LOCUS       sequence-292437-        8133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human EZH2 in insect cells, , under a C3-cleavable 
            (PreScission Protease) N-terminal hexahistidine-MBP tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8133)
  AUTHORS   Zhang Q, McKenzie NJ, Warneford-Thomson R, Gail EH, Flanigan SF, 
            Owen BM, Lauman R, Levina V, Garcia BA, Schittenhelm RB, Bonasio R, 
            Davidovich C
  TITLE     RNA exploits an exposed regulatory site to inhibit the enzymatic 
            activity of PRC2.
  JOURNAL   Nat Struct Mol Biol. 2019 Mar;26(3):237-247. doi: 
            10.1038/s41594-019-0197-y. Epub 2019 Mar 4.
  PUBMED    30833789
REFERENCE   2  (bases 1 to 8133)
  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: 
            "10.1038/s41594-019-0197-y"; journalName: "Nat Struct Mol Biol"; 
            date: "2019-03"; volume: "26"; issue: "3"; pages: "237-247"
COMMENT     Sequence Label: pFB1.HMBP.PrS.EZH2
FEATURES             Location/Qualifiers
     source          1..8133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        113..204
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     129..146
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             265..282
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             289..1386
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1243..1252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1360..1383
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1414..1437
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    3862..3996
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3899..3918)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3953..3972
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4025..4190
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4374..4828
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4461..4480)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4670..4691
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4854..4958
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4959..5027,5028..5819)
                     /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     complement(5177..5196)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5990..6578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6479..6498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6721..6738
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6872..7096
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(7163..7696)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7866..7871,7872..7888,7889..7894))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 cggatgaagt ggttcgcatc ctcggttttc tggaaggcga gcatcgtttg ttcgcccagg
       61 actctagcta tagttctagt ggttggctac gtatactccg gaatattaat agatcatgga
      121 gataattaaa atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt
      181 tttgtaataa aaaaacctat aaatattccg gattattcat accgtcccac catcgggcgc
      241 ggatctcggt ccgaaaccat gggtcatcac catcatcatc acggtaccaa aactgaagaa
      301 ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt
      361 aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa
      421 gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac
      481 gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg
      541 ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt
      601 gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac
      661 ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag
      721 agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac
      781 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat
      841 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg
      901 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg
      961 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta
     1021 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca
     1081 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg
     1141 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg
     1201 aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccaccat ggaaaacgcc
     1261 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt
     1321 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg
     1381 cagactggta ccgattacga tatcccaacg accctggaag ttctgttcca ggggcccctg
     1441 ggatccggcc agactgggaa gaaatctgag aagggaccag tttgttggcg gaagcgtgta
     1501 aaatcagagt acatgcgact gagacagctc aagaggttca gacgagctga tgaagtaaag
     1561 agtatgttta gttccaatcg tcagaaaatt ttggaaagaa cggaaatctt aaaccaagaa
     1621 tggaaacagc gaaggataca gcctgtgcac atcctgactt ctgtgagctc attgcgcggg
     1681 actagggagt gttcggtgac cagtgacttg gattttccaa cacaagtcat cccattaaag
     1741 actctgaatg cagttgcttc agtacccata atgtattctt ggtcacccct acagcagaat
     1801 tttatggtgg aagatgaaac tgttttacat aacattcctt atatgggaga tgaagtttta
     1861 gatcaggatg gtactttcat tgaagaacta ataaaaaatt atgatgggaa agtacacggg
     1921 gatagagaat gtgggtttat aaatgatgaa atttttgtgg agttggtgaa tgcccttggt
     1981 caatataatg atgatgacga tgatgatgat ggagacgatc ctgaagaaag agaagaaaag
     2041 cagaaagatc tggaggatca ccgagatgat aaagaaagcc gcccacctcg gaaatttcct
     2101 tctgataaaa tttttgaagc catttcctca atgtttccag ataagggcac agcagaagaa
     2161 ctaaaggaaa aatataaaga actcaccgaa cagcagctcc caggcgcact tcctcctgaa
     2221 tgtaccccca acatagatgg accaaatgct aaatctgttc agagagagca aagcttacac
     2281 tcctttcata cgcttttctg taggcgatgt tttaaatatg actgcttcct acatcgtaag
     2341 tgcaattatt cttttcatgc aacacccaac acttataagc ggaagaacac agaaacagct
     2401 ctagacaaca aaccttgtgg accacagtgt taccagcatt tggagggagc aaaggagttt
     2461 gctgctgctc tcaccgctga gcggataaag accccaccaa aacgtccagg aggccgcaga
     2521 agaggacggc ttcccaataa cagtagcagg cccagcaccc ccaccattaa tgtgctggaa
     2581 tcaaaggata cagacagtga tagggaagca gggactgaaa cggggggaga gaacaatgat
     2641 aaagaagaag aagagaagaa agatgaaact tcgagctcct ctgaagcaaa ttctcggtgt
     2701 caaacaccaa taaagatgaa gccaaatatt gaacctcctg agaatgtgga gtggagtggt
     2761 gctgaagcct caatgtttag agtcctcatt ggcacttact atgacaattt ctgtgccatt
     2821 gctaggttaa ttgggaccaa aacatgtaga caggtgtatg agtttagagt caaagaatct
     2881 agcatcatag ctccagctcc cgctgaggat gtggatactc ctccaaggaa aaagaagagg
     2941 aaacaccggt tgtgggctgc acactgcaga aagatacagc tgaaaaagga cggctcctct
     3001 aaccatgttt acaactatca accctgtgat catccacggc agccttgtga cagttcgtgc
     3061 ccttgtgtga tagcacaaaa tttttgtgaa aagttttgtc aatgtagttc agagtgtcaa
     3121 aaccgctttc cgggatgccg ctgcaaagca cagtgcaaca ccaagcagtg cccgtgctac
     3181 ctggctgtcc gagagtgtga ccctgacctc tgtcttactt gtggagccgc tgaccattgg
     3241 gacagtaaaa atgtgtcctg caagaactgc agtattcagc ggggctccaa aaagcatcta
     3301 ttgctggcac catctgacgt ggcaggctgg gggattttta tcaaagatcc tgtgcagaaa
     3361 aatgaattca tctcagaata ctgtggagag attatttctc aagatgaagc tgacagaaga
     3421 gggaaagtgt atgataaata catgtgcagc tttctgttca acttgaacaa tgattttgtg
     3481 gtggatgcaa cccgcaaggg taacaaaatt cgttttgcaa atcattcggt aaatccaaac
     3541 tgctatgcaa aagttatgat ggttaacggt gatcacagga taggtatttt tgccaagaga
     3601 gccatccaga ctggcgaaga gctgtttttt gattacagat acagccaggc tgatgccctg
     3661 aagtatgtcg gcatcgaaag agaaatggaa atcccttgat ctagagcctg cagtctcgag
     3721 gcatgcggta ccaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt
     3781 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3841 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3901 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3961 tccaaactca tcaatgtatc ttatcatgtc tggatctgat cactgatatc gcctaggaga
     4021 tccgaaccag ataagtgaaa tctagttcca aactattttg tcatttttaa ttttcgtatt
     4081 agcttacgac gctacaccca gttcccatct attttgtcac tcttccctaa ataatcctta
     4141 aaaactccat ttccacccct cccagttccc aactattttg tccgcccaca gcggggcatt
     4201 tttcttcctg ttatgttttt aatcaaacat cctgccaact ccatgtgaca aaccgtcatc
     4261 ttcggctact ttttctctgt cacagaatga aaatttttct gtcatctctt cgttattaat
     4321 gtttgtaatt gactgaatat caacgcttat ttgcagcctg aatggcgaat gggacgcgcc
     4381 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4441 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4501 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4561 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4621 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttcttaatag tggactcttg
     4681 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4741 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4801 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     4861 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4921 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4981 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5041 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5101 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5161 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5221 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5281 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5341 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5401 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5461 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5521 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5581 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5641 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5701 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5761 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5821 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5881 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5941 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6001 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6061 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6121 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6181 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6241 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6301 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6361 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6421 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6481 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6541 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6601 tacggttcct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6661 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6721 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     6781 ctgtgcggta tttcacaccg catagaccag ccgcgtaacc tggcaaaatc ggttacggtt
     6841 gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga
     6901 acaaaataga tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg
     6961 aaatcagtcc agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac
     7021 tcttcatttt ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ccaagggcat
     7081 ggtaaagact atattcgcgg cgttgtgaca atttaccgaa caactccgcg gccgggaagc
     7141 cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca
     7201 cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg
     7261 cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga
     7321 gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga
     7381 tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg
     7441 cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt
     7501 aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa
     7561 gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga
     7621 tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt
     7681 tgctgctgcg taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc
     7741 gaggcataga ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt
     7801 taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc
     7861 attacagttt acgaaccgaa caggcttatg tcaactgggt tcgtgccttc atccgtttcc
     7921 acggtgtgcg tcacccggca accttgggca gcagcgaagt cgaggcattt ctgtcctggc
     7981 tggcgaacga gcgcaaggtt tcggtctcca cgcatcgtca ggcattggcg gccttgctgt
     8041 tcttctacgg caaggtgctg tgcacggatc tgccctggct tcaggagatc ggaagacctc
     8101 ggccgtcgcg gcgcttgccg gtggtgctga ccc
//
