LOCUS       sequence-252142-        3200 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Donor to generate i-WT-r system .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3200)
  AUTHORS   Oksuz O, Narendra V, Lee CH, Descostes N, LeRoy G, Raviram R, 
            Blumenberg L, Karch K, Rocha PP, Garcia BA, Skok JA, Reinberg D
  TITLE     Capturing the Onset of PRC2-Mediated Repressive Domain Formation.
  JOURNAL   Mol Cell. 2018 Jun 21;70(6):1149-1162.e5. doi: 
            10.1016/j.molcel.2018.05.023.
  PUBMED    29932905
REFERENCE   2  (bases 1 to 3200)
  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.molcel.2018.05"; journalName: "Mol Cell"; date: 
            "2018-06-21- 21"; volume: "70"; issue: "6"; pages: "1149-1162.e5"
COMMENT     Sequence Label: A365Y-GFP Donor for iWTr EED
FEATURES             Location/Qualifiers
     source          1..3200
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     34..50
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     35..51
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(654..687)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    688..895
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     884..901
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(929..1639,1640..1642))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(929..1639,1640..1642))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(971..992)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1320..1339
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1579..1600
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1643..1696)
                     /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             complement(1712..1738)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1748..1771)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    2316..2349
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3178..3194)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3178..3194)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 cactataggg cgaattgggt accgggcccc ccctcgaggt cgacggtatc gataagcttg
       61 atgttgaaac agggtgtctc accatctacc caagccagcc ttaaacttgc tatagcccat
      121 gctaacttca aacttctcct cagtctccca agtacagaaa ctatgtgaca cttaatttga
      181 gaatcacctc tttatgtaca gcttatctgg cttacttttt agtaatgtgt atgtgtgtct
      241 gtgtatgcag gtgcttgtgt gctgctgcac tgtgtggaag tcagaggcta ccttcgaggg
      301 tcttttctct tttgcataga atctgggcat tgcactcagc ttgcaaggct tgtgcatgga
      361 ggacttttgc ctgtgtgcca tcttgctggc ccgtgtggag catatttctt tgtgtagcag
      421 ctttagaggt tgctttgctt tagatgtcat atttcgaata tgtaaccttt cagactacca
      481 tatgttggca tattggtaat cagaggaagt ataggaccct tcccttgtca ttgctttgtc
      541 cccagcttat gatctaatta tcttaagtat ttaacttata tttgagaacc atattgggtt
      601 gtgtaatttt tgcagtgaaa caaatttgga aagtgaacat gggactgtct gctataactt
      661 cgtatagcat acattatacg aacggtatcc ccagcatgcc tgctattctc ttcccaatcc
      721 tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa
      781 tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc
      841 aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag
      901 gctgatcagc gagctctagt tagaattcct tgtacagctc gtccatgccg agagtgatcc
      961 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     1021 tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     1081 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc
     1141 ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt
     1201 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     1261 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     1321 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     1381 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca
     1441 cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     1501 agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     1561 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     1621 acagctcctc gcccttgctc actgggccag gattctcctc gacgtcaccg catgttagca
     1681 gacttcctct gccctctcca ctgccgaatt cagcgtaatc tggcacatcg tatgggtatc
     1741 cggagccctt atcgtcgtca tccttgtaat cgcctgatcc tcgaagtcga tcccatcgcc
     1801 aaatgctggc atcatcgcag acagctatga ggatgctgct atccctactg aaactggttt
     1861 gtcgaatagc cgcgccacat ttatgatggg tcagtgttgt gcatttggct ttatgaggat
     1921 cttctacttc taaatcccaa acatacagtt tgccaacctg attgcccaat gcaagcatct
     1981 tttgccagaa atccatagaa aacctcatgt accaaatgtc acactggctg taatcaaatc
     2041 gcccaagaat agtcacatta gactcactag gtttaatttt atctatatca tcctccattt
     2101 tgccaggttt ccagcataca atggcatttt cacaagacta aaagagaaat aagagacaca
     2161 taattagaac tgcatttcaa aaagtaaaat taaaactgtt accattttag aatagctcag
     2221 agagcctact ttccaagact ataaactgtt ttgcaagtca aggcttacac ctgaggacaa
     2281 gaaatctgac tagcaataga attaagcaat tcagaataac ttcgtataat gtatgctata
     2341 cgaacggtat cctgcagcta ttcattagag gtctactagt aacaaattct cttgtttttt
     2401 tcttagtttt tctctctact ctagaatata tctattaaat acatatttct ggctttacag
     2461 tgccacccct ctccccagac ttgaaaagta tttgcccttt ctctggcctg catagcttct
     2521 aatgtgtaat attttgtcat taacctccca accccaaact tatttgtagt tcattgatct
     2581 ggaatatgtg gtggaattat atattagaga aatattttgt ctcatacttt gaactataac
     2641 actaatgaca ctgttttctt cccttatttt gagacactaa tcctcattat taatatatta
     2701 gatatttttg gtgcgcacac acagtgctgg tcttattttg tccctactct aacactaaga
     2761 tttttgtcag agatcttgtc agatggctgt cttcctcctc catttgtttc ttgtttgttg
     2821 gatgatattc tatcgggttg tcttgattgg ggattctcct ctttgctcgt ccagccggtc
     2881 agttaagctt tcattgtgtt tactgcatta gttacttggt gctgagacag aatgcccagc
     2941 aaggcaattt aagagttaat ttggctccag tttgatggac agtctcatgg catggaagtc
     3001 gtggcagaaa gcagtgtaag gcagctagtg catgcatctt tattgagaaa cagagcagtg
     3061 aattctgctg cccattcact ttcttaagtc ccacgacgta agttggtagt caagattagg
     3121 tatcacagtt atatttttac tggggcctga atgttatcga attcctgcag cccgggggat
     3181 ccactagttc tagagcggcc
//
