LOCUS       sequence-222279-        3252 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 3252)
  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 3252)
  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-cmlc2-mRFP
FEATURES             Location/Qualifiers
     source          1..3252
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1798..1818
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1822..2499
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1822..2499
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1952..1970)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2220..2239
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2414..2437
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2509..2733
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2803..2884
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3008..3089
                     /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 cggcggcatt catccatcct
     1261 tttcatccct caaatctctc attcacgtcc ccctccccat ctgcacactt tatctcattt
     1321 tccaccctgc tggaatctga gcacttgtgc agttatcagg gctcctgtat ttaggaggct
     1381 ctgggtgtcc atgtagggga cgaacagaaa cactgcagac ctttatagaa gaacaaatga
     1441 taagagtcct catacataaa gactccatta gaaacgtcag tgacccagga gcccagacca
     1501 acagcaaagc agacagtgaa ggatccacag cctgactaac tcgacaaacc cccccaaacc
     1561 taaggtgagt tgatctttaa gctttttaca ttttcagctc gcatatatca attcgcgcgc
     1621 attgtcacac tagcacccac tagcggtcag actgcagatt gcagcacgaa acaggaagct
     1681 gactccacat ggtcacatgc tcactgaagt gttgacttcc ctgacagctg tgcactttct
     1741 aaaccggttt tctcattcat ttacagttca gccggaattc acctcgagac catggctcca
     1801 aagaagaagc gtaaggtaac catggcctcc tccgaggacg tcatcaagga gttcatgcgc
     1861 ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1921 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     1981 ctgcccttcg cctgggacat cctgtcccct cagttccagt acggctccaa ggcctacgtg
     2041 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     2101 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     2161 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     2221 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcggat gtaccccgag
     2281 gacggcgccc tgaagggcga gatcaagatg aggctgaagc tgaaggacgg cggccactac
     2341 gacgccgagg tcaagaccac ctacatggcc aagaagcccg tgcagctgcc cggcgcctac
     2401 aagaccgaca tcaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2461 tacgagcgcg ccgagggccg ccactccacc ggcgcctaaa gctgagatct gtgccttcta
     2521 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2581 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2641 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2701 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accaaagggc
     2761 gaattaattc attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2821 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2881 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2941 tttttaaagc aagtaaaacc tctacaaatg tggtcattga tgagtttgga caaaccacaa
     3001 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3061 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3121 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggtc
     3181 attcagatca attaaccctc acccgggcta gcgatgcatg ggagctctga cgtacgttgt
     3241 ctagcaagga ag
//
