LOCUS       sequence-222270-        3363 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 3363)
  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 3363)
  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-gcry-mRFP
FEATURES             Location/Qualifiers
     source          1..3363
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1909..1929
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1933..2610
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1933..2610
                     /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(2063..2081)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2331..2350
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2525..2548
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2620..2844
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2914..2995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3119..3200
                     /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 cggcgttcct agttctttaa
     1261 actgttacaa agtgtttgga ccagcaactg aattgcactg taaattacca catcagggtc
     1321 tccggggtca acgctgctaa gcaatggact ggaaaatttg tgacattgag tttttggaat
     1381 aactgggaat gctggataac tctttgggta aagctatgga acaaaagtac cagcatgaac
     1441 taagaaatga ctgtcacagc acaacatcaa acacaatgca agacaaaggt tctctgcttt
     1501 tgccttactt gacagaaaaa gtaagctcta ggttcatcgc atgcagacag cagcaataca
     1561 tgacatatat aaaggctggc tcttttctcc cttccattga actgaaactt ccactcagtc
     1621 agacttgcgg gggatccaca gcctgactaa ctcgacaaac ccccccaaac ctaaggtgag
     1681 ttgatcttta agctttttac attttcagct cgcatatatc aattcgcgcg cattgtcaca
     1741 ctagcaccca ctagcggtca gactgcagat tgcagcacga aacaggaagc tgactccaca
     1801 tggtcacatg ctcactgaag tgttgacttc cctgacagct gtgcactttc taaaccggtt
     1861 ttctcattca tttacagttc agccggaatt cacctcgaga ccatggctcc aaagaagaag
     1921 cgtaaggtaa ccatggcctc ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg
     1981 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2041 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
     2101 gcctgggaca tcctgtcccc tcagttccag tacggctcca aggcctacgt gaagcacccc
     2161 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2221 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     2281 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     2341 cagaagaaga ccatgggctg ggaggcctcc accgagcgga tgtaccccga ggacggcgcc
     2401 ctgaagggcg agatcaagat gaggctgaag ctgaaggacg gcggccacta cgacgccgag
     2461 gtcaagacca cctacatggc caagaagccc gtgcagctgc ccggcgccta caagaccgac
     2521 atcaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc
     2581 gccgagggcc gccactccac cggcgcctaa agctgtgatc tgtgccttct agttgccagc
     2641 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2701 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2761 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2821 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccaaaggg cgaattaatt
     2881 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2941 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3001 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     3061 caagtaaaac ctctacaaat gtggtcattg atgagtttgg acaaaccaca actagaatgc
     3121 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3181 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     3241 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt cattcagatc
     3301 aattaaccct cacccgggct agcgatgcat gggagctctg acgtacgttg tctagcaagg
     3361 aag
//
