LOCUS       sequence-417390-        8040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AviTag counterpart binding domain, Traptavidin, 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 8040)
  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 8040)
  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: LLP788_Traptavidin-EZH2-FL-CO
FEATURES             Location/Qualifiers
     source          1..8040
                     /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="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2745..2786
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2787..2828
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    5666..5787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5703..5722)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5889..5905)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5889..5905)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5902..5924)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5913..5929
                     /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(5937..5943,5944..5961,5962..5967))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5982..6003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6120..6137)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6291..6879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6371..6390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7050..7841,7842..7910))
                     /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     7673..7692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7911..8015)
                     /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 tgggtaagcc tatccctaac cctctcctcg gtctcgattc tacgggcaag cccatcccca
     2761 accccctgct gggcctggac agcaccggta aacctattcc taatccactc ctcggactcg
     2821 attccacaca tatggctgaa gctggtatca ccggcacctg gtacaaccag ctgggatcca
     2881 ccttcatcgt taccgctggt gctgacggtg ctctgaccgg tacctacgaa tccgctgttg
     2941 gtaacgctga aggcgattac gttctgaccg gtcgttacga ctccgctccg gctaccgacg
     3001 gttccggaac cgctctgggt tggaccgttg cttggaaaaa caactaccgt aacgctcact
     3061 ccgctaccac ctggtctggc cagtacgttg gtggtgctga agctcgtatc aacacccagt
     3121 ggttgttgac ctccggcacc accgaagcta acgcgtggaa atccaccctg gttggtcacg
     3181 acaccttcac caaagttaaa ccgtccgctg cttccagcgc tggagggggt ggctccggcg
     3241 gaggaggctc tggtggcgga gggagtggag cggccgctat gggccagact gggaagaaat
     3301 ctgagaaggg accagtttgt tggcggaagc gtgtaaaatc agagtacatg cgactgagac
     3361 agctcaagag gttcagacga gctgatgaag taaagagtat gtttagttcc aatcgtcaga
     3421 aaattttgga aagaacggaa atcttaaacc aagaatggaa acagcgaagg atacagcctg
     3481 tgcacatcct gacttctgtg agctcattgc gcgggactag ggagtgttcg gtgaccagtg
     3541 acttggattt tccaacacaa gtcatcccat taaagactct gaatgcagtt gcttcagtac
     3601 ccataatgta ttcttggtct cccctacagc agaattttat ggtggaagat gaaactgttt
     3661 tacataacat tccttatatg ggagatgaag ttttagatca ggatggtact ttcattgaag
     3721 aactaataaa aaattatgat gggaaagtac acggggatag agaatgtggg tttataaatg
     3781 atgaaatttt tgtggagttg gtgaatgccc ttggtcaata taatgatgat gacgatgatg
     3841 atgatggaga cgatcctgaa gaaagagaag aaaagcagaa agatctggag gatcaccgag
     3901 atgataaaga aagccgccca cctcggaaat ttccttctga taaaattttt gaagccattt
     3961 cctcaatgtt tccagataag ggcacagcag aagaactaaa ggaaaaatat aaagaactca
     4021 ccgaacagca gctcccaggc gcacttcctc ctgaatgtac ccccaacata gatggaccaa
     4081 atgctaaatc tgttcagaga gagcaaagct tacactcctt tcatacgctt ttctgtaggc
     4141 gatgttttaa atatgactgc ttcctacatc gtaagtgcaa ttattctttt catgcaacac
     4201 ccaacactta taagcggaag aacacagaaa cagctctaga caacaaacct tgtggaccac
     4261 agtgttacca gcatttggag ggagcaaagg agtttgctgc tgctctcacc gctgagcgga
     4321 taaagacccc accaaaacgt ccaggaggcc gcagaagagg acggcttccc aataacagta
     4381 gcaggcccag cacccccacc attaatgtgc tggaatcaaa ggatacagac agtgataggg
     4441 aagcagggac tgaaacgggg ggagagaaca atgataaaga agaagaagag aagaaagatg
     4501 aaacttcgag ctcctctgaa gcaaattctc ggtgtcaaac accaataaag atgaagccaa
     4561 atattgaacc tcctgagaat gtggagtgga gtggtgctga agcctcaatg tttagagtcc
     4621 tcattggcac ttactatgac aatttctgtg ccattgctag gttaattggg accaaaacat
     4681 gtagacaggt gtatgagttt agagtcaaag aatctagcat catagctcca gctcccgctg
     4741 aggatgtgga tactcctcca aggaaaaaga agaggaaaca ccggttgtgg gctgcacact
     4801 gcagaaagat acagctgaaa aaggacggct cctctaacca tgtttacaac tatcaaccct
     4861 gtgatcatcc acggcagcct tgtgacagtt cgtgcccttg tgtgatagca caaaattttt
     4921 gtgaaaagtt ttgtcaatgt agttcagagt gtcaaaaccg ctttccggga tgccgctgca
     4981 aagcacagtg caacaccaag cagtgcccgt gctacctggc tgtccgagag tgtgaccctg
     5041 acctctgtct tacttgtgga gccgctgacc attgggacag taaaaatgtg tcctgcaaga
     5101 actgcagtat tcagcggggc tccaaaaagc atctattgct ggcaccatct gacgtggcag
     5161 gctgggggat ttttatcaaa gatcctgtgc agaaaaatga attcatctca gaatactgtg
     5221 gagagattat tagccaggac gaggccgaca ggaggggcaa ggtgtacgac aagtacatgt
     5281 gctccttcct gttcaacctg aacaacgact tcgtggtgga cgccaccaga aagggcaaca
     5341 agatcagatt cgccaaccac agcgtgaacc ccaactgcta cgccaaggtg atgatggtga
     5401 acggcgacca cagaatcggc atcttcgcca agagggccat ccagaccggc gaggagctgt
     5461 tcttcgacta caggtacagc caggccgacg ccctgaaata cgtgggcatc gagagggaga
     5521 tggagatccc ctgacgttga gtctctgttt gtggtgactg atcataatca gccataccac
     5581 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     5641 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     5701 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5761 tttgtccaaa ctcatcaatg tatcttaatg cgtcgagtgc attctagttg tggtttgtcc
     5821 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5881 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5941 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6001 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6061 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6121 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6181 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6241 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6301 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6361 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6421 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6481 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6541 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6601 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6661 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6721 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6781 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6841 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6901 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6961 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7021 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7081 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7141 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7201 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7261 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7321 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7381 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7441 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7501 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7561 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7621 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7681 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7741 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7801 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7861 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7921 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7981 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
//
