LOCUS       sequence-222315-        3922 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 3922)
  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 3922)
  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-Fuse-cmlc2-mRFP
FEATURES             Location/Qualifiers
     source          1..3922
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             31..705
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     CDS             31..705
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     CDS             2468..2488
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2492..3169
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             2492..3169
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     polyA_signal    3179..3403
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    3473..3554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3678..3759
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cttctaggag gaggtagcgg aggtagcgga gcctcctccg aggacgtcat caaggagttc
       61 atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      121 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
      181 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tccagtacgg ctccaaggcc
      241 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      301 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      361 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
      421 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gcggatgtac
      481 cccgaggacg gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa ggacggcggc
      541 cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca gctgcccggc
      601 gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta caccatcgtg
      661 gaacagtacg agcgcgccga gggccgccac tccaccggag cctaagacta gtaattaagt
      721 ctcagccacc gttaactgaa catgtcaaaa cctgtggaga ctgttgagat ttgatgttct
      781 gaaaagataa agcctataaa taaaatgttg cccaaatttc ctgcctgatg tttttctttg
      841 tctttgctac atggctttgc tgctcggatc ggctcactct gtgtatgcca cgttcacttt
      901 gtactctcct tctcacggta ggtttattat ttttagatgt gcagttagtt tctgtgaaat
      961 aacacaccac acactgatat tgtctgtgca ttgacttggt gagtgcacat tgtttttgat
     1021 cttgacatat ttatatttga ttgatcaggt gaactgtgtg aatctaaagt gctccataca
     1081 gatgttctgc attgaaaata ttctcatttt attagtggaa gtgagtgtat gccacatcca
     1141 atcaatttca gcaaacaccc cagtatgatt taatgcaaaa aaatgaaggt atcaaacacg
     1201 cattactact ttgcagttaa atatttaaca tttattccaa cacgaaaaaa agcagtaaat
     1261 aacactttga caaacacgtc aggacatctt atttttgtca ccctcacagg caatttagta
     1321 taatatatta tatatatata tatatcatat aataatattc agtataatat atatatatat
     1381 atcatattat aatattcagt ataatataaa acacaaacac atatatgtat aatataatat
     1441 aacattttta tttattgaga tgcctctatg gaccgtgtta taagaagtaa agatcaggag
     1501 aagtaaacat gaagtgtaat tatgaatact gatgttaaat taagctatga tgagttttca
     1561 ctgttaattt accatctcaa ttaaatgttg atgcctccat gaccaagtta agcagatgag
     1621 actgagacaa ctgtagaaga caagatgttc actttgctga atatagctgg cttgacagtt
     1681 atctatgact ctataaatat atatatattt ttttttttat aaaatgattt atttataact
     1741 atatatccat ttctcagaca ggtgcttcat atccctcact cccgtagctg tccatgctgg
     1801 atctgtcccc gttgttttta aaaagctaaa taagttatta acatgactgc atccagcgag
     1861 ccaaacctgt ctggtgtaca gctaccagag aagcttgaga tcctagtcac cggcggcatt
     1921 catccatcct tttcatccct caaatctctc attcacgtcc ccctccccat ctgcacactt
     1981 tatctcattt tccaccctgc tggaatctga gcacttgtgc agttatcagg gctcctgtat
     2041 ttaggaggct ctgggtgtcc atgtagggga cgaacagaaa cactgcagac ctttatagaa
     2101 gaacaaatga taagagtcct catacataaa gactccatta gaaacgtcag tgacccagga
     2161 gcccagacca acagcaaagc agacagtgaa ggatccacag cctgactaac tcgacaaacc
     2221 cccccaaacc taaggtgagt tgatctttaa gctttttaca ttttcagctc gcatatatca
     2281 attcgcgcgc attgtcacac tagcacccac tagcggtcag actgcagatt gcagcacgaa
     2341 acaggaagct gactccacat ggtcacatgc tcactgaagt gttgacttcc ctgacagctg
     2401 tgcactttct aaaccggttt tctcattcat ttacagttca gccggaattc acctcgagac
     2461 catggctcca aagaagaagc gtaaggtaac catggcctcc tccgaggacg tcatcaagga
     2521 gttcatgcgc ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt tcgagatcga
     2581 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     2641 gggcggcccc ctgcccttcg cctgggacat cctgtcccct cagttccagt acggctccaa
     2701 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     2761 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     2821 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     2881 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcggat
     2941 gtaccccgag gacggcgccc tgaagggcga gatcaagatg aggctgaagc tgaaggacgg
     3001 cggccactac gacgccgagg tcaagaccac ctacatggcc aagaagcccg tgcagctgcc
     3061 cggcgcctac aagaccgaca tcaagctgga catcacctcc cacaacgagg actacaccat
     3121 cgtggaacag tacgagcgcg ccgagggccg ccactccacc ggcgcctaaa gctgagatct
     3181 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3241 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3301 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3361 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3421 accaaagggc gaattaattc attgatgagt ttggacaaac cacaactaga atgcagtgaa
     3481 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     3541 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     3601 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtcattga tgagtttgga
     3661 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3721 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3781 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     3841 aaatgtggtc attcagatca attaaccctc acccgggcta gcgatgcatg ggagctctga
     3901 cgtacgttgt ctagcaagga ag
//
