LOCUS       sequence-222258-        4024 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 4024)
  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 4024)
  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-cmlc2-TagBFP
FEATURES             Location/Qualifiers
     source          1..4024
                     /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             2546..2566
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2570..3271
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    3281..3505
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    3575..3656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3780..3861
                     /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 gcggcattca tccatccttt tcatccctca aatctctcat tcacgtcccc
     2041 ctccccatct gcacacttta tctcattttc caccctgctg gaatctgagc acttgtgcag
     2101 ttatcagggc tcctgtattt aggaggctct gggtgtccat gtaggggacg aacagaaaca
     2161 ctgcagacct ttatagaaga acaaatgata agagtcctca tacataaaga ctccattaga
     2221 aacgtcagtg acccaggagc ccagaccaac agcaaagcag acagtgaagg atccacagcc
     2281 tgactaactc gacaaacccc cccaaaccta aggtgagttg atctttaagc tttttacatt
     2341 ttcagctcgc atatatcaat tcgcgcgcat tgtcacacta gcacccacta gcggtcagac
     2401 tgcagattgc agcacgaaac aggaagctga ctccacatgg tcacatgctc actgaagtgt
     2461 tgacttccct gacagctgtg cactttctaa accggttttc tcattcattt acagttcagc
     2521 cggaattcac ctcgagacca tggctccaaa gaagaagcgt aaggtaacca tgagcgagct
     2581 gattaaggag aacatgcaca tgaagctgta catggagggc accgtggaca accatcactt
     2641 caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa
     2701 ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacta gcttcctcta
     2761 cggcagcaag accttcatca accacaccca gggcatcccc gacttcttca agcagtcctt
     2821 ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc
     2881 tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt
     2941 gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggccttcac
     3001 cgagacgctg taccccgctg acggcggcct ggaaggcaga aacgacatgg ccctgaagct
     3061 cgtgggcggg agccatctga tcgcaaacat caagaccaca tatagatcca agaaacccgc
     3121 taagaacctc aagatgcctg gcgtctacta tgtggactac agactggaaa gaatcaagga
     3181 ggccaacaac gagacctacg tcgagcagca cgaggtggca gtggccagat actgcgacct
     3241 ccctagcaaa ctggggcaca agctcaatta aagctgagat ctgtgccttc tagttgccag
     3301 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3361 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3421 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3481 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccaaagg gcgaattaat
     3541 tcattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3601 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     3661 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     3721 gcaagtaaaa cctctacaaa tgtggtcatt gatgagtttg gacaaaccac aactagaatg
     3781 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3841 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3901 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tcattcagat
     3961 caattaaccc tcacccgggc tagcgatgca tgggagctct gacgtacgtt gtctagcaag
     4021 gaag
//
