LOCUS       sequence-254798-        4529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter to quantify elicitation of the strand annealing
            mediated repair pathway.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4529)
  AUTHORS   Wierson WA, Simone BW, WareJoncas Z, Mann C, Welker JM, Kar B, Emch 
            MJ, Friedberg I, Gendron WAC, Barry MA, Clark KJ, Dobbs DL, McGrail 
            MA, Ekker SC, Essner JJ
  TITLE     Expanding the CRISPR Toolbox with ErCas12a in Zebrafish and Human 
            Cells.
  JOURNAL   CRISPR J. 2019 Dec;2(6):417-433. doi: 10.1089/crispr.2019.0026. Epub
            2019 Nov 19.
  PUBMED    31742435
REFERENCE   2  (bases 1 to 4529)
  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"; doi: 
            "10.1089/crispr.2019.0026"; journalName: "CRISPR J"; date: 
            "2019-12"; volume: "2"; issue: "6"; pages: "417-433"
COMMENT     Sequence Label: pMini-CAAGs::RFP-DR48
FEATURES             Location/Qualifiers
     source          1..4529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(10..21,22..209)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(231..250)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(232..250)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        336..639
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        641..916
                     /label=chicken beta-actin promoter
     primer_bind     1172..1191
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1219..1228
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2040..2059)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2105..2160
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2159..2178)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        2521..2539
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(2559..2721,2722..2733)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2763..2780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2934..3522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3014..3033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3665..4474)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4388..4407
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4466..4485
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gctggatccc agaggtgtaa agtacttgag taattttact tgattactgt acttaagtat
       61 tatttttggg gatttttact ttacttgagt acaattaaaa atcaatactt ttacttttac
      121 ttaattacat ttttttagaa aaaaaagtac tttttactcc ttacaatttt atttacagtc
      181 aaaaagtact tattttttgg agatcacttc attctatttt cccttggtct ccctatagtg
      241 agtcgtatta gatcccggga ttactagttt aaacctagtt attaatagta atcaattacg
      301 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      361 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      421 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      481 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      541 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      601 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      661 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      721 taattatttt gtgcagcgat gggggcgggg gggggggggg cgcgcgccag gcggggcggg
      781 gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
      841 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
      901 aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc
      961 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
     1021 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt
     1081 tctgtggctg cgtgaaagcc ttgaggggct ccgggagcta gagcctctgc taaccatgtt
     1141 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
     1201 cattttggca aagaattagc caccatggtc gaggtgtcta agggcgagga actgattaag
     1261 gagaacatgc acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc
     1321 acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc
     1381 gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagc
     1441 agaaccttca tcaaccacac ccagggcatc cccgacttct ttaagcagtc cttccctgag
     1501 ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag
     1561 gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgatttag
     1621 gggaggcgtt cgggccacag cggcttgcgg agggatcaag cctccaggac ggctgcctca
     1681 tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga
     1741 aaacactcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc ggcctggaag
     1801 gcagaagcga catggccctg aagctcgtgg gcgggggcca cctgatctgc aacttcaaga
     1861 ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg
     1921 accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgagg
     1981 tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt aattgaagat
     2041 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     2101 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     2161 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     2221 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
     2281 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     2341 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     2401 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     2461 gctgtccctc ttctcttatg gagatccctc gacctgcata ccggtcaagc tagcgatatc
     2521 aattaaccct cactaaaggg agaccactct agttttcttt cttgctttta cttttacttc
     2581 cttaatactc aagtacaatt ttaatggagt acttttttac ttttactcaa gtaagattct
     2641 agccagatac ttttactttt aattgagtaa aattttccct aagtacttgt actttcactt
     2701 gagtaaaatt tttgagtact ttttacacct ctgctcggta ccatttaaat ctgttccgct
     2761 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2821 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2881 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2941 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3001 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3061 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aaccgtggcg
     3121 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3181 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3241 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3301 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3361 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3421 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3481 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3541 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3601 ttatcaaaaa ggatcttcac ctagatcctt tttgccagtg ttacaaccaa ttaaccaatt
     3661 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     3721 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     3781 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     3841 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     3901 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     3961 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     4021 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     4081 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     4141 gatattcttc taatacctgg aatgctgttt ttccggggat cgcagtggtg agtaaccatg
     4201 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     4261 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     4321 gaaacaactc tggcgcatcg ggcttcccat acaagcgata gattgtcgca cctgattgcc
     4381 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     4441 gcggcctcga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     4501 aagcagacag ttttattgtt catgatgca
//
