LOCUS       sequence-417377-        7947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SnoopTag counterpart binding domain, SnoopCatcher, fused to polycomb
            repressive complex subunit EZH2, with GFP selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7947)
  AUTHORS   Swain T, Pflueger C, Freytag S, Poppe D, Pflueger J, Nguyen TV, Li 
            JK, Lister R
  TITLE     A modular dCas9-based recruitment platform for combinatorial 
            epigenome editing.
  JOURNAL   Nucleic Acids Res. 2023 Nov 24:gkad1108. doi: 10.1093/nar/gkad1108.
  PUBMED    38000387
REFERENCE   2  (bases 1 to 7947)
  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: "Nucleic 
            Acids Res. 2023 Nov 24:gkad1108. doi: 10.1093/nar/gkad1108."
COMMENT     Sequence Label: LLP787_SnoopCatcher-EZH2-FL-CO
FEATURES             Location/Qualifiers
     source          1..7947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    6..230
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(join(240..950,951..953,954..956))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(240..950,951..953,954..956))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(282..303)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     631..650
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     890..911
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      953..962
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(981..1191)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      987..1122
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1041..1060)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(1248..1267)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1485..2663
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1716..2654
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2611..2631
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2670..2679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2682..2702
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2703..2744
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2745..2786
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2787..3122
                     /label=SnoopCatcher
     polyA_signal    5573..5694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5610..5629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5796..5812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5796..5812)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5809..5831)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5820..5836
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5844..5850,5851..5868,5869..5874))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5889..5910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6027..6044)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6198..6786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6278..6297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6957..7748,7749..7817))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7580..7599
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7818..7922)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtccata gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc
       61 cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga
      121 tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga
      181 aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcaactgccc
      241 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
      301 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg
      361 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
      421 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc
      481 ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
      541 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
      601 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
      661 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
      721 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg
      781 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
      841 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
      901 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg
      961 gccgtacggg tggcgcgcgt agcctaggcc tccaaaaaag cctcctcact acttctggaa
     1021 tagctcagag gcagaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     1081 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     1141 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg cctgaattca
     1201 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     1261 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     1321 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     1381 ctgcttaggg ttaggcgttt tgcgtgcaaa gatggataaa gttttaaaca gagaggaatc
     1441 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
     1501 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     1561 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     1621 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     1681 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
     1741 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc
     1801 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt
     1861 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc
     1921 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
     1981 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
     2041 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
     2101 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
     2161 tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt
     2221 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
     2281 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag
     2341 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
     2401 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt
     2461 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
     2521 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
     2581 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
     2641 ttcttccatt tcaggtgtcg tgacgtacgg ccaccatggg tcctaagaaa aagaggaagg
     2701 tgtccggctt tgcgaacgaa cttggacctc gattgatggg caagagcggc ttcgccaacg
     2761 agctgggccc cagactgatg ggcaaaaagc cgctgcgtgg tgccgtgttt agcctgcaga
     2821 aacagcatcc cgactatccc gatatctatg gcgcgattga tcagaatggg acctatcaaa
     2881 atgtgcgtac cggcgaagat ggtaaactga cctttaagaa tctgagcgat ggcaaatatc
     2941 gcctgtttga aaatagcgaa cccgctggct ataaaccggt gcagaataag ccgattgtgg
     3001 cgtttcagat tgtgaatggc gaagtgcgtg atgtgaccag cattgtgccg caggatattc
     3061 cggctacata tgaatttacc aacggtaaac attatatcac caatgaaccg ataccgccga
     3121 aaagcgctgg agggggtggc tccggcggag gaggctctgg tggcggaggg agtggagcgg
     3181 ccgctatggg ccagactggg aagaaatctg agaagggacc agtttgttgg cggaagcgtg
     3241 taaaatcaga gtacatgcga ctgagacagc tcaagaggtt cagacgagct gatgaagtaa
     3301 agagtatgtt tagttccaat cgtcagaaaa ttttggaaag aacggaaatc ttaaaccaag
     3361 aatggaaaca gcgaaggata cagcctgtgc acatcctgac ttctgtgagc tcattgcgcg
     3421 ggactaggga gtgttcggtg accagtgact tggattttcc aacacaagtc atcccattaa
     3481 agactctgaa tgcagttgct tcagtaccca taatgtattc ttggtctccc ctacagcaga
     3541 attttatggt ggaagatgaa actgttttac ataacattcc ttatatggga gatgaagttt
     3601 tagatcagga tggtactttc attgaagaac taataaaaaa ttatgatggg aaagtacacg
     3661 gggatagaga atgtgggttt ataaatgatg aaatttttgt ggagttggtg aatgcccttg
     3721 gtcaatataa tgatgatgac gatgatgatg atggagacga tcctgaagaa agagaagaaa
     3781 agcagaaaga tctggaggat caccgagatg ataaagaaag ccgcccacct cggaaatttc
     3841 cttctgataa aatttttgaa gccatttcct caatgtttcc agataagggc acagcagaag
     3901 aactaaagga aaaatataaa gaactcaccg aacagcagct cccaggcgca cttcctcctg
     3961 aatgtacccc caacatagat ggaccaaatg ctaaatctgt tcagagagag caaagcttac
     4021 actcctttca tacgcttttc tgtaggcgat gttttaaata tgactgcttc ctacatcgta
     4081 agtgcaatta ttcttttcat gcaacaccca acacttataa gcggaagaac acagaaacag
     4141 ctctagacaa caaaccttgt ggaccacagt gttaccagca tttggaggga gcaaaggagt
     4201 ttgctgctgc tctcaccgct gagcggataa agaccccacc aaaacgtcca ggaggccgca
     4261 gaagaggacg gcttcccaat aacagtagca ggcccagcac ccccaccatt aatgtgctgg
     4321 aatcaaagga tacagacagt gatagggaag cagggactga aacgggggga gagaacaatg
     4381 ataaagaaga agaagagaag aaagatgaaa cttcgagctc ctctgaagca aattctcggt
     4441 gtcaaacacc aataaagatg aagccaaata ttgaacctcc tgagaatgtg gagtggagtg
     4501 gtgctgaagc ctcaatgttt agagtcctca ttggcactta ctatgacaat ttctgtgcca
     4561 ttgctaggtt aattgggacc aaaacatgta gacaggtgta tgagtttaga gtcaaagaat
     4621 ctagcatcat agctccagct cccgctgagg atgtggatac tcctccaagg aaaaagaaga
     4681 ggaaacaccg gttgtgggct gcacactgca gaaagataca gctgaaaaag gacggctcct
     4741 ctaaccatgt ttacaactat caaccctgtg atcatccacg gcagccttgt gacagttcgt
     4801 gcccttgtgt gatagcacaa aatttttgtg aaaagttttg tcaatgtagt tcagagtgtc
     4861 aaaaccgctt tccgggatgc cgctgcaaag cacagtgcaa caccaagcag tgcccgtgct
     4921 acctggctgt ccgagagtgt gaccctgacc tctgtcttac ttgtggagcc gctgaccatt
     4981 gggacagtaa aaatgtgtcc tgcaagaact gcagtattca gcggggctcc aaaaagcatc
     5041 tattgctggc accatctgac gtggcaggct gggggatttt tatcaaagat cctgtgcaga
     5101 aaaatgaatt catctcagaa tactgtggag agattattag ccaggacgag gccgacagga
     5161 ggggcaaggt gtacgacaag tacatgtgct ccttcctgtt caacctgaac aacgacttcg
     5221 tggtggacgc caccagaaag ggcaacaaga tcagattcgc caaccacagc gtgaacccca
     5281 actgctacgc caaggtgatg atggtgaacg gcgaccacag aatcggcatc ttcgccaaga
     5341 gggccatcca gaccggcgag gagctgttct tcgactacag gtacagccag gccgacgccc
     5401 tgaaatacgt gggcatcgag agggagatgg agatcccctg acgttgagtc tctgtttgtg
     5461 gtgactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     5521 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     5581 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5641 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaatgcgt
     5701 cgagtgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5761 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5821 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5881 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5941 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6001 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6061 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6121 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6181 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6241 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6301 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6361 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6421 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6481 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6541 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6601 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6661 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6721 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6781 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6841 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6901 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6961 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7021 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7081 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7141 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7201 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7261 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7321 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7381 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7441 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7501 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7561 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7621 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7681 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7741 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7801 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7861 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7921 cgcacatttc cccgaaaagt gccacct
//
