LOCUS       sequence-222309-        4070 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 4070)
  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 4070)
  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-eGFP
FEATURES             Location/Qualifiers
     source          1..4070
                     /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"
     primer_bind     complement(158..176)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     426..445
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     620..643
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2579..2599
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2603..2605,2606..2608,2609..3322)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2603..2605,2606..2608,2609..3319)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2648..2669)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2909..2928)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3256..3277
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3327..3551
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    3621..3702
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3826..3907
                     /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 ccatggtgag caagggcgag gagctgttca ccggggtggt
     2641 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2701 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2761 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2821 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2881 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2941 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3001 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3061 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     3121 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3181 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     3241 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3301 catggacgag ctgtacaagt aaagatctgt gccttctagt tgccagccat ctgttgtttg
     3361 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3421 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3481 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3541 gggctctatg gcttctgagg cggaaagaac caaagggcga attaattcat tgatgagttt
     3601 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3661 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3721 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     3781 tacaaatgtg gtcattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3841 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3901 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3961 ggttttttaa agcaagtaaa acctctacaa atgtggtcat tcagatcaat taaccctcac
     4021 ccgggctagc gatgcatggg agctctgacg tacgttgtct agcaaggaag
//
