LOCUS       sequence-204849-        1353 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Donor scafold with dTOMATO positive selection module, and tagBFP 
            negative selection module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1353)
  AUTHORS   Arias-Fuenzalida J, Jarazo J, Qing X, Walter J, Gomez-Giro G, 
            Nickels SL, Zaehres H, Scholer HR, Schwamborn JC
  TITLE     FACS-Assisted CRISPR-Cas9 Genome Editing Facilitates Parkinson's 
            Disease Modeling.
  JOURNAL   Stem Cell Reports. 2017 Nov 14;9(5):1423-1431. doi: 
            10.1016/j.stemcr.2017.08.026. Epub 2017 Oct 5.
  PUBMED    28988985
REFERENCE   2  (bases 1 to 1353)
  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: 
            "10.1016/j.stemcr.2017.08.026"; journalName: "Stem Cell Reports"; 
            date: "2017-11-14- 14"; volume: "9"; issue: "5"; pages: "1423-1431"
COMMENT     Sequence Label: pDONOR-tagBFP-PSM-dTOMATO
FEATURES             Location/Qualifiers
     source          1..1353
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..702
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(134..152)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     402..421
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     596..619
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             703..756
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             757..1353
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     1250..1270
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 atggtgagca agggcgagga ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag
       61 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
      121 ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac
      181 atcctgtccc cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc
      241 cccgattaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
      301 gaggacggcg gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc
      361 tacaaggtga agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag
      421 accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc
      481 gagatccacc aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagacc
      541 atctacatgg ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg
      601 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc
      661 cgccaccacc tgttcctgta cggcatggac gagctgtaca aggagggcag aggaagtctt
      721 ctaacatgcg gtgacgtgga ggagaatccc ggccctaccg agtacaagcc cacggtgcgc
      781 ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac
      841 taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg
      901 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
      961 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     1021 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     1081 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     1141 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     1201 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     1261 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     1321 acctggtgca tgacccgcaa gcccggtgcc tga
//
