LOCUS       sequence-292443-        7203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human EED 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 7203)
  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 7203)
  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.EED
FEATURES             Location/Qualifiers
     source          1..7203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        98..189
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     114..131
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             250..267
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             274..1371
                     /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      1228..1237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1345..1368
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1399..1422
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    2917..3051
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2954..2973)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3008..3027
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3080..3245
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3429..3883
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3516..3535)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3725..3746
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3909..4013
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4014..4082,4083..4874)
                     /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(4232..4251)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5045..5633
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5534..5553
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5776..5793
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5927..6151
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6218..6751)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6921..6926,6927..6943,6944..6949))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc ccaggactct agctatagtt
       61 ctagtggttg gctacgtata ctccggaata ttaatagatc atggagataa ttaaaatgat
      121 aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt aataaaaaaa
      181 cctataaata ttccggatta ttcataccgt cccaccatcg ggcgcggatc tcggtccgaa
      241 accatgggtc atcaccatca tcatcacggt accaaaactg aagaaggtaa actggtaatc
      301 tggattaacg gcgataaagg ctataacggt ctcgctgaag tcggtaagaa attcgagaaa
      361 gataccggaa ttaaagtcac cgttgagcat ccggataaac tggaagagaa attcccacag
      421 gttgcggcaa ctggcgatgg ccctgacatt atcttctggg cacacgaccg ctttggtggc
      481 tacgctcaat ctggcctgtt ggctgaaatc accccggaca aagcgttcca ggacaagctg
      541 tatccgttta cctgggatgc cgtacgttac aacggcaagc tgattgctta cccgatcgct
      601 gttgaagcgt tatcgctgat ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg
      661 gaagagatcc cggcgctgga taaagaactg aaagcgaaag gtaagagcgc gctgatgttc
      721 aacctgcaag aaccgtactt cacctggccg ctgattgctg ctgacggggg ttatgcgttc
      781 aagtatgaaa acggcaagta cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa
      841 gcgggtctga ccttcctggt tgacctgatt aaaaacaaac acatgaatgc agacaccgat
      901 tactccatcg cagaagctgc ctttaataaa ggcgaaacag cgatgaccat caacggcccg
      961 tgggcatggt ccaacatcga caccagcaaa gtgaattatg gtgtaacggt actgccgacc
     1021 ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc
     1081 agtccgaaca aagagctggc aaaagagttc ctcgaaaact atctgctgac tgatgaaggt
     1141 ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa
     1201 gagttggcga aagatccacg tattgccgcc accatggaaa acgcccagaa aggtgaaatc
     1261 atgccgaaca tcccgcagat gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac
     1321 gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac tggtaccgat
     1381 tacgatatcc caacgaccct ggaagttctg ttccaggggc ccctgggatc ctccgagagg
     1441 gaagtgtcga ctgcgccggc gggaacagac atgcctgcgg ccaagaagca gaagctgagc
     1501 agtgacgaga acagcaatcc agacctctct ggagacgaga atgatgacgc tgtcagtata
     1561 gaaagtggta caaacactga acgccctgat acacctacaa acacgccaaa tgcacctgga
     1621 aggaaaagtt ggggaaaggg aaaatggaag tcaaagaaat gcaaatattc tttcaaatgt
     1681 gtaaatagtc tcaaggaaga tcataaccaa ccattgtttg gagttcagtt taactggcac
     1741 agtaaagaag gagatccatt agtgtttgca actgtaggaa gcaacagagt taccttgtat
     1801 gaatgtcatt cacaaggaga aatccggttg ttgcaatctt acgtggatgc tgatgctgat
     1861 gaaaactttt acacttgtgc atggacctat gatagcaata cgagccatcc tctgctggct
     1921 gtagctggat ctagaggcat aattaggata ataaatccta taacaatgca gtgtataaag
     1981 cactatgttg gccatggaaa tgctatcaat gagctgaaat tccatccaag agatccaaat
     2041 cttctcttgt cagtaagtaa agatcatgct ttacgattat ggaatatcca gacggacact
     2101 ctggtggcaa tatttggagg cgtagaagga cacagagatg aagttctaag tgctgattat
     2161 gatcttttgg gtgaaaaaat aatgtcctgt ggtatggatc attctcttaa actttggagg
     2221 atcaattcaa agagaatgat gaatgcaatt aaggaatctt atgattataa tccaaataaa
     2281 actaacaggc catttatttc tcagaaaatc cattttcctg atttttctac cagagacata
     2341 cataggaatt atgttgattg tgtgcgatgg ttaggcgatt tgatactttc taagtcttgt
     2401 gaaaatgcca ttgtgtgctg gaaacctggc aagatggaag atgatataga taaaattaaa
     2461 cccagtgaat ctaatgtgac tattcttggg cgatttgatt acagccagtg tgacatttgg
     2521 tacatgaggt tttctatgga tttctggcaa aagatgcttg cattgggcaa tcaagttggc
     2581 aaactttatg tttgggattt agaagtagaa gatcctcata aagccaaatg tacaacactg
     2641 actcatcata aatgtggtgc tgctattcga caaaccagtt ttagcaggga tagcagcatt
     2701 cttatagctg tttgtgatga tgccagtatt tggcgctggg atcgacttcg atgatctaga
     2761 gcctgcagtc tcgaggcatg cggtaccaag cttgtcgaga agtactagag gatcataatc
     2821 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2881 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2941 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3001 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg
     3061 atatcgccta ggagatccga accagataag tgaaatctag ttccaaacta ttttgtcatt
     3121 tttaattttc gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc
     3181 cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc
     3241 ccacagcggg gcatttttct tcctgttatg tttttaatca aacatcctgc caactccatg
     3301 tgacaaaccg tcatcttcgg ctactttttc tctgtcacag aatgaaaatt tttctgtcat
     3361 ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttgca gcctgaatgg
     3421 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3481 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3541 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3601 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3661 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3721 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3781 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3841 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     3901 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3961 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4021 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4081 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4141 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4201 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4261 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4321 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4381 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4441 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4501 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4561 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4621 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4681 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4741 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4801 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4861 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4921 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4981 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5041 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5101 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5161 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5221 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5281 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5341 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5401 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5461 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5521 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5581 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5641 gcaacgcggc ctttttacgg ttcctggcct tttgctcaca tgttctttcc tgcgttatcc
     5701 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5761 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     5821 tttctcctta cgcatctgtg cggtatttca caccgcatag accagccgcg taacctggca
     5881 aaatcggtta cggttgagta ataaatggat gccctgcgta agcgggtgtg ggcggacaat
     5941 aaagtcttaa actgaacaaa atagatctaa actatgacaa taaagtctta aactagacag
     6001 aatagttgta aactgaaatc agtccagtta tgctgtgaaa aagcatactg gacttttgtt
     6061 atggctaaag caaactcttc attttctgaa gtgcaaattg cccgtcgtat taaagagggg
     6121 cgtggccaag ggcatggtaa agactatatt cgcggcgttg tgacaattta ccgaacaact
     6181 ccgcggccgg gaagccgatc tcggcttgaa cgaattgtta ggtggcggta cttgggtcga
     6241 tatcaaagtg catcacttct tcccgtatgc ccaactttgt atagagagcc actgcgggat
     6301 cgtcaccgta atctgcttgc acgtagatca cataagcacc aagcgcgttg gcctcatgct
     6361 tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg gtgctcgccg gagactgcga
     6421 gatcatagat atagatctca ctacgcggct gctcaaactt gggcagaacg taagccgcga
     6481 gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga
     6541 gcaagttccc gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga
     6601 actcacgacc gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc
     6661 tacatgtgcg aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct
     6721 gctgcgtaac atcgttgctg ctgcgtaaca tcaaacatcg acccacggcg taacgcgctt
     6781 gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa
     6841 ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc
     6901 gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac tgggttcgtg
     6961 ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg
     7021 catttctgtc ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat
     7081 tggcggcctt gctgttcttc tacggcaagg tgctgtgcac ggatctgccc tggcttcagg
     7141 agatcggaag acctcggccg tcgcggcgct tgccggtggt gctgaccccg gatgaagtgg
     7201 ttc
//
