LOCUS       sequence-292459-        7009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human EPOP 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 7009)
  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 7009)
  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.EPOP
FEATURES             Location/Qualifiers
     source          1..7009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        159..250
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     175..192
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             311..328
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             335..1432
                     /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      1289..1298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1406..1429
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1469..1492
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    2786..2920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2823..2842)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2877..2896
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2949..3114
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3298..3752
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3385..3404)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3594..3615
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3778..3882
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3883..3951,3952..4743)
                     /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(4101..4120)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4914..5502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5403..5422
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5645..5662
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5796..6020
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6087..6620)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6790..6795,6796..6812,6813..6818))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 agatcggaag acctcggccg tcgcggcgct tgccggtggt gctgaccccg gatgaagtgg
       61 ttcgcatcct cggttttctg gaaggcgagc atcgtttgtt cgcccaggac tctagctata
      121 gttctagtgg ttggctacat actccggaat attaatagat catggagata attaaaatga
      181 taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg taataaaaaa
      241 acctataaat attccggatt attcataccg tcccaccatc gggcgcggat ctcggtccga
      301 aaccatgggt catcaccatc atcatcacgg taccaaaact gaagaaggta aactggtaat
      361 ctggattaac ggcgataaag gctataacgg tctcgctgaa gtcggtaaga aattcgagaa
      421 agataccgga attaaagtca ccgttgagca tccggataaa ctggaagaga aattcccaca
      481 ggttgcggca actggcgatg gccctgacat tatcttctgg gcacacgacc gctttggtgg
      541 ctacgctcaa tctggcctgt tggctgaaat caccccggac aaagcgttcc aggacaagct
      601 gtatccgttt acctgggatg ccgtacgtta caacggcaag ctgattgctt acccgatcgc
      661 tgttgaagcg ttatcgctga tttataacaa agatctgctg ccgaacccgc caaaaacctg
      721 ggaagagatc ccggcgctgg ataaagaact gaaagcgaaa ggtaagagcg cgctgatgtt
      781 caacctgcaa gaaccgtact tcacctggcc gctgattgct gctgacgggg gttatgcgtt
      841 caagtatgaa aacggcaagt acgacattaa agacgtgggc gtggataacg ctggcgcgaa
      901 agcgggtctg accttcctgg ttgacctgat taaaaacaaa cacatgaatg cagacaccga
      961 ttactccatc gcagaagctg cctttaataa aggcgaaaca gcgatgacca tcaacggccc
     1021 gtgggcatgg tccaacatcg acaccagcaa agtgaattat ggtgtaacgg tactgccgac
     1081 cttcaagggt caaccatcca aaccgttcgt tggcgtgctg agcgcaggta ttaacgccgc
     1141 cagtccgaac aaagagctgg caaaagagtt cctcgaaaac tatctgctga ctgatgaagg
     1201 tctggaagcg gttaataaag acaaaccgct gggtgccgta gcgctgaagt cttacgagga
     1261 agagttggcg aaagatccac gtattgccgc caccatggaa aacgcccaga aaggtgaaat
     1321 catgccgaac atcccgcaga tgtccgcttt ctggtatgcc gtgcgtactg cggtgatcaa
     1381 cgccgccagc ggtcgtcaga ctgtcgatga agccctgaaa gacgcgcaga ctgcggccgc
     1441 aggcaccgat tacgatatcc caacgaccct ggaagttctg ttccaggggc ccggggaaac
     1501 actgtgtcct gcgcctcgcc tggcagtgcc ggcgtccccg cgagggtcgc cctgctcccc
     1561 tacgcctcgt aaaccatgtc gggggacaca ggaattctct ccgctgtgcc tgcgtgccct
     1621 agccttctgc gcccttgcca agcccagggc ttcctctctt ggcccgggac ctggggaact
     1681 ggcggctcgg tcaccagtgc tgcggggtcc tcaggctcct ctgcgacctg gaggctgggc
     1741 tccggatggc ctgaagcatc tctgggcacc aaccgggcgg cctggagttc ctaacaccgc
     1801 cgccggcgag gatgcggacg tcgcagcgtg tccacgccgc ggagaggaag aagagggagg
     1861 aggcggtttc ccgcatttcg gcgttcgctc ctgtgcacct cctggccgct gccctgcgcc
     1921 cccgcaccct agggaatcta caaccagctt cgcctcggcc ccacctcgcc ctgcacctgg
     1981 tctagagcct cagcgtggac cagccgccag cccgcctcaa gaaccaagtt ccaggcctcc
     2041 gtcgccacca gcgggcctat ccaccgagcc cgcaggtccc ggaacggcac cacgtccgtt
     2101 cctgcccgga cagcctgctg aagtagatgg aaaccctccg ccggcagccc cagaggctcc
     2161 agcggccagt ccatcgacgg ccagtccggc tccggccgca ccaggagatc tccgccagga
     2221 acatttcgat cgtctgatcc gccggtcgaa actttggtgt tacgcgaagg gcttcgcctt
     2281 ggacactccg agtttgcgcc gtgggccaga gcgtcctcct gcgaaagggc ctgctcgagg
     2341 agcagccaag aaacgccggc tgccggcgcc tcctccgcga acagcacaac ctcgccgccc
     2401 tgcaccgacg ctacccacca cgagcacctt cagcctcctc aactgcttcc cctgcccccc
     2461 ggccctggtg gtgggggaag acggtgacct aaagccggca tcctcgcttc gactccaggg
     2521 agattctaag cccccgcccg cccacccgct gtggaggtgg cagatggggg gtcccgctgt
     2581 ccctgagccc cctggcctca aattctgggg gatcaacatg gatgaaagct gataatagct
     2641 cgaggcatgc ggtaccaagc ttgtcgagaa gtactagagg atcataatca gccataccac
     2701 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2761 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2821 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2881 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatcactga tatcgcctag
     2941 gagatccgaa ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg
     3001 tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc
     3061 cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg
     3121 catttttctt cctgttatgt ttttaatcaa acatcctgcc aactccatgt gacaaaccgt
     3181 catcttcggc tactttttct ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat
     3241 taatgtttgt aattgactga atatcaacgc ttatttgcag cctgaatggc gaatgggacg
     3301 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3361 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3421 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3481 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3541 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttctta atagtggact
     3601 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3661 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3721 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     3781 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3841 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3901 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3961 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4021 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4081 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4141 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4201 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4261 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4321 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4381 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4441 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4501 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4561 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4621 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4681 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4741 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4801 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4861 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4921 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4981 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5041 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5101 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5161 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5221 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5281 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5341 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5401 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5461 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5521 tttttacggt tcctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5581 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5641 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     5701 gcatctgtgc ggtatttcac accgcataga ccagccgcgt aacctggcaa aatcggttac
     5761 ggttgagtaa taaatggatg ccctgcgtaa gcgggtgtgg gcggacaata aagtcttaaa
     5821 ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa actagacaga atagttgtaa
     5881 actgaaatca gtccagttat gctgtgaaaa agcatactgg acttttgtta tggctaaagc
     5941 aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt aaagaggggc gtggccaagg
     6001 gcatggtaaa gactatattc gcggcgttgt gacaatttac cgaacaactc cgcggccggg
     6061 aagccgatct cggcttgaac gaattgttag gtggcggtac ttgggtcgat atcaaagtgc
     6121 atcacttctt cccgtatgcc caactttgta tagagagcca ctgcgggatc gtcaccgtaa
     6181 tctgcttgca cgtagatcac ataagcacca agcgcgttgg cctcatgctt gaggagattg
     6241 atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg agactgcgag atcatagata
     6301 tagatctcac tacgcggctg ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca
     6361 accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct tactacggag caagttcccg
     6421 aggtaatcgg agtccggctg atgttgggag taggtggcta cgtctccgaa ctcacgaccg
     6481 aaaagatcaa gagcagcccg catggatttg acttggtcag ggccgagcct acatgtgcga
     6541 atgatgccca tacttgagcc acctaacttt gttttagggc gactgccctg ctgcgtaaca
     6601 tcgttgctgc tgcgtaacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat
     6661 gcccgaggca tagactgtac aaaaaaacag tcataacaag ccatgaaaac cgccactgcg
     6721 ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac
     6781 ttgcattaca gtttacgaac cgaacaggct tatgtcaact gggttcgtgc cttcatccgt
     6841 ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg aagtcgaggc atttctgtcc
     6901 tggctggcga acgagcgcaa ggtttcggtc tccacgcatc gtcaggcatt ggcggccttg
     6961 ctgttcttct acggcaaggt gctgtgcacg gatctgccct ggcttcagg
//
