LOCUS       sequence-149765-        8029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Untagged EZH2-TALEN_Left to target the N-terminus of human EZH2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8029)
  AUTHORS   Dalvai M, Loehr J, Jacquet K, Huard CC, Roques C, Herst P, Cote J, 
            Doyon Y
  TITLE     A Scalable Genome-Editing-Based Approach for Mapping Multiprotein 
            Complexes in Human Cells.
  JOURNAL   Cell Rep. 2015 Oct 7. pii: S2211-1247(15)01020-7. doi: 
            10.1016/j.celrep.2015.09.009.
  PUBMED    26456817
REFERENCE   2  (bases 1 to 8029)
  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: "Cell Rep. 
            2015 Oct 7. pii: S2211-1247(15)01020-7. doi: 
            10.1016/j.celrep.2015.09.009."
COMMENT     Sequence Label: EZH2-TALEN_Left_No_FLAG
FEATURES             Location/Qualifiers
     source          1..8029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             953..973
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3290..3877
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     CDS             3893..3934
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3944..3961
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3984..4001)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3990..4214
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4260..4688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4347..4366)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4557..4578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4697..4717)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4702..5032
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4883..5018
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4945..4964
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5080..5127
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5146..5544
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5702..5823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5739..5758)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5793..5812
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5872..5888)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5872..5888)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5885..5907)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5896..5912
                     /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(5920..5926,5927..5944,5945..5950))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5965..5986
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6103..6120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6274..6862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6354..6373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7033..7824,7825..7893))
                     /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     7656..7675
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7894..7998)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagctgatcc actagtccag tgtggtggaa ttcgccatgg cccccaagaa
      961 gaagaggaag gtgggcatcc acggggtacc tatggtggac ttgaggacac tcggttattc
     1021 gcaacagcaa caggagaaaa tcaagcctaa ggtcaggagc accgtcgcgc aacaccacga
     1081 ggcgcttgtg gggcatggct tcactcatgc gcatattgtc gcgctttcac agcaccctgc
     1141 ggcgcttggg acggtggctg tcaaatacca agatatgatt gcggccctgc ccgaagccac
     1201 gcacgaggca attgtagggg tcggtaaaca gtggtcggga gcgcgagcac ttgaggcgct
     1261 gctgactgtg gcgggtgagc ttagggggcc tccgctccag ctcgacaccg ggcagctgct
     1321 gaagatcgcg aagagagggg gagtaacagc ggtagaggca gtgcacgcct ggcgcaatgc
     1381 gctcaccggg gcccccttga acctgacccc agaccaggta gtcgcaatcg cgaacaataa
     1441 tgggggaaag caagccctgg aaaccgtgca aaggttgttg ccggtccttt gtcaagacca
     1501 cggccttaca ccggagcaag tcgtggccat tgcatcccac gacggtggca aacaggctct
     1561 tgagacggtt cagagacttc tcccagttct ctgtcaagcc cacgggctga ctcccgatca
     1621 agttgtagcg attgcgtcca acggtggagg gaaacaagca ttggagactg tccaacggct
     1681 ccttcccgtg ttgtgtcaag cccacggttt gacgcctgca caagtggtcg ccatcgcctc
     1741 gaatggcggc ggtaagcagg cgctggaaac agtacagcgc ctgctgcctg tactgtgcca
     1801 ggatcatgga ctgaccccag accaggtagt cgcaatcgcg tcaaacggag ggggaaagca
     1861 agccctggaa accgtgcaaa ggttgttgcc ggtcctttgt caagaccacg gccttacacc
     1921 ggagcaagtc gtggccattg caagcaatgg gggtggcaaa caggctcttg agacggttca
     1981 gagacttctc ccagttctct gtcaagccca cgggctgact cccgatcaag ttgtagcgat
     2041 tgcgtcgaac attggaggga aacaagcatt ggagactgtc caacggctcc ttcccgtgtt
     2101 gtgtcaagcc cacggtttga cgcctgcaca agtggtcgcc atcgccaaca acaacggcgg
     2161 taagcaggcg ctggaaacag tacagcgcct gctgcctgta ctgtgccagg atcatggact
     2221 gaccccagac caggtagtcg caatcgcgtc gaacattggg ggaaagcaag ccctggaaac
     2281 cgtgcaaagg ttgttgccgg tcctttgtca agaccacggc cttacaccgg agcaagtcgt
     2341 ggccattgca agcaacatcg gtggcaaaca ggctcttgag acggttcaga gacttctccc
     2401 agttctctgt caagcccacg ggctgactcc cgatcaagtt gtagcgattg cgtccaacgg
     2461 tggagggaaa caagcattgg agactgtcca acggctcctt cccgtgttgt gtcaagccca
     2521 cggtttgacg cctgcacaag tggtcgccat cgcctccaat attggcggta agcaggcgct
     2581 ggaaacagta cagcgcctgc tgcctgtact gtgccaggat catggactga ccccagacca
     2641 ggtagtcgca atcgcgtcga acattggggg aaagcaagcc ctggaaaccg tgcaaaggtt
     2701 gttgccggtc ctttgtcaag accacggcct tacaccggag caagtcgtgg ccattgcaag
     2761 caatgggggt ggcaaacagg ctcttgagac ggttcagaga cttctcccag ttctctgtca
     2821 agcccacggg ctgactcccg atcaagttgt agcgattgcg tcgcatgacg gagggaaaca
     2881 agcattggag actgtccaac ggctccttcc cgtgttgtgt caagcccacg gtttgacgcc
     2941 tgcacaagtg gtcgccatcg cctccaatat tggcggtaag caggcgctgg aaacagtaca
     3001 gcgcctgctg cctgtactgt gccaggatca tggactgaca cccgaacagg tggtcgccat
     3061 tgcttctaat gggggaggac ggccagcctt ggagtccatc gtagcccaat tgtccaggcc
     3121 cgatcccgcg ttggctgcgt taacgaatga ccatctggtg gcgttggcat gtcttggtgg
     3181 acgacccgcg ctcgatgcag tcaaaaaggg tctgcctcat gctcccgcat tgatcaaaag
     3241 aaccaaccgg cggattcccg agagaacttc ccatcgagtc gcgggatccc aactagtcaa
     3301 aagtgaactg gaggagaaga aatctgaact tcgtcataaa ttgaaatatg tgcctcatga
     3361 atatattgaa ttaattgaaa ttgccagaaa ttccactcag gatagaattc ttgaaatgaa
     3421 ggtaatggaa ttttttatga aagtttatgg atatagaggt aaacatttgg gtggatcaag
     3481 gaaaccggac ggagcaattt atactgtcgg atctcctatt gattacggtg tgatcgtgga
     3541 tactaaagct tatagcggag gttataatct gccaattggc caagcagatg aaatgcaacg
     3601 atatgtcgaa gaaaatcaaa cacgaaacaa acatatcaac cctaatgaat ggtggaaagt
     3661 ctatccatct tctgtaacgg aatttaagtt tttatttgtg agtggtcact ttaaaggaaa
     3721 ctacaaagct cagcttacac gattaaatca tatcactaat tgtaatggag ctgttcttag
     3781 tgtagaagag cttttaattg gtggagaaat gattaaagcc ggcacattaa ccttagagga
     3841 agtcagacgg aaatttaata acggcgagat aaacttttaa gggcccttcg aaggtaagcc
     3901 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
     3961 ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4021 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4081 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4141 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     4201 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     4261 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4321 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4381 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     4441 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4501 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4561 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4621 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4681 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4741 ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4801 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4861 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4921 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4981 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     5041 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     5101 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagcctttg
     5161 tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc
     5221 tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt
     5281 gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct
     5341 gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc
     5401 atcttgagcc cctgcggacg gtgtcgacag gtgcttctcg atctgcatcc tgggatcaaa
     5461 gcgatagtga aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc
     5521 tctggttatg tgtgggaggg ctaagcactt cgtggccgag gagcaggact gacacgtgct
     5581 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5641 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5701 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5761 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5821 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5881 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5941 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6001 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6061 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6121 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6181 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6241 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6301 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6361 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6421 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6481 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6541 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6601 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6661 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6721 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6781 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6841 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6901 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6961 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7021 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7081 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7141 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7201 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7261 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7321 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7381 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7441 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7501 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7561 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7621 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7681 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7741 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7801 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7861 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7921 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7981 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
