LOCUS       sequence-222249-        4135 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 4135)
  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 4135)
  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-TagRFP-gcry-TagBFP
FEATURES             Location/Qualifiers
     source          1..4135
                     /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..783
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDM
                     ALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVA
                     RYCDLPSKLGHKLN"
     CDS             2657..2677
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2681..3382
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    3392..3616
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    3686..3767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3891..3972
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cttcccggag ccacgaactt ctctctgtta aagcaagcag gagacgtgga agaaaacccc
       61 ggtcctctcg aggtgtctaa gggcgaggaa ctgattaagg agaacatgca catgaagctg
      121 tacatggagg gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag
      181 ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc
      241 gccttcgaca tcctggctac cagcttcatg tacggcagca gaaccttcat caaccacacc
      301 cagggcatcc ccgacttctt taagcagtcc ttccctgagg gcttcacatg ggagagagtc
      361 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc
      421 tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc catccaacgg ccctgtgatg
      481 cagaagaaaa cactcggctg ggaggccaac accgagatgc tgtaccccgc tgacggcggc
      541 ctggaaggca gaagcgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac
      601 ttcaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac
      661 tatgtggacc acagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag
      721 cacgaggtgg ctgtggccag atactgcgac ctccctagca aactggggca caaacttaat
      781 tgaaactagt aattaagtct cagccaccgt taactgaaca tgtcaaaacc tgtggagact
      841 gttgagattt gatgttctga aaagataaag cctataaata aaatgttgcc caaatttcct
      901 gcctgatgtt tttctttgtc tttgctacat ggctttgctg ctcggatcgg ctcactctgt
      961 gtatgccacg ttcactttgt actctccttc tcacggtagg tttattattt ttagatgtgc
     1021 agttagtttc tgtgaaataa cacaccacac actgatattg tctgtgcatt gacttggtga
     1081 gtgcacattg tttttgatct tgacatattt atatttgatt gatcaggtga actgtgtgaa
     1141 tctaaagtgc tccatacaga tgttctgcat tgaaaatatt ctcattttat tagtggaagt
     1201 gagtgtatgc cacatccaat caatttcagc aaacacccca gtatgattta atgcaaaaaa
     1261 atgaaggtat caaacacgca ttactacttt gcagttaaat atttaacatt tattccaaca
     1321 cgaaaaaaag cagtaaataa cactttgaca aacacgtcag gacatcttat ttttgtcacc
     1381 ctcacaggca atttagtata atatattata tatatatata tatcatataa taatattcag
     1441 tataatatat atatatatat catattataa tattcagtat aatataaaac acaaacacat
     1501 atatgtataa tataatataa catttttatt tattgagatg cctctatgga ccgtgttata
     1561 agaagtaaag atcaggagaa gtaaacatga agtgtaatta tgaatactga tgttaaatta
     1621 agctatgatg agttttcact gttaatttac catctcaatt aaatgttgat gcctccatga
     1681 ccaagttaag cagatgagac tgagacaact gtagaagaca agatgttcac tttgctgaat
     1741 atagctggct tgacagttat ctatgactct ataaatatat atatattttt ttttttataa
     1801 aatgatttat ttataactat atatccattt ctcagacagg tgcttcatat ccctcactcc
     1861 cgtagctgtc catgctggat ctgtccccgt tgtttttaaa aagctaaata agttattaac
     1921 atgactgcat ccagcgagcc aaacctgtct ggtgtacagc taccagagaa gcttgagatc
     1981 ctagtcaccg gcgttcctag ttctttaaac tgttacaaag tgtttggacc agcaactgaa
     2041 ttgcactgta aattaccaca tcagggtctc cggggtcaac gctgctaagc aatggactgg
     2101 aaaatttgtg acattgagtt tttggaataa ctgggaatgc tggataactc tttgggtaaa
     2161 gctatggaac aaaagtacca gcatgaacta agaaatgact gtcacagcac aacatcaaac
     2221 acaatgcaag acaaaggttc tctgcttttg ccttacttga cagaaaaagt aagctctagg
     2281 ttcatcgcat gcagacagca gcaatacatg acatatataa aggctggctc ttttctccct
     2341 tccattgaac tgaaacttcc actcagtcag acttgcgggg gatccacagc ctgactaact
     2401 cgacaaaccc ccccaaacct aaggtgagtt gatctttaag ctttttacat tttcagctcg
     2461 catatatcaa ttcgcgcgca ttgtcacact agcacccact agcggtcaga ctgcagattg
     2521 cagcacgaaa caggaagctg actccacatg gtcacatgct cactgaagtg ttgacttccc
     2581 tgacagctgt gcactttcta aaccggtttt ctcattcatt tacagttcag ccggaattca
     2641 cctcgagacc atggctccaa agaagaagcg taaggtaacc atgagcgagc tgattaagga
     2701 gaacatgcac atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac
     2761 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga
     2821 gggcggccct ctccccttcg ccttcgacat cctggctact agcttcctct acggcagcaa
     2881 gaccttcatc aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg
     2941 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga
     3001 caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac
     3061 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacgct
     3121 gtaccccgct gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg
     3181 gagccatctg atcgcaaaca tcaagaccac atatagatcc aagaaacccg ctaagaacct
     3241 caagatgcct ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa
     3301 cgagacctac gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa
     3361 actggggcac aagctcaatt aaagctgaga tctgtgcctt ctagttgcca gccatctgtt
     3421 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3481 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3541 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3601 gcggtgggct ctatggcttc tgaggcggaa agaaccaaag ggcgaattaa ttcattgatg
     3661 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     3721 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3781 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     3841 acctctacaa atgtggtcat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3901 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3961 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     4021 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtcattcaga tcaattaacc
     4081 ctcacccggg ctagcgatgc atgggagctc tgacgtacgt tgtctagcaa ggaag
//
