LOCUS       sequence-293821-        6053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human EED in insect cells, under a TEV-cleavable 
            N-terminal polyhistidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6053)
  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 6053)
  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: pFBOH-MHL.EED
FEATURES             Location/Qualifiers
     source          1..6053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        184..275
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     200..217
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             334..351
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             364..384
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1842..1976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1879..1898)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1933..1952
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2005..2170
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2354..2808
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2441..2460)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2650..2671
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2834..2938
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2939..3007,3008..3799)
                     /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(3157..3176)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3970..4558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4459..4478
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4712..4729
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4863..5087
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5154..5687)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5857..5862,5863..5879,5880..5885))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt
       61 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
      121 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggaatattaa
      181 tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt
      241 ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca
      301 ccatcgggcg cggatctcgg tccgaaaacc atgcatcatc atcatcatca cagcagcggc
      361 agagaaaact tgtatttcca gggctccgag agggaagtgt cgactgcgcc ggcgggaaca
      421 gacatgcctg cggccaagaa gcagaagctg agcagtgacg agaacagcaa tccagacctc
      481 tctggagacg agaatgatga cgctgtcagt atagaaagtg gtacaaacac tgaacgccct
      541 gatacaccta caaacacgcc aaatgcacct ggaaggaaaa gttggggaaa gggaaaatgg
      601 aagtcaaaga aatgcaaata ttctttcaaa tgtgtaaata gtctcaagga agatcataac
      661 caaccattgt ttggagttca gtttaactgg cacagtaaag aaggagatcc attagtgttt
      721 gcaactgtag gaagcaacag agttaccttg tatgaatgtc attcacaagg agaaatccgg
      781 ttgttgcaat cttacgtgga tgctgatgct gatgaaaact tttacacttg tgcatggacc
      841 tatgatagca atacgagcca tcctctgctg gctgtagctg gatctagagg cataattagg
      901 ataataaatc ctataacaat gcagtgtata aagcactatg ttggccatgg aaatgctatc
      961 aatgagctga aattccatcc aagagatcca aatcttctct tgtcagtaag taaagatcat
     1021 gctttacgat tatggaatat ccagacggac actctggtgg caatatttgg aggcgtagaa
     1081 ggacacagag atgaagttct aagtgctgat tatgatcttt tgggtgaaaa aataatgtcc
     1141 tgtggtatgg atcattctct taaactttgg aggatcaatt caaagagaat gatgaatgca
     1201 attaaggaat cttatgatta taatccaaat aaaactaaca ggccatttat ttctcagaaa
     1261 atccattttc ctgatttttc taccagagac atacatagga attatgttga ttgtgtgcga
     1321 tggttaggcg atttgatact ttctaagtct tgtgaaaatg ccattgtgtg ctggaaacct
     1381 ggcaagatgg aagatgatat agataaaatt aaacccagtg aatctaatgt gactattctt
     1441 gggcgatttg attacagcca gtgtgacatt tggtacatga ggttttctat ggatttctgg
     1501 caaaagatgc ttgcattggg caatcaagtt ggcaaacttt atgtttggga tttagaagta
     1561 gaagatcctc ataaagccaa atgtacaaca ctgactcatc ataaatgtgg tgctgctatt
     1621 cgacaaacca gttttagcag ggatagcagc attcttatag ctgtttgtga tgatgccagt
     1681 atttggcgct gggatcgact tcgatgatga cgaagcttgt cgagaagtac tagaggatca
     1741 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     1801 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     1861 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1921 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat
     1981 cactgatatc gcctaggaga tccgaaccag ataagtgaaa tctagttcca aactattttg
     2041 tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac
     2101 tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc aactattttg
     2161 tccgcccaca gcggggcatt tttcttcctg ttatgttttt aatcaaacat cctgccaact
     2221 ccatgtgaca aaccgtcatc ttcggctact ttttctctgt cacagaatga aaatttttct
     2281 gtcatctctt cgttattaat gtttgtaatt gactgaatat caacgcttat ttgcagcctg
     2341 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2401 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2461 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2521 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2581 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2641 ttcttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2701 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2761 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     2821 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2881 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2941 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3001 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3061 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3121 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3181 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3241 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3301 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3361 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3421 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3481 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3541 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3601 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3661 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3721 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3781 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3841 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3901 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3961 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4021 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4081 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4141 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4201 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4261 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4321 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4381 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4441 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4501 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4561 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4621 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     4681 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     4741 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagacca
     4801 gccgcgtaac ctggcaaaat cggttacggt tgagtaataa atggatgccc tgcgtaagcg
     4861 ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta tgacaataaa
     4921 gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc
     4981 atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg
     5041 tcgtattaaa gaggggcgtg gccaagggca tggtaaagac tatattcgcg gcgttgtgac
     5101 aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg
     5161 gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag
     5221 agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc
     5281 gcgttggcct catgcttgag cagattgatg agcgcggtgg caatgccctg cctccggtgc
     5341 tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc
     5401 agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg
     5461 aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag
     5521 gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact
     5581 tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt
     5641 ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc
     5701 acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca
     5761 taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg
     5821 gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat
     5881 gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc
     5941 agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc
     6001 acgcatcgtc aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtg
//
