LOCUS       sequence-222306-        4033 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 4033)
  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 4033)
  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-gcry-mRFP
FEATURES             Location/Qualifiers
     source          1..4033
                     /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             2579..2599
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2603..3280
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             2603..3280
                     /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    3290..3514
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    3584..3665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3789..3870
                     /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 cggcgttcct
     1921 agttctttaa actgttacaa agtgtttgga ccagcaactg aattgcactg taaattacca
     1981 catcagggtc tccggggtca acgctgctaa gcaatggact ggaaaatttg tgacattgag
     2041 tttttggaat aactgggaat gctggataac tctttgggta aagctatgga acaaaagtac
     2101 cagcatgaac taagaaatga ctgtcacagc acaacatcaa acacaatgca agacaaaggt
     2161 tctctgcttt tgccttactt gacagaaaaa gtaagctcta ggttcatcgc atgcagacag
     2221 cagcaataca tgacatatat aaaggctggc tcttttctcc cttccattga actgaaactt
     2281 ccactcagtc agacttgcgg gggatccaca gcctgactaa ctcgacaaac ccccccaaac
     2341 ctaaggtgag ttgatcttta agctttttac attttcagct cgcatatatc aattcgcgcg
     2401 cattgtcaca ctagcaccca ctagcggtca gactgcagat tgcagcacga aacaggaagc
     2461 tgactccaca tggtcacatg ctcactgaag tgttgacttc cctgacagct gtgcactttc
     2521 taaaccggtt ttctcattca tttacagttc agccggaatt cacctcgaga ccatggctcc
     2581 aaagaagaag cgtaaggtaa ccatggcctc ctccgaggac gtcatcaagg agttcatgcg
     2641 cttcaaggtg cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     2701 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc
     2761 cctgcccttc gcctgggaca tcctgtcccc tcagttccag tacggctcca aggcctacgt
     2821 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     2881 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     2941 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     3001 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgga tgtaccccga
     3061 ggacggcgcc ctgaagggcg agatcaagat gaggctgaag ctgaaggacg gcggccacta
     3121 cgacgccgag gtcaagacca cctacatggc caagaagccc gtgcagctgc ccggcgccta
     3181 caagaccgac atcaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     3241 gtacgagcgc gccgagggcc gccactccac cggcgcctaa agctgagatc tgtgccttct
     3301 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3361 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3421 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3481 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccaaaggg
     3541 cgaattaatt cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     3601 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     3661 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     3721 ttttttaaag caagtaaaac ctctacaaat gtggtcattg atgagtttgg acaaaccaca
     3781 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3841 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3901 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     3961 cattcagatc aattaaccct cacccgggct agcgatgcat gggagctctg acgtacgttg
     4021 tctagcaagg aag
//
