctttttacat tttcagctcg
     2401 catatatcaa ttcgcgcgca ttgtcacact agcacccact agcggtcaga ctgcagattg
     2461 cagcacgaaa caggaagctg actccacatg gtcacatgct cactgaagtg ttgacttccc
     2521 tgacagctgt gcactttcta aaccggtttt ctcattcatt tacagttcag ccggaattca
     2581 cctcgagacc atggctccaa agaagaagcg taaggtaacc atgagcgagc tgattaagga
     2641 gaacatgcac atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac
     2701 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga
     2761 gggcggccct ctccccttcg ccttcgacat cctggctact agcttcctct acggcagcaa
     2821 gaccttcatc aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg
     2881 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga
     2941 caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac
     3001 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacgct
     3061 gtaccccgct gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg
     3121 gagccatctg atcgcaaaca tcaagaccac atatagatcc aagaaacccg ctaagaacct
     3181 caagatgcct ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa
     3241 cgagacctac gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa
     3301 actggggcac aagctcaatt aaagctgaga tctgtgcctt ctagttgcca gccatctgtt
     3361 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3421 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3481 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3541 gcggtgggct ctatggcttc tgaggcggaa agaaccaaag ggcgaattaa ttcattgatg
     3601 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     3661 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3721 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     3781 acctctacaa atgtggtcat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3841 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3901 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     3961 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtcattcaga tcaattaacc
     4021 ctcacccggg ctagcgatgc atgggagctc tgacgtacgt tgtctagcaa ggaag
//
